Quantitative measurements of performance parameters has the potential to increase consistency and enhance performance of the surfaces as well as to contribute to the safety of horses and riders. This study investigates how factors known to influence the performance of the surface, incorpo-ration of a drainage package, control of the moisture control, and introduction of a geotextile reinforcement, affect quantitative measurements of arena materials. The measurements are made by using affordable lightweight testing tools which are readily available or easily constructed. Sixteen boxes with arena materials at a consistent depth were tested with the Going Stick (GS), both penetration resistance and shear, the impact test device (ITD), and the rotational peak shear device (RPS). Volumetric moisture content (VMC %) was also tested with time-domain reflectometry (TDR). Results obtained using GS, RPS, ITD, and TDR indicate that the presence of the drainage package, moisture content, and geotextile addition were detected. Alterations due to combinations of treatments could also be detected by GS, ITD, and TDR. While the testing showed some limi-tations of these devices, the potential exists to utilize them for quality control of new installations as well as for the monitoring of maintenance of the surfaces.

Understanding the factors which control endothelial cell (EC) function and angiogenesis is crucial for developing the horse as a disease model, but equine ECs remain poorly studied. In this study we have optimised methods for the isolation and culture of equine aortic endothelial cells (EA-oECs) and characterised their angiogenic functions in vitro. Mechanical dissociation, followed by magnetic purification using an anti-VE-cadherin antibody, resulted in EC-enriched cultures suitable for further study. Fibroblast growth factor 2 (FGF2) increased EAoEC proliferation rate and stimulated scratch wound closure and tube formation by EAoECs on extracellular matrix. Pharmacological inhibitors of FGFR1 (SU5402) or MEK (PD184352) blocked FGF2-induced ERK1/2 phosphorylation and functional responses, suggesting that these are dependent on FGFR1/MEK-ERK signalling. In marked contrast, VEGF-A had no effect on EAoEC proliferation, migration or tubulogenesis and did not promote ERK1/2 phosphorylation, indicating a lack of sensitivity to this classical pro-angiogenic growth factor. Gene expression analysis showed that, unlike human ECs, FGFR1 is expressed by EAoECs at a much higher level than both VEGFR1 and VEGFR2. These results suggest a predominant role for FGF2 versus VEGF-A in controlling the angiogenic functions of equine ECs. Collectively, our novel data provide a sound basis for stud-ying angiogenic processes in the horse and lay the foundations for comparative studies of EC biology in horses versus humans.

Polo is an equestrian team sport, consisting of Open and Women’s only handicapping systems. As cumulative player handicap increases in Open Polo, distance covered, average speeds and high intensity work performed per chukka also increase. These activities may differ in terms of distribution of, and their affect upon, match outcome in Women’s Polo, and thus have implications for equine preparation and management. This study aimed to quantify spatiotemporal differences between Open and Women’s Polo when matched for handicap and assess their affect upon chukka and match outcome using a prospective cohort design. Distance, speed and high intensity activity data were collected via player worn global positioning system (GPS) units during 16-goal Open and Women’s Polo tournaments. Notational analysis quantified chukka duration and chukka and game outcomes. Between group differences were assessed by independent samples t-tests, and two factor mixed effects ANOVA for within group analyses. Between group differences were analysed using an independent samples t-test with alpha defined a priori as p<0.05. Open and Women’s Polo differed by a small to large extent (ES: 0.54 – 1.81) for all spatiotemporal metrics. In Open Polo, players covered moderately more distance (429.0m; 238.9m to 619.0m), with small to large increases in high intensity activities performed in games won. Whereas in Women’s Polo, moderately higher maximum speeds were attained in games won (17.13 km/h; 11.86 km/h to 22.40 km/h) and a small increase in accelerations performed (5.1; 0.2 to 10.0). Open and Women’s Polo, when matched for handicap, present with small to large spatiotemporal differences that are likely of practical significance, and influence game outcome differently between codes. These differences do not necessarily mean that Polo ponies need to be trained differently for each code.

Equine hepacivirus (EqHV, Flaviviridae, Hepacivirus) is a small, enveloped, RNA virus generally causing sub-clinical hepatitis with occasional fatalities. EqHV is reported in equids worldwide, but for Italy data are limited. To address this, a survey study was set up to estimate prevalence at national level and among different production categories (equestrian, competition, work and meat and reproduction) and national macro-regions (North, Central, South and Islands). Data obtained testing 1801 horse serum samples by Real-Time RT PCR were compared within the categories and regions. The NS3 fragment of the PCR-positive samples was sequenced by Sanger protocol for phylogenetic and mutational analysis. The tertiary structure of the NS3 protein was also assessed. The estimated national prevalence was 4.27% [1.97-6.59, 95%CI] and no statistical differences were detected among production categories and macro-regions. The phylogenesis confirmed the distribution in Italy of the three known EqHV subtypes, suggesting also a possible fourth sub-type that however requires further confirmation. Mutational profiles that could also affect the NS3 binding affinity to the viral RNA were detected. The present paper demonstrates that EqHV should be included in diagnostic protocols when investigating causes of hepatitis and in quality control protocols for blood derived products, due to its parental transmission.

Leishmaniosis is a zoonoses caused by Leishmania spp., an intracellular protozoan parasite. This parasite is transmitted by sandflies and the disease is endemic in Mediterranean basin. In the last years, the number of species which could be reservoir of parasite is increased. One of the most relevant is the horses, due to contact with humans and its ability to control the disease, being a possible silent reservoir. In this study, we have analyzed the prevalence and factors related to L. infantum infection in healthy horses in Mediterranean region. Epidemiological data and serum samples were obtained to 167 apparently healthy horses and presence of L. infantum was evaluated by ELISA method and qPCR. The results show 27,5% of prevalence and the main factors related to infection are equine breed, outdoor living, use and season. In conclusion, the prevalence of L. infantum infection in apparently horses from Eastern Spain (Mediterranean basin) is elevated. To control this zoonosis, it would be advisable to carry out more studies on this and other species that could be silent reservoirs of the parasite, as well as carry out measures such as the use of repellents on a regular basis.

Passive dynamics is an aspect of locomotion which is entirely dependent on the mechanical configuration and linkages of adjacent body segments. Tension distribution along mechanical linkages enables execution of movement patterns with reduced need for complex neurological pathways and may play a role in reestablishing postural stability following external disturbances. Here we demonstrate a uni-directional mechanical relationship between the equine forelimb, head and neck, which may have implications for balance and forelimb loading in the horse. These observations suggest that forelimb, head and neck movement coordination (observed in the horse during unrestrained locomotion) is largely controlled by the mechanical linkages between body segments, rather than being entirely dependent on neurological input as previously thought. This highlights the potential significance of research directed at investigating passively induced movements in understanding common locomotory patterns. Additionally, it suggests a mode of postural control which may provide instantaneous adjustments to postural disturbances, thus promoting rapid and efficient locomotion.

Control of Theileria equi, the primary cause of equine theileriosis, is largely reliant on acaracide use and chemosterilization with imidocarb dipropionate (ID). However, it is currently unknown if ID is effective against Theileria haneyi, the recently identified second causative agent of equine theileriosis, or if the drug maintains effectiveness against T. equi in the presence of T. haneyi co-infection. The purpose of this study was address these questions using ID treatment of the following three groups of horses: 1. Five T. haneyi infected horses; 2. Three T. haneyi-T. equi infected horses; and 3. Three T. equi-T. haneyi infected horses. Clearance was first evaluated using nPCR for each Theileria sp. on peripheral blood samples. ID failed to clear T. haneyi in all three groups of horses, and failed to clear T. equi in 2/3 horses in group two. For definitive confirmation of infection status, horses in groups two and three underwent splenectomy post-treatment. The T. equi-nPCR-positive horses in group two developed severe clinical signs and were euthanized. Remaining horses exhibited moderate signs consistent with T. haneyi. Our results demonstrate that ID therapy lacks efficacy against T. haneyi, and T. haneyi-T. equi co-infection may interfere with ID clearance of T. equi.

Control of Theileria equi, the primary cause of equine theileriosis, is largely reliant on acaracide use and chemosterilization with imidocarb dipropionate (ID). However, it is currently unknown if ID is effective against Theileria haneyi, the recently identified second causative agent of equine theileriosis, or if the drug maintains effectiveness against T. equi in the presence of T. haneyi co-infection. The purpose of this study was address these questions using ID treatment of the following three groups of horses: 1. Five T. haneyi infected horses; 2. Three T. haneyi-T. equi infected horses; and 3. Three T. equi-T. haneyi infected horses. Clearance was first evaluated using nPCR for each Theileria sp. on peripheral blood samples. ID failed to clear T. haneyi in all three groups of horses, and failed to clear T. equi in 2/3 horses in group two. For definitive confirmation of infection status, horses in groups two and three underwent splenectomy post-treatment. The T. equi-nPCR-positive horses in group two developed severe clinical signs and were euthanized. Remaining horses exhibited moderate signs consistent with T. haneyi. Our results demonstrate that ID therapy lacks efficacy against T. haneyi, and T. haneyi-T. equi co-infection may interfere with ID clearance of T. equi.

Globally, rotavirus group A (RVA) remains a major cause of severe childhood diarrhea, despite the use of vaccines in &gt; 100 countries. RVA sequencing for local outbreaks facilitates investigation into strain composition, origins, spread, and vaccine failure. In 2018, we collected 248 stool samples from children aged &lt;13 years admitted with diarrheal illness to Kilifi County Hospital, coastal Kenya. Antigen screening detected RVA in 55 samples (22.2%). Of these, VP7 (G) and VP4 (P) segments were successfully sequenced in 48 (87.3%) and phylogenetic analysis based on the VP7 sequences identified seven genetic clusters with six different GP combinations; G3P[8], G1P[8], G2P[4], G2P[8], G9P[8] and G12P[8]. The G3P[8] strains predominated the season (n=37, 67.2%) and comprised three G3 genetic clusters that fell within Lineage I and IX (the latter also known as equine-like G3 lineage). Both two G3 lineages have been recently detected in several countries. Our study is the first to document African children infection with G3 lineage IX. These data highlight the global nature of RVA transmission and the importance of increasing global rotavirus vaccine coverage.

Equine herpesvirus-1 (EHV-1) causes respiratory diseases, abortion, and encephalomyelitis in horses. EHV-1 immediate-early (IE) protein, essential for viral replication, is transactivated by the binding of a multiprotein complex including the open reading frame 12 (ORF12) and some host factors to the IE promoter region. Promoter-associated non-coding RNAs (pancRNAs), which are transcribed from bidirectional promoters, regulate the transcription of neighboring genes in mammals and pathogens. In this study, we identified a novel pancRNA transcribed from across the areas of the 5’-untranslated region and promoter of EHV-1 IE and named it IE pancRNA. IE pancRNA and mRNA were simultaneously expressed in EHV-1-infected RN33B-A68B2M cells. This pancRNA was also transcribed in RK13 and E. Derm cells, which are highly susceptible to EHV-1 infection. Furthermore, IE pancRNA upregulated IE gene expression in the presence of ORF12, and stable expression of IE pancRNA increased the number of EHV-1-infected RN33B-A68B2M cells. These results suggest that IE pancRNAs facilitate EHV-1 proliferation by promoting IE gene expression.

Equine Asthma Syndrome (EAS) is a common respiratory problem that affects horses of any age. In the severe EAS form (historically referred to as recurrent airway obstruction or RAO) the diagnosis, based mainly on history and clinical signs, it is definitively confirmed by the cytological examination of the bronchoalveolar lavage (BAL), also very useful in monitoring lower airway inflammation in response to environmental management and medication. Cytocentrifugated preparation is usually the staging method for the BAL cytological interpretation. To evaluate whether the BAL cytology in horses with severe EAS under different environmental conditions and before and after treatment can undergo significant changes, and at the same time to investigate whether in the cytological evaluation of the BAL there are interpretative differences between two methods of smear preparation of the collected fluid, a study was carried out on a series of BAL samples (n=48) collected in 8 withdrawals from 6 EAS-affected horses subjected to process exacerbation through environmental stimuli and then to pharmacological treatment. Each BAL fluid collected was equally divided into duplicate portions: one set up by cytocentrifugation and one by sediment from simple centrifugation. The cytological examination revealed any significant difference between the EAS-affected horses in all experimental phases (asymptomatic, early exacerbation phase, late exacerbation phase, and remission phase). Diagnostic interpretive comparison between the two BAL preparations then showed no significant differences in results, suggesting how they can be used indifferently in the evaluation of BAL under conditions of airway inflammation in the horse. Thus, the concentrated smear preparation appears to be an equally diagnostically useful method in conditions where there is no possibility of using an appropriate cytocentrifuge.

Alphavirus species are globally distributed zoonoses, primarily transmitted by arthropods. In Costa Rica, Venezuelan equine encephalitis virus (VEEV) and Eastern equine encephalitis virus (EEEV) are endemic. The objective of this study is to detect these viruses in brain samples from equines displaying nervous signs. For this purpose, four published universal RT-PCR methods were compared. The most sensitive and specific RT-PCR method was used to test a total of 70 brain samples, including 40 from bovines and 30 from equines, all exhibiting nervous signs. In the positive cases, eight different brain regions were extracted and tested using this RT-PCR. Positive cases were confirmed through sequencing. Among the four universal RT-PCR assays, Torii RT-PCR demonstrated the highest sensitivity and specificity for diagnosing VEEV and EEEV. Not all assessed brain regions showed DNA amplification. None of the bovine brains was positive and out of the 30 equine brain samples, only four tested positive, and sequencing confirmed two of these samples as VEEV subtype IE. Torii RT-PCR successfully detected VEEV in pools of the hip-pocampus, spinal cord, and basal nuclei, making these brain regions suitable for diagnosing this virus. None of the samples were positive for EEEV.

Alphaviruses are arboviruses that cause arthritis and encephalitis in humans. Eastern Equine Encephalitis Virus (EEEV) is a mosquito transmitted alphavirus that is implicated in severe encephalitis in humans with high mortality. However, limited insights are available into its fundamental biology of EEEV and residue-level details of its interactions with host proteins. In recent years, outbreaks of EEEV have been reported mainly in the United States, raising concerns about public safety. This review article summarizes recent advances in the structural biology of EEEV based mainly on recent single particle cryogenic electron microscopy (cryoEM) structures. Together with functional analyses of EEEV and related alphaviruses, these structural investigations provide clues to how EEEV interacts with host proteins, which may open avenues for the development of therapeutics.

Redox regulation and oxidative stress have become areas of major interest in spermatology. Alteration of redox homeostasis is recognized as a significant cause of male factor infertility and is behind the damage that spermatozoa experience after freezing and thawing or conservation in a liquid state. While for a long time, oxidative stress was just considered an overproduction of ROS, nowadays it is considered as a consequence of redox deregulation. Many essential aspects of spermatozoa functionality are redox regulated, with reversible oxidation of thiols in cysteine residues of key proteins acting as an “on-off” switch controlling spermatic function. However, if deregulation occurs, these residues may experience irreversible oxidation and oxidative stress leading to spermatic malfunction and ultimately death. Stallion spermatozoa are “professional producers” of ROS due to their intense mitochondrial activity, and thus sophisticated systems to control redox homeostasis are also characteristic of this species. As a result, combined with the fact that embryos can easily be collected in this species, horses are a good model for the study of redox biology in the spermatozoa and its impact on the embryo.

Gastric pH seems to be a key factor for the development of Equine Gastric Ulcer Syndrome (EGUS). It is influenced by the amount and duration of roughage uptake, as chewing increases the production of alkaline saliva. The protone-pump-inhibitor omeprazole is currently recommended by the American College of Veterinary Internal Medicine/European College of Equine Internal Medicine (ACVIM/ECEIM) consensus statement for therapy of both Equine Squamous Gastric Disease (ESGD) and Equine Glandular Gastric Disease (EGGD). We hypothesized that dental disorders decrease saliva production due to altered chewing and in consequence gastric pH, therefore predisposing horses to EGUS. In 54 horses, a gastroscopy with measurement of the intragastric pH after 12 hours of fasting and an examination of the oral cavity were carried out. Omeprazole therapy was recommended (n=32), dental disorders were corrected (n=22) and gastroscopy was repeated 4 weeks later (n=9). No influence of moderate-severe dental disorders on ESGD (grade ≥2/4) (p = 0.394) and EGGD (grade ≥2/3) (p = 0.857) could be determined. Nevertheless, there was a trend that moderate- severe dental disorders were associated with a low gastric pH (1-4) (p = 0.100). In conclusion, we found no evidence that gastroscopy should be recommend in horses with dental disorders and vice versa.

Colic, a primary cause of illness and death in horses, necessitates improved prognostic tools. We assessed the predictive value of physiological parameters in colic prognosis. A comprehensive clinical evaluation encompassed heart rate, mucous membranes, capillary refill time, rectal temperature, respiratory rate, gut motility, reflux, and limb pulse strength. Results reveal key predictors of fatal outcomes in colic-affected horses. Stomach reflux, absence of gut noises, and increased heart rate (mean increase of 12 bpm) strongly correlate with a poor prognosis. Prolonged capillary refill (mean increase of 3 seconds), elevated packed cell volume (mean increase of 4%), and blood lactate levels (mean increase of 5 mmol/L) underscore the significance of these markers. Notably, blood lactate (p < 0.001), gut noises (p < 0.05), and heart rate (p < 0.001) exhibit the highest predictive significance based on statistical analysis. Future research should explore the prognostic potential of additional parameters and assess the impact of recommended treatments on colic prognosis. This data-driven study emphasizes the critical role of early recognition and thorough assessment in colic cases, offering vital insights into improving equine healthcare and mitigating mortality rates.

Equines play an important role throughout the world, whether for work, culture, or leisure, providing an ever-growing significant contribution on the economy. The increase in importation and movement of horses, both nationally and internationally, has inevitably allowed for the global equine industry to grow. Subsequently however, the potential for transmission of fatal equine bacterial diseases has also escalated, and devasting outbreaks continue to occur. To prevent such events, disease surveillance and diagnosis must be heightened throughout the industry. Current common, or "gold-standard" techniques, have shown to be inadequate at times, thus requiring newer technology to impede outbreaks. Loop-mediated isothermal amplification (LAMP) has proven to be a reliable, rapid, and accessible tool in both diagnostics and surveillance. This review will discuss equine bacterial diseases of biosecurity relevance and their current diagnostic approaches, as well as their respective LAMP assay developments. Additionally, we will provide insight regarding newer technology and advancements associated with this technique, and their potential use for the outlined diseases.

Training has a significative effect on physiology of blood coagulation both in humans and in horses. Several hemostatic changes have been reported after exercise in the horse but data available are inconclusive. The aim of this study was to investigate the platelet activation and primary platelet-related hemostasis modifications in young untrained Thoroughbred racehorses in the first 4-month timeframe incremental training period in order to improve knowledge on this topic. Twenty-nine clinically healthy, untrained, 2-year-old Thoroughbred racehorses. Never trained Thoroughbreds were followed during their incremental 4-month sprint exercise schedule. Blood collection was performed once a month (T-30, T0, T30, T60, and T90). Platelet aggregation was measured by light transmission aggregometer in response to various agonists: adenosine diphosphate (ADP), collagen and calcium ionophore A23187. Platelet function was evaluated using the Platelet Function Analyzer (PFA-100®) using collagen/ADP and collagen/adrenaline cartridges. Nitrite-nitrate (NOx) plasma concentrations were measured by a colorimetric assay to assess in vivo nitric oxide bioavailability. Platelet activation was investigated also through gene expression analyses (selectin P-SELP, ectonucleotidase CD39-ENTPD1, prostaglandin I2 synthase-PTGIS, endothelial nitric oxide synthase 3-NOS3). Differences among the time points were analyzed and mean±SEM were calculated. Significant modifications were identified compared with T-30 with increase of platelet aggregation (collagen:32.6±4.8 vs 21.6±4.9%; ADP: 35.5±2.0 vs 24.5±3.1%; A23187: 30±4.7 vs 23.8±4%) and a shorter closure time of C-ADP cartridges (75.6±4.4 vs 87.7±3.4 sec) that tended to return to baseline value at T90. NOx concentrations in plasma significantly increased after 30 days of training program compared with baseline. The first long-term training period induce platelet hyperactivity already after 30 days in untrained Thoroughbreds. Regular physical training blunts the negative effects of acute efforts on platelets activation.

The Receptor Binding Domain (RBD) of SARS-CoV-2, the virus responsible for the COVID-19 pandemic, is the functional region of the viral Spike protein (S), which is involved in cell attachment to target cells. The virus has accumulated progressively mutations in its genome, particularly in the RBD region, many of them associated with immune evasion to the host neutralizing antibodies. Some of the viral lineages derived from this evolution, have been classified as Variant of Interest (VOI) or Concern (VOC). The neutralizing capacity of a F(ab’)2 preparation from sera of horses immunized with viral RBD was evaluated, by lytic plaque reduction assay, against different SARS-CoV-2 variants. A F(ab’)2 preparation of an hyperimmune serum with 15 immunizations with RBD, exhibited a high titer of neutralizing antibodies against the ancestral-like strain (1/18,528). A reduction in the title of the F(ab’)2 preparation was observed against the different variants tested. The highest reduction was observed for the Omicron VOC (4.7 fold), followed by the Mu VOI (2.6), Delta VOC (1.8 fold) and Gamma VOC (1.5). Even if a progressive reduction in the neutralizing antibodies titer against the different variants evaluated was observed, the serum still exhibited a significant neutralizing titer, against the Mu VOI and the Omicron VOC (1/7113 and 1/3918 respectively), the strains evaluated most resistant to neutralization.

The apicomplexan parasite Theileria haneyi is one of two known causative agents of equine theileriosis. It causes milder clinical disease than its more virulent counterpart, Theileria equi, in experimentally infected horses, and can superinfect T. equi-positive horses. The current EMA1-based competitive ELISA used in the U.S. to detect equine theileriosis detects T. equi but not T. haneyi, and the complexity of molecular assays precludes widespread use for epidemiologic studies. In order to facilitate urgently needed studies on the prevalence of T. haneyi, the goal of this study was to develop a sensitive and specific serologic assay for the diagnosis of T. haneyi based on the equi mero-zoite antigen 11 (ThEMA11). To achieve this objective, ThEMA11 was recombinantly expressed in eukaryotic cells and its antigenicity assessed using sera from T. haneyi-experimentally infected horses. Confirmation of sera reactiv-ity enabled design and optimization of an indirect ELISA. Specificity of the ELISA for T. haneyi was assessed using a cohort of sera from horses experimentally infected and confirmed PCR-positive for either T. equi or T. haneyi. Data from field samples further demonstrate that the ThEMA11 ELISA is capable of identifying T. haneyi antibodies in horses from multiple continents around the world.

The mechanisms of the innate immunity control on equine infectious anaemia virus in asymptomatic horses are not yet widely described. Horse monocytes from peripheral blood can be differentiated in vitro into macrophages, that generally gives rise to mixed populations, morphologically referable to M1 and M2. The addition of specific cytokines or of virus, results in a more specific differentiation. As stated for other species (Crespo et al, 2013), stimulation with equine recombinant cytokine IFNγ polarized horse macrophages towards the M1 phenotype while equine recombinant cytokine IL4 polarized towards the M2 phenotype. EIAV Wyoming and Miami strains infection resulted in morphological transformation of macrophages compatible with the M1 pattern of differentiation. In this study MMP13 represented a reliable target gene to evaluate proinflammatory status of macrophages in horses since IFNγ and EIAV infection considerably increased its expression. A more in-depth study of the expression genes of both cytokine-induced and virus-induced markers of eMDM polarization may help us to understand whether these are the same in the horse as those found in other animal species with similar modes of activation of innate immunity. The identification of the markers of each population of macrophages would allow to analyze the differentiation profiles, and to check the control of virus infectivity in equid populations envisioning therapeutic strategies through the use of this information.

In this paper, we report a comprehensive and consistent annotation of the locus encoding the horse T cell receptor b-chain (TRB), as inferred from the most recent genome assembly. The horse TRB locus spans approximately 1 Mb, making it the largest locus among the mammalian species stud-ied to date, with a significantly higher number of genes related to extensive duplicative events. In the region, 136 TRBV, belonging to 29 subgroups, 2 TRBD, 13 TRBJ and 2 TRBC genes, were iden-tified. The general genomic organization resembles that of other mammals, with a V-cluster of 135 TRBV genes, located upstream of two in-tandem aligned TRBD-J-C clusters, and an inverted TRBV gene at the 3’ end of the last TRBC gene. However, the horse b-chain repertoire would be affected by a high number of non-functional TRBV genes. Thus, we queried a transcriptomic da-taset derived from splenic tissue of a healthy adult horse, using each TRBJ gene as a probe to ana-lyse clonotypes encompassing the V(D)J junction. This analysis provided insights into the usage of the TRBV, TRBD and TRBJ genes and the variability of the juxtaposition region. Our results clearly demonstrated that the horse β-chain constitutes a complex level of variability broadly like that described in other mammalian species.

Eastern (EEEV) and Venezuelan (VEEV) equine encephalitis viruses (EEVs) are related, (+)ssRNA arboviruses that can cause severe, sometimes fatal, encephalitis in humans. EEVs are highly infectious when aerosolized, raising concerns for potential use as biological weapons. No licensed medical countermeasures exist; given the severity/rarity of natural EEV infections, efficacy studies require animal models. Cynomolgus macaques exposed to EEV aerosols develop fever, encephalitis, and other clinical signs similar to humans. Fever is nonspecific for encephalitis in macaques. Electrocardiography (ECG) metrics may predict onset, severity, or outcome of EEV-attributable disease. Macaques were implanted with thermometry/ECG radiotransmitters and exposed to aerosolized EEV. Data was collected continuously, and repeated-measures ANOVA and frequency-spectrum analyses identified differences between courses of illness and between pre-exposure and post-exposure states. EEEV-infected macaques manifested widened QRS-intervals in severely ill subjects post-exposure. Moreover, QT-intervals and RR-intervals decreased during the febrile period. VEEV-infected macaques suffered decreased QT-intervals and RR-intervals with fever onset. Frequency-spectrum analyses revealed differences in the fundamental frequencies of multiple metrics in the post-exposure and febrile periods compared to baseline and confirmed circadian dysfunction. Heart rate variability (HRV) analyses revealed diminished variability post-exposure. These analyses support using ECG data alongside fever and clinical laboratory findings for evaluating medical countermeasure efficacy.

Recently, we isolated and characterized a new pool of mesenchymal stem cells- HPC (Hoof Progenitor Cells), which reside in the coronary corium of equine hooves. We hypothesize, that due to their location, HPC may be involved in the pathogenesis of laminitis, and could possibly be utilized in its treatment. The aim of this study was to compare the newly described HPC to a well-established cell pool-ASC (Adipose Stem Cells). The two cell populations were maintained either in standard culture conditions or under a hyperinsulinemic milieu as hyperinsulinemia is often observed alongside laminitis. Cell cycle dynamics, mitochondrial membrane potential and oxidative stress were analyzed with microcapillary cytometry. Furthermore, the expression of key markers of mitochondrial metabolism, oxidative stress, apoptosis, immunomodulation and insulin signalling were analyzed with the RT-qPCR method. Additionally, a selected panel of miRNA was analyzed with the RT-qPCR method. In standard culture conditions, ASC exhibit higher proliferation than HPC while in hyperinsulinemia a shift towards S phase was observed in both populations. As for the mitochondrial membrane potential, in standard culture conditions, HPC was characterized by a higher percent of live cells. Hyperinsulinemia did not strongly affect the mitochondrial membrane potential of the analyzed populations. Our findings indicate that HPC are characterized by higher activity in terms of mitochondrial dynamics, mitophagy, and mitochondrial biogenesis under standard culture conditions. Yet in both populations, a diminished mitochondrial dynamic and metabolism was observed in hyperinsulinemia. The analysis of oxidative stress revealed, that HPC might possess somewhat higher resistance towards oxidative stress than ASC. Minimal change was observed in the expression of key markers of apoptosis in ASC and HPC under hyperinsulinemia. The analysis of key immunomodulatory markers indicated that HPC may help the cytokine storm for a more effective phenotype change to an immunosuppressing pool of cells with better migratory and healing abilities under hyperinsulinemia than ASC. As for insulin signalling, the obtained data suggests that HPC might be more resistant to hyperinsulinemia and may have greater therapeutic potential in reducing laminitis. Obtained results demonstrated that HPC possesses better potential to modulate the PTEN-AKT pathway by miR-21 and reverse high glucose and high insulin-induced insulin resistance. Taken together, the obtained results indicate that HPC may be a very promising cell pool with therapeutic potential in laminitis treatment, and could be more effective than ASC.

This study was conducted to investigate bit use in equestrian sports, focusing on welfare and performance as bit use has been subject of debate. The multi-faceted study combines a literature review, survey, and video analyses. Research on physiological and behavioral effects of bit use, discussing concerns and benefits, was evaluated. The survey involved 250 equestrians, gathering data on experiences, opinions, and practices regarding bit usage. Results demonstrate that different bits are used during competitions and that riders prioritize factors like horse satisfaction, consistent connection, and chewing behavior when riding at home, while show use focusses on factors like control. Video analyses showed high conformity between veterinary expert and show jumper, indicating reliability and validity. Pelham and loose ring bits were most used, while 3-ring and full cheek bits were associated with the most aversive movements, particularly opening of the mouth. Other defense movements included putting ears back, tail swishing, and head tilting. The study highlighted the need for understanding bit preferences in various settings and cautioned against generalizing bit usage based solely on show observations. It also emphasized the potential for developing new designs that prioritize horse comfort and responsiveness, to enhance communication and partnership between horses and riders.

Equine-assisted services (EAS) are being increasingly used as complementary interventions for military veterans who have experienced trauma. However, there is limited evidence of benefit for this population and almost no literature describing desired potential outcomes and possible mechanisms of action. The aim of this article is to address these gaps by reviewing the extant literature of animal-assisted interventions in general, and equine-assisted services in particular, with the goal of providing guidance for future investigations in the field. Currently, the field is in the early stage of scientific development, but published results are promising. Interventions that enhance treatment compliance and/or outcomes could benefit this population. Preliminary results, reviewed herein, indicate that EAS interventions might benefit the military veteran population by enhancing treatment engagement and therapeutic alliance, as well as contributing to symptom reduction and resulting in various transdiagnostic benefits. It is recommended that future studies include exploration of potential beneficial outcomes discussed herein as well as investigate suggested mechanisms of action.

Equine Assisted Activities (EAA) are considered a suitable innovative rehabilitative practice for children with neurodevelopmental disorders, including Autism Spectrum Disorder (ASD). While standardized scales have been previously used as a tool to evaluate the effects of EAA on different domains of functioning in ASD children, few studies have considered an ethological approach as a means to describe human-horse interactions in the context of ASD. In this study, we aimed to evaluate the behaviour of 19 children with ASD - in comparison with 19 typically developing children (TD) - during EAA sessions. We developed an ethogram from the video-recordings, to assess spatial relationships, social interactions and communicative behaviours displayed by the child towards the horse, as well as the occurrence of problem behaviours. Results indicate that children’s behaviours during EAA sessions are modulated by sex and age, while previous children’s experience with EAA appeared to improve interpersonal distance and horse handling. Results from this study highlight the importance of exploring children's behavioural response during animal-assisted activities through direct measurements. This may allow linking the quality/strength of the child-horse relationship to the benefits obtained by the child, particularly in the social/communicative domain, a core symptom of ASD.

The irregular timing and spatial variation in zoonotic arbovirus spillover from vertebrate hosts to humans and livestock present challenges to predicting their occurrence from year to year and within their broader geographic range, compromising effective prevention and control strategies. The objective of this study was to quantify effects of landscape composition and configuration and dynamic temperature and precipitation values on the 2018 spatiotemporal distribution of eastern equine encephalitis virus (EEEV) (Togaviridae, Alphavirus) and West Nile virus (WNV) (Flaviviridae, Flavivirus) sentinel chicken seroconversion in northeastern Florida using Earth Observation (EO) data and a modeling framework that incorporated joint spatial and temporal effects. We investigated environmental effects using Bernoulli generalized linear mixed effects models (GLMMs) including a site level random effect, and then added spatial random effects and spatiotemporal random effects in subsequent runs. Models were executed using integrated nested Laplace approximation (INLA) and a stochastic partial differential equation (SPDE) approach in R-INLA. GLMMs that included a spatiotemporal random effect performed better relative to models that included only spatial random effects and better than non-spatial models. Results indicated strong spatiotemporal structure in seroconversion for both viruses, but EEEV exhibited more punctuated and compact structure at the beginning of the sampling season, while WNV exhibited more gradual and diffuse structure across the study area toward the end of the sampling season. Percentage of cypress/tupelo wetland land cover within 3500 m of coop sites and edge density of forest land cover within 500 m had a strong positive effect on EEEV seroconversion, while the best fitting model for WNV was the intercept only model with spatiotemporal random effects. Lagged temperature and precipitation variables included in our study did not have a strong effect on seroconversion for either virus when accounting for temporal autocorrelation, demonstrating the utility of capturing this structure to avoid Type I errors. Predictive accuracy on out-of-sample data for EEEV seroconversion demonstrates the potential to develop a temporally dynamic framework to predict arbovirus transmission.

Musculoskeletal injuries such as equine osteoarthritis, osteoarticular defects, tendonitis/desmitis, and muscular disorders are prevalent among sport horses, with a fair prognosis for returning to exercise or previous performance levels. The field of equine medicine has witnessed rapid and fruitful development, resulting in a diverse range of therapeutic options for musculoskeletal problems. Staying abreast of these advancements can be challenging, prompting the need for a comprehensive review of commonly used and recent treatments. The aim is to compile current therapeutic options for managing these injuries, spanning from simple to complex physiotherapy techniques, conservative treatments including steroidal and non-steroidal anti-inflammatory drugs, hyaluronic acid, and polyacrylamides, to the promising regenerative therapies such as hemoderivatives and stem cell-based therapies. Each therapeutic modality is scrutinized for its benefits, limitations, and potential synergistic actions to facilitate their most effective application for intended healing/regeneration of the injured tissue/organ and subsequent patient recovery. While stem cell-based therapies emerge as particularly promising for equine musculoskeletal injuries, a multidisciplinary approach is underscored throughout the discussion, emphasizing the importance of considering various therapeutic modalities in tandem.

Children’s beneficial relationships with animals are well known. Companion animals, particularly dogs have become an integral part of family life and children’s material culture. Aside the proven physiological benefits there is little research about what children say about their relationships with animals and how they describe them. In this paper we bring together both horse-human and dog-human interactions, finding common ground for understanding the complexity of human development, well-being and flourishing. Dogs in schools are fast becoming a trend in helping support and enhance children’s learning as well as their social and emotional well-being. Studies have shown that the very presence of a dog can increase children’s concentration, executive function and behavior. Also, equine therapy is gaining momentum and empirical studies are showing noteworthy benefits to children and young people. However, the lack of children’s voices means that the mechanisms for these benefits are somewhat unknown and unclear. In seeking to explore this, the authors utilize a visual, sensory and diffractive ethnographic approach to illuminate and illustrate, experiment and re-enact, how the children relate, share spaces and multiple subjectivities with their classroom canine, “Ted” and companion horse “Henry”. ”Henry” is part of a programme in which youngsters care for and engage in activities with horses.

Fatigue-related subchondral bone injury of the third metacarpal/ metatarsal (McIII/ MtIII) bone is a common cause of wastage and welfare concern in racehorses. Better understanding of bone health and strength would improve animal welfare and be of benefit for the racing industry. Porosity index (PI) of bone is an indirect measure of osseous pore size and number and is therefore an interesting indicator of bone strength. MRI of compact bone using traditional methods even with short echo times fails to generate enough signal to assess bone architecture as the water protons are tightly bound. Ultrashort echo time (UTE) sequences aim to increase the amount of signal detected in the equine McIII/MtIII condyle. Cadaver specimens were imaged using a novel dual-echo UTE MRI technique and PI was calculated and validated against quantitative CT-derived bone mineral density (BMD). BMD and PI are inversely correlated in the equine distal Mc/ MtIII with a mean r value of -0.29. The r value is significantly different between fore- and hindlimbs. Further work is needed to assess how correlation patterns behave in different areas of the bone and to evaluate PI in horses with and without clinically relevant stress injury.

Articular cartilage healing and repair is one of the biggest clinical challenges in trauma and orthopaedic medicine, leading to high economic loss due to athletic impairment in sport horses. Multipotent Mesenchymal stem/stromal cells are part of novel therapeutic arsenal aimed at tissue regeneration and restoring organ function. These cells release biomolecules and factors in the culture media, producing the so called conditioned medium, which have a pro-regenerative effect through paracrine mechanisms. Considering this, a therapeutic product combining equine synovial membrane derived Mesenchymal stem/stromal cells and equine umbilical cord stroma-derived Mesenchymal stem/stromal cells conditioned medium was employed. In this work the treatment of two articular defects, in the first phalanx and third metacarpus of the left front limb of a five-year-old horse, is described. Following surgical debridement of the lesions, the treatment comprised two intra-articular administrations of the above-mentioned therapeutic combination, 15 days apart. This was followed by a thorough rehabilitation program. A second-round treatment was performed 6 months later. Diagnostic imaging demonstrated significant improvement of the articular defects. The clinical outcome allowed a full return to the athletic performance demonstrating the safety and potential advantages of this approach when compared with other therapeutic options.

Parasite control strategies such as blanket anthelmintic treatment of all horses in a herd or interval treatment regimen based on egg reappearance remain widely followed practices for the last several decades. These practices have resulted in the fast emergence of resistance in equine strongyles to all major classes of drugs on the market. In this regard, the guidelines set by the American Association of Equine Practitioners advocate evidence-based strongyle control in horses. It recommends targeted treatment of all heavy egg shedders [&gt; 500 eggs per gram (EPG) of feces] while the low shedders (0-200 EPG) are left untreated to maintain susceptible parasite refugia in a herd. As 50-75% of adult horses in a herd are low shedders, preventing them from unnecessary anthelmintic exposure is critical to tackling resistance. There are various fecal egg count (FEC) techniques with many modifications and variations in use, but none is identified as a gold standard and methods comparison studies are lacking. We hypothesized that the diagnostic performance of commonly used quantitation methodologies differs and the FEC methods must be evaluated to determine the gold standard. In this regard, we performed methods comparison studies using polystyrene beads with size and specific gravity comparable to strongyle eggs, as proxy. The linearity of 12 commonly used FEC methodologies (3 techniques with 4 different solutions) was studied using bead standards in the clinically applicable range (63 to 1,000) to determine their suitability for strongyle egg counts. Deming regression analysis identified Mini-FLOTAC with sugar (specific gravity 1.33) as the test with the greatest coefficient of determination. All tests underestimated the true bead count, and we determined a correction factor (CF) for each test to estimate the true count. CF negates the preference for a gold standard test. Finally, we analyzed the validity of CF for 5 tests with R2&gt;0.95 to accurately quantify strongyle eggs from 40 different horses. Overall, this study identified methodologies with the highest diagnostic performance and the limitations to standardizing routine FEC tests to promote uniformity in implementing AAEP parasite control guidelines.