Invitro antibacterial activity of bark, leaf and root extracts of combretum molle plant against streptococcus equi isolated from clinical cases of strangles in donkeys and horses.

BACKGROUND: Effective therapy for many infections is becoming difficult due to the evolutionary development of drug resistance, and hence, the development of alternative treatment options mainly from herbs is crucial. The objective of this study was to investigate the antibacterial effects of ethanol extracts of stem bark, leaves and roots of Combretum molle against Streptococcus equi isolated from clinical cases of strangles using in vitro tests. METHODS: Plant extraction was performed using a maceration technique with 80% ethanol. The mean zone of inhibition was determined using the agar well diffusion method. Six serial dilutions with different concentrations (10%, 5%, 2.5%, 1.25%, 0.625% and 0.3125%) of each plant extract were prepared using dimethyl sulfoxide (DMSO). A modified agar microdilution method was used to determine the minimum inhibitory concentration (MICs) of the extracts. RESULTS: The results revealed that all plant extracts showed significant antibacterial activity. The root extract showed the best antibacterial effect compared to the others at all concentrations, with MZI values of 27.5, 23.225, 20.5, 17.9, 15.65 and 12.25 for the respective concentrations mentioned above and an MIC of 250 µg/ml. It was followed by the stem bark extract, which had MZI values of 24.67, 22.35, 18.225, 16.175, 11.125 and 8.2 millimeters and an MIC of 375 µg/ml. The leaf extract also had significant activity, with MZI values of 20.175, 18.25, 15.7, 13.125, 9.4 and 6.75 in millimeters and an MIC of 500 µg/ml. There was a direct relationship between the concentrations of the plant extracts and the level of inhibition. CONCLUSION: The test plant extracts were compared with the conventional antibiotic penicillin G, and the results indicated that the parts of the test plant have significant antibacterial activity, which may support traditional claims and could be candidates for alternative drug discoveries.

Characterizing the relationship between gonadotropin releasing hormone (GnRH), kisspeptin, and RFamide related peptide 3 (RFRP-3) neurons in the equine hypothalamus across the estrous cycle and in the anovulatory seasons.

To understand better the role that kisspeptin plays in regulating seasonal and estrous cycle changes in the mare, this study investigated the number, location and interactions between GnRH, kisspeptin and RFRP-3 neurons in the equine hypothalamus. Hypothalami were collected from mares during the non-breeding season, vernal transition and various stages of the breeding season. Fluorescent immunohistochemistry was used to label the neuropeptides of interest. GnRH cells were observed primarily in the arcuate nucleus (ARC), while very few labeled cells were identified in the pre-optic area (POA). Kisspeptin cells were identified primarily in the ARC, with a small number of cells observed dorsal to the ARC, surrounding the third ventricle (3V). The mean number of kisspeptin cells varied between animals and typically showed no pattern associated with season or stage of estrous cycle, but a seasonal difference was identified in the ARC population. Small numbers of RFRP-3 cells were observed in the ARC, ventromedial hypothalamus (VMH) and dorsomedial hypothalamus (DMH). The mean number of RFRP-3 cells appeared higher in pre-ovulatory animals compared to all other stages. The percentage of GnRH cell bodies with kisspeptin appositions did not change with season or stage of estrous cycle. The percentage of kisspeptin cells receiving inputs from RFRP-3 fibers did not vary with season or stage of estrous cycle. These interactions suggest the possibility of the presence of an ultra-short loop feedback system between these three peptides. The changes in RFRP-3 neurons suggest the possibility of a role in the regulation of reproduction in the horse, but it is unlikely to be as a gonadotropin inhibitory factor.

Development and pilot of equine facilitated physical therapy outcome measure tool for chronic low back pain patients.

Equine Facilitated Physical Therapy (EFPT) lacks consistent documentation due to being an unconventional physical therapy treatment to chronic low back pain patients (LBP) and lacking rehabilitation outcome measure tools for a stable (equestrian) environment. The objectives were to develop an online evaluation tool as well as to define inter- and intra-rater reliability to validate the outcome measurement tool "Evaluation of maintaining sitting position (on a horse) and walking (short distances)" designed for LBP patients in EFPT". A total of 48 movement related functions (n = 48), were derived from the International Classification of Functioning (ICF) and organized to an online evaluation tool. Depending on the state of validation two to six (2-6) raters scored randomized patient (n = 22) video material, recorded during a 12-week EFPT intervention, with the designed tool. Inter-rater agreement level between the experts reached good (α = 87) reliability for the scoring of the items and calculated per patient excellent (α = 100). Intra-rater reliability reached good (α = 87) and per patient good (α = 80) repeatability. For the healthy adults the reliability between raters reached acceptable (α = 72) levels and per rated excellent (α = 100). The developed assessment tool was found satisfactory to fulfil the requirement for the therapeutic practice. With the use of the tool physical therapist may detect postural changes for LBP patients as outcome report in EFPT. The tool may be used to identify treatment progress and to help design home exercises. The created tool will help to collect similar outcome measures from LBP patients in EFPT and to validate the treatment within industry.

Experiences of a Therapeutic Equine-Assisted Group Intervention for People with Common Mental Disorders on Sick-Leave.

Common mental disorders such as anxiety, depression and stress-related disorders are increasing worldwide, resulting in long-term sick leave and lower quality of life. Traditional treatment is often insufficient to facilitate the recovery process, and the need for holistic interventions that enable successful recovery is evident. Equine-assisted interventions have shown promising results in health promotion among people with mental disorders, and further research is needed to implement them within the range of available care. The aim of the study is therefore to describe experiences of a therapeutic equine-assisted group intervention for people with common mental disorders on sick leave. The study has a descriptive qualitative design with an inductive approach. Ten participants with common mental disorders on sick leave were interviewed after participating in a 12-week equine-assisted intervention and the data were analysed with conventional content analysis. The analysis generated the overall theme "The equine-assisted group intervention facilitates recovery," which summons the four categories "The environment of the horses contributes to relaxation and an ability to be in the present," "The presence of the horses facilitates supportive relationships in the group," "Interplay with the horses entails physical activity and bodily improvements" and "Emotional interactions with the horses strengthen inner power." In conclusion, equine-assisted interventions have the potential to facilitate recovery for people with common mental disorders by providing relaxation, promoting the ability to be in the present, cultivating supportive relationships and providing physical activity and bodily improvements as well as emotional interactions that strengthen the participants' inner power.

Effectiveness of photo-ozone therapy against equine Pythium insidiosum.

Cutaneous pythiosis is a life-threatening infectious disease. Low-level laser therapy (LLLT) and ozone (O3) have been used individually in the treatment of infected wounds. The goals of the study were a) to characterize the antimicrobial action of the photo-ozone therapy (LLLT-O3) against equine Pythium insidiosum, and b) to assess the cytotoxic potential of the LLLT-O3 in keratinocytes. Specimens of pathogen were isolated from 10 horses. After culturing, 120 hyphae plugs were distributed among four groups (n=30 hyphae plugs/group): LLLT (laser irradiation for 160 sec;), O3 (exposition to O3 for 15 min;), LLLT-O3 (LLLT and O3 treatments in sequence) and control (untreated plugs). The hyphae growth was measured during the first 14 days post-treatment. Where there was an absence of hyphae growth, the plug was recultured for an additional 7 days. The cytotoxic potential of the treatments against HaCaT keratinocytes was assessed by colorimetric assays. The LLLT-O3 and O3 treatments inactivated, respectively, 92.3% (28/30) and 30% (9/30) of the samples. No growth was detected after 7 days reculture of inactivated hyphae plugs on new media. Hyphae growth was visualized in 100% of the control and LLLT hyphae plugs. The viability of HaCaT cells was not affected by the isolated treatments (LLLT and O3), while the LLLT-O3 showed slight cytotoxic effect (20%) when compared to the control group (P<0.05). Photo-ozone therapy inactivated equine P. insidiosum hyphae with minimal cytotoxicity in skin cells in vitro.

The effect of casing and gypsum on the yield and psychoactive tryptamine content of Psilocybe cubensis (Earle) Singer.

Psychedelic fungi have experienced a surge in interest in recent years. Most notably, the fungal secondary metabolite psilocybin has shown tremendous promise in the treatment of various psychiatric disorders. The mushroom species that produce this molecule are poorly understood. Here we sought to examine for the first time, the response of a psilocybin-producing species Psilocybe cubensis to casing (peat moss and vermiculite) and supplementation with gypsum (calcium sulfate dihydrate), two common practices in commercial mushroom cultivation. Mycelial samples of genetically authenticated P. cubensis were used to inoculate popcorn grain bags. The fully colonized bags of popcorn grain (0.15 kg) were transferred to bins of 0.85 kg pasteurized horse manure, with or without 1 cm thick layer of casing and/or 5 % gypsum. Our results indicate that the use of a casing layer significantly increases the biological efficiency (161.5 %), by approximately four fold, in comparison to control (40.5 %), albeit with a slight delay (∼2 days) for obtaining fruiting bodies and a somewhat reduced total tryptamine content (0.85 %) as gauged by High Performance Liquid Chromatography measurements. Supplementation with both casing and gypsum, however, appears to promote maximal yields (896.6 g/kg of dried substrate), with a biological efficiency of 89.6 %, while also maintaining high total tryptamine expressions (0.95 %). These findings, revealing methods for maximizing yield of harvest and expressions of psychoactive tryptamines, may prove useful for both home growers and commercial cultivators of this species, and ultimately support the growth of a robust industry with high quality natural products.

Multiple Equine Therapies for the Treatment of Gross Motor Function in Children with Cerebral Palsy: A Systematic Review.

PURPOSE: This study aimed to evaluate the impact of three equine therapy approaches on gross motor function in children with cerebral palsy. METHODS: The studies were retrieved from PubMed, Web of Science, Science Direct, and the Cochrane Library, in accordance with the style commonly found in scientific journal publications:(1) peer-reviewed articles written in English; (2) experimental or quasi-experimental; (3) three Equine Therapy Interventions as experiment's independent variable; (4) children with cerebral palsy; and (5) measurement of outcomes related to Gross Motor Function. RESULTS: The study examined 596 patients with cerebral palsy, whose average age was 8.03 years. The three types of horse therapy interventions had a significant impact on gross motor function in children with cerebral palsy (SMD = 0.19, 95% CI 0.02-0.36, p = 0.031). Additionally, the interventions positively affected dimensions C (SMD = 0.31, 95% CI 0.00-0.62, p = 0.05), D (SMD = 0.30, 95% CI 0.06-0.56, p = 0.017), and B (SMD = 0.72, 95% CI 0.10-1.34, p = 0.023). The Gross Motor Function Measure (GMFM) consists of 88 or 66 items, which are divided into five functional dimensions: GMFM-A (lying down and rolling), GMFM-B (sitting), GMFM-C (crawling and kneeling), GMFM-D (standing), and GMFM-E (walking, running, and jumping). Each subsection of the GMFM can be used separately to evaluate motor changes in a specific dimension of interest. Subgroup analysis revealed that different horse-assisted therapy approaches, types of cerebral palsy, exercise duration, frequency, and intervention periods are important factors influencing treatment outcomes. CONCLUSION: The intervention period ranged from 8 to 12 weeks, with session durations of 30 to 45 minutes, 2 to 3 times per week. Equine-assisted therapy (EAT) demonstrated significant improvements in the overall gross motor function score, Dimension B, Dimension C, and Dimension D among children with cerebral palsy. The most effective treatment is provided by Equine-Assisted Therapy, followed by Horseback Riding Simulator (HRS). Due to its economic practicality, HRS plays an irreplaceable role. CLINICAL EVIDENCE: Equine-Assisted Therapy (EAT) demonstrates the most effective treatment outcomes, suggesting that hospitals and healthcare professionals can form specialized teams to provide rehabilitation guidance. 2. Within equine-assisted therapy, Horseback Riding Simulator (HRS) exhibits treatment efficacy second only to Equine-Assisted Therapy (EAT), making it a cost-effective and practical option worthy of promotion and utilization among healthcare institutions and professionals. 3. In equine-assisted therapy, Therapeutic Horseback Riding (THR) holds certain value in rehabilitation due to its engaging and practical nature.

Growth inhibitory effect of selected medicinal plants from Southern Ethiopia on the mycelial phase of Histoplasma capsulatum var. farciminosum.

BACKGROUND: Epizootic lymphangitis is an infectious and chronically debilitating disease of the equines. Histoplasma capsulatum var. farciminosum, a thermally dimorphic fungi, is the causative agent for the disease. In Ethiopia, the disease significantly affects carthorses, posing threats to animal welfare, and resulting in substantial economic losses. Limited availability of widely accessible antifungals in addition to the chronic nature of the disease is the major challenge against management of epizootic lymphangitis. This study aimed to assess the in vitro efficacy of specific local medicinal plant extracts against the mycelial phase development of H. capsulatum var. farciminosum in southern Ethiopia. The leaves of Xanthium strumarium, Kanda (Family Rubiaceae), Croton macrostachyus (Bisana in Amharic), and Centella Asiatica (Echere waye as a local name in Zeyissegna) that are traditionally used for the treatment of different skin ailments were collected and extracted for the in vitro trial. RESULTS: The study revealed that methanol extracts of Xanthium strumarium, Kanda, Croton macrostachyus, and Centella Asiatica, at minimum inhibitory concentrations of 1.25 mg/ml, 2.5 mg/ml, 2.5 mg/ml, and 5 mg/ml, respectively, inhibited the growth of H. capsulatum var. farciminosum. CONCLUSION: This in vitro finding could serve as significant preliminary data in the exploration of effective alternative treatment options for epizootic lymphangitis. This study provides a crucial foundation for further research aimed at determining the chemical components and in vivo effectiveness of these plant extracts against both the mycelial and yeast forms of Histoplasma capsulatum var. farciminosum.

α-Casozepine supplementation shows no effect on foals' growth and cortisol levels before and after weaning.

OBJECTIVE: The goal of this study was to determine whether oral administration of α-casozepine (Zylkene) supplementation decreases the risk of disease in foals, as measured through salivary cortisol levels, temperature, clinical signs of disease, and body weight before and after weaning. ANIMALS: 20 Thoroughbred foals were observed over 10 weeks, beginning 14 days prior to the time of weaning. METHODS: Experimental foals were randomly selected and given α-casozepine daily for 10 weeks, while the controls received a placebo with the same frequency as the experimental group. The average daily height and weight gain were calculated to examine the effects of α-casozepine before and after weaning. Salivary cortisol levels were also compared before versus after weaning. RESULTS: There was no statistically significant difference in average daily height/weight gain or cortisol levels between the α-casozepine group and the control before and after weaning. However, the α-casozepine group had numerically less of a decline in height and growth compared to the control. This suggests that with further study Zylkene may decrease losses in foal growth rate. CLINICAL RELEVANCE: The weaning process is one of the largest stressors in a young horse's life. This abrupt process can contribute to an increased risk of disease and even morbidity. Other major stressors including separation from the mare, diet change, and new social groups can also lead to animal health and welfare concerns, as well as economic losses in weanling foals. However, further study is required to determine the benefits of α-casozepine on equine stress and health.

A systematic analysis of anthocyanins inhibiting human, murine, and equine herpesviruses.

BACKGROUND: Herpesviruses are common animal and human pathogens that cause severe health problems in children, immunocompromised patients, and infected animals with a host range from fish to mammals. Anthocyanin-containing plant extracts have been described as potent antivirals, which might cause fewer harmful side effects than direct-acting antivirals. Here, we report that an extract of Aristotelia chilensis (Molina) Stuntz (Elaeocarpaceae) (MBE) with a high content of the anthocyanin delphinidin suppresses lytic replication of equine, murine and human herpesviruses of replication in vitro. METHODS: We treated cultured cells with MBE and purified individual anthocyanins present in the extract to determine the most active compound at different concentrations. We subsequently infected the cultures with human herpesviruses 1 (HSV-1) or 8 (HHV-8), murine cytomegalovirus (CMV), or equine herpesviruses 1 (EHV-1) and determined the number of infected cells and viral infectivity. RESULTS: MBE inhibited the HSV-1, murine CMV, and EHV-1 by up to 2 orders of magnitude. In the presence of the stabilizing randomly methylated-beta-cyclodextrin, the inhibitory concentration could be lowered significantly. We identified delphinidin as an active antiviral compound and showed that the non-glycosylated delphinidin solved and stabilized with sulfobutylether-beta-cyclodextrin allowed usage of approximately 50 times lower concentrations. CONCLUSION: Glycosylated delphinidin derivatives were identified as active antiviral compounds of MBE. This suggests that plant extracts rich in delphinidin-anthocyanins have potent antiviral properties that could be used in treatment and prevention.

Impact of apitherapy on canine, equine, and chicken lymphocytes, in vitro.

Apitherapy is a form of alternative medicine that utilizes products from the western honeybee (Apis mellifera), including honey, propolis, and honeybee venom, to improve the health status of human patients by altering host immunity. An added benefit of these products is that they are nutraceuticals and relatively inexpensive to aquire. Currently, little is known about the use of honeybee products in veterinary species, as well as their impact on host immunity. In the present in vitro study, honey, propolis, and honeybee venom were co-cultured with enriched canine, equine, and chicken peripheral blood lymphocytes (PBLs) with cell proliferation, cell viability/apoptosis, and cellular morphology evaluated. Concanavalin A (Con A) and dexamethasone were used as stimulatory and suppressive controls, respectively. Honeybee products' effects on the three veterinary species varied by product and the species. Honey stimulated the PBLs proliferation in all three species but also displayed some increased cytotoxicity. Propolis stimulated proliferation in canine and equine PBLs, however, it suppressed proliferation in the chicken PBLs. Honeybee venom was the strongest PBL stimulant for all three species and in the equine, surpassed the stimulant response of Con A and yet, enhanced PBL cell viability post culture. In summary, the results of this preliminary in vitro study show that these three honeybee products do impact lymphocyte proliferation and viability in dogs, horses, and chickens, and that more research both in vitro and in vivo will be necessary to draw conclusions regarding their future use as immune stimulants or inhibitors.

Efficacy of treatment of equine anhidrosis with acupuncture and Chinese herbs is low but higher in treated horses compared with placebo.

OBJECTIVE: To assess the therapeutic efficacy of acupuncture in combination with Chinese herbs for treatment of horses affected with anhidrosis. ANIMALS: 44 horses affected with anhidrosis for up to 3 years' duration were enrolled. Inclusion required both compatible clinical signs and results of a quantitative intradermal terbutaline sweat test. METHODS: Study horses were randomly allocated into 2 groups. Group 1 (n = 19) was treated with daily Chinese herbs and 4 weekly acupuncture sessions. Group 2 (n = 25) was given daily hay powder as a placebo and 4 weekly sham acupuncture sessions. Horses were tested by quantitative intradermal terbutaline sweat test within 2 days after treatment completion and again 4 weeks following treatment. RESULTS: Terbutaline-induced sweat responses (mg) were not different between groups within 2 days and 4 weeks after treatment. Two days after treatment, ratios of sweat responses (compared to baseline) were higher (P < .05) in the treatment group compared to the placebo group at terbutaline concentrations of 1.0, 100, and 1,000 µg/mL. The number of horses responding to treatment was higher in the treatment group (5/19 [26%]), compared to horses in the placebo group (1/25 [4%]) for 1 of 5 terbutaline concentrations 2 days (10 µg/mL) or 4 weeks (0.1 µg/mL) after treatment. CLINICAL RELEVANCE: Ratios of sweat responses were higher in treatment horses 2 days after treatment, compared to baseline, but not 4 weeks later. The efficacy of a traditional Chinese veterinary medicine protocol for anhidrosis treatment with acupuncture and Chinese herbs was low but higher in treated horses compared with placebo.

Effects of intravenous administration of ascorbic acid (vitamin C) on oxidative status in healthy adult horses.

BACKGROUND: Ascorbic acid (AA) is an antioxidant that might be beneficial for adjunctive treatment of sepsis in horses. The optimal dose and effects on oxidative status are unknown. HYPOTHESIS: Ascorbic acid administration will increase plasma AA concentrations and decrease determinants of reactive oxygen metabolites (dROM), basal and stimulant-induced intraerythrocytic reactive oxygen species (ROS) concentrations, and stimulant-induced neutrophil ROS production, and increase plasma antioxidant capacity (PAC) in a dose-dependent manner. ANIMALS: Eight healthy horses. METHODS: Randomized placebo-controlled crossover study. Each horse received 4 single-dose IV treatments including AA at 25, 50, and 100 mg/kg and saline (placebo) with each treatment separated by ≥1 week. Blood was collected at baseline, 2 and 6 hours for assessment of plasma dROM and PAC via photometer, intraerythrocytic ROS by flow cytometry, and stimulant-induced neutrophil ROS by a fluorometric assay. Plasma AA concentrations were measured by high-performance liquid chromatography/electrochemical detection. RESULTS: Ascorbic acid at 100 mg/kg resulted in decreased dROM 2 hours after treatment (P = .03, 95% CI 5.51-121.2, point estimate 63.3). There was no effect of AA on basal or stimulant-induced intraerythrocytic ROS (P = .88, 95% CI -0.156 to 0.081, point estimate -0.037; P = .93, 95% CI -0.123 to 0.112, point estimate -0.006, respectively), basal or stimulant-induced neutrophil ROS (P ≥ .12, 95% CI -644.9 to 56.2, point estimate -294.4), or PAC (P ≥ .64, 95% CI -1567 to 463.4, point estimate -552.0) at any dose or timepoint. Plasma AA concentrations increased in a dose-dependent manner. CONCLUSIONS AND CLINICAL IMPORTANCE: High-dose administration of AA might provide antioxidant benefits in horses.

[Hippotherapy versus hippotherapy simulators as a treatment option in children with cerebral palsy: A systematic review]. / Hipoterapia versus simuladores de hipoterapia como opción de tratamiento en niños con parálisis cerebral: revisión sistemática.

INTRODUCTION AND OBJECTIVE: Hippotherapy (HPOT) and hippotherapy simulators (SHPOT) are used in children with cerebral palsy to achieve their maximum functionality and independence. The aim is to find out if HPOT and SHPOT produce the same effects on balance, gross motor function, and postural control in children under 18 years old with cerebral palsy. MATERIALS AND METHODS: The keywords used were: hippotherapy, equine-assisted therapy and cerebral palsy. The databases used were PeDro, Scopus, LILACS, ScienceDirect, Cochrane Library, Web of Science and CINAHL Complete (Ebsco). Studies were included if they were randomized clinical trials that studied the effect of HPOT and/or SHPOT on the variables mentioned in these patients. RESULTS: Four studies assessed balance, 4 studied gross motor function, and 2 investigated postural control. Both HPOT and SHPOT produced benefits in all of them. CONCLUSIONS: According to the studied variables both interventions produce similar improvements. Although, they increase with HPOT possibly due to greater sensory stimulation.

A pilot study of skin stretching and blood circulation effects of dermal suctioning in dogs.

Dermal suctioning has been reported to yield effects similar to those of cupping therapy in humans and horses, including pain reduction, increased blood circulation, improved flexibility, and healing. However, there is a dearth of reported outcomes concerning cupping or dermal suctioning in dogs. In this study, we examined the physiological effects of dermal suctioning in dogs. Employing the Medicell MINI pro8 device, dermal suctioning was applied to the dorsal surfaces of eight healthy beagle dogs for 20 min. Metrics such as body surface temperature, rectal temperature, pulse rate, respiratory rate, and skin pinch test results were gauged. Seven healthy beagle dogs were used as controls, and the same measurements were performed without dermal suctioning. The results showed a significant increase in the body surface temperature and skin pinch test results after dermal suctioning. We believe that the elimination of torsion in the blood vessels and nerves in the shallow fascia positively affected the thermoregulatory mechanism, resulting in an increase in body surface temperature, and also improving skin flexibility. Thus, dermal suctioning promotes subcutaneous blood circulation and improves skin flexibility in dogs. Further research is needed to identify the mechanisms underlying the effects of dermal suctioning and evaluate the stress in dogs caused by the implementation of the process.

Heterologous Seminal Plasma Reduces the Intracellular Calcium and Sperm Viability of Cryopreserved Stallion Spermatozoa.

Despite the vital role of seminal plasma (SP) in maintaining sperm function and aiding gamete interaction in many species, SP is usually removed before cryopreservation of stallion sperm to improve cryosurvival of sperm. The present study assessed if the vital sperm functional parameters of genetically superior stallions producing poor quality semen can be enhanced by the supplementation of heterologous SP from the stallion producing high quality semen. Spermatozoa from poor quality semen producing stallions were divided into three aliquots: two aliquots were supplemented with SP obtained from good quality semen producing stallions at the rate of 20% and 30%, respectively, whereas the third aliquot remained as control (0% SP) and incubated at 37°C for 30 minutes. Sperm membrane integrity, mitochondrial membrane potential (MMP), mitochondrial superoxide (mtROS) generation, and intracellular calcium status were assessed at different time intervals during incubation by flow cytometry. It was observed that the dead sperm population increased (p < 0.01) during incubation in both the 20% and 30% SP-supplemented groups. However, no significant changes were observed in MMP in both the control and treatment groups at different time intervals. Interestingly, it was found that sperm mtROS production increased (p < 0.01) during incubation in the SP-supplemented groups compared with the control group. The proportion of live spermatozoa with high intracellular calcium was reduced (p < 0.01) during incubation in the SP-incubated groups. Collectively, heterologous SP addition could not repair the damages caused by the cryopreservation and further resulted in deterioration of semen quality as observed in our study by reducing viability, increasing reactive oxygen species (ROS) production possibly due to high proportion of dead cells, or some factors (yet to be identified) that are inducive of oxidative stress in stallion spermatozoa.

Examining the Potential of Vitamin C Supplementation in Tissue-Engineered Equine Superficial Digital Flexor Tendon Constructs.

Because equine tendinopathies are slow to heal and often recur, therapeutic strategies are being considered that aid tendon repair. Given the success of utilizing vitamin C to promote tenogenesis in other species, we hypothesized that vitamin C supplementation would produce dose-dependent improvements in the tenogenic properties of tendon proper (TP) and peritenon (PERI) cells of the equine superficial digital flexor tendon (SDFT). Equine TP- and PERI-progenitor-cell-seeded fibrin three-dimensional constructs were supplemented with four concentrations of vitamin C. The gene expression profiles of the constructs were assessed with 3'-Tag-Seq and real-time quantitative polymerase chain reaction (RT-qPCR); collagen content and fibril ultrastructure were also analyzed. Moreover, cells were challenged with dexamethasone to determine the levels of cytoprotection afforded by vitamin C. Expression profiling demonstrated that vitamin C had an anti-inflammatory effect on TP and PERI cell constructs. Moreover, vitamin C supplementation mitigated the degenerative pathways seen in tendinopathy and increased collagen content in tendon constructs. When challenged with dexamethasone in two-dimensional culture, vitamin C had a cytoprotective effect for TP cells but not necessarily for PERI cells. Future studies will explore the effects of vitamin C on these cells during inflammation and within the tendon niche in vivo.

Changes in the nutrient profile and the load of mycotoxins, phytoestrogens, and pesticides in horse pastures during spring and summer in Austria.

Pastures are used for grazing and the production of conserved roughage in horses. Yet, the nutritional profile of the forage varies from spring to late summer, affecting equine nutrient supply and health. In addition, environmental factors may also favor plant contaminants such as mycotoxins. This study aimed to determine the nutritional profile and contaminant load of selected horse pastures from early spring till late summer. The nutrient composition (main macronutrients, macro elements and trace elements), as well as mycotoxins, metabolites, pesticides, and plant-derived compounds of seven horse pastures were analyzed. Each pasture was sampled three times and the samples were categorized according to the status of the pasture plants: ear emergence, early- till full bloom, and drought-damaged vegetation. Drought-damaged pastures demonstrated a rise in the acid to neutral detergent fiber ratio, calcium, iron, and magnesium but lower potassium contents. Mycotoxins and other contaminants were found in the pastures including 64 fungal compounds (ergot alkaloids (13) and metabolites from Fusarium (21), Aspergillus (2), Penicillium (8), Alternaria (8) and other fungal species (12), one bacterial metabolite (cereulide), twelve plant metabolites (including eight phytoestrogens and three cyanogenic glycosides (linamarin, lotaustralin and prunasin)), 11 nonspecific metabolites and six pesticides. Fusarium metabolites showed the highest concentrations among the fungal metabolites and drought-induced stress increased the contamination levels (range: 123-3873 µg/kg DM). In conclusion, there was a dominant effect of the developmental stages of the plants, botanical composition of the pastures and weather conditions on the nutritional composition and presence of contaminants on pastures.

Effect of a blend of magnesium oxide on Equine Squamous Gastric Disease in young trotter horses under training.

BACKGROUND: Equine squamous gastric disease (ESGD), as part of the equine gastric ulcer syndrome (EGUS), are common in racing horses. The use of buffering feed supplements to treat and/or prevent gastric ulcers is an option to control this condition. OBJECTIVE: The purpose of this study was to evaluate the effect of a 30-day supplementation with a blend of magnesium oxide (MgO) on ESGD scores in trotters under training. METHODS: Forty-two young trotters were submitted to a gastroscopic evaluation to assess their ESGD score and were randomly assigned in a group supplemented with MgO or in a control group. After 30 days, a second evaluation by gastroscopy was performed. The effect of the MgO supplementation was assessed by comparing the evolution of the ESGD score in supplemented and control groups between day 0 and day 30. RESULTS: The results confirm the high prevalence of EGUS in young Trotters. The supplementation significantly decreased the ESGD scoring in the supplemented group whereas the control group remain unchanged. CONCLUSION: The oral MgO supplementation was efficient to control ESGD in the population studied.

Pupillometry to show stress release during equine sports massage therapy.

Anecdotal reports state that wellness treatments for horses, such as massage therapy, relaxes the treated animal. Massage therapists and horse owners typically report an "improvement" without verifying or quantifying the treatment results. This paper shows that the effect of wellness treatment and stress release can be measured with pupillometry. One of the horse's pupils was photographed at the beginning and end of the treatment to determine the changes in the balance between the sympathetic and parasympathetic system activities. The owners assigned horses to two experimental groups: animals receiving a massage (N = 18) and horses standing with a person next to the horse for the time of a massage in the stable lane (N = 10). Six animals in the experimental group were excluded from the analysis because the pupils could not be traced. We opened the images of the pupil with Fiji (ImageJ) and used the elliptical selection tool to measure the pupils' and iris' areas. The ratio between the pupils' aperture and the iris' area was a normalized measure for pupil size. At the end of the experiment, we compared the normalized size of the pupils with a two-tailed paired t-test within groups and a two-tailed t-test between groups. For the experimental group, it was before and after the treatment, and for the control group, before and after the horse was placed in the stable lane. Comparisons between the experimental and control groups were made at the procedure's beginning and end. The treatment significantly decreased the normalized pupil area in the experimental group, on average, by a factor of 0.78 ± 0.15 (P = 0.042). For the horses in the control group, the pupil size increased, on average, by a factor of 1.14 ± 018. Changes were statistically not significant (P = 0.19). The initial pupil size of the horses in the experimental group was 1.88 times larger than that in the control group. After the treatment, the difference was reduced to a factor of 1.25. At the beginning of the experiment, the horses in the experimental group had, on average, larger pupil sizes than the horses in the control group, indicating that the horses in the experimental group were more stressed before the treatment than the control animals. The observed changes in pupil size in the experimental group likely resulted from enhanced parasympathetic and diminished sympathetic activity resulting from the treatment. Observed changes in pupil size agree with the anecdotal horse owner reports and the therapist's treatment notes.