Healthy Equine Research Articles

Artificial mitochondrial transplantation (AMT) reverses aging of mesenchymal stromal cells and improves their immunomodulatory properties in LPS-induced synoviocytes inflammation

Nowadays, regenerative medicine techniques are usually based on the application of mesenchymal stromal cells (MSCs) for the repair or restoration of injured damaged tissues. However, the effectiveness of autologous therapy is limited as therapeutic potential of MSCs declines due to patient's age, health condition and prolonged in vitro cultivation as a result of decreased growth rate. For that reason, there is an urgent need to develop strategies enabling the in vitro rejuvenation of MSCs prior transplantation in order to enhance their in vivo therapeutic efficiency.In presented study, we attempted to mimic the naturally occurring mitochondrial transfer (MT) between neighbouring cells and verify whether artificial MT (AMT) could reverse MSCs aging and improve their biological properties. For that reason, mitochondria were isolated from healthy donor equine adipose-derived stromal cells (ASCs) and transferred into metabolically impaired recipient ASCs derived from equine metabolic syndrome (EMS) affected horses, which were subsequently subjected to various analytical methods in order to verify the cellular and molecular outcomes of the applied AMT.Mitochondria recipient cells were characterized by decreased apoptosis, senescence and endoplasmic reticulum stress while insulin sensitivity was enhanced. Furthermore, we observed increased mitochondrial fragmentation and associated PARKIN protein accumulation, which indicates on the elimination of dysfunctional organelles via mitophagy. AMT further promoted physioxia and regulated autophagy fluxes. Additionally, rejuvenated ASCs displayed an improved anti-inflammatory activity toward LPS-stimulated synoviocytes. The presented findings highlight AMT as a promising alternative and effective method for MSCs rejuvenation, for potential application in autologous therapies in which MSCs properties are being strongly deteriorated due to patients' condition.

Micro-CT and histological examination of accessory canals in 34 equine cheek teeth.

Accessory canals and apical deltas have been extensively studied in human dentistry. Their clinical role as a difficult to clean reservoir for bacteria during endodontic treatments has been well described. Many papers describe in detail the pulp anatomy of equine dentition but little attention has been given to their apical ramifications. The goal of this paper is to describe the presence and anatomy of these accessory canals and apical deltas in healthy equine cheek teeth and discuss their possible relevance in the light of equine endodontics. To accomplish this, 15 maxillary and 19 mandibular healthy cheek teeth were collected ranging from Triadan 06 s to 11 s with eruption ages from 4 to 9 years. Their root anatomy was documented in detail based on micro computed tomography images. A subset of 3 teeth also underwent histological examination. Accessory canals were found in all but two teeth examined. Up to 44 accessory canals per tooth have been found with locations ranging from the root furcation down to the apex of the root and with highly variable diameters. Apical deltas in different stages of development were found in 84% of the roots. The presence of accessory canals identified on microCT images could be confirmed using histological examination although some of them were obliterated by reparative dentin. Accessory canals can be found in most equine cheek teeth and add complexity to their endodontic anatomy. This could have important implications for their treatment in case of pulp pathology. In humans, failure to remove bacterial biofilm from such canals has been associated with failure of endodontic treatment. Research on diseased equine teeth is required to gain a better understanding of their clinical relevance in horses.

Método de avaliação para identificação de bloqueio da cauda de equinos

ABSTRACT This study aims to standardize the clinical and complementary examination using surface electromyography (EMG) to assess the neuromuscular function of the equine tail. Firstly, clinical examinations of the tails and EMG measurements were performed on ten healthy animals without any tail interference or manipulation. This initial experiment aimed to establish a baseline for clinical examination and EMG results. The animals underwent tail nerve plexus blockade in the second experiment using an anesthetic agent. The objective compares the clinical examination and EMG findings between the anesthetized and non-manipulated tails. Finally, two animals were used in the third experiment, where peripheral infiltration of the tail nerve plexuses was performed using a neurolytic agent. The study successfully obtained a pattern for the clinical examination of a healthy equine tail and established a pattern for the clinical examination of a tail after infiltration with anesthetic and neurolytic agents. Significant differences were observed in the clinical examination before and after the application of local anesthesia, as well as in the EMG values before and after local infiltration with anesthetic (P=0.04) and neurolytic agent (P=0.03). The presented study introduces a clinical examination protocol that utilizes surface electromyography as a diagnostic tool for cauda equina block.

Procalcitonin and carbonylated protein concentrations in equine synovial fluid.

Early diagnosis of joint diseases is fundamental for prompt and appropriate management, particularly in septic arthritis. Procalcitonin (PCT) and protein carbonylated content (PCC) have been investigated in both human and veterinary medicine. An increase in PCT has been shown in infectious bacterial diseases, while higher levels of PCC have been shown in inflammatory pathologies characterized by oxidative damage. This study evaluated PCT and PCC in plasma and synovial fluid (SF), in healthy and pathological equine joints, affected by different types of arthropathy. Twenty-nine joints were evaluated and underwent orthopedic, radiographic, ultrasonographic and SF evaluation. The joints were divided in three groups: healthy, septic, and non-septic arthritis. PCT and PCC were measured in horse plasma and SF. Data distribution was evaluated and results were expressed as median, quartile values. Statistical differences in SF values among groups and correlations were assessed between plasma and SF of both PCT and PCC. The groups of joint disease included: 8/29 healthy, 13/29 non-septic and 8/29 septic. Significant differences were obtained for SF PCC and plasma PCT between healthy and septic joints, while no differences were found for plasma PCC and SF PCT. A positive correlation was found between plasma and SF PCT. To the best of our knowledge, this is the first study reporting PCT in equine SF. SF PCC could be a useful biomarker to differentiate between septic and healthy joints.

Cationic tantalum oxide nanoparticle contrast agent for micro computed tomography reveals articular cartilage proteoglycan distribution and collagen architecture alterations

ObjectiveCationic tantalum oxide nanoparticles (Ta2O5-cNPs), as a newly introduced contrast agent for computed tomography (CT) of cartilage, offer quantitative evaluation of proteoglycan content and biomechanical properties. However, knowledge on the depth-wise impact of cartilage constituents on NP diffusion, particularly the influence of the collagen network, is lacking. In this study, we aim to establish the depth-dependent relationship between Ta2O5-cNP diffusion and cartilage constituents (proteoglycan content, collagen content and network architecture). MethodsOsteochondral samples (n = 30) were harvested from healthy equine stifle joints (N = 15) and the diffusion of 2.55 nm diameter cationic Ta2O5-cNPs into the cartilage was followed with µCT imaging for up to 96 hours. The diffusion-related parameters, Ta2O5-cNP maximum partition (Pmax) and diffusion time constant, were compared against biomechanical and depth-wise structural properties. Biomechanics were assessed using a stress-relaxation and sinusoidal loading protocols, whereas proteoglycan content, collagen content and collagen network architecture were determined using digital densitometry, Fourier-transform infrared spectroscopy and polarized light microscopy, respectively. ResultsThe Pmax correlates with the depth-wise distribution of proteoglycans (bulk Spearman’s ρ = 0.87, p < 0.001). More open collagen network architecture at the superficial zone enhances intake of Ta2O5-cNPs, but collagen content overall decreases the intake. The Pmax values correlate with the equilibrium modulus (ρ = 0.80, p < 0.001) of articular cartilage. ConclusionThis study establishes the feasibility of Ta2O5-cNPs for the precise and comprehensive identification of biomechanical and structural changes in articular cartilage via contrast-enhanced µCT.

Bacteria in the healthy equine vagina during the estrous cycle

An understanding of the normal bacterial microbiota of any organ is essential to provide the background to conditions and interventions that might cause the microbiota to change. In the vagina of the mare, a change of bacterial microbiota could be induced by introduction of semen, treatment with antibiotics, discharge from an unhealthy uterus etc. Previous studies on equine vaginal bacteria are not all conducted in the same way and results are not altogether consistent. Therefore, this study was designed to provide a deeper understanding of the bacterial microbiota of the mare vagina, and possible changes throughout the estrous cycle. The cranial portion of the vagina was sampled on day 0 (ovulation), day 3, day 7, and day 14 of the estrous cycle. The vaginal sampling was conducted with double-guarded occluded swabs from the cranial floor of the vagina. Ovulation was determined by rectal palpation and ultrasonic examination, and the day 0 samples were taken within ±24 h of ovulation. Swabs were brought to the laboratory in Amies medium within 2–3 h and were plated out immediately on both selective and non-selective agars. Results were registered as amount of growth (qualitatively), bacterial species and number of isolates. Bacterial growth was highest on day 3 and 7, representing the beginning and middle of diestrus. The dominant bacteria were Escherichia coli and Streptococcus zooepidemicus. Escherichia coli was especially dominant in maiden mares, compared to the mares that had foaled. An increase in bacterial diversity throughout the estrous cycle was observed, being highest on day 14. These results suggest that there are changes in the bacterial microbiota of the mare vagina throughout the normal estrous cycle.

Alterations of the bacterial ocular surface microbiome are found in both eyes of horses with unilateral ulcerative keratitis.

Next generation sequencing (NGS) studies in healthy equine eyes have shown a more diverse ocular surface microbiota compared to culture-based techniques. This study aimed to compare the bacterial ocular surface microbiota in both eyes of horses with unilateral ulcerative keratitis (UK) with controls free of ocular disease. Conjunctival swabs were obtained from both ulcerated eyes and unaffected eyes of 15 client-owned horses with unilateral UK following informed consent, as well as from one eye of 15 healthy horses. Genomic DNA was extracted from the swabs and sequenced on an Illumina platform using primers that target the V4 region of bacterial 16S rRNA. Data were analyzed using Quantitative Insights Into Molecular Ecology (QIIME2). The ocular surface of ulcerated eyes had significantly decreased species richness compared with unaffected fellow eyes (Chao1 q = 0.045, Observed ASVs p = 0.045) with no differences in evenness of species (Shannon q = 0.135). Bacterial community structure was significantly different between either eye of horses with UK and controls (unweighted UniFrac: control vs. unaffected, p = 0.03; control vs. ulcerated, p = 0.003; unaffected vs. ulcerated, p = 0.016). Relative abundance of the gram-positive taxonomic class, Bacilli, was significantly increased in ulcerated eyes compared with controls (q = 0.004). Relative abundance of the taxonomic family Staphylococcaceae was significantly increased in ulcerated and unaffected eyes compared with controls (q = 0.030). The results suggest the occurrence of dysbiosis in infected eyes and reveal alterations in beta diversity and taxa of unaffected fellow eyes. Further investigations are necessary to better understand the role of the microbiome in the pathophysiology of ocular surface disease.

Topical ropivacaine hydrochloride 0.5% and lidocaine hydrochloride 2% significantly reduce corneal sensitivity without short-term negative side effects in horses.

To evaluate corneal sensitivity and acute side effects following application of ropivacaine hydrochloride 0.5% and lidocaine hydrochloride 2% on the healthy equine cornea. Eight healthy adult horses. A randomized, masked, crossover study design was utilized. Baseline Semiquantitative Preclinical Ocular Toxicology (SPOT) scores and corneal touch thresholds (CTT) using a Cochet-Bonnet esthesiometer were recorded and measured, respectively, for eight healthy adult horses before medication application. Commercially available eyewash was used as a negative control. Ropivacaine hydrochloride 0.5% or lidocaine hydrochloride 2% solution was sprayed on a randomly selected eye, and the contralateral eye received eyewash. CTT was measured in both eyes at 1, 5, 15, 25, 35, 45, 55, 65, and 75 min post-application. Post-application SPOT scores were recorded immediately following the trial. Linear mixed model statistical analyses (mean ± standard error) were performed (p < .05). Mean eyewash CTT (3.41 cm ± 0.464) was significantly different from ropivacaine-treated (1.44 cm ± 0.562) (p = .008) and lidocaine-treated eyes (1.75 cm ± 0.562) (p = .024); CTT was not significantly different between drug groups (p = .88). Time to maximum anesthesia was not significantly different between ropivacaine (13.25 min ± 3.353) and lidocaine (16.25 min ± 3.353) (p = .40). No side effects were appreciated as confirmed by SPOT. Ropivacaine and lidocaine similarly decreased corneal sensitivity when applied topically without clinically evident short-term ocular side effects. Lidocaine may be preferable in clinical settings due to its large, multi-use vials and similar effects to ropivacaine.

Platelet-rich plasma: lyophilization as an alternative for conservation and transport

Platelet-rich plasma (PRP) has been used in equine reproduction to improve pregnancy rates due to its immune-modulating effect. Platelet granules contain numerous proteins, such as growth factors, cytokines, and antimicrobial peptides. However, the shelf life of platelets is 5 days and used at a temperature above 20°C, where there is the concern of bacterial contamination. Also, preparation is a time-consuming procedure. Therefore, studies to find an effective method of PRP storage would be of great benefit, saving costs, and could also guarantee a faster and more effective treatment. This work compared the platelet levels between fresh and lyophilized PRP. Peripheral blood from 8 healthy equine males with a mean age of 4±2years was collected in 4.5-mL vacutainer tubes containing 3.2% sodium citrate. The PRP preparation was performed by double centrifugation in vacutainer tubes. Blood was homogenized and centrifuged at 120g for 10 minutes. After the first centrifugation, the upper 30% of the plasma was discarded. The remaining plasma (70%) was submitted to another centrifugation at 240g for 10 minutes. After the second centrifugation, 30% of plasma at the bottom of each tube was considered PRP. Platelet count was carried out with an automated hematology analyzer (HematoClin 2.6VET - Bioclin IL, USA). The lyophilization process was carried out at the Research Support Center of the Federal RuralUniversity of Pernambuco. Before being lyophilized, samples were frozen at -80°C for 24 h and then processed in an Alpha 1-4 LD Plus Christ apparatus, a drying chamber temperature of -27°C and vacuum of 0.52 mbar, for 24 hours. The lyophilized PRP obtained was stored in well-sealed microtubes and kept frozen at -4°C for further analysis. The results were subjected to analysis of variance (ANOVA) to determine the effects of lyophilization on the platelet concentrations. Significance was declared at P<0.05. The median platelet count in the fresh PRP was 108±35.2 × 103/uL. The platelet count after lyophilization was 46.6% lower (p<0.05) when compared with the initial count (56±17.7 × 103/uL). Although there was a significant reduction in platelet concentration after lyophilization, 53.4% of platelets were viable after the process. Our study showed the viability of lyophilizing PRP using a standard and easy system. Despite several published studies on PRP standardization, to the best of our knowledge, this is the first work on equine PRP lyophilization. In addition, lyophilized PRP appears to be a possible alternative to fresh PRP; with minor variability, a large-scale production with increased shelf life could be achieved following a single production process. Further studies are needed, mainly on growth factors after lyophilization and the use of PRP in vivo.

Effect of end-inspiratory pause on airway and physiological dead space in anesthetized horses

ObjectiveTo evaluate the impact of a 30% end-inspiratory pause (EIP) on alveolar tidal volume (VTalv), airway (VDaw) and physiological (VDphys) dead spaces in mechanically ventilated horses using volumetric capnography, and to evaluate the effect of EIP on carbon dioxide (CO2) elimination per breath (Vco2br–1), PaCO2, and the ratio of PaO2-to-fractional inspired oxygen (PaO2:FiO2). Study designProspective research study. AnimalsA group of eight healthy research horses undergoing laparotomy. MethodsAnesthetized horses were mechanically ventilated as follows: 6 breaths minute–1, tidal volume (VT) 13 mL kg–1, inspiratory-to-expiratory time ratio 1:2, positive end-expiratory pressure 5 cmH2O and EIP 0%. Vco2br–1 and expired tidal volume (VTE) of 10 consecutive breaths were recorded 30 minutes after induction, after adding 30% EIP and upon EIP removal to construct volumetric capnograms. A stabilization period of 15 minutes was allowed between phases. Data were analyzed using a mixed-effect linear model. Significance was set at p < 0.05. ResultsThe EIP decreased VDaw from 6.6 (6.1–6.7) to 5.5 (5.3–6.1) mL kg–1 (p < 0.001) and increased VTalv from 7.7 ± 0.7 to 8.6 ± 0.6 mL kg–1 (p = 0.002) without changing the VTE. The VDphys to VTE ratio decreased from 51.0% to 45.5% (p < 0.001) with EIP. The EIP also increased PaO2:FiO2 from 393.3 ± 160.7 to 450.5 ± 182.5 mmHg (52.5 ± 21.4 to 60.0 ± 24.3 kPa; p < 0.001) and Vco2br–1 from 0.49 (0.45–0.50) to 0.59 (0.45–0.61) mL kg–1 (p = 0.008) without reducing PaCO2. Conclusions and clinical relevanceThe EIP improved oxygenation and reduced VDaw and VDphys, without reductions in PaCO2. Future studies should evaluate the impact of different EIP in healthy and pathological equine populations under anesthesia.

Pneumatic compression therapy using the EQ Press accelerates lymphatic flow in healthy equine forelimbs as determined by lymphoscintigraphy.

Limb lymphedema in horses can be debilitating and painful. Pneumatic compression therapy has shown significant benefits for people suffering from lymphedema. The objective of this study was to determine the effect of a novel, equine-specific pneumatic compression device on the lymphatic flow of healthy horse forelimbs as determined by Tc-99m sulfur colloid lymphoscintigraphy. 6 healthy Thoroughbreds. In a randomized crossover design, horses underwent bilateral forelimb lymphoscintigraphy following subcutaneous injection of Tc-99m sulfur colloid at the coronary band as untreated control or with pneumatic compression therapy using the EQ Press. Lateral, static images were obtained of the distal limb (time 0 to 60 minutes) and proximal limb (time 30 to 60 minutes) using a standard gamma camera. Lymphatic flow was determined by assigning a score to the time point at which Tc-99m sulfur colloid was first visualized at the level of the accessory carpal bone (1 to 7) in the distal limb and the cubital lymph node (1 to 4) in the proximal limb. EQ Press treatment led to a significantly faster lymphatic flow of Tc-99m sulfur colloid to the predetermined anatomic locations of the accessory carpal bone (P = .002) in the distal limb and the cubital lymph node (P = .001) in the proximal limb. Pneumatic compression therapy as provided by an equine-specific device encouraged lymphatic flow in healthy, nonedematous equine forelimbs. These data support further study of the EQ Press for pneumatic compression therapy in horses clinically affected by lymphedema and lymphatic drainage disorders.

Characterization of Myeloperoxidase in the Healthy Equine Endometrium.

Myeloperoxidase (MPO), as a marker of neutrophil activation, has been associated with equine endometritis. However, in absence of inflammation, MPO is constantly detected in the uterine lumen of estrous mares. The aim of this study was to characterize MPO in the uterus of mares under physiological conditions as a first step to better understand the role of this enzyme in equine reproduction. Total and active MPO concentrations were determined, by ELISA and SIEFED assay, respectively, in low-volume lavages from mares in estrus (n = 26), diestrus (n = 18) and anestrus (n = 8) in absence of endometritis. Immunohistochemical analysis was performed on 21 endometrial biopsies randomly selected: estrus (n = 11), diestrus (n = 6) and anestrus (n = 4). MPO, although mostly enzymatically inactive, was present in highly variable concentrations in uterine lavages in all studied phases, with elevated concentrations in estrus and anestrus, while in diestrus, concentrations were much lower. Intracytoplasmic immunoexpression of MPO was detected in the endometrial epithelial cells, neutrophils and glandular secretions. Maximal expression was observed during estrus in mid and basal glands with a predominant intracytoplasmic apical reinforcement. In diestrus, immunopositive glands were sporadic. In anestrus, only the luminal epithelium showed residual MPO immunostaining. These results confirm a constant presence of MPO in the uterine lumen of mares in absence of inflammation, probably as part of the uterine mucosal immune system, and suggest that endometrial cells are a source of uterine MPO under physiological cyclic conditions.

Integrated multi-omics reveals novel microbe-host lipid metabolism and immune interactions in the donkey hindgut.

Evidence has shown that gut microbiota play a key role in host metabolism and health; however, little is known about the microbial community in the donkey hindgut as well as the interactions that occur between gut microbes and the host. This study aimed to explore the gut microbiome differences by analyzing the microbial community and differentially expressed genes (DEGs) related to lipid metabolism and the immune system along the donkey hindgut. The hindgut tissues (cecum, ventral colon, and dorsal colon) were separated, and the contents of each section were collected from six male donkeys for multi-omics analysis. There were significant differences in terms of dominant bacteria among the three sections, especially between the cecum and dorsal colon sites. For instance, species belonging to Prevotella and Treponema were most abundant in the cecum, while the Clostridiales_bacterium, Streptococcus_equinus, Ruminococcaceae_bacterium, etc., were more abundant in the dorsal colon. Apart from propionate, the concentrations of acetate, isobutyrate, valerate and isovalerate were all lower in the cecum than in the dorsal colon (p < 0.05). Furthermore, we identified some interesting DEGs related to lipid metabolism (e.g., ME1, MBOAT1, ACOX1, ACOX2 and LIPH) and the immune system (e.g., MUC3B, mucin-2-like, IL17RC, IL1R2, IL33, C1QA, and MMP9) between the cecum and dorsal colon and found that the PPAR pathway was mainly enriched in the cecum. Finally, we found a complex relationship between the gut microbiome and gene expression, especially with respect to the immune system, and combined with protein-protein interaction (PPI) data, suggesting that the PPAR pathway might be responsible, at least in part, for the role of the hindgut microbiota in the donkeys' gut homeostasis. Our data provide an in-depth understanding of the interaction between the microbiota and function in the healthy equine hindgut and may also provide guidance for improving animal performance metrics (such as product quality) and equine welfare.

Immunohistochemical staining of immunoglobulin G in healthy equine, canine, and feline corneas.

ObjectiveEstablishing an immunohistochemical approach for semi‐quantitative assessment of the presence of immunoglobulin G (IgG) in equine, canine, and feline corneas.ProceduresHealthy corneas of horses, dogs, and cats, euthanized because of a fatal disease or an unrecoverable trauma unrelated to and without a history of ophthalmic disease were formalin‐fixed, paraffin‐embedded, and determined to be pathomorphologically healthy by light microscopy. Automated immunohistochemistry was performed using primary antibodies against IgG, biotin‐conjugated secondary antibodies, and streptavidin‐peroxidase, as well as diaminobenzidine for visualization. After counterstaining with hematoxylin, epithelium, stroma, Descemet´s membrane (DM), and endothelium were semi‐quantitatively scored for the presence of IgG on a 4‐grade scale (0 = no, 1 = faint, 2 = medium, 3 = strong staining) by light microscopy.ResultsCorneal specimens of 20 horses (40 eyes) with a median age of 15.5 years (range 2–31 years), 12 dogs (21 eyes) with a median age of 10.0 years (range 4–16), and 13 cats (24 eyes) with a median age of 10.0 years (range 2–18) were included in the study. Different sexes and breeds were represented. In all corneas (100%), significant medium signal intensity in the stroma was observed. Variable immunosignal was obtained in epithelium, DM, and endothelium.ConclusionThis method reproducibly allows for the detection of IgG in healthy equine, canine, and feline corneas, particularly stroma. Semi‐quantitative results evidence medium presence of IgG in the corneal stroma. Further research is needed to evaluate IgG presence in diseased corneas.

The Prevalence and Molecular Biology of Staphylococcus aureus Isolated from Healthy and Diseased Equine Eyes in Egypt

This work aimed to characterize S. aureus isolates from the eyes of healthy and clinically affected equines in the Kafrelsheikh Governorate, Egypt. A total of 110 animals were examined for the presence of S. aureus, which was isolated from 33 animals with ophthalmic lesions and 77 healthy animals. We also investigated the antimicrobial resistance profile, oxacillin resistance mechanism, and the major virulence factors implicated in many studies of the ocular pathology of pathogenic S. aureus. The association between S. aureus eye infections and potential risk factors was also investigated. The frequency of S. aureus isolates from clinically affected equine eyes was significantly higher than in clinically healthy equids. A significant association was found between the frequency of S. aureus isolation from clinically affected equine eyes and risk factors including age and season but not with sex or breed factors. Antimicrobial resistance to common antibiotics used to treat equine eyes was also tested. Overall, the isolates showed the highest sensitivity to sulfamethoxazole (100%) and the highest resistance to cephalosporin (90.67%) and oxacillin (90.48%). PCR was used to demonstrate that mecA was present in 100% of oxacillin- and β-lactam-resistant S. aureus strains. The virulence factor genes Spa (x region), nuc, and hlg were identified in 62.5%, 100%, and 56%, of isolates, respectively, from clinically affected equines eyes. The severity of the eye lesions increased in the presence of γ-toxin-positive S. aureus. The phylogenetic tree of the Spa (x region) gene indicated a relationship with human reference strains isolated from Egypt as well as isolates from equines in Iran and Japan. This study provides insight into the prevalence, potential risk factors, clinical pictures, zoonotic potential, antimicrobial resistance, and β-lactam resistance mechanism of S. aureus strains that cause eye infection in equines from Egypt.

Mesenchymal Stromal Cells Adapt to Chronic Tendon Disease Environment with an Initial Reduction in Matrix Remodeling

Tendon lesions are common sporting injuries in humans and horses alike. The healing process of acute tendon lesions frequently results in fibrosis and chronic disease. In horses, local mesenchymal stromal cell (MSC) injection is an accepted therapeutic strategy with positive influence on acute lesions. Concerning the use of MSCs in chronic tendon disease, data are scarce but suggest less therapeutic benefit. However, it has been shown that MSCs can have a positive effect on fibrotic tissue. Therefore, we aimed to elucidate the interplay of MSCs and healthy or chronically diseased tendon matrix. Equine MSCs were cultured either as cell aggregates or on scaffolds from healthy or diseased equine tendons. Higher expression of tendon-related matrix genes and tissue inhibitors of metalloproteinases (TIMPs) was found in aggregate cultures. However, the tenogenic transcription factor scleraxis was upregulated on healthy and diseased tendon scaffolds. Matrix metalloproteinase (MMPs) expression and activity were highest in healthy scaffold cultures but showed a strong transient decrease in diseased scaffold cultures. The release of glycosaminoglycan and collagen was also higher in scaffold cultures, even more so in those with tendon disease. This study points to an early suppression of MSC matrix remodeling activity by diseased tendon matrix, while tenogenic differentiation remained unaffected.

Effects of Lactic Acid Bacteria Isolated from Equine on Salmonella-Infected Gut Mouse Model.

The aim of this study was to evaluate the antibacterial potential of lactic acid bacteria (Weissella confuse, Pediococcus acidilactici, and Ligilactobacillus equi) isolated from healthy equine in Wuhan against Salmonella Typhimurium CVCC542-induced mice model on intestinal microflora. In previous studies, these isolated strains showed good probiotic potentials in vitro. In this study, fifty healthy mice were randomly divided into five groups, the blank control group, the control group, the Pediococcus acidilactici group (1 × 108CFU/day), the Ligilactobacillus equi group (1 × 108CFU/day), and the Weissella confuse group (1 × 108CFU/day). The body weight in control group and Weissella confuse group showed significant decreased (P < 0.05, P < 0.01), while Pediococcus acidilactici group and Ligilactobacillus equi group showed good recovering after treatments. The lowest diarrhea rate was shown in Ligilactobacillus equi group after treatment. In histopathology, Ligilactobacillus equi group showed the least structural damage in duodenum, and all probiotic treatment groups showed less damage in cecum. The sequence data and optical transform unit showed that Pediococcus acidilactici group and Ligilactobacillus equi group had higher number than control group, while the diversity data showed that the control group and Weissella confuse group had lower diversity in cecum. Microbial community analysis showed increased abundance of Firmicutes, Bacteroidetes, uncultured_bacterium_f_Muribaculaceae, and Lactobacillus in treatment groups, while potential microbes that can induce intestinal diseases such as Verrucomicrobia, Akkermansia, and Lachnospiraceae_NK4A136_group decreased in the treatment groups. In conclusion, lactic acid bacteria isolated from the healthy horses could alleviate the infection of Salmonella and regulate intestinal flora.

Fracture Resistance of Equine Cheek Teeth With and Without Occlusal Fissures: A Standardized ex vivo Model.

Background:Ex vivo fracture models are frequently used in human dentistry to provide insights in the fracture mechanisms of teeth. Equine cheek teeth fractures are an important dental pathology, but there has been no research performed to examine the fracture resistance ex vivo.Objective: To evaluate the fracture resistance of equine cheek teeth and identify anatomical predictors that might influence fracture resistance in healthy teeth. It was further evaluated if the presence of a fissure caused a decrease in fracture resistance.Study design:Ex vivo experimental design.Methods: Individual cheek teeth were subjected to a compression load in a universal testing machine until fracture occurred. Testing was performed in two study groups. A first group of healthy cheek teeth was tested to examine anatomical predictors on fracture resistance. A second group comprised cheek teeth with occlusal fissures and an equal number of age- and size-matched fissure-free teeth as controls. The effect of possible predictors on fracture resistance was investigated by regression analysis.Results: In the first group, fracture resistance was significantly influenced by the location on the tooth where testing was performed in both maxillary (p < 0.001) and mandibular teeth (p < 0.001). Additional significantly associated factors were Triadan number in mandibular (p = 0.009) and the mesiodistal length of the occlusal surface of maxillary teeth (p = 0.01). Experimentally induced crown fractures that extended below the simulated bone level were more frequently associated with pulp horn exposure (p < 0.001). In the second group, significant lower fracture loads were recorded in teeth with fissures (mandibular p = 0.006; maxillary p < 0.001), compared to fissure-free teeth.Main limitations: This ex vivo model does not imitate the in vivo masticatory forces and lacks the shock-absorbing properties of the periodontal ligament.Conclusions: The methodology used in this study provides an ex vivo experimental set-up to test fracture resistance of equine cheek teeth enabling evidence-based research to examine the potentially weakening effects of tooth pathology and its treatments. Crown resistance to fracture differed along the occlusal surface of healthy equine cheek teeth, and the presence of fissures further decreased fracture resistance.

Use of Polyamide (Nylon) Cable Ties for Vascular Ligation of Healthy Equine Jejunal Mesentery.

Jejunal vascular ligation is an essential step in performing jejunojejunostomy. Hand sewn ligation is typically used and can increase operative time with long sections of bowel to be removed. Nylon cable ties (NCT) have been used for vascular ligation in horses but are yet to be investigated for application on the mesenteric vasculature of the gastrointestinal tract. Our objective was to evaluate the efficacy and short-term safety of NCT jejunal mesenteric vessel ligation in healthy horses. Eight healthy adult horses underwent midline celiotomy. A segment of jejunal mesentery was identified (≥4 arcades). Briefly, three fenestrations (proximal, middle, distal) were made 5–10 mm apart adjacent to the first and last vascular arcade to be ligated. Two sterilized NCT were passed to encircle the mesentery through the proximal and middle fenestrations, separated by intact mesentery. NCT were closed tightly and the vascular pedicle transected with Mayo scissors through the distal fenestration. Jejunojejunostomy was then performed and the mesentery sutured closed. The number of vascular arcades and time to ligate using NCT were recorded. At 2 weeks, horses underwent repeat celiotomy to assess the healing of the NCT ligation site and an equal number of vascular arcades were hand sewn double ligated using 2-0 Polyglactin 910 as a timed comparison. NCT mesenteric ligation was significantly faster than hand sewn methods (P < 0.01). Effective hemostasis was achieved in all cases. There was no evidence of local infection or adhesions at 14 days post-operatively. Further investigation in the long-term effects in horses as well as horses with strangulating jejunal lesions are needed for clinical application.

Metabolism and global protein glycosylation are differentially expressed in healthy and osteoarthritic equine carpal synovial fluid.

Carpal osteochondral fragmentation and subsequent post-traumatic osteoarthritis (PTOA) are leading causes of wastage in the equine athlete. Identification of synovial fluid biomarkers could contribute to the diagnosis and understanding of osteoarthritis (OA) pathophysiology. The aim of this study was to identify differentially expressed metabolic and glycosylation pathways in synovial fluid from healthy horses and horses with naturally occurring carpal OA. Cross-sectional, in vivo metabolomics and glycomics study. In cohort 1, carpal synovial fluid (n=12 horses; n=6 healthy, n=6 OA) was analysed using high-resolution liquid chromatography mass spectrometry (LC-MS). In cohort 2 (n=40 horses; n=20 healthy, n=20 OA), carpal synovial fluid was analysed using lectin microarrays and a lubricin sandwich ELISA. Metabolomic analysis identified >4900 LC-MS features of which 84 identifiable metabolites were differentially expressed (P<.05) between healthy and OA joints, including key pathways related to inflammation (histidine and tryptophan metabolism), oxidative stress (arginine biosynthesis) and collagen metabolism (lysine metabolism). Principle Component Analysis and Partial Least Squares Discriminant Analysis demonstrated separation between healthy and OA synovial fluid. Lectin microarrays identified distinct glycosylation patterns between healthy and OA synovial fluid, including increased Core 1/Core 3 O-glycosylation, increased α-2,3 sialylation and decreased α-1,2 fucosylation in OA. O-glycans predominated over N-glycans in all synovial fluid samples, and synovial fluid lubricin was increased in OA joints as compared to controls. The sample size in cohort 1 was limited, and there is inherent variation in severity and duration of joint injury in naturally occurring OA. However, LC-MS identified up to 5000 unique features. These data suggest new potential diagnostic and therapeutic targets for equine OA. Future targeted metabolomic and glycomic studies should be performed to verify these results. Lectin microarrays could be investigated as a potential screening tool for the diagnosis and therapeutic monitoring of equine OA.