Equine melanomas are a common neoplasm in gray horses. However, scientific knowledge about their progression over time is quite scarce. Some owners and veterinarians still believe that early intervention is not necessary, stating that tumors evolve very slowly and intervention could worsen the animal’s condition. This work aims to identify clinical and histological differences that may exist between equine melanomas with different excision intervals (time between tumor detection and surgical excision). A total of 42 tumors (13 benign and 29 malignant) from 34 horses were included in this study. There was a statistically significant association between excision interval and tumor size (p = 0.038), with tumors excised later being significantly larger than the ones excised sooner. The excision interval was also statistically associated with the number of tumors (p = 0.011), since the horses that carried a tumor for longer seemed to be prone to have multiple tumors. Furthermore, there was an association between excision interval and malignancy (p = 0.035), with tumor excised later being fives times more likely to be malignant. This study provides evidence of delayed excision’s effect on the progression of equine melanomas. Additionally, it reinforces the importance of the early excision of these tumors.

Currently available treatments for equine melanocytic tumors have limitations, mainly due to mass localization and dimension, or the presence of metastases. Therefore, a search for new therapies is necessary. Programmed cell death-ligand 1 (PD-L1) is expressed by several tumors, blocking T cell-mediated elimination of the tumor cells by binding to programmed cell death protein 1 (PD-1). A novel therapeutic approach using PD-1/PD-L1 blockade in human melanoma resulted in tumor regression and prolonged tumor-free survival. This study aimed to evaluate the immunohistochemical expression of PD-L1 in equine melanocytic tumors. A total of 77 melanocytic tumors were classified as benign or malignant and evaluated by extension of labeling. A total of 59.7% of the tumors showed >50% of immunolabeled cells. Regarding malignant tumors, 24/38 tumors presented >50% of labeled cells, 13 tumors presented between 25–50% and one tumor presented <10%. Regarding benign tumors, 22/39 tumors presented >50% of labeled cells, nine tumors presented 25–50%, three tumors presented 10–25%, two tumors presented <10% and three tumors did not present expression. Our results suggest that PD-L1 blockade may be a potential target for immunotherapy in equine melanocytic tumors and that future clinical research trials into the clinical efficacy of the anti-PD-L1 antibody are necessary.

Background:Sarcoids and squamous cell carcinomas (SCCs) are the most concerning equine oncological diseases. Both tumors are challenging to manage due to their invasive behavior and high prevalence of recurrences. Furthermore, SCCs have a propensity to metastasize. Programed cell-death ligand 1 (PD-L1) has been one of the main therapeutic targets for immunotherapy in various human tumors. PD-L1 research in equine tumors is scarce and more efforts are necessary to understand the potential of this biomarker as a therapeutical target. Aim:Evaluate the immunohistochemical expression of PD-L1 in equine sarcoids and SCC. Methods:Thirteen equine tumors (seven sarcoids and 6 SCCs) were tested by immunohistochemistry and evaluated semi quantitatively to assess the percentage of positive cells. Results:None of the sarcoids presented PD-L1 expression. Regarding SCC, 2 tumors presented <10% of labeled cells; 2 tumors presented 10%-25% of labeled cells and 2 tumors presented 25%-50% of labeled cells. There were statistically significant differences between sarcoids and SCC regarding the expression of PD-L1. Conclusion:Our results point to the fact that PD-L1 could be a potential therapeutic target against SCC, and also encourage in-depth studies in this area, with larger sample sizes.

Equine melanocytic tumors are common and have an unusual benign behavior with low invasiveness and metastatic rates. However, tumoral mass growth is usually a concern that can have life-threatening consequences. COX-2 is related to oncogenesis, promoting neoplastic cell proliferation, invasion, and metastasis. The aim of this study was to evaluate the immunohistochemical expression of COX-2 in equine melanocytic tumors. Through extension and intensity of labeling, 39 melanocytomas and 38 melanomas were evaluated. Of the malignant tumors, 13.2% were negative and 63.2% presented a low COX-2 expression. Only 6 malignant tumors presented >50% of labeled cells, 18 malignant and 8 benign had an expression between 21 and 50%, 8 malignant and 3 benign tumors had an expression between 6 and 20%, 1 malignant tumor had an expression between 1 and 5%, and 5 malignant and 28 benign tumors had no expression. Malignant tumors showed higher COX-2 expression than did benign tumors, with statistically significant differences. The low levels of COX-2 may be one of the molecular reasons for the presence of expansive mass growth instead of the invasive pattern of other species, which is related to high COX-2 levels.

Neonatal encephalopathy represents a broad neurological syndrome which encompasses newborn foals presenting a variety of non-infectious neurologic signs and/or abnormal behaviors in the immediate postpartum period. It is recognized as the most predominant neurological disorder in neonatal foals. Prognostic factors can guide clinicians in medical decision-making, aiding in the establishment of survival probabilities. The aim of this work was to scrutinize the prognostic value of clinical and laboratorial findings upon admission and posterior comorbidities developed during hospitalization, assessing their influence on the outcome of foals with NE. For this purpose, the medical records of 61 neonatal foals with a primary diagnosis of NE were retrospectively analyzed. The overall survival rate was 57.4%. Most foals presented recumbency at admission, which was associated with higher mortality rates (p = 0.002). Hypothermic foals at admission were 4.85 times more likely to succumb (p = 0.015). The presence of hypoglycemia at admission was associated with higher mortality rates (p = 0.002). Foals with hypercreatinemia at admission had 6.67 times greater odds of dying. The development of seizures contributed to 4.14 greater odds of dying. Foals that developed comorbidities during hospitalization had 40.1 times greater odds of dying, with pneumonia and sepsis being the most relevant comorbidities. In foals with NE, rectal temperature, hematocrit, blood glucose, and creatinine concentrations are simple, quick-to-measure parameters that may have prognostic value during admission.

The Lusitano horse is gaining popularity in the equestrian world, and as a result, the significance of applied sports medicine for this breed is growing. As cardiology plays a crucial role in this field, numerous studies have been conducted to establish electrocardiographic reference values in various breeds to ensure a more accurate evaluation. However, studies regarding healthy Lusitano horses are lacking. So, this study aimed to establish electrocardiographic reference values for Lusitano horses, utilizing a sample of 82 clinically healthy animals. The evaluation involved lead II and base–apex lead measurements, with a median heart rate of 39 beats per minute being recorded. The P wave demonstrated a predominantly bifid configuration, while the QRS complex exhibited various forms. The most common QRS configurations were QR and R in lead II, and RS in the base–apex lead. Additionally, most T waves displayed a biphasic shape in both methods. Furthermore, statistically significant differences were noted based on age and gender. Some of the electrocardiographic values obtained differed from those previously published for other breeds. Given the relevance of electrocardiogram in cardiovascular evaluation, these findings bring valuable insights regarding the specific parameters for Lusitano horse and emphasize the importance of obtaining breed-specific electrocardiographic reference values.

Horses are considered as reservoirs of multidrug resistant bacteria that can be spread through the environment and possibly to humans. The aim of this study was to characterize the oral Gram-negative microbiota of healthy horses and evaluate their antimicrobial susceptibility profile in a One Health approach. For this purpose, samples were collected from the gingival margin of healthy horses, free of antimicrobial therapy, cultured in selective mediums, identified, and tested for antimicrobial susceptibility. Fifty-five Gram-negative isolates were identified, with 89.5% being zoonotic and 62% affecting humans, which were also found commonly in the environment. Forty-eight isolates (96%) were MDR. The phenotypic resistance presented as higher to macrolides (81.8%), β-lactams (55.4%), and quinolones (50%), and lower to sulfonamides (27.3%), tetracyclines, and amphenicols (both with 30.9%). In total, 51.5% of the isolates presented resistance to carbapenems. In addition to being the first report on the commensal oral microbiota of horses and respective susceptibility profile, this study highlights the horse as a valuable sentinel that can control the evolution and transmission of multidrug-resistant bacteria between the "One Health triad" since it is in contact with humans, other animals, and the environment, in different geographic locations.

Considering the increasing antibiotics resistance, there has been a propensity to replace them with antiseptics when it comes to wound management and treatment. Nevertheless, in recent years, there have been reports regarding resistance to antiseptics by some bacterial strains. There is also concern about the environmental impact of these substances. The aim of this study was to compare the antimicrobial efficacy of antiseptics and eucalyptus essential oils on bacterial strains from horse's wounds. We used twelve Escherichia coli, eight Staphylococcus aureus, two Staphylococcus pseudintermedius, one Staphylococcus vitulinus and one Staphylococcus saprophyticus strains from equine wounds. The effect of Eucalyptus radiata essential oil, Eucalyptus globulus essential oil, povidone-iodine and chlorhexidine gluconate against the isolated strains was evaluated applying the Kirby-Baüer method. Regarding the Escherichia coli strains, E. radiata and the mixture of E. radiata and E. globulus had a better inhibitory effect than antiseptics. E. globulus had a better effect against most Staphylococcus spp. compared to E. radiata. For both Gram-negative and Gram-positive strains tested, chlorhexidine gluconate had a better inhibitory effect than povidone-iodine. The antibacterial efficacy of essential oils highlights their potential to substitute or complement the use of antiseptics and so reduce resistance to antiseptics.

As in human medicine, in veterinary medicine, chronic wounds are often related to polymicrobial infections and the presence of a biofilm, which compromises the effectiveness of therapeutic approaches. In this study, a Lusitano mare presented a 21-day-old chronic wound that was only being treated with an antiseptic. A swab sample was collected, and three isolates of Staphylococcus aureus and one of Pseudomonas aeruginosa were isolated. S. aureus did not show resistance to a panel of antibiotics. However, the P. aeruginosa isolate showed a resistance profile to carbapenems and fluoroquinolones, which may suggest a cross-resistance between antiseptic and antibiotics, given that no antibiotic therapy was applied to the wound or the mare in the previous year. Further experiments were conducted to assess the ability of the isolates to form biofilms, and to ascertain their susceptibility to gentamicin. The results demonstrated that the isolates produced biofilms. Gentamicin at the minimum inhibitory concentration (MIC) and 10× MIC caused biofilm removal between 59.3% and 85.7%, with the highest removal percentage being obtained for the P. aeruginosa isolate (at 10× MIC concentration). This study reveals that an equine wound was colonized by antibiotic resistant bacteria, and that all the wound colonizers could form biofilms, demonstrating the relevance of an adequate diagnosis and treatment when there is a suspicion of a biofilm-infected wound. It also highlights the possibility of resistance transmission between animals, animals and humans, or animals and the environment.

Adult grey horses have a high incidence of melanocytic tumors. This article narratively reviews the role of some genetic features related to melanoma formation in horses, such as STX17 mutation, ASIP or MITF alterations, and the link between the graying process and the development of these tumors. A clear system of clinical and pathological classification of melanocytic tumors in naevus, dermal melanoma, dermal melanomatosis and anaplastic malignant melanoma is provided. Clinical and laboratorial methods of diagnosing are listed, with fine needle aspiration and histopathology being the most relevant. Relevance is given to immunohistochemistry, describing potentially important diagnostic biomarkers such as RACK1 and PNL2. Different therapeutical options available for equine practitioners are mentioned, with surgery, chemotherapy and electroporation being the most common. This article also elucidatesnew fields of research, perspectives, and new therapeutic targets, such as CD47, PD-1 and COX-2 biomarkers.

Melanocytic tumors are an important neoplastic disease in human and veterinary medicine, presenting large differences regarding tumor behavior between species. In horses, these tumors present a prolonged benign behavior, with rare invasiveness and metastases. In humans and small animals, invasion and metastasis have been associated with an Epithelial-Mesenchymal Transition, where the loss of E-cadherin expression plays a key role in tumor progression. This process and the role of E-cadherin have not yet been evaluated in equine melanocytic tumors. This study aimed to assess the immunolabeling of E-cadherin in equine melanocytic tumors and relate this with clinicopathological variables. A total of 72 equine melanocytic tumors were classified as benign and malignant and evaluated by immunohistochemistry for E-cadherin expression. A different pattern of immunostaining was found, contrasting with other species. A total of 69.4% of tumors presented raised immunolabeling of E-cadherin, with 70.7% of melanomas remaining with high expression. The typical loss of immunostaining was not seen in malignant melanomas and no differences were found between benign and malignant melanomas regarding E-cadherin immunostaining. The high immunolabeling of E-cadherin may contribute to the low invasiveness of these tumors, and it is in accordance with the benign behavior of equine melanoma and with the genetic factors associated with its development.

Background: Many emergent pathogenic agents are cross-transmitted from animals to humans.Horses are considered as potential reservoirs of commensal, zoonotic, and multidrug-resistant bacteria.Equine bites could lead to infections caused by these agents, considering equine species as a public health concern.The more it is known about the equine oral microbiota the best secondary problems created by their commensal flora can be controlled.There are very few reports of Serratia rubidaea, a zoonotic and opportunistic bacterium, both in human and veterinary medicine.Aim: This study aimed to evaluate the Gram-negative microbiota of healthy equine oral cavities and their antimicrobial susceptibility.Methods: During equine routine oral procedures, eight healthy horses were selected for this study, after discarding any abnormal dental conditions.Samples were collected from the subgingival space and gingival margin from the tooth 406 and both the identification and antimicrobial susceptibility test of Gram-negative bacteria were performed.Results: This study reports the isolation of 32 Gram-negative agents, 27 of which were multidrug-resistant to the antimicrobial classes tested.High resistance rates were obtained to commonly used antimicrobial drugs, particularly macrolides and aminoglycosides as to carbapenems that are specific to human medicine.Two multi-drug resistance strains of S. rubidaea were found in the mouth of two healthy horses.Conclusion: Most Gram-negative isolates found in healthy horses were zoonotic and multi-drug resistant.This is a strong reason to consider the horse as an animal with a major place in the "One Health" concept.Equine clinicians should take precautions when working with horses' mouths.Antimicrobial sensitivity tests should be taken into consideration when finding the appropriate antimicrobial therapy protocol.To the authors' best knowledge, this is the first report about isolation of S. rubidaea from the mouth of the equine species.

Background: Complete fractures of the third metacarpal/metatarsal bones (Mc/t3) are not common, but can occur in various situations and all types of horses, contributing for approximately one third of all long bone fractures in the horse, mostly related to external trauma or to high energy injuries. To stabilize Mc/t3 fractures in the horse, conservative management with walking casts and/or open reduction and internal fixation techniques are referred in the scientific literature, these last one generally by double platting application to bone surface in 90 degrees. In the present case will be described a Mt3 complete diaphyseal fracture stabilization on an adult pony from a circus by applying only one bone plate. This work pony could return to its previous activity fully recovered 3 months after fracture stabilization. Case Description: A 7-year-old, male, Shetland pony (103 kg), used for performance work at a circus was admitted to the Veterinary Teaching Hospital of University of Tras-os-Montes and Alto Douro, presenting on lateral recumbency after being struck by a truck. On physical examination, a penetrating wound of small dimension was detected at the medial aspect of the right hindlimb. After radiographic examination, a complete diaphyseal fracture with slight obliquity of the right Mt3 was confirmed. The owner opted for surgical repair thus fracture stabilization was achieved by the placement of just a single 8-hole 4.5 mm broad dynamic compression plate applied to the lateral face of Mt3 in compression function with 7 cortical screws (4.5 mm). Post-operatively a full limb cast was performed and maintained for 4 weeks, while the animal was kept under movement limitation at the hospital facilities for this period of time although the animal stayed until his full recovery 3 months’ post-surgery. Conclusion: A complete recovery was achieved and the animal returned to the previous level of exercise without any significant postoperative complications or degree of lameness. In the authors knowledge, this is the first case report where stabilization of a complete Mt3 diaphyseal fracture was attain using a single compression plate contoured to the lateral face of the bone, instead of double plating, on an adult work pony from a circus subjected to a very demanding and intense physical activity.

Neospora spp. are intracellular protozoa with worldwide distribution and closely related to Toxoplasma gondii, which can infect a variety of mammals including horses. From September 2013 to June 2014, 185 horses from northern, central and southern parts of mainland Portugal were randomly sampled and tested for detection of immunoglobulin (Ig) G antibodies to Neospora spp. using an indirect multi-species enzyme-linked immunosorbent assay (ELISA) commercial test (ID Screen® Neospora caninum Indirect Multi-species; ID.vet Innovative Diagnostics, Grabels, France). Two horses (1.1%; CI: 0.1-3.8%), one male and one female, were found to be seropositive for Neospora spp. Both seropositive animals were horses housed indoors but with access to outdoors, used for leisure activities and were apparently healthy, with good body condition and with no alterations at physical examination. This was the first serologic survey of antibodies to Neospora spp. carried out in horses from Portugal.

The Lusitano horse is an autochthonous Portuguese breed with a growing worldwide expansion. Our objective was to establish reference intervals for haematological parameters using the haematological cell counter LaserCyte (IDEXX). For this purpose, blood samples from 100 healthy adult horses (13 females and 87 males, ranging from 3 to 25 years of age) were analysed. The reference intervals were estimated following the ASVCP guidelines with the Reference Value Advisor software. The obtained reference intervals were 6.4–10.1 × 1012/L for red blood cells, 30.6–45.1% for haematocrit, 11.6–17.1 g/dL for haemoglobin, 42.8–53.2 fL for mean corpuscular volume (MCV), 15.5–20.8 pg for mean corpuscular haemoglobin (MCH), 33.7–39.4 g/dL for mean corpuscular haemoglobin concentration, 17.8–20.3% for red cell distribution width (RDW), 4.5–10.1 × 109/L for white blood cells, 2.2–6.0 × 109/L for neutrophils, 0.9–4.9 × 109/L for lymphocytes, 0.2–0.5 × 109/L for monocytes, 0.1–0.6 × 109/L for eosinophils, 0.0–0.1 × 109/L for basophils, 78.5–172.2 K/mL for platelets, 4.3–9.4 fL for mean platelet volume, 18.8–24.2% for platelet distribution width, and 0.06–0.12% for plateletcrit. LaserCyte equine reference intervals are transferable to the Lusitano horse for 18 of the 22 analytes studied. Regarding age, significant statistical differences were observed for MCV, RDW, neutrophils and lymphocytes between the mean values of young (3–6 years old), middle-aged (7–14 years old) and old (< 15 years old) age groups. MCH means were statistically significantly different between the three age groups. The haematological reference intervals established in this study might represent a valuable and applicable tool for haematological assessment of adult Lusitano horses, providing useful information that helps clinicians to interpret clinical data.

Background Disseminated intravascular coagulation ( DIC ) is frequent in horses with severe gastrointestinal disorders. Postmortem studies have found fibrin microthrombi in tissues of these horses, but studies relating these histopathological findings with antemortem hemostatic data are lacking. Hypothesis Antemortem classification of coagulopathy is related to the presence and severity of fibrin deposits observed postmortem in horses with severe gastrointestinal disorders. Animals Antemortem hemostatic profile data and postmortem tissue samples (kidney, lung, liver) from 48 horses with colic. Methods Tissue samples were stained with phosphotungstic acid hematoxylin and immunohistochemical methods for histological examination. A fibrin score (grades 0–4) was assigned for each technique, tissue and horse, as well as the presence or absence of DIC at postmortem examination. D‐dimer concentration, prothrombin time ( PT ), activated partial thromboplastin time ( aPTT ), and antithrombin ( AT ) activity, as well as the clinicopathological evidence of coagulopathy, were determined from plasma samples collected 0–24 hours before death or euthanasia. Histologic and clinicopathologic data from the same horses were compared retrospectively. Results No association was found between antemortem classification of coagulopathy and postmortem diagnosis of DIC based on tissue fibrin deposition. None of the hemostatic parameters was significantly different between horses with or without postmortem diagnosis of DIC . There was no association between horses with fibrin in tissues or different cut‐offs for D‐dimer concentration and postmortem evidence of DIC . Conclusions and Clinical Importance Abnormalities of the routine clotting profile, including D‐dimer concentration, were not useful in predicting histologic evidence of DIC at necropsy in horses with severe gastrointestinal disorders.

Leishmania infantum and Toxoplasma gondii are protozoa with zoonotic and economic importance. Prevalences of antibodies to these agents were assessed in 173 horses from the north of Portugal.