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Mammalian spermatogenesis is a temperature-sensitive process, and an increase in testicular temperature impairs spermatogenesis. Leptin modulates testicular activity, but the effect of leptin or its synthetic analogue on heat-induced testicular impairment is unclear. We investigated the effects of synthetic leptin peptide (116-130 amides) on testicular activity in heat-stressed mice model. 15 adult mice (25.54 ± 1.43 g) were selected for the study. Ten mice were subjected to a single heat stress treatment (HS) at 43 °C for 15 min by submerging the lower half of the body in a thermostatic water bath. After heat treatment, mice were divided into two groups, the heat-stressed HS group (n = 5) and the second group as HSL, treated with leptin peptide (116-130 amide) for 14 days. The HS group showed a significant (p

Made available in DSpace on 2022-04-29T08:36:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2022-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The neonatal period in dogs remains associated with high mortality rates. Sepsis is the main cause of neonatal losses during the first three weeks of life. Additionally, failure in the clinical assessment and early diagnosis of sick newborns is still common, leading to inadequate care, which contributes to a high mortality rate. Thus, the objective of this study was to describe the incidence of sepsis in canine newborns, the clinical aspects involved, the main isolated bacterial agents and mortality rates to facilitate clinicians’ early recognition of this condition. Of the 152 litters and 762 neonates evaluated, 14.8% (113/762) had sepsis or septic shock, and the mortality rate among affected puppies was 25.6% (29/113). Among the puppies with sepsis that died, early mortality (0–2 days of age) occurred in 69% (20/29) of affected neonates, and late mortality (3–30 days of age) occurred in 31% (9/29) of affected neonates. Significant differences (p < 0.0001) in clinical parameters (heart and respiratory rates, blood glucose, body temperature, peripheral oxygen saturation and reflexes) were noted among healthy neonates and neonates with sepsis and septic shock. The main and most relevant clinical signs were apathy, a reduced sucking reflex, diarrhea, the neonatal triad, failure to gain weight, bradycardia, dyspnea, cyanotic mucous membranes, body erythema, reduced peripheral oxygen saturation, cyanosis and tissue necrosis in the extremities. The mother may have been the main source of infection for 87.6% (99/113) of neonates with sepsis. Most infections were transmitted during pregnancy (68%, 77/113) in cases of neonatal sepsis. The major source of infection for neonates was the uterus, followed by breast milk and maternal oropharyngeal secretions. The most frequently isolated bacterial agent was Escherichia coli, accounting for 25.6% (29/113) of sepsis cases. The morbidity and mortality of neonatal sepsis in dogs is high. The clinical evaluation and diagnosis of sepsis in neonates differ from those in adult animals. Thus, knowledge of the neonatal particularities of sepsis is essential for proper clinical management and greater survival of these patients. Veterinary Neonatology Research Group São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinics São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Animal Improvement and Nutrition São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Surgery and Animal Reproduction Veterinary Neonatology Research Group São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Clinics São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Animal Improvement and Nutrition São Paulo State University (Unesp) School of Veterinary Medicine and Animal Science Department of Veterinary Surgery and Animal Reproduction

Many disorders affect the equine foot, and many hoof problems have multiple predisposing causes. Surgery may be necessary after conservative management has failed. Diseases of the hoof capsule may seem simple, but their effect on performance can be long-lasting and healing is often prolonged. Diagnosis of problems within the hoof capsule is enhanced with the use of computed tomography and MRI. The prognosis of fractures has improved with strategic placement of lag screws across fracture planes using aiming devices and advanced intraoperative imaging techniques. Collaboration between the clinician and a skilled farrier is important for successful management of hoof disorders.

The multiple topics summarized in this article and discussed in detail in this issue of VCNA are comprehensive and in-depth, exploring concepts and clinical experiences for state-of-the-art care of the equine foot. The research on the equine foot will translate to the clinical setting and with this the compassionate care of the horse.

Cryptorchidism is a common disorder in the canine population with some aspects still unclear. Although the bilateral condition is known to lead to fertility problems and predisposition to testicular cancer, the neoplastic risk for scrotal testis in unilateral cryptorchid dog is controversial. Therefore, the therapeutic approach to the canine unilateral cryptorchid is arbitrary so far. This study aimed to investigate precancerous testicular lesions, such as immaturity and atrophy, and compare them in scrotal and undescended testes using an in-depth diagnostic analysis based on immunophenotypic patterns. With this purpose, 26 adult male dogs of different ages and breeds, affected by unilateral or bilateral cryptorchidism were enrolled. After surgical removal, testes were examined immunohistochemically to assess their positivity for specific markers of the canine foetal/neonatal period, that is vimentin (VIM), cytokeratin (CK), desmin (DES), inhibin-α (INH), and anti-Müllerian hormone (AMH) in Sertoli cells, and placental alkaline phosphatase (PLAP) in germ cells. Except for the ubiquitous VIM, all the markers were more expressed in neoplastic gonads compared to healthy ones (P 0.05). Similarly, testes detected with Sertoli cell-only tubules as well as with Sertoli cells hyperplasia showed higher expression than gonads without such alterations for CK, DES, AMH and PLAP, and for CK and DES, respectively (P 0.05). The same trend was observed in undescended respect to scrotal testes even though their positivity was significant only for DES, INH and AMH (P 0.05). Immunohistochemical positivity found in scrotal testes of unilateral cryptorchid dogs, even in absence of detectable anatomical abnormalities, was suggestive of precancerous lesions. Despite the limited sample size, this study could help to clarify the predisposition to neoplastic development in normally descended testes. These markers expression in adult life could allow identifying the early stages of the testicular carcinogenesis process besides suggesting a precautionary bilateral surgical approach in unilateral cryptorchid dogs.

Treatment of equine laminitis continues to be a challenge despite recent advancements in knowledge of the pathophysiology of laminitis. With more evidence supporting its use, distal limb hypothermia or cryotherapy has become a standard of care for both prevention of laminitis and treatment of the early stages of acute laminitis. Recent studies have demonstrated that cryotherapy reduces the severity of sepsis-related laminitis and hyperinsulinemic laminitis in experimental models and reduces the incidence of laminitis in clinical colitis cases. This article reviews the recent literature supporting the use of distal limb cryotherapy in horses.

A healthy foot requires a well-balanced foot capable of shock absorption, traction, and normal proprioception. Radiographs and venograms are helpful in assessing health of the external and internal structures of the foot and in early diagnosis. Other techniques to assess foot mechanics include force plate and inertial sensors. When foot pathology ensues, early recognition and emergency mechanical treatment can improve prognosis and overall outcome. Sheared heels, under-run heels, and clubfeet are common problems that need to be corrected early. Successful management and results require he veterinarians and farriers establishing a professional, collaborative, and respectful relationship.

Rate‐adaptive single chamber pacemakers with accelerometer, closed loop stimulation (CLS), and remote monitoring functionality (Eluna 8 SR‐T, Biotronik, SE & Co, Germany) were implanted in 3 miniature donkeys with third‐degree atrioventricular block and syncope. After recovery, different pacemaker programming modes were tested at rest, during stress without physical exercise and during physical exercise. Pacing rates were compared to actual atrial rates and showed that CLS functionality allowed physiological heart rate adaptation. A transmitter installed in the stable provided wireless connection of the pacemaker to the internet. Home monitoring was activated which performed daily wireless transmission of pacemaker functional measurements to an online server allowing diagnosis of pathological arrhythmias and pacemaker malfunction from a distance. Closed loop stimulation and remote monitoring functionality resulted in nearly physiological rate adaptation and allowed remote “from‐the‐stable” patient follow‐up.

Most old, intact male dogs usually have prostate disorders, especially benign prostatic hypertrophy and prostatitis with or without abscesses, and concurrent cystitis. The successful treatment of dogs with prostatitis concurrent with cystitis has relied on choosing an appropriate antimicrobial drug based on a bacterial culture and drug sensitivity testing. The objective of the study was to compare the prevalence of bacterial species and results of drug susceptibility testing of bacteria that were isolated from the prostatic fluids and urine samples that were collected from dogs with both prostatitis and cystitis. One hundred and sixty intact male dogs, who presented with both diseases, were recruited for the study. The disease diagnoses were based on clinical history notes, physical examinations, abdominal ultrasonography, prostatic fluid cytology, urinalysis and bacterial cultures from both prostatic fluid and urine samples. The bacterial culture results demonstrated that the major species that were detected in either the prostatic fluid or urine samples were Staphylococcus spp., Escherichia coli, Pseudomonas spp., Streptococcus spp., Proteus mirabilis and Klebsiella pneumoniae. Staphylococcus spp. (26.5 %, 43/162) and Escherichia coli (26.1 %, 12/46) were the most prevalent species from the prostatic fluid and urine samples, respectively. Statistical tests revealed that there were no significantly different prevalence levels among the isolated bacteria between the prostatic fluid and urine samples. Imipenem and gentamicin were the most potent antimicrobial drugs tested against the bacterial isolates in the present study. However, the administration of imipenem to treat prostatitis and cystitis in dogs was of concern. Interestingly, there were no significant differences in the antimicrobial drug susceptibility trends between the prostatic fluid and urine samples. Based on these results, a urine sample might be considered as an optional sample for bacterial cultures and antimicrobial drug susceptibility testing when it is not possible to collect a prostatic fluid sample.

Glucocorticoids (GCs) are known to play an important role in maintaining basal and stress-related homeostasis by interacting with endocrine mediators and prostaglandins (PGs). Although a growing body of evidence shows that GCs exert their regulatory action at a multitude of sites in the reproductive axis through corticosteroid receptors, little is known about the direct role of cortisol, an active form of GCs, in the equine endometrium. Thus, the study aimed to determine the effect of cortisol on PGF2α synthesis in the endometrial tissue and cells in vitro. In Exp.1, the immunolocalization and the expression of the glucocorticoid receptor (GCR) in the endometrium throughout the estrous cycle were established. In Exp. 2 and 3, the effects of cortisol on PGF2α secretion and transcripts associated with the arachidonic acid (AA) cascade in endometrial tissues, and cells were defined. Endometrial tissues obtained from the early, mid, and late luteal phases and the follicular phase of the estrous cycle were exposed to cortisol (100, 200, and 400 nM) for 24 h. Endometrial epithelial and stromal cells (early phase of estrous cycle) were exposed to cortisol (100 nM) for 24 h. Then, PGF2α secretion and transcripts associated with the AA cascade (PLA2G2A, PLA2G4A, PTGS2, and PGFS) were assessed. GCR was expressed in the cytoplasm and the nucleus in the luminal and glandular epithelium as well as in the stroma. Endometrial GCR protein abundance was up-regulated at the late luteal phase compared to the mid-luteal phase of the estrous cycle. Cortisol dose-dependently decreased PGF2α secretion, PLA2G2A and PLA2G4A transcripts in endometrial tissues. Additionally, cortisol treatment decreased PGF2α secretion from endometrial epithelial and stromal cells. Moreover, it affected PLA2G2A, PLA2G4A, and PTGS2 transcripts in endometrial stromal cells. These findings suggest that cortisol suppresses the synthesis of PGF2α by affecting the AA cascade in the equine endometrium during the estrous cycle.

Oxidative stress, caused by extreme accumulation of un-scavenged reactive oxygen species, plays an integral role in the Ischemia-Reperfusion (I/R) injury to the testicles following testicular torsion. The current research aimed to examine the protective effects of crocin as a natural antioxidant on testicular I/R injury in rats. Animals were divided randomly into five groups (seven each): (1) sham group, (2) torsion/detorsion (T/D) group, (3) intact group with 100 mg/kg crocin, (4) and (5) T/D groups followed by treatment with two different doses of crocin (50 and 100 mg/kg (IP)). I/R injury was induced by 720° clockwise torsion of the left testicles for 2 h. After 24 h of reperfusion, blood samples and epididymal sperms were collected to measure biochemical (GPx, SOD, and MDA), hormonal (testosterone), and sperm parameters (total sperm recovery, motility, viability, and morphology). Moreover, affected testicles were subjected to histopathology examination. I/R injury caused a significant reduction in sperm characteristics (except for morphology) (P 0.05), which could not be significantly improved by crocin administration at either dose (P 0.05). Johnsen's testicular score, mean seminiferous tubular diameter, and germinal epithelial cell thickness were significantly decreased in the T/D group compared to the intact and sham groups. However, crocin could significantly improve the histopathological parameters in both treatment groups compared to the T/D group (P 0.05). T/D reduced SOD and GPx activity and testosterone level significantly (except for GPx) compared to the sham group (P 0.05). However, crocin administration could significantly reverse them. Also, crocin reduced the amount of MDA significantly in the high-dose treatment group in comparison to T/D group (P 0.05). The results of the current study revealed that crocin could be a promising agent to protect against I/R injury following surgical correction of the testicular torsion.

The circadian clock system plays an important role in regulating testosterone synthesis in mammals. Male Bmal1-/- mice are infertile with low serum testosterone levels and decreased expression of testicular steroidogenic genes, suggesting that circadian clock genes regulate testosterone biosynthesis by activating steroidogenic gene transcription. However, whether the circadian clock regulates testosterone production via other genes remains unknown. Using Bmal1-/- mice and their wild-type (WT) siblings, we aimed to identify additional genes by which the circadian clock regulates testosterone synthesis. WT and Bmal1-/- mouse testes sections had similar normal morphologies, although there was a decrease in testicular spermatozoa in the Bmal1-/- mice. Low serum testosterone levels were detected in the Bmal1-/- mice. RNA sequencing identified 37 and 48 genes that were differentially expressed between WT and Bmal1-/- mouse testes at circadian time (CT2 and CT14), respectively. The cholesterol metabolism pathway was significantly enriched in the KEGG pathway analysis, and there was lower expression of three apolipoprotein genes (Apoa1, Apoa2, and Apoc3) at CT2 in the testes of Bmal1-/- mice than in those of WT mice. These decreases in Apoa1, Apoa2, and Apoc3 expression were verified by quantitative polymerase chain reaction analysis, which also revealed downregulation of the expression of the circadian clock (Per2, Dbp, and Nr1d1) and steroidogenic (StAR, Cyp11a1, and Hsd17b3) genes. The expression of circadian clock genes was relatively stable in WT mice over a 20-h period, whereas there was clear circadian rhythmic expression of Apoa1, Apoa2, Apoc3, StAR, Cyp11a1, Hsd3b2, and Hsd17b3. Bmal1-/- mice showed severely reduced expression of testicular circadian clock genes at three time points (CT4, CT12, and CT20), and a reduction in mRNA expression levels of Apo (Apoa1, Apoa2, and Apoc3) and steroidogenic (StAR, Cyp11a1, Hsd3b2, and Hsd17b3) genes. Oil Red O staining showed decreased lipid aggregation in the Leydig cells of Bmal1-/- mouse testes. Considering the vital role of Apo genes in high-density lipoprotein formation and cholesterol transport, the present data suggest that the circadian clock system regulates testosterone production by orchestrating the rhythmic expression of Apo genes. These data extend our understanding of the role of the circadian clock in regulating testosterone production in mammals.

The aim of this study was to investigate the effects of maternal, hormonal, and fetal factors on early fetal volume (FV) measurements in mares obtained by three-dimensional (3D) ultrasound. Furthermore, postpartum parameters were explored in regard to their association with early FV. For this purpose, 149 German warmblood mares that were artificially inseminated and confirmed to be pregnant between days 14–16 of gestation, were examined transrectally at day 45 ± 1 of gestation with the portable 3D ultrasound device Voluson® i (GE, Zipf, Austria). FV was calculated by using the extension software Virtual Organ Computer-aided AnaLysis (VOCAL™). Two different mixed linear models were used to analyze associations between the investigated maternal and fetal factors and the FV. Explanatory variables investigated in the first model were: maternal age, parity, maternal weight, and body condition score, type of pregnancy (recipient or biological mother), barren status (lactating or non-lactating), fetal sex, progesterone (P4) and equine chorionic gonadotropin (eCG) concentrations; and in the second model outcome variables such as gestational length, birth weight, placental weight, fetal sex, and abortion were included in the analysis. The final models revealed a significant relation between FV and eCG (b = 0.011, P = 0.030), as well as with P4 (b = −0.053, P = 0.016), but interestingly P4 was negatively related to FV. Fetal sex showed the most prominent effect on FV (b = −0.256, P = 0.039), with female fetuses being smaller than male fetuses. In the second model none of the investigated parameters were related to early FV except for fetal sex (b = -0.328, P = 0.047), again with female fetuses being smaller. In summary, it was found that FV is related with eCG, P4 and fetal sex, but was no suitable predicting factor for the investigated outcome parameters. Furthermore, the findings suggest that sex specific growth differences exist already in early gestation. The detailed biological mechanism by which P4 and eCG affect fetal size has to be investigated in prospective studies.

In this paper, we described the incidence of early pregnancy loss (EPL) both after natural mating and embryo transfer, evaluated risk factors, and summarized the outcome of twin pregnancies throughout gestation in dromedaries under reproductive care. Data were collected over seven breeding seasons at the world's largest camel dairy farm (study 1). In addition, we determined the timing of EPL and monitored serum progesterone (P4) concentration between Days 13-70 of gestation during one breeding season (study 2). In the first study, out of 2970 pregnancies, 507 cases (17.1%) of EPL were diagnosed with transrectal ultrasonography. The rate of EPL after natural mating and embryo transfer was 16.1% (n = 422 out of 2616) and 24.0% (n = 85 out of 354), respectively. Twin pregnancies were detected in 215 cases (7.2% of all gestations), and 57 of those (26.5%) underwent complete EPL. Almost half of the early losses (n = 243; 47.9%) occurred before 30 d of gestation. Another 43.2% (n = 219) of EPL was diagnosed during the next month, and 8.9% (n = 45) occurred after 60 d of gestation. Multivariable mixed effects logistic regression models revealed that the breeding season (year) and twin pregnancy were the most important exposure variables affecting the rate of EPL (P 0.001). The effect of some male camels was also demonstrated while other factors, such as type of breeding, age category, month of mating, breed/ecotype and reproductive history did not prove to have a significant influence. In the second study, the overall rate of EPL was 24.5% (n = 34 of 139). There was no difference in the incidence of EPL between ET recipient (24.2%, n = 23 of 95) and mated (25%, n = 11 of 44) camels. Weekly rate of EPL ranged from 0.9% to 4.8% with a decreasing tendency, and approx. 41% of the animals (n = 14 of 34) had some ultrasonographic signs of impending EPL 1 week before the final diagnosis. Mean serum P4 concentration in camels with subsequent EPL was 5.3 ± 0.1 ng/ml compared to 5.6 ± 0.04 ng/ml in normal pregnant dromedaries. Day of gestation and future EPL influenced serum P4 levels (P 0.001) with an interaction between the two fixed factors (P 0.05). At the time of the final diagnosis of EPL, mean serum P4 concentration was 2.8 ± 0.44 ng/ml. Although twinning had an unfavorable prognosis with a total pregnancy loss of 36.7%, it was not entirely detrimental for the final outcome of gestation as two-thirds of twin pregnancies (n = 136 out of 212) resulted in the birth of a live calf.

The aim of this study was to determine the cellular and molecular mechanisms of leptin (LEP) and the leptin receptor (LEPR) in testicular development of prepubertal ganders. In an in vivo animal experiment, active immunization against LEPR severely depressed prepubertal testicular development by significantly reducing testicular weights at 200 and 227 days of age. The number of elongated spermatids in the seminiferous tubules was also significantly decreased by immunization with LEPR at ages of 200 and 227 days. Inhibition of testicular development by LEPR immunization was associated with decreases in LHR, StAR, 3β-HSD, CYP11A1, CYP17A1, and PRLR mRNA expression levels in testicular tissue, which resulted in a significant decrease in testosterone synthesis. In the in vitro experiments, the addition of LEP combined with anti-LEPR antibodies strengthened LEPR signal transduction, and inhibited significantly testosterone production in cultured Leydig cells isolated from prepubertal gander testes. The mRNA expression of LHR, StAR, 3β-HSD, CYP11A1, CYP17A1 also decreased significantly after treatment with LEP combined with anti-LEPR antibodies in cultured Leydig cells. These results suggest that anti-LEPR antibodies strengthen LEPR signaling transduction in the presence of LEP, and immunization against LEPR inhibited testes development and testosterone secretion in prepubertal ganders.

Kisspeptins (KISSs) and RFamide-related peptide-3 (RFRP-3) affect the synthesis and secretion of luteinizing hormone (LH) and modulate female reproductive processes. The presence of KISS and RFRP-3 in the porcine pituitary gland and their contribution to the regulation of follicle-stimulating hormone (FSH) synthesis and secretion is unknown. This study analyzed the presence of KISS and RFRP-3 in the pituitary of estrous-cyclic pigs on days 2 to 3, 10 to 11, 12 to 13, 15 to 16 and 19 to 20 and early pregnant pigs on days 10 to 11, 12 to 13 and 15 to 16, and evaluated the effect of KISS and RFRP-3 on β-Fsh mRNA expression and FSH secretion in vitro by pituitary cells collected on selected days of the estrous cycle. The cells were cultured in vitro and treated with KISS (10−6 M, 10−7 M) and RFRP-3 (10−6 M, 10−7 M) or gonadotropin-releasing hormone (GnRH; 100 ng/mL) alone and in combinations (4 h or 24 h). The relative abundance of Kiss and Rfrp-3 and their receptor mRNA transcripts, as well as the KISS and RFRP-3 proteins, were found in the pituitaries of estrous-cyclic and early pregnant pigs. KISS after 4 h increased the secretion of FSH in estrous cyclic pigs mostly during the early-luteal phase and luteolysis. RFRP-3 inhibited the synthesis and secretion of FSH in estrous-cyclic pigs on days 19 to 20 and the secretion of FSH on days 2 to 3 and 10 to 12 of the estrous cycle compared with GnRH-treated cells. KISS in co-treatment with GnRH after 24 h enhanced FSH release on days 2 to 3 and 15 to 16 of the estrous cycle. In conclusion, KISS and RFRP-3 systems are present in the pituitary of estrous-cyclic and pregnant pigs. In estrous-cyclic pigs, KISS and RFRP-3 may affect the synthesis and secretion of FSH by pituitary cells.

Background Rates of detecting ≥1 potential enteric pathogens (PEP) or toxins (PEP-T) in feces, blood, or both of horses ≥6 months of age with enteric disease and impact of multiple detections on outcome of horses with colitis has not been reported. Objective To determine detection rates of PEP/PEP-T in feces, blood, or both of horses with enteric disease and effect of detecting multiple agents on outcome of horses with colitis. Animals Thirty-seven hundred fifty-three fecal samples submitted to IDEXX Laboratories and 239 fecal and blood samples submitted to Michigan State University's Veterinary Diagnostic Laboratory (MSUVDL). Methods Retrospective evaluation of PEP/PEP-T testing results was performed to determine rates of detection of 1 or more PEP/PEP-T. Impact of detecting multiple agents on outcome was assessed in 239 horses hospitalized for colitis. Results One or more PEP/PEP-T was detected in 1175/3753 (31.3%) and 145/239 (60.7%) of samples submitted to IDEXX Laboratories and MSUVDL, respectively. In a hospitalized cohort, survival to discharge was lower (76%) in horses with 1 agent, compared to horses with either no (88%) or multiple (89%) agents. There was no difference (P = .78) in days of hospitalization between horses with 0 (1-17), 1 (1-33), and > 1 positive (1-20) result. There was no difference in cost of hospitalization (P = .25) between horses with 0 ($2357, $1110-15 553), 1 ($2742, $788-11 005), and >1 positive ($2560, $1091-10 895) result. Conclusions and clinical importance Detection rates of PEP/PEP-T in horses with colitis vary with cohorts and tests performed. Detection of more than 1 PEP or PEP-T did not affect outcome.

Background The role of the gastric microbiome in development or persistence of equine glandular gastric disease (EGGD) remains to be investigated. Hypothesis/objectives The objective was to characterize the glandular mucosal and gastric fluid microbiomes of horses with and without EGGD. It was hypothesized that differences in the mucosal microbiome are associated with EGGD. Animals Twenty-four horses were enrolled. Methods Gastroscopy was performed and EGGD scores recorded (score 0, n = 6; score 1, n = 8; score ≥2, n = 10). Gastric fluid and pinch biopsies of healthy glandular mucosa and EGGD lesions were collected via gastroscope. 16S rRNA amplicon sequencing of the gastric fluid and glandular mucosal biopsies was performed. Relationships between gastric fluid and mucosal microbial community composition were evaluated among EGGD score groups (EGGD 0-BX, EGGD 1-BX, EGGD ≥2-BX) and among endoscopic appearances: controls from horses without EGGD and normal areas, hyperemic areas, and lesions from horses with EGGD. Results Microbial community structure of mucosal biopsies differed among EGGD score groups (Jaccard similarity index; P = .009). Principal coordinate analysis showed separate clusters for EGGD 0-BX and EGGD ≥2-BX. Conclusions and clinical importance A modest difference was detected in the community structure of the gastric glandular mucosal microbiome in association with EGGD score.

Respiratory distress in the horse and foal is an emergency. Managing equine respiratory distress in the field starts with appropriate assessment of the patient to determine whether the breathing obstruction stems from the upper or lower respiratory tract or is nonrespiratory in origin. From a thorough, but efficient, physical examination to point-of-care ultrasound and endoscopy, the veterinarian has many tools in the field to help diagnose the patient.

Background Antemortem definitive diagnosis of lymphoma in horses is often difficult. Thymidine kinase 1 (TK1) assay is a potentially useful biomarker for lymphoma in horses. Hypothesis/objectives To report the clinical performance of a commercially available TK1 assay for diagnosis of lymphoma in horses. We hypothesized that there would be no association between serum TK1 activity and a diagnosis of lymphoma in horses. Animals Forty‐two hospitalized horses, 14 with a definitive diagnosis of lymphoma, 4 with other neoplasia, and 24 with inflammatory disease. Methods Retrospective medical record review, groups were compared via Kruskal‐Wallis and Mann‐Whitney tests, and logistic regression was performed. Results Median (range) TK1 was 3 U/L (0.4‐17.7 U/L) in horses with lymphoma and 3.9 U/L (0.8‐94 U/L) in horses without lymphoma (P = .59). There was no significant difference in total protein between horses with and without lymphoma (6.6 g/dL [5.5‐8.3 g/dL] vs 6.6 g/dL [4.7‐10.4 g/dL]; P = .83). There was no significant difference in fibrinogen between horses with and without lymphoma (447 [100‐1364] mg/dL vs 433 [291‐2004] mg/dL; P = .47). On logistic regression, serum TK1 activity was not associated with a diagnosis of lymphoma (odds ratio, 0.97; 95% confidence interval, 0.9‐1.05, P = .48). Conclusion and Clinical Importance Serum TK1 values were not predictive of lymphoma diagnosis in this cohort of horses.

Background Bilateral sinus disease is relatively uncommon in horses, accounting for 3%–4.5% of horses with sinonasal disease, but may require bilateral paranasal surgery for complete resolution. Complications and recurrence following bilateral sinusotomy have not been reported or compared to those following unilateral procedures. Objective To describe clinical features and outcomes in horses undergoing standing single, caudally based bilateral frontonasal sinusotomy compared to unilateral frontonasal surgery. Methods Records of horses (n = 37) undergoing surgical treatment for sinus disease (five bilateral, 32 unilateral) were retrospectively reviewed (2010–2017) for signalment, presenting complaint, duration of signs preoperatively, diagnostic imaging, treatments administered, duration hospitalization, complications, and owner satisfaction with the procedure. Mann–Whitney testing was used to compare age, duration of hospitalization, and follow‐up time in horses undergoing unilateral or bilateral procedures. Fisher's exact testing was used to determine if sex predilection was present for unilateral or bilateral disease. Survival time and time to recurrence were compared by Kaplan–Meier survival curves and log‐rank curve comparison testing. Significance was assessed at p, Bilateral paranasal sinus disease was safely and effectively treated in standing, sedated horses through a single, caudally based bilateral front‐nasal bone flap approach, resulting in return to intended use in all cases. The bilateral approach may be considered to treat bilateral paranasal sinus disease without concern for increased risk of life‐threatening complications or longer hospitalization than would be typical for unilateral sinus bone flap procedures.

Background Left-sided cardiac volume overload (LCVO) can cause fluid accumulation in lung tissue changing the distribution of ventilation, which can be evaluated by electrical impedance tomography (EIT). Objectives To describe and compare EIT variables in horses with naturally occurring compensated and decompensated LCVO and compare them to a healthy cohort. Animals Fourteen adult horses, including university teaching horses and clinical cases (healthy: 8; LCVO: 4 compensated, 2 decompensated). Methods In this prospective cohort study, EIT was used in standing, unsedated horses and analyzed for conventional variables, ventilated right (VAR) and left (VAL) lung area, linear-plane distribution variables (avg-max VΔZLine, VΔZLine), global peak flows, inhomogeneity factor, and estimated tidal volume. Horses with decompensated LCVO were assessed before and after administration of furosemide. Variables for healthy and LCVO-affected horses were compared using a Mann-Whitney test or unpaired t-test and observations from compensated and decompensated horses are reported. Results Compared to the healthy horses, the LCVO cohort had significantly less VAL (mean difference 3.02; 95% confidence interval .77-5.2; P = .02), more VAR (−1.13; −2.18 to −.08; P = .04), smaller avg-max VΔZLLine (2.54; 1.07-4.00; P = .003) and VΔZLLine (median difference 5.40; 1.71-9.09; P = .01). Observation of EIT alterations were reflected by clinical signs in horses with decompensated LCVO and after administration of furosemide. Conclusions and Clinical Importance EIT measurements of ventilation distribution showed less ventilation in the left lung of horses with LCVO and might be useful as an objective assessment of the ventilation effects of cardiogenic pulmonary disease in horses.

Management of fractures in the field starts with successful assessment and stabilization of the patient by the practitioner on the front lines. A careful examination is vital to succesful patient management. This includes identifying the fracture location and severity, evaluating skin integrity and potential contamination of the fracture, and treating any ongoing hemorrhage, hypovolemia or stress. Appropriate application of splints in the field will minimize ongoing tissue damage and improve patient comfort. This ultimately aids further assessment, facilitates referral, and improves opportunities for successful fracture repair.

Selected conditions affecting broodmares are discussed, including arterial rupture, dystocia, foal support with ex utero intrapartum treatment, uterine prolapse, postpartum colic, the metritis/sepsis/systemic inflammatory response syndrome complex, and retained fetal membranes. Postpartum colic beyond third-stage labor contractions should prompt comprehensive evaluation for direct injuries to the reproductive tract or indirect injury of the intestinal tract. Mares with perforation or rupture of the uterus are typically recognized 1 to 3 days after foaling, with depression, fever, and leukopenia; laminitis and progression to founder can be fulminant. The same concerns are relevant in mares with retention of fetal membranes.

Background Acute colitis is a serious cause of morbidity and death in horses. Recent studies have compared clinical features of coronavirus and salmonellosis, but no study has compared clinical features of enteric salmonellosis, coronavirus, and neorickettsiosis. Hypothesis/Objectives To identify risk factors for laminitis and nonsurvival to discharge in horses with enteric salmonellosis, coronavirus, or neorickettsiosis. Animals Eighty‐five horses hospitalized for acute colitis from 2011 to 2019. Methods Retrospective case series. Medical record review (2011‐2019) of adult (≥2 years) horses with colitis. Primary outcomes were laminitis and survival to discharge. Multivariable logistic regression was performed to assess association between variables and the development of laminitis. Stepwise Cox regression was performed to assess association between variables and survival. Results Seventeen of 85 (20%) horses developed laminitis during hospitalization. Neorickettsiosis cases (11/26, 42%) were more likely to develop laminitis than coronavirus (0/16, 0%) cases (odds ratio [OR] 24.48; 95% confidence interval [CI]: 1.33‐451.74, P = .03). There was no significant difference in laminitis between salmonellosis and neorickettsiosis cases (OR 0.27; 95% CI: 0.07‐1.07, P = .06). Admission heart rate (OR 1.08; 95% CI: 1.02‐1.15, P = .01), total solids (OR 0.17; 95% CI: 0.06‐0.54, P = .003), band neutrophils (OR 1248.47; 95% CI: 6.62‐235 540, P = .008), and bicarbonate concentration (OR 0.68; 95% CI: 0.5‐0.92, P = .01) were predictive of development of laminitis during hospitalization. Sixty‐three of 85 (74%) horses survived to discharge: 16/16 (100%) coronavirus cases, 17/26 (65%) neorickettsiosis cases, 14/20 (70%) salmonellosis cases, and 16/23 (70%) unknown cases. Packed cell volume (hazard ratio [HR] 1.17; 95% CI: 1.09‐1.26, P

Despite substantial research efforts to improve the treatment and outcome of horses with asthma, glucocorticoids (GC) remain the cornerstone of drug treatment of this prevalent disease. The high efficacy of GC to relieve airway obstruction explains their extensive use despite potential deleterious effects. However, much is yet to be uncovered concerning GC use in horses with asthma, including the comparative efficacy of the different drugs, the determination of minimal effective doses and the mechanisms underlying their variable modulation of airway inflammation. The objectives of this structured review were to report and compare the plethora of effects of the various GC used in asthmatic horses with a focus on impact on lung function, airway inflammation, and bronchial remodeling. Adverse effects are also briefly described, with an emphasis on those that have been specifically reported in horses with asthma. Ultimately, we aimed to highlight gaps in the literature and to identify future research areas.

Background Sphingolipids modulate insulin sensitivity in mammals. Increased synthesis of ceramides is linked to decreased insulin sensitivity of tissues. Conversely, activation of the insulin signaling pathway can downregulate ceramide synthesis. Elucidating the association between sphingolipid metabolism and insulin response during oral glucose testing may help explain the pathophysiology of insulin dysregulation in horses. Hypotheses Horses with insulin dysregulation will have a plasma sphingolipid profile characterized by increased ceramide concentrations. The plasma sphingolipid profile will have decreased ceramide concentrations after acute activation of the insulin signaling pathway by oral glucose testing. Animals Twelve Icelandic horses. Methods Horses were subjected to an oral glucose test (0.5 g/kg body weight glucose), with plasma insulin concentrations measured at 0, 30, 60, 120, 180, and 240 minutes postglucose administration. Plasma samples were collected at 0 and 120 minutes for sphingolipid profiling using a liquid chromatography‐mass spectrometry‐based metabolomics analysis. Eighty‐three species of sphingolipids were detected, including 3‐ketosphinganines, dihydroceramides, ceramides, dihydrosphingomyelins, sphingomyelins, galatosylceramides, glucosylceramides, lactosylceramides, and ceramide‐1‐phosphates. Results Glucose administration did not significantly alter plasma sphingolipid profiles. C22:0‐ceramide, C24:1‐ceramide, C23:0‐ceramide, C16:1‐sphingomyelin, C22:0‐dihydroceramide, and C24:0‐ceramide were positively correlated with the insulin response (area under the curve). Conclusion and Clinical Importance Positive correlation between the insulin response and sphingolipid concentrations implies upregulated sphingolipid metabolism in insulin dysregulated horses. A high plasma ceramide concentration can indicate insulin dysregulation in horses.

The secretion of prolactin and growth hormone in response to exercise and emotional stresses was investigated in Thoroughbreds. Two experiments were performed: one with loading of only exercise stress and one with simultaneous loading of exercise and emotional stresses. Exercise stress was loaded in 4 steps using a treadmill for horses: pre-exercise period (5 min), walking period (6.5 min), galloping period (3 min), and cooling down period (10 min). Emotional stress was loaded by showing a loud video of an audience at a racetrack during the walking period. The results clearly demonstrated that exercise stress rapidly increased the secretion of prolactin and growth hormone and that secretion of them persisted for a specific period after the exercise. In addition, emotional stress promoted prolactin secretion.

The physiological and biochemical responses of the equine athlete, before, during and after physical exercise, are studied in the exercise physiology and, for this to start and muscle contraction occurs, for this it is necessary that the organism produces energy through the production of ATP, which occur in three processes, being alactic or lactic, the alactic process occurs through the metabolism of creatine phosphokinase (anaerobic form) and glucose through the Krebs cycle (aerobic form), whereas the lactic process occurs through the metabolism of lactate. During exercise, in an attempt to re-establish equine parameters, physiological changes occur. The physiology of exercise has been studying, through stress tests, performed on an ergometric treadmill or in the field, ways of obtaining parameters to evaluate the species. This study aimed to evaluate an essential tool that is the physiology of exercise on the physical conditioning of equine athletes, through parameters that observe the performance of these animals. For the methodology, a bibliographic survey of articles was made and directed to the physiology of equine exercise, in addition, necessary tools for the evaluation of equine athletes were addressed, being these neurological, cardiovascular, skeletal, and electrolytic responses. It is concluded that the use of tests for the evaluation of the athletic performance in horses performed by treadmill or in the field together with the physiological responses obtained by the action of exercise and training, is considered a valuable tool for equine sports medicine.

Background Severe asthma in horses is characterized by structural changes that thicken the lower airway wall, a change that is only partially reversible by current treatments. Increased vascularization contributes to the thickening of the bronchial wall in humans with asthma and is considered a potential new therapeutic target. Objective To determine the presence of angiogenesis in the bronchi of severely asthmatic horses, and if present, to evaluate its reversibility by treatment with corticosteroids. Animals Study 1: Bronchial samples from asthmatic horses in exacerbation (7), in remission (7), and aged‐matched healthy horses. Study 2: Endobronchial biopsy samples from asthmatic horses in exacerbation (6) and healthy horses (6) before and after treatment with dexamethasone. Methods Blinded, randomized controlled study. Immunohistochemistry was performed using collagen IV as a marker for vascular basement membranes. Number of vessels, vascular area, and mean vessel size in the bronchial lamina propria were measured by histomorphometry. Reversibility of vascular changes in Study 2 was assessed after 2 weeks of treatment with dexamethasone. Results The number of vessels and vascular area were increased in the airway walls of asthmatic horses in exacerbation (P = .01 and P = .02, respectively) and in remission (P = .02 and P = .04, respectively) when compared to controls. In Study 2, the differences observed between groups disappeared after 2 weeks of treatment with corticosteroids because of the increased number of vessels in healthy horses. Conclusions and Clinical Importance Angiogenesis contributes to thickening of the airway wall in asthmatic horses and was not reversed by a 2‐week treatment with corticosteroids.

Background Blood l-lactate concentrations are higher in people with developing or established diabetes mellitus and insulin resistance. Objectives To investigate whether blood l-lactate concentrations are positively correlated with measures of insulin dysregulation (ID) or increased autumnal ACTH concentrations in ponies. Animals Systemically healthy client-owned ponies (n = 101). Methods Prospective case-control study. Blood samples were obtained from 101 clinically healthy ponies. Breed, weight, height, and subjective and objective measures of body condition were recorded. Blood l-lactate, glucose, triglyceride, total adiponectin, and ACTH concentrations were measured and an oral sugar test (OST) was carried out. Correlations between blood l-lactate and variables of endocrine health were determined. Results Using a seasonal cutoff point of ACTH concentrations ≥47 pg/mL, 55 ponies had increased autumnal ACTH concentrations and 45 did not. Using a basal insulin concentration of >50 μiU/mL, 42 ponies were diagnosed with ID and 58 were not. Using a 60 minutes after OST cutoff point of >45 μiU/mL, 57 ponies had ID and 37 did not. Blood l-lactate concentrations were significantly lower in obese (average body condition score ≥ 7/9) compared to nonobese ponies (0.6 mmol/L; range, 0.0-1.9 mmol/L vs 0.8 mmol/L; range, 0.3-2.7 mmol/L; P = .01). No other significant correlations were detected. No differences were detected between ponies with and without increased autumnal ACTH concentrations (0.7 mmol/L; range, 0.0-2.7 mmol/L vs 0.7 mmol/L; range, 0.3-1.8 mmol/L; P = .84) and with and without ID (0.7 mmol/L; range, 0.3-2.7 mmol/L vs 0.8 mmol/L; range, 0.0-1.6 mmol/L; P = .63). Conclusions and clinical importance Results do not support an effect of endocrine status on l-lactate concentrations in blood of ponies.

Nasal polyps are not very common in horses, comprising of just about 2,5 % of equine sinonasal disease (Tremaine and Dixon, 2001). However, nasal polyps can develop into extensive masses, obstructing partially or totally air flow thus giving dramatic clinical presentations. The aim of this report was to present a surgical technique used to successfully remove a massive nasal polyp in a standing mare. A 9 year old mixed-breed mare was referred to the equine teaching hospital of the Faculty of Veterinary Medicine in Cluj –Napoca for the evaluation of an extensive, unilateral nasal mass. Upon inspection, a cauliflower-appearing mass was protruding from the right nostril. A decision was made to try to surgically remove the mass using a direct approach through a nasal bone flap. Classic approach through the opening of the nostril was impossible due to the extent of the mass. The surgery was performed with the mare standing under sedation and regional anesthesia. The mass was approximately 30 cm by 7 cm and was described histologically as a nasal polyp. The mare made a full recovery, and there was no recurrence in the next three years after surgery.

Background Ventricular septal defects (VSDs) are the most common congenital cardiac defect in horses. Objectives To identify prevalence, age, breed, and sex distribution of VSD and to describe associated clinical and ultrasonographic findings. Animals Hospital‐based population of 21 136 horses presented to the equine internal medicine department. Methods Medical records over a 12‐year period were reviewed for VSD confirmed by ultrasonography. Age, breed, sex, sport discipline, murmur, clinical signs, outcome, VSD type, VSD size, shunt velocity, cardiac dimensions, concomitant cardiac anomalies, and valvular regurgitations were recorded. Results From 1894 horses that underwent echocardiography, 54 had a VSD: 42 as an isolated lesion and 12 as part of complex congenital heart disease (CHD). Median age was 5 years (range, 0‐26) and 1 year (range, 0‐8), respectively. Warmbloods and males were overrepresented. In the isolated VSD group, only 15% had associated clinical signs and most horses had a perimembranous VSD (pmVSD; 36/42). Horses with a pmVSD and clinical signs showed a significantly lower maximal shunt velocity (3.77 vs 5.20 m/s; P

Tracheal fungal infections in horses are rare. This case report describes surgical and clinical management of a filly with a Curvularia sp. infection within the trachea and skin that caused severe intraluminal granulomas and cutaneous nodules, respectively. The patient was successfully treated with itraconazole and surgical excision. RESUMO Infecções fúngicas traqueais em equinos são raras. Este relato de caso descreve condutas clínicas e cirúrgicas em uma égua com infecção por Curvularia sp. na traqueia e na pele, causando granulomas intraluminais severos e nódulos cutâneos, respectivamente. O animal foi tratado com sucesso com itraconazol e exérese cirúrgica.

Background Nicoletella semolina was identified in the airways of horses and its low prevalence could be because of its difficult differentiation from other Pasteurellaceae. Objectives To develop a molecular method for the identification of N. semolina and to evaluate its prevalence in the mouth and the airways of healthy and severe asthmatic horses. Animals Six healthy and 6 severely asthmatic horses in phase I, 10 severely asthmatic horses in phase II, and 10 healthy horses in phase III. Methods Cohort (phases I and II) and cross‐sectional (phase III) studies. Quantitative polymerase chain reaction primers targeting the sodA gene were optimized. N. semolina was quantified in oral and nasal washes and in bronchoalveolar lavage fluid (BALF; phase I, sampled twice), in nasal washes and BALF (phase II, sampled twice), and in nasal washes (phase III). Results N. semolina was found in the nose of 5, 10, and 9 horses in phases I, II, and III, respectively (first sampling for phases I and II). Six BALF from 5 different horses were positive for N. semolina in phase II. In phase I, there was no significant difference in the nasal loads of healthy horses (median (range): 2.04 × 104 copies/mL (0‐2.44 × 105)) and asthmatic horses in exacerbation (3.75 × 102 (0‐4.84 × 106); Wilcoxon's rank sum test, P = .57). Conclusions and Clinical Importance N. semolina is commonly found in the airways of horses. The potential pathogenicity of N. semolina remains to be elucidated, but the molecular technique we developed will facilitate future studies.

Background A study reported low systemic availability of injectable dexamethasone nebulized to healthy horses using the Flexineb mask. When used in horses with severe asthma and a different nebulizer, lack of efficacy and cortisol suppression were observed. Hypothesis Nebulized dexamethasone is as effective as PO administration for the treatment of severe asthma in horses. Animals Twelve horses with severe asthma from a research herd. Methods Randomized clinical trial. Horses were divided into 2 groups and received 5 mg of dexamethasone sodium phosphate by nebulization using a Flexineb mask (NE, n = 6) or PO (OR, n = 6) q24h for 7 days. Lung function and serum cortisol concentrations were evaluated at baseline, after 4 days of treatment (D4) and 1 day after the last treatment (D8). Data were analyzed using linear mixed models with Benjamini‐Hochberg adjustments. Results Lung resistance significantly improved at D4 (mean decrease ± SD, −1.5 ± 0.45 cm H₂O/L/s; 95% confidence interval [CI], −2; −0.6) and D8 (−1.4 ± 0.45 cm H₂O/L/s; 95% CI, −2.4; −0.5) compared to baseline in the OR group only (P = .004 and .01, respectively). Serum cortisol concentration was significantly decreased at D4 and D8 for both groups (maximum decrease, −1.2 ± 0.3 μg/dL; 95% CI, −1.9; −0.6 at D4 for NE group and −2.2 ± 0.3 μg/dL; 95% CI, −2.8; −1.6 at D8 for OR group; P