Energy hormone response to fasting-induced dyslipidemia in obese and non-obese donkeys.

Metabolic and endocrine disorders, such as dyslipidemia, are common in donkeys. Negative energy balance due to fasting, stressful conditions, or disease is a major trigger for fat mobilization often leading to dyslipidemia. The hormonal response to fasting has not been well characterized in donkeys. Therefore, this work aimed to study variations in insulin, glucagon, leptin, total adiponectin, ghrelin, and insulin-like growth factor-1 concentrations, insulin-to-glucagon (IGR) and glucagon-to-insulin (GIR) molar ratios, and lipid and carbohydrate parameters during a 66 h fasting period in 8 adult donkeys, and to determine differences depending on body condition. Obese donkeys developed earlier lipid mobilization (increased plasma total triglyceride and total cholesterol concentrations) compared to non-obese donkeys. Plasma glucose and leptin concentrations decreased in obese animals. After 60 h fasting, obese donkeys showed a significant increase in glucagon and decrease in leptin. GIR significantly increased, while insulin and IGR decreased in both groups. These findings support faster lipid mobilization in response to negative energy status in obese donkeys during fasting, which could be linked to greater glucagonemia and could explain the predisposition of these animals to dyslipidemia.

Using dried orange pulp in the diet of dairy goats: effects on milk yield and composition and blood parameters of dams and growth performance and carcass quality of kids.

Although dried orange pulp (DOP) may conveniently replace cereals in ruminant diets, few studies have considered similar diet substitution for goats. We hypothesised that DOP could replace cereal-based concentrate in goat diets without detrimental effects on growth performance and carcass quality of suckling kids and milk performance and blood biochemical parameters of dams in early lactation. We also hypothesised that DOP substitution may increase the levels of antioxidants, such as phenolic compounds and vitamin E, in milk and improve its total antioxidant capacity (TAC). Therefore, 44 primiparous Payoya dairy goats were allocated to three experimental groups, each fed a different diet: control (CD, n = 14) based on a commercial concentrate with alfalfa hay as forage; and DOP40 (n = 16) in which 40% and DOP80 (n = 14) in which 80% of the cereal in the concentrate were replaced by DOP. The experiment lasted from the final month of pregnancy to 55 days postpartum. The DOP diets did not affect suckling kids' carcass quality, but at 28 days, led to improvement in live weight (LW) and average daily gain (ADG) from birth, although no differences were found between DOP40 and DOP80 (for CD, DOP40 and DOP80, LW at 28 days was 8.00, 8.58 and 8.34 kg and ADG was 184, 199 and 195 g/day, respectively). Diet had no significant effect on milk yield (average daily milk yield and total yield at 55 days were 1.66 l/day and 90.6 l, respectively) and commercial and fatty acid composition. Nevertheless, α-tocopherol, total phenolic compound (TPC) and TAC concentration in milk increased with substitution of cereals by DOP (for CD, DOP40 and DOP80, concentration of α-tocopherol was 21.7, 32.8 and 42.3 µg/100 g, TPCs was 63.5, 84.1 and 102 mg gallic acid equivalents/l, and TAC was 6.63, 11.1 and 12.8 µmol Trolox equivalents/ml, respectively). Every plasma biochemistry parameter considered was within reference values for healthy goats; therefore, no pathological effect was detected for these variables due to dietary treatment. However, DOP diets caused a reduction in plasmatic creatine kinase and aspartate aminotransferase, implying reduced oxidative damage to muscles. In conclusion, DOP may be an interesting alternative to cereals in early lactation goat diets for increasing farmers' income and the healthy antioxidant capacity of milk.

Evaluation of dynamic testing for pituitary pars intermedia dysfunction diagnosis in donkeys.

BACKGROUND: Endocrine disorders are common in donkeys. Pituitary pars intermedia dysfunction (PPID) is thought to be a frequent disturbance in donkeys due to their longevity. However, information on PPID dynamic testing in donkeys is lacking. OBJECTIVES: The objective of this study was to evaluate the previously described guidelines for PPID diagnosis in horses in donkeys with suspicion of PPID. STUDY DESIGN: Prospective experimental study. METHODS: Eighty donkeys were evaluated for PPID suspicion based on clinical signs and baseline adrenocorticotropic hormone (ACTH) concentrations. Six mix-breed donkeys (one jack and five non-pregnant jennies) fulfilling inclusion criteria were subjected to dexamethasone suppression test (DST), thyrotropin-releasing hormone stimulation test (TRH) and combined DST-TRH challenge. Tests were interpreted according to guidelines for PPID diagnosis in horses. RESULTS: Donkeys fulfilling inclusion criteria were diagnosed with PPID by TRH stimulation test (six of six). Both DST (three of six) and DST-TRH (4/6) challenges failed to detect those animals and showed conflicting results. Similarly, cortisol basal concentrations were not consistent with PPID suspicion. MAIN LIMITATIONS: Characterisation of seasonal and geographical location effect on baseline ACTH concentrations and response to TRH is compelling in this species. Further studies with a larger number of donkeys are needed. CONCLUSIONS: This is the first study in donkeys to evaluate common dynamic tests used for PPID diagnosis in horses. Preliminary results agree with the guidelines for PPID diagnosis in horses and baseline ACTH measurement followed by TRH challenge are recommended tests for diagnosis of PPID in donkeys.

Effect of intravenous glucose and combined glucose-insulin challenges on energy-regulating hormones concentrations in donkeys.

Metabolic disorders are highly prevalent in donkeys. Differences in energy regulatory hormones and glucose dynamic testing, including the intravenous glucose tolerance test (IVGTT) and combined glucose-insulin test (CGIT), have been documented between donkeys and horses. The aims of this study were to characterise the insulin:glucagon (IGR) and glucagon:insulin (GIR) molar ratios, at baseline and in response to the IVGTT and CGIT in healthy donkeys, and to determine their correlation with endocrine (leptin, ghrelin and adiponectin) and morphometric variables. Median values and interquartile ranges (IQRs) for IGR and GIR in 49 healthy adult donkeys were 1.5 (IQR, 1.0-1.8) and 0.7 (IQR 0.5-0.9), respectively. IVGTT and CGIT were each performed on eight donkeys, while dynamic testing was performed on six donkeys due to loss of two donkeys from the study. IVGTT induced an increase in IGR (and a decrease in GIR) from 15 to 180min after the onset of the test, but had no effect on leptin, adiponectin or ghrelin concentrations. CGIT resulted in a significant elevation in IGR (and a decrease in GIR) from 15 to 120min after the onset of the test. Plasma leptin concentrations increased significantly at 240min. No correlations were found between ratios, hormones and morphometric measurements. The findings support differences between donkeys and horses, which are likely to be related to proportionally higher glucagon compared to insulin concentrations in donkeys, and may be relevant to disorders related to energy dysregulation in donkeys, including metabolic syndrome and dyslipidaemias.

Epidemiological survey of equine influenza in Andalusia, Spain.

Equine influenza is a highly contagious respiratory disease considered the most important respiratory disease in equids. Although influenza A virus (IAV) has caused outbreaks in equids worldwide, surveillance in these species in Spain has not been conducted. A cross-sectional study was carried out to determine the individual and herd prevalence of antibodies against H3N8 and H7N7 IAV in equids in Andalusia (southern Spain). Antibodies againsts IAV were measured by the single radial haemolysis assay. A spatial scan statistical analysis was carried out using a Bernoulli model. Risk factors associated with IAV infection were assessed by multivariate analysis. Antibodies to H3N8 IAV were detected in 241 out of 464 unvaccinated equids (51.9%; 95% CI: 47.4-56.5). Seropositivity against the H7N7 subtype IAV was not found in any of the analysed animals. Significantly higher seropositivity was found in geriatric (OR = 6.1, P = 0.008, 95% CI = 1.6-23.1) and adult (OR = 4.8, P < 0.001, 95% CI = 2.5-9.0) equids compared to young animals. Specific antibodies against A/equine/Shropshire/2010 (H3N8) or A/equine/Newmarket/5/2003 (H3N8) only were confirmed in 11 and 45 of the animals, respectively. The spatial analysis showed a statistically significant cluster centred in the west part of Andalusia. The results confirmed widespread H3N8 subtype IAV exposure in equine species in Andalusia. Conversely, the absence of seropositivity against H7N7 IAV obtained in the present study suggests that this subtype has not circulated in southern Spain in recent years. Because of the animal health and economic consequences of IAV in equids, further surveillance and molecular studies are required to monitor and characterize the most prevalent IAV circulating in these species in Spain.

Characterisation of clotting factors, anticoagulant protein activities and viscoelastic analysis in healthy donkeys.

BACKGROUND: Studies have demonstrated differences in commonly measured haemostatic parameters between donkeys and horses. Whether clotting factors, anticoagulant protein activities and thromboelastography parameters also differ between species is still unknown. OBJECTIVES: To characterise haemostatic parameters in healthy donkeys and to compare these with those in horses. STUDY DESIGN: Cross-sectional study. METHODS: Clotting factors (V, VII, VIII, IX, X, XI and XII), and antithrombin III, Protein C and Protein S activities were measured in 80 healthy Andalusian and crossbred donkeys and 40 healthy Andalusian crossbred horses with assays based on human deficient plasmas. Thromboelastography was performed in 34 donkeys using a coagulation and platelet function analyser. RESULTS: Donkeys had shorter activated partial thromboplastin time (mean ± s.d. 33.4 ± 5.2 s vs. 38.8 ± 4.2 s; P<0.001) and higher Factor VII (1825 ± 206 vs. 1513 ± 174; P<0.001), IX (142 ± 41 vs. 114 ± 28; P<0.05) and XI (59.4 ± 14.0 vs. 27.2 ± 6.3; P<0.001) activities, whereas horses showed higher Factor X (130 ± 32 vs. 145 ± 23; P>0.05) and XII (96 ± 21 vs. 108 ± 15; P<0.001) activities. Antithrombin III (204 ± 26 vs. 174 ± 29; P<0.001), Protein C (33.16 ± 10.0 vs. 7.57 ± 1.70; P<0.001) and Protein S (median [interquartile range]: 7.8 [5.8-9.3] vs. 6.2 [5.2-7.0]; P<0.001) activities were higher in donkeys. Activated clot time (175 [159-189]), time to peak (6.5 [5.8-7.8]) and clot formation rate (26.9 [16.9-36.4]) in donkeys were shorter than reported values in horses. MAIN LIMITATIONS: Haemostatic pathways could not be fully evaluated in donkeys because some tests are unavailable. Certain fibrinolytic parameters (plasmin, plasminogen, etc.) have not been characterised in donkeys and this may have affected our results. CONCLUSIONS: The haemostatic system in donkeys differs from that in horses and extrapolation of reference values between these species is not appropriate.

Characterization of the intravenous glucose tolerance test and the combined glucose-insulin test in donkeys.

Glucose-insulin dynamic challenges such as the intravenous glucose tolerance test (IVGTT) and combined glucose-insulin test (CGIT) have not been described in donkeys. The objectives of this study were (1) to characterize the IVGTT and CGIT in healthy adult donkeys, and (2) to establish normal glucose-insulin proxies. Sixteen donkeys were used and body morphometric variables obtained each. For the IVGTT, glucose (300 mg/kg) was given IV. For the CGIT, glucose (150 mg/kg) followed by recombinant insulin (0.1 IU/kg) were administered IV. Blood samples for glucose and insulin determinations were collected over 300 min. In the IVGTT the positive phase lasted 160.9 ± 13.3 min, glucose concentration peaked at 323.1 ± 9.2 mg/dL and declined at a rate of 1.28 ± 0.15 mg/dL/min. The glucose area under the curve (AUC) was 21.4 ± 1.9 × 10(3) mg/dL/min and the insulin AUC was 7.2 ± 0.9 × 10(3) µIU/mL/min. The positive phase of the CGIT curve lasted 44 ± 3 min, with a glucose clearance rate of 2.01 ± 0.18 mg/dL/min. The negative phase lasted 255.9 ± 3 min, decreasing glucose concentration at rate of -0.63 ± 0.06 mg/dL/min, and reaching a nadir (33.1 ± 3.6 mg/dL) at 118.3 ± 6.3 min. The glucose and insulin AUC values were 15.2 ± 0.9 × 10(3) mg/dL/min and 13.2 ± 0.9 × 10(3) µIU/mL/min. This is the first study characterizing CGIT and IVGTT, and glucose-insulin proxies in healthy adult donkeys. Distinct glucose dynamics, when compared with horses, support the use of species-specific protocols to assess endocrine function.

Energy-related parameters and their association with age, gender, and morphometric measurements in healthy donkeys.

Donkeys are commonly afflicted by endocrine and metabolic disturbances but few studies have investigated endocrine variables involved in energy regulation and their association with morphometric indices, age or gender in this species. Hemostatic and clinical differences have been demonstrated between horses and donkeys, so to consider both species as metabolically and endocrinologically similar could lead to misdiagnosis. In this study, plasma concentrations of glucose, triglycerides and endocrine factors involved in energy homeostasis (insulin, glucagon, leptin, adiponectin, ghrelin and insulin-like growth factor [IGF]-1) were measured and their association with morphometric variables (body condition score, neck scoring and body mass index), gender and age was determined in 62 healthy donkeys. In addition, a neck scoring system specific for donkeys was developed. Insulin, glucagon, leptin and IGF-1 concentrations were found to be similar between donkeys and other species, but adiponectin and active ghrelin were lower in donkeys than horses. Donkeys with larger neck scores and body mass indices had higher triglyceride, leptin and IGF-1 concentrations. A sexual dimorphism was observed on all morphometric measurements and plasma glucose concentrations independent of adiposity. Younger animals had lower morphometric measurements and triglyceride and leptin concentrations.

Verrucous hemangioma with pseudoepitheliomatous epidermal hyperplasia in an adult horse.

Verrucous hemangiomas are a rare specific variant of equine skin tumors not well described in the literature. An 8-year-old gelding presented a unilateral lesion on the pastern. Macroscopically, the mass showed a warty and verrucous surface with focal ulcerations. The histology showed a dermal proliferation of endothelial-layered capillaries and venules separated by a delicate stroma of scant fibroblasts and collagen deposition, with pseudoepitheliomatous hyperplasia (exuberant reactive irregular epithelial hyperplasia with tongue-like projections extending into the dermis, mimicking downgrowth of squamous cell carcinoma) and orthokeratotic hyperkeratosis of the overlying epidermis. The immunohistochemical study confirmed the endothelial origin of the tumor, and a final diagnosis of verrucous hemangioma with pseudoepitheliomatous hyperplasia was confirmed. To the knowledge of the authors, this is the first detailed description of this entity in adult horses. Moreover, the clinical progression and epidermal changes have not been previously reported, emphasizing the importance of a pathological study of any epithelial preneoplastic lesion to rule out an underlying dermal neoplasia.

Humoral immune response in goats immunised with cathepsin L1, peroxiredoxin and Sm14 antigen and experimentally challenged with Fasciola hepatica.

The humoral immune response was analysed in goats immunised with FhCL1, FhPrx, Sm14, and experimentally challenged with Fasciola hepatica. All immunised animals developed significant levels of anti-fluke specific antibodies and those immunised with FhCL1 showed the highest antibody titre. After experimental infection, an increase in the antibody level was detected only in goats immunised with FhCL1. In the adjuvant-control animals, the experimental challenge induced significant production of specific antibodies against FhCL1, FhPrx and Sm14. While liver fluke specific humoral responses were seen in all groups, no significant protection in any of the vaccinated groups was found.

Immune response of goats immunised with glutathione S-transferase and experimentally challenged with Fasciola hepatica.

Glutathione S-transferase (FhGST) purified from Fasciola hepatica adult worms was used to immunise goats against F. hepatica in an experimental infection; the level of protection, in terms of fluke burden, faecal egg counts and hepatic damage was determined, as well as the humoral and cellular immune response elicited. Animals were allocated into three groups of six animals each: group 1 (immunised with FhGST and infected), group 2 (unimmunised and infected), and group 3 (unimmunised and uninfected). There was no significant reduction of fluke burden (9.3%) or faecal egg counts; hepatic damage was also similar in both infected groups. However, immunisation with FhGST induced the development of a well-defined immune response, characterized by the production of specific-FhGST antibodies as well as the appearance of circulating IL-4.

A study of the liver of goats immunized with a synthetic peptide of the Sm14 antigen and challenged with Fasciola hepatica.

A synthetic peptide of the Sm14 antigen of Schistosoma mansoni was used to immunize goats against experimental challenge with Fasciola hepatica. The goats used consisted of: group 1 (unimmunized and uninfected [controls]); group 2 (unimmunized and infected); group 3 (immunized and infected). Compared with group 2, the animals of group 3 showed at necropsy a reduction in hepatic worm burden; however, because of variability in the two groups this reduction was not statistically significant. A gross morphometric study of the hepatic changes revealed a correlation between lesions and worm burdens in group 3. No significant differences in damaged areas were found between groups 2 and 3, except that group 2 had more severe bile duct hyperplasia. A striking hepatic inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells was found in both groups 2 and 3, especially the latter.