Utility of immature platelet fraction in the Sysmex XN-1000V for the differential diagnosis of central and peripheral thrombocytopenia in dogs and cats.

BACKGROUND: The immature platelet fraction (IPF), a parameter obtained by the Sysmex XN-1000V analyzer, is used in humans to differentiate between central (CEN) and peripheral (PER) thrombocytopenia (TP) but has not been evaluated in small animals. OBJECTIVES: Compare IPF between healthy, clinical non-TP and TP dogs and cats, study IPF in different causes of TP in dogs and cats and, establish IPF reference intervals (RIs), and study the effect of age and sex on IPF in healthy dogs and cats. ANIMALS: A total of 3281 dogs and 726 cats. METHODS: Retrospective review of medical records. Animals were classified as nonthrombocytopenic (healthy group and group of clinical patients without TP [NTP]) or TP. These latter animals were subclassified as pseudothrombocytopenia (PSE), CEN and PER, based on evaluation of platelet clumps, estimated platelet count in blood smears and final diagnosis. Blood samples were evaluated using a Sysmex XN-1000V with a specific platelet channel (PLT-F). RESULTS: The IPF was significantly different between each subtype of TP in both species. Immature platelet fractions <6.9% in dogs or 13.6% in cats, once PSE has been eliminated by review of blood smears, are indicative of CEN. Reference intervals for IPF were 0.5%-8% in healthy dogs and 1%-40.3% in healthy cats. CONCLUSIONS AND CLINICAL IMPORTANCE: We determined that IPF can differentiate between CEN and PER in dogs and cats, guiding additional testing and avoiding more invasive procedures (bone marrow sampling). A blood smear always should be evaluated to rule out platelet clumping.

Metabolic and Endocrine Insights in Donkeys.

Donkey medicine is gaining attention due to their increased use as companion animals, in shows, asinotherapy, etc. The increasing demand and unique aspects call for specialized care, requiring new information (physiology, infectious disorders, pharmacology, etc.). Since obesity is common in this species, hyperlipemia, metabolic syndrome and insulin dysregulation (ID) are common disorders in donkeys, in some cases with high mortality, either directly (multiorgan dysfunction) or indirectly due to poor quality of life (chronic laminitis). Donkeys have long-life expectancy and are often afflicted with pituitary pars intermedia dysfunction (PPID), a neurodegenerative and endocrine disease. Hyperlipemia is diagnosed based on high plasma triglyceride concentration in association with clinical findings and laboratory abnormalities from affected tissues (liver, kidney and pancreas). The measurement of resting serum insulin and plasma ACTH concentrations is the first step in ID and PPID diagnosis. In donkeys with clinical signs of ID (obesity or recurrent laminitis) or PPID (hypertrichosis, regional adiposity, laminitis and weight loss), where these hormones are in the normal or non-diagnostic range (donkey-specific cut-off values and reference ranges need to be established), dynamic tests are recommended (oral sugar test or thyrotropin-releasing hormone, respectively). Equine treatment protocols apply to donkeys, although pharmacological studies for most drugs, except pergolide, are lacking.

The Immature Reticulocyte Fraction (IRF) in the Sysmex XN-1000V Analyzer Can Differentiate between Causes of Regenerative and Non-Regenerative Anemia in Dogs and Cats.

The Sysmex XN-1000V analyzer can identify those reticulocytes with high RNA content and fluorescence, providing the immature reticulocyte fraction (IRF). While this parameter has been used in human medicine to identify the cause of anemia, few studies have focused on its use in veterinary medicine. In this study, we determined the IRF and related reticulocyte parameters in a large population of non-anemic and anemic dogs and cats (subclassified depending on the origin of their anemia). The IRF was significantly higher in hemolytic anemias compared to hemorrhagic ones in both species. Moreover, the IRF was significantly lower in dogs and cats with bone marrow failure than in other non-regenerative anemias and in both groups compared to pre-regenerative anemias. The accurate cut-off values for the differential in regenerative anemias and reference ranges for both species using the Sysmex XN-1000V are also reported. The measurement of the IRF in this analyzer can help clinicians to further classify the type of anemia in both species.

Evaluation of the combined glucose-insulin and intravenous glucose tolerance tests for insulin dysregulation diagnosis in donkeys.

BACKGROUND: Insulin dysregulation (ID) and donkey metabolic syndrome (DMS) are common in this species. Contrary to horses, diagnostic guidelines compiling insulin cut-offs values and dynamic testing interpretations have not been reported for this species. OBJECTIVES: To evaluate resting serum insulin concentrations, the combined glucose-insulin test (CGIT) and the glucose intravenous tolerance test (IVGTT) for the diagnosis of DMS with ID suspicion. STUDY DESIGN: Diagnostic test comparison. METHODS: Six of 80 mix-breed adult donkeys fulfilled the inclusion criteria for DMS based on history or clinical evidence of recurrent laminitis, body condition >6 and neck score >2 or baseline insulin and leptin concentrations >20 µIU/mL and >12 ng/mL respectively. CGIT and IVGTT were performed in all donkeys within a week and interpreted following guidelines reported for equine metabolic syndrome (EMS). Insulin and glucose curves were analysed, proxies calculated and correlations and multivariate analysis assessed. RESULTS: Following EMS guidelines, CGIT classified 2 (using glucose-positive phase duration) or 3 (using insulin concentration) and IVGTT classified 5 donkeys as ID. ID donkeys showed a lower glucose/insulin ratio, QUICKI and RISQI, and a higher insulin/glucose ratio, MIRG and HOMA-B%. MAIN LIMITATIONS: Comparison of these tests with additional dynamic testing including a larger number of ID donkeys is necessary. CONCLUSIONS: This is the first study evaluating dynamic tests to assess ID/DMS in DMS-suspected donkeys. IVGTT detected more ID donkeys than CGIT. EMS recommendations could also be used for DMS diagnosis, although a baseline insulin cut-off value is needed.

Characterisation of the oral glucose and sugar tolerance tests and the enteroinsular axis response in healthy adult donkeys.

BACKGROUND: Insulin dysregulation (ID) is diagnosed in horses and ponies using oral glucose (OGTT) and oral sugar (OSTT) tolerance tests. The enteroinsular axis plays a major role in postprandial glucose disposal and insulin response in horses, ponies and foals. The insulin and incretin response to oral carbohydrate challenges has not been characterised in donkeys. OBJECTIVES: (a) To characterise OGTT and OSTT, and (b) to assess the plasma incretin response to OGTT and OSTT in healthy donkeys. STUDY DESIGN: In vivo experiments. METHODS: Six healthy adult female Andalusian donkeys were challenged with OGTT (1 g/kg glucose, 20% solution by nasogastric tube) and OSTT (0.45 mL/kg corn syrup orally by syringe) with a 1-week washout. Blood samples were collected for glucose (spectrophotometry), insulin (radioimmunoassay), glucose-dependent insulinotropic polypeptide (GIP, ELISA) and active glucagon-like peptide-1 (aGLP-1, ELISA) determination over 6 hours. Curves were analysed and proxies calculated. RESULTS: Glucose and insulin concentrations peaked at 180 minutes in OGTT, but at 300 and 150 minutes in OSTT, respectively. Plasma GIP concentrations increased in the OGTT and OSTT (peaked at 180 and 360 minutes, respectively), but aGLP-1 increased only in OGTT (240 minutes). MAIN LIMITATIONS: Single breed, narrow age and sample, diet, season and not having donkeys with evidence of ID to provide clinical validation. CONCLUSIONS: Donkeys have a functional enteroinsular axis that is activated by enteral carbohydrates. Donkeys have evident endocrine differences with horses, supporting the validation of the OSTT and OGTT to assess insulin sensitivity in this species to avoid extrapolation from horses.

Supplementing the Diet of Dairy Goats with Dried Orange Pulp throughout Lactation: I. Effect on Milk Performance, Nutrient Utilisation, Blood Parameters and Production Economics.

Dried orange pulp (DOP) can be incorporated into ruminant diets, but no reports have considered this strategy during the entire lactation period in goats. Two experiments were performed using lactating Payoya goats. In experiment 1, to study the effect, over 180 days, of DOP on milk yield and composition, blood metabolites and economic values, 44 primiparous goats were allocated into three groups: control diet (concentrate plus lucerne) and DOP40 and DOP80 diets, in which DOP replaced 40% and 80%, respectively, of the cereals. Nutrient digestibility and rumen fermentation were also studied (experiment 2). The DOP diets did not affect milk yield and composition. DOP triggered lower intake and digestibility of ether extract and crude protein. Ruminal fermentation was unaffected by DOP, except for a decrease in butyrate for DOP80. The energy balance was unaltered by diet while the balance and retention of nitrogen decreased. Regarding plasma biochemistry, DOP supplementation caused changes that could indicate an improvement in hepatic function and reduced muscular damage and oxidative muscular stress. Moreover, DOP80 provided a profit increase of EUR 3.27/goat. In conclusion, the partial replacement of cereals by DOP is a profitable and healthy nutritional strategy in dairy goats and is suitable for the entire lactation period without compromising productivity.

Reference intervals of acute phase proteins in healthy Andalusian donkeys and response to experimentally induced endotoxemia.

BACKGROUND: Assessment of acute phase proteins (APPs) may allow prompt detection of diseases in donkeys, that otherwise may be missed because of the stoic behavior of donkeys. Reference intervals (RIs) of APPs measured using immunoassays and a comparison of the response of these biomarkers to a controlled inflammatory insult are lacking in donkeys. OBJECTIVES: (a) To describe the RIs for APPs in healthy Andalusian donkeys, (b) to study the effects of sex and age on APPs, and (c) to assess the early response of APPs to experimentally induced endotoxemia. ANIMALS: Seventy-three healthy Andalusian donkeys (67 for RIs and 6 for endotoxemia). METHODS: Serum amyloid A (SAA), haptoglobin (Hp), C-reactive protein (CRP), ceruloplasmin (Cp), α1-acid glycoprotein (AGP), procalcitonin (PCT), ferritin (Ft), and fibrinogen (Fb) RIs were determined. Endotoxemia was induced and samples for APP determination were obtained at regular intervals for 4 hours. RESULTS: The RIs in Andalusian donkeys were: SAA (0.1-0.6 mg/L), Hp (75-2261 mg/L), CRP (1.3-7.0 mg/L), Cp (0-745 mg/L), AGP (0-884 mg/L), PCT (0-504 pg/mL), Ft (26.9-31.8 µg/L), and Fb (115-466 mg/dL). Concentrations of SAA were higher (P < .05) in jacks. Donkeys <5 years old had higher Cp, AGP, and PCT compared to older donkeys. Concentrations of SAA and Hp were significantly increased in endotoxemic donkeys from 2 hours postinduction. CONCLUSIONS AND CLINICAL IMPORTANCE: We illustrated the importance of using species-specific RIs for APPs in donkeys and the effect of age and sex on APP concentrations. Concentrations of SAA and Hp appear to be the most useful biomarkers in donkeys in the early stages of endotoxemia.

Blood Storage Conditions Affect Hematological Analysis in Samples From Healthy Donkeys and Donkeys With Experimentally-Induced Endotoxemia.

Preanalytical factors such as storage time and temperature are proved to induce marked artifactual changes in hematological parameters in horses, small animals and humans. These errors can mirror findings typical of endotoxemia, leading to dangerous misdiagnosis. Since donkeys are common in warm climates and remote regions, blood samples from this species can be subjected to long lasting travels from the farm to the nearest laboratory, frequently under suboptimal conditions. Moreover, as other equids, donkeys are prone to suffer endotoxemia. Nonetheless, stability has not been evaluated in samples for hematology in this species. The aim of this study was to characterize the effect of temperature and storage time in hematological parameters from healthy donkeys and donkeys with induced endotoxemia. Blood samples were collected from six healthy female Andalusian donkeys and stored for 6, 12, 24, and 48 h at several temperatures (4, 24, and 35°C). Endotoxemia was induced in the same animals by an intravenous LPS infusion and samples obtained 30 min post-infusion were handled similarly. Hematological analysis was performed using a laser-based analyzer and blood smear examination. Storage at 24°C caused significant neutropenia after 48 h as well as morphological changes typical of endotoxemia in blood from healthy donkeys as soon as 24 h post-storage. Samples kept at 35°C displayed more profound and earlier artifactual variations. Conservation at 4°C did not cause any significant change in blood parameters. Prolonged (48 h) storage of samples from animals with induced endotoxemia at 24 and 35°C accentuated pre-existing leukopenia and neutropenia. These findings highlight that donkey samples should be stored at 4°C, instead of 24°C as recommended for horses. Moreover, blood smear interpretation should be cautious in samples stored for longer than 24 h and could be misleading when blood is kept at 35°C.

Meloxicam ameliorates the systemic inflammatory response syndrome associated with experimentally induced endotoxemia in adult donkeys.

BACKGROUND: Little information is available about endotoxemia in donkeys. Characterizing the systemic inflammatory response (SIRS) to lipopolysaccharide (LPS) in donkeys would provide valuable clinical and therapeutic information. The effects of meloxicam on endotoxemia have not been studied in this species. OBJECTIVES: To study the pathophysiology and gene expression associated with experimentally induced endotoxemia, and evaluate the effects of meloxicam on experimentally induced endotoxemia in donkeys and in equine monocyte cultures. ANIMALS: Six healthy adult female donkeys. METHODS: Endotoxemia was induced by an IV infusion of LPS for 30 minutes. Animals either received 20 mL of saline or 0.6 mg/kg of meloxicam IV after LPS infusion. The experiments lasted 6 hours. Blood samples were collected serially for hematology, serum biochemistry, interleukin measurement, and leukocyte gene expression analysis. Vital signs were recorded throughout the study. Monocyte cultures were used to test the effects of meloxicam on LPS-activated monocytes. RESULTS: Lipopolysaccharide induced fever, leukopenia, and neutropenia of similar magnitude in both groups, but meloxicam attenuated increases in plasma lactate, tumor necrosis factor-alpha (TNFα), and interleukin 1ß concentrations compared to controls. No differences were detected between groups for cytokine mRNA expression. Furthermore, meloxicam decreased TNFα release in LPS-activated monocyte cultures. CONCLUSIONS AND CLINICAL IMPORTANCE: Meloxicam could be a feasible option for the treatment of endotoxemia and SIRS in donkeys. Additional studies are necessary to investigate possible meloxicam-related posttranscriptional regulation and to compare this drug with other nonsteroidal anti-inflammatory drugs (NSAIDs) in animals with endotoxemia.

Clinical Pharmacology in Donkeys and Mules.

Donkeys and mules show several pharmacodynamic and pharmacokinetic idiosyncrasies that have to be fully considered by any clinician dealing with these species. Because they possess an increased metabolic rate and cellular water content compared with horses, higher doses (or shorter dosing intervals) are usually recommended for those drugs where pharmacologic studies have been performed. Nonetheless, owing to the lack of species-specific information, this assumption cannot be arbitrarily applied. Thus, when a drug protocol published for horses is extrapolated to a donkey or a mule, a close monitoring is required to detect any secondary effect or subdosing.

Metabolic and Endocrine Disorders in Donkeys.

The donkey evolved under harsh and arid environmental conditions, developing unique energy-efficiency traits, with an efficiency to rapidly mobilize fat in situations of increased energy demands or when food is scarce. This evolution has led to an inherent predisposition of donkeys to obesity, dyslipidemias, insulin dysregulation/metabolic syndrome, pituitary pars intermedia dysfunction, and endocrinopathic laminitis. Marked differences have been described in hormone dynamics and testing protocols for the diagnosis of these endocrine and metabolic diseases in donkeys compared with horses, underlining the necessity of a species-specific approach in order to avoid misdiagnosis, unnecessary or inadequate treatments, and additional costs.

Pharmacokinetics of meloxicam after oral administration of a granule formulation to healthy horses.

BACKGROUND: Nonsteroidal anti-inflammatory drugs are administered in horses for several systemic diseases. Selective cyclooxygenase-2 inhibitors are preferred because of lower risk of adverse effects. Several meloxicam formulations have been tested in horses, but a recently marketed granule oral formulation has not been studied. OBJECTIVE: To characterize the pharmacokinetics of a novel granule meloxicam formulation in fasted and fed horses, and to compare pharmacokinetic features with oral suspension and tablets. ANIMALS: Seven healthy adult horses. METHODS: Meloxicam was administered at 0.6 mg/kg in fasted or fed horses. Blood samples were collected for pharmacokinetic analysis, and vital signs, hematology, and biochemistry variables were monitored for 72 hours. RESULTS: No adverse effects were detected. Volume of distribution and clearance after intravenous administration of meloxicam were 0.36 L/kg and 29.12 mL/h/kg, respectively, with a 12.39 hours of terminal half-life. Protein binding was of 97%. Bioavailability was high for every oral formulation, ranging 70%-110%, without feed effect. Because of a slower absorption, meloxicam after administration of granules had a longer half-life (24 and 34 hours, fasted and fed, respectively) and mean residence time (31 and 47 hours), than suspension and tablets (ranging 10-13 and 13-15 hours, respectively). In addition, the time above therapeutic concentration was higher for the granule formulation than other formulations. CONCLUSIONS AND CLINICAL IMPORTANCE: Granule formulation has different PK parameters compared to other oral formulations, which could enable this formulation to be used for different dosage regimens in order to reach a desired clinical effect or decrease the risk of adverse effects.

Pharmacokinetics of the anti-androgenic drug flutamide in healthy stallions.

Alternatives to surgical castration are necessary for controlling the sexual behaviour of stallions with breeding potential in training and competition. Flutamide is a potent selective non-steroidal androgen receptor competitive antagonist that has been used in human beings as an anti-androgenic drug. In this study, the pharmacokinetics and bioavailability of flutamide and its main active metabolite, 2-hydroflutamide, were determined in seven healthy mature stallions. Single doses of flutamide (1mg/kg intravenously, 1mg/kg orally in fasted horses, 5mg/kg orally in fasted horses and 5mg/kg orally in fed horses) were administered randomly at intervals of 2 weeks. All horses had full physical examinations and blood samples were collected for pharmacokinetics, complete blood counts and biochemistry before and after drug administration. Administration of flutamide did not result in any abnormalities on physical examination or in blood parameters. After intravenous administration of flutamide, the volume of distribution was 0.83L/kg and clearance was 1.20L/h/kg. Flutamide and its metabolite had high protein binding values (93-97%). After oral administration, flutamide was rapidly transformed to 2-hydroxyflutamide, with areas under the concentration-time curve ratios of metabolite:drug ∼7. Oral bioavailability was 6.63% after 1mg/kg flutamide in fasted horses, 6.50% after 5mg/kg flutamide in fasted horses and 6.95% after 5mg/kg in fed horses. Half lives of flutamide were close to 1h after intravenous administration and 2h after oral administration. Half lives of 2-hydroxyflutamide were 4.79-6.84h for all routes and doses. After oral administration, oral flutamide reached plasma concentrations that could be effective as an anti-androgenic agent in horses, but further studies are needed to determine whether flutamide has clinical value as an alternative to castration for controlling sexual behaviour in stallions.

Equid herpesvirus 1 and rhodococcus equi coinfection in a foal with bronchointerstitial pneumonia.

A 2-month-old foal with septic shock and severe respiratory distress was referred to the Veterinary Teaching Hospital. Due to poor prognosis, the foal was euthanized. Histopathology showed lesions suggestive of Rhodococcus equi infection associated with a diffuse interstitial infiltrate of foamy macrophages and syncytial cells presenting large acidophilic intranuclear inclusion bodies, fibrin exudates and hyaline membranes. Bacteriological examination from lung and respiratory exudates confirmed R. equi infection, whilst immunohistochemistry and PCR yielded a positive result for Equid herpesvirus type 1 (EHV-1). Several etiologies have been proposed for bronchointerstitial pneumonia in foals, although a multifactorial origin for this lesional pattern could be possible. This work is the first one describing a combined EHV-1 and R. equi infection in a foal affected with bronchointerstitial pneumonia.

Comparison of donkey hemogram using the LaserCyte hematology analyzer, an impedance system, and a manual method.

BACKGROUND: Donkeys are becoming increasingly important worldwide; therefore a reliable and accurate method of diagnosing disease is necessary. Flow cytometry-based hematologic analyzers are present in veterinary laboratories, but performance of LaserCyte has not been evaluated in donkeys. OBJECTIVES: The objective of the study was to compare the results of donkey blood obtained from the LaserCyte with impedance and manual methods. METHODS: Blood samples were collected from 84 healthy donkeys (1-20 years old) and measured with LaserCyte, Sysmex F-820 and manually. Agreement between methods was studied using Passing-Bablok test and Bland-Altman plots. Influence of blood abnormalities found on blood smears on LaserCyte counts was examined using Mann-Whitney or Kruskal-Wallis test. Intraassay precision was calculated. RESULTS: Hematologic variables obtained from the LaserCyte were significantly different from those obtained with impedance or manual methods; numerous values were flagged. Agreement between LaserCyte and manual method was poor for the majority of variables, but agreement between LaserCyte and impedance was only poor for HCT, MCH, and MCHC. LaserCyte had an intraassay precision < 10% for RBC and platelet variables, and > 10% for WBC variables. CONCLUSIONS: LaserCyte results were not interchangeable with results from other methods due to poor agreement. LaserCyte provided no additional hematologic variables or clinically relevant indices for donkey blood analysis. A large number of results were flagged, requiring the evaluation of blood smears. No benefits were found for the use of LaserCyte analyzer over the use of impedance or manual methods in this study. Specific software for LaserCyte for donkey blood would be beneficial.

Granulomatous giant cell submandibular sialadenitis in a dog.

A 4-month-old dog was presented with a progressive swelling of the submandibular area. The history, course, cytological, and sialographic findings were consistent with an aseptic pyogranulomatous sialadenitis with concurrent duct blockage. This rare entity, responsive to medical treatment, appears to be similar to the granulomatous giant cell sialadenitis of humans.

Sialadénite submandibulaire à cellules géantes granulomateuses chez un chien. Un chien âgé de quatre mois a été présenté avec une enflure progressive de la région submandibulaire. L'anamnèse, l'évolution, les résultats cytologiques et sialographiques étaient conformes à une sialadénite pyogranulomateuse avec un blocage concomitant des canaux.. Cette entité rare, qui a réagi au traitement médical, semble être semblable à la sialadénite à cellules géantes granulomateuses des humains.(Traduit par Isabelle Vallières).