Effect of Dietary Nonstructural Carbohydrate Content on Activation of 5′‐Adenosine Monophosphate‐Activated Protein Kinase in Liver,Skeletal Muscle,and Digital Laminae of Lean and Obese Ponies

Background

In EMS‐associated laminitis, laminar failure may occur in response to energy failure related to insulin resistance (IR) or to the effect of hyperinsulinemia on laminar tissue. 5′‐Adenosine‐monophosphate‐activated protein kinase (AMPK) is a marker of tissue energy deprivation, which may occur in IR.

Hypothesis/Objectives

To characterize tissue AMPK regulation in ponies subjected to a dietary carbohydrate (CHO) challenge.

Animals

Twenty‐two mixed‐breed ponies.

Methods

Immunohistochemistry and immunoblotting for total AMPK and phospho(P)‐AMPK and RT‐qPCR for AMPK‐responsive genes were performed on laminar, liver, and skeletal muscle samples collected after a 7‐day feeding protocol in which ponies stratified on body condition score (BCS; obese or lean) were fed either a low‐CHO diet (ESC + starch, approximately 7% DM; n = 5 obese, 5 lean) or a high‐CHO diet (ESC + starch, approximately 42% DM; n = 6 obese, 6 lean).

Results

5′‐Adenosine‐monophosphate‐activated protein kinase was immunolocalized to laminar keratinocytes, dermal constituents, and hepatocytes. A high‐CHO diet resulted in significantly decreased laminar [P‐AMPK] in lean ponies (P = .03), but no changes in skeletal muscle (lean, P = .33; obese, P = .43) or liver (lean, P = .84; obese, P = .13) [P‐AMPK]. An inverse correlation existed between [blood glucose] and laminar [P‐AMPK] in obese ponies on a high‐CHO diet.

Conclusions and Clinical Importance

Laminar tissue exhibited a normal response to a high‐CHO diet (decreased [P‐AMPK]), whereas this response was not observed in liver and skeletal muscle in both lean (skeletal muscle, P = .33; liver, P = .84) and obese (skeletal muscle, P = .43; liver, P = .13) ponies.     

Adaptation of thermal threshold analgesiometry for NSAIDs in cats: effects of ketoprofen

Nonsteroidal anti‐inflammatory drugs (NSAIDs) are widely used to provide analgesia in clinical veterinary medicine, but there are few objective data evaluating this effect under controlled conditions in cats. Analgesia is more difficult to detect with acute analgesiometry after NSAIDs than after opioids. This investigation aimed to adapt the feline thermal analgesiometry method previously employed with opioids ( Dixon et al. 2002 ) for use with NSAIDs. Ketoprofen, a COX1 inhibitor licensed for cats was chosen. Six cats (2 neutered, four entire females, weighing 2.2–5.4 kg) were studied in two blinded randomized crossover trials each at least 2 weeks apart. Thermal thresholds (TT) were measured using the thermal threshold‐testing device previously developed for cats. A heater element and temperature sensor in a small probe were held at constant pressure against the cats' shaved thorax with an elasticized band. Skin temperature was recorded before each test, then the heater activated. When the cat responded by flinching, turning or jumping the heater was turned off and the temperature recorded. In the first study TT were measured following subcutaneous (SC) injection of ketoprofen (2 mg kg^?1) or a similar volume of saline. In the second study, prior to TT, and under isoflurane restraint, a mild inflammatory focus was produced at the probe site by five SC injections of 5 mg kaolin in 0.1 mL saline at each corner and in the center of a 1.5‐cm square. Saline or ketoprofen as in the first study were injected at the same time. Three baseline temperatures were recorded before any injections were given. Thermal thresholds were measured at 1 and 2 hours and then two‐hourly for 24 hours. Data were analysed using anova . Baseline skin temperature increased (37.3 ± 0.5–38.1 ± 0.8 °C) 24 hours after saline injection in study 2 (p < 0.05) but did not change after any other treatment. Thermal thresholds decreased (40.0 ± 1.3 to 39.1 ± 0.4 °C) 16 hours after ketoprofen in study 1 (p < 0.05) and increased (41.6 ± 1.5–44.8 ± 6.1 °C) 16–24 hours after ketoprofen in study 2 (p < 0.05), with no significant changes after saline. No obvious increase in sensitivity to thermal stimulation after kaolin injection was detected although obvious inflammation was present for up to 36 hours and the cats responded to digital pressure at the treated site. The method detected some effects of a COX1 selective NSAID and may be suitable for future NSAID studies in cats. However, a pressure stimulus ( Dixon et al. 2000) may prove better than thermal, and it requires investigation.     

First Steps Towards an Understanding of a Mode ofCarcinogenic Action for Vanadium Pentoxide

Inhalation of vanadium pentoxide clearly increases the incidence of alveolar/bronchiolar neoplasms in male and female B6C3F1 mice at all concentrations tested (1, 2 or 4 mg/m³), whereas responses in F344/N rats was, at most, ambiguous. While vanadium pentoxide is mutagenic in vitro and possibly in vivo in mice, this does not explain the species or site specificity of the neoplastic response. A nose-only inhalation study was conducted in female B6C3F1 mice (0, 0.25, 1 and 4 mg/m³, 6 h/day for 16 days) to explore histopathological, biochemical (α-tocopherol, glutathione and F2-isoprostane) and genetic (comet assays and 9 specific DNA-oxo-adducts) changes in the lungs. No treatment related histopathology was observed at 0.25 mg/m³. At 1 and 4 mg/m³, exposure-dependent increases were observed in lung weight, alveolar histiocytosis, sub-acute alveolitis and/or granulocytic infiltration and a generally time-dependent increased cell proliferation rate of histiocytes. Glutathione was slightly increased, whereas there were no consistent changes in α-tocopherol or 8-isoprostane F2α. There was no evidence for DNA strand breakage in lung or BAL cells, but there was an increase in 8-oxodGuo DNA lesions that could have been due to vanadium pentoxide induction of the lesions or inhibition of repair of spontaneous lesions. Thus, earlier reports of histopathological changes in the lungs after inhalation of vanadium pentoxide were confirmed, but no evidence has yet emerged for a genotoxic mode of action. Evidence is weak for oxidative stress playing any role in lung carcinogenesis at the lowest effective concentrations of vanadium pentoxide.     

Early harvest and ensilage of forage sorghum infected with ergot (Claviceps africana) reduces the risk of livestock poisoning

Sorghum ergot produces dihydroergosine (DHES) and related alkaloids, which cause hyperthermia in cattle. Proportions of infected panicles (grain heads), leaves and stems were determined in two forage sorghum crops extensively infected 2 to 4 weeks prior to sampling and the panicles were assayed for DHES. Composite samples from each crop, plus a third grain variety crop, were coarsely chopped and half of each sealed in plastic buckets for 6 weeks to simulate ensilation. The worst-infected panicles contained up to 55 mg DHES/kg, but dilution reduced average concentrations of DHES in crops to approximately 1 mg/kg, a relatively safe level for cattle. Ensilation significantly (P = 0.043) reduced mean DHES concentrations from 0.85 to 0.46 mg/kg.     

Modeling Bare Ground With Classification Trees in Northern Spain

Bare ground abundance is an important rangeland health indicator and its detection is a fundamental part of range management. Remote sensing of bare ground might offer solutions for land managers but also presents challenges as modeling in semiarid environments usually involves a high frequency of spectral mixing within pixels. Classification tree analysis (CTA) and maximum likelihood classifiers were used to model bare ground in the semiarid steppes of the middle Ebro valley, Aragon, Spain using Satellite Pour l'Observation de la Terre 4 (SPOT 4) imagery and topographic data such as elevation, slope, aspect, and a morphometric characterization model. A total of 374 sample points of bare-ground fraction from sixteen 500-m transects were used in the classification and validation process. Overall accuracies were 85% (Kappa statistic = 0.70) and 57% (Kappa statistic = 0.13) from the CTA and maximum likelihood classifiers, respectively. Although spectral attributes were essential in bare-ground classification, the topographic and morphometric properties of the landscape were equally critical in this modeling effort. Although the specific layers best suited for each specific model will vary from region to region, this study provided an important insight on both bare-ground modeling and the potential advantages of CTA.     

Diagnostic orientation values for ACTH and other parameters for clinically healthy donkeys and mules (insulin,triglycerides, glucose,fructosamines, and ɣ-GT)

Pituitary pars intermedia dysfunction is the most prevalent endocrine disease in horses. Although donkeys and mules may also be affected, only a few data have been published. Reference values for diagnostic parameters, such as adrenocorticotropic hormone (ACTH), are especially scarce or even lacking. Therefore, in the present study, available data from the literature have been verified and completed to facilitate a reliable diagnosis. Clinical inspections and haematological and biochemical examinations were carried out four times in a three-month interval (February to November) in 44 donkeys and 31 mules. Data from clinically healthy animals were used as an orientation. Plasma ACTH concentrations showed seasonal changes in both animal groups. However, it was generally higher in donkeys than mules. Although blood glucose (EDTA plasma) showed no difference between groups, serum insulin concentrations were consistently higher in donkeys. Serum fructosamine levels were slightly higher in mules, whereas, in some cases, serum triglyceride levels were considerably higher in donkeys. Serum gamma-glutamyltransferase showed a striking peak in mules in August, whereas the remaining gamma-glutamyltransferase values were lower compared to donkeys. By comparing donkeys and mules, the present work reveals differences in various blood parameters which should be considered for diagnoses and future studies.     

Clostridium septicum myositis in a western lowland gorilla (Gorilla gorilla gorilla).

A 10-yr-old male gorilla (Gorilla gorilla gorilla) with a history of conspecific bite wounds was evaluated for acute onset of depression, anorexia, and right hemiparesis. The animal was immobilized for diagnostic examination and treatment for suspected toxic shock from a necrotizing, emphysematous wound infection, but was euthanized due to complications during recovery. Gross and histopathologic examination revealed acute necrotizing myositis, fasciitis, cellulitis, and emphysema in the affected wound area, with large numbers of large Gram-positive rods among necrotic muscle fibers. Severe pulmonary edema with airways containing fibrin, acute hemorrhage in multiple body sites, thrombosis in blood vessels in the skeletal muscle, liver, and lung, and lymph node hyperplasia with lymphoid necrosis and hemorrhage. Immunohistochemical fluorescent antibody staining of muscle from the wound site was positive for