Ovine rumen papillae biopsy via oral endoscopy; a rapid and repeatable method for serial sampling

AIMS: To explore and validate the utility of rumen endoscopy for collection of rumen papillae for gene expression measurement.

METHODS: Four adult Coopworth ewes were fasted for either 4 or 24 hours. Animals were sedated, placed in a dorsally recumbent position at 45 degrees with the head upright, and an endoscope inserted via a tube inserted into the mouth. Biopsies of rumen papillae were taken from the ventral surface of the rumen atrium under visual guidance. Two biopsies were collected from one of the animals that had been fasted for 4 hours, and three from one of the animals that had been fasted for 24 hours. Video of the rumen atrium and reticulum was also collected. The animals recovered uneventfully. Biopsies were subsequently used for extraction and sequencing of mRNA.

RESULTS: The ventral surface of the rumen atrium was accessible after 4 hours off pasture, but a larger region was accessible after 24 hours of fasting. Sedation allowed access for endoscope use for around 5 to 10 minutes after which increased saliva flow was noted. Rumen papillae biopsies were easily collected, with samples from a variety of sites collected in the ～10 minute time window. High quality RNA was obtained for stranded mRNA sequencing. Of the resulting reads, 69–70% mapped uniquely to version 3.1 of the ovine genome, and 48–49% to a known gene. The rumen mRNA profiles were consistent with a previously reported study.

CONCLUSIONS: This method for obtaining rumenal tissue was found to be rapid and resulted in no apparent short or long term effects on the animal. High quality RNA was successfully extracted and amplified from the rumen papillae biopsies, indicating that this technique could be used for future gene expression studies. The use of rumen endoscopy could be extended to collection of a variety of rumen and reticulum anatomical measurements and deposition and retrieval of small sensors from the rumen. Rumen endoscopy offers an attractive and cost effective approach to repeated rumen biopsies compared with serial slaughter or use of cannulated animals.     

LOW‐FIELD MAGNETIC RESONANCE IMAGING APPEARANCE OF POSTARTHROSCOPIC MAGNETIC SUSCEPTIBILITY ARTIFACTS IN HORSES

An awareness of magnetic susceptibility artifacts is important for interpreting prepurchase and postoperative magnetic resonance imaging (MRI) studies in horses. These artifacts occur when a metallic or a paramagnetic substance creates a local magnetic field deformity. Aims of the current experimental study were to determine prevalence of these artifacts after arthroscopy in a sample of nonlame horses, and to describe effects of time and type of pulse sequence on low‐field MRI signal intensity and detection of the artifacts. Ten, nonlame Standardbred horses were prospectively recruited. All horses underwent arthroscopy of both metacarpophalangeal joints for purposes unrelated to the study. Serial low‐Field MRI examinations were performed on each horse and each joint (before, and 6 and 12 weeks postsurgery). In two horses, more detailed longitudinal evaluations were performed with additional MRI examinations. Magnetic susceptibility artifacts were detected postoperatively at the surgical access sites in eight metacarpophalangeal joints at both 6 and 12 weeks after surgery (40% prevalence). Neither of the two longitudinally followed horses had artifacts at any time. Artifacts were only detected on gradient echo (GRE) sequences. Findings indicated that magnetic susceptibility artifacts can be present in postarthroscopy MRI studies in horses and can persist up to 12 weeks after arthroscopy. For this sample of horses, the artifacts did not interfere with evaluation of the joint. Further longitudinal studies are needed to determine the full duration of magnetic susceptibility artifact persistence in affected tissues.     

Use of the Anemon Index to evaluate the quality of analgesia during fentanyl and sevoflurane anaesthesia in pigs

Anemon I is a new monitoring system that can be used to evaluate autonomic nervous system reactivity in real time by showing a simple, easily interpretated quantitative index (0–200), the Anemon Index (AI) ( Junke et al. 2000 ). This study used the AI to evaluate the quality of analgesia during sevoflurane and fentanyl anaesthesia in pigs. Six healthy pigs, weighing 24.76 ± 3.40 kg, were induced to anaesthesia with 5% sevoflurane (SEVO) in 5 L minute^?1 oxygen. After endotracheal intubation SEVO was given at 1 MAC (2.66%) in 3 L minute^?1 oxygen. Fentanyl was infused IV at 50 µg kg^?1 hour^?1 for the first 30 minutes of anaesthesia, discontinued for 30 minutes, and then infused at 100 µg kg^?1 hour^?1 for another 30 minutes. Three mechanical noxious stimuli (needle prick, pin‐prick and pressure on the abdomen) were applied for 15 seconds at 30, 60 and 90 minutes. The AI, ECG, invasive mean arterial blood pressure (MAP), heart rate (HR), SpO₂ by pulse oximetry, tidal volume, Fe′sevo , Fe ′CO₂ and respiratory rate were recorded before induction (baseline), after induction, after intubation and extubation, and before and during noxious stimulation at 30, 60 and 90 minutes. Recovery times were recorded. Statistically significant differences were determined by anova . Spearman rank‐correlation was used to evaluate the relationship between AI and hemodynamic variables. A p‐value of < 0.05 was considered significant. A significant (p < 0.01) decrease in AI was recorded after anaesthetic induction, from 82.3 ± 21.1 to 52.7 ± 20.3. After intubation, AI increased slightly, but not significantly, to 71.7 ± 27.1. A significant (p < 0.05) increase of AI occurred after extubation. Nociceptive stimuli did not have any measurable effect either on AI or on recorded cardiovascular variables. There was no movement, respiratory changes, or any other visible response to noxious stimulation. The AI did not change significantly with the different doses of fentanyl. Respiratory depression and apnoea were seen in all animals during the fentanyl infusion; therefore, pigs received intermittent positive pressure ventilation. Anaesthesia with sevoflurane and fentanyl resulted in a significant (p < 0.001) decrease in MAP. Heart rate did not change significantly. There was no correlation between AI and cardiovascular variables (HR and MAP). Endotracheal intubation caused an increase and extubation a greater significant increase in the AI. This suggests that intubation and extubation may represent stressful events during general anaesthesia, although further studies are needed to validate the use of the AI in pigs. Sevoflurane anesthetic induction may not prevent the sympathetic stimulus caused by endotracheal intubation in pigs, as indicated by the increased AI values.     

Effect of day and night temperatures during short photoperiods on growth and flowering of Chrysanthemum morifolium Ramat

The effect of day and night temperatures of 10, 14 and 18°C on growth and flowering under short days was studied with six cultivais of chrysanthemum. A high day temperature resulted in earlier flowering and taller stems, but did not influence flower number and final total fresh weight, and only slightly influenced the distribution of fresh matter over stem, leaves and flowers. A high night temperature resulted in earlier flowering, more flowers and reduced stem and leaf weight. It did not affect leaf number and it influenced height and total fresh weight only slightly. Except for height, the day temperature acted independently from the night temperature. The cultivars responded similarly, except for two cultivars which generally did not flower at 10/10,10/14 and 14/10°C D/N. One cul-tivar produced more flowers at 14 than at 18°C.     

Biochemical markers of bone metabolism in draught andwarmblood horses

Concentrations of the cross-linked carboxyterminal telopeptide of type I collagen (ICTP) and osteocalcin (OC) have been determined in the serum of one hundred clinically healthy adult Draught or Warmblood horses. The correlation between these two markers has been evaluated and the influence of gender, age and type of horse described. No significant variations were observed between animals of different sex, but a significant inverse correlation (P<0.0001) with age was observed for both measured parameters. After correction for age, serum levels of OC were found to be lower in Draught [adjusted least square mean (LSM)=6.612μg. L⁻¹] than in Warmblood horses (adjusted LSM=8.596μg.L⁻¹), whereas levels of ICTP were higher in Draughts (adjusted LSM=8.035pg.L⁻¹) than in Warmbloods (adjusted LSM=6.643μg.L⁻¹). A significant correlation (P<0.0001) was observed between OC and ICTP. This correlation was stronger if the type of horse was taken into account in the statistical model. The ratio OC:ICTP was independent of gender and age. A higher OC:ICTP ratio in Warmbloods compared to the Draught horses might reflect a higher bone remodelling level of horses submitted to regular daily work. It was concluded that ICTP and OC are influenced by the type of horse, and probably reflect a physiological difference in bone remodelling between these animals.     

Equine third metacarpal bone assessment by quantitative ultrasound and dual energy X-ray absorptiometry: an ex vivo study

The purpose of this ex vivo study was to analyse two commonly established methods of mechanical bone property assessment for application in horses: Quantitative ultrasound (QUS), which depends on the bone's density and Young's modulus, and dual energy X-ray absorptiometry (DXA), which depends on the areal bone mineral density (BMD). The third metacarpal bone (MC III) of horses was selected as examination region for practical reasons. An interrelationship between QUS- and DXA-values was examined. Both MC III of eleven randomly selected equine cadavers were divided in nine regions of interest (ROI). A multi-site QUS device was used for axial transmission speed of sound (SOS) measurements and a DXA device was used for BMD evaluation. Full cortical thickness BMD (FcBMD), overall aspect BMD and 4 mm cortical border slice BMD (4 mmBMD) were evaluated. In addition, each ROI of one MC III was measured 10 times to determine QUS- and DXA-measurement precision. SOS values and BMD values obtained at different aspects of MC III were different (P < 0.001). FcBMD and overall BMD obtained at different levels were different (P < 0.001). SOS data correlated with FcBMD-, overall BMD- and 4 mmBMD-data at various ROI. FcBMD-, overall BMD- and 4 mmBMD-data were strongly correlated. The intra-operator coefficient of variation was 1.3% for SOS-measurements and ranged between 1.94 and 10.3% for BMD-measurements. Multi-site axial transmission QUS as well as DXA can be used to precisely measure bone characteristics of MC III in horses. However, both techniques do not measure the same bone properties. It is therefore concluded, that QUS and DXA techniques are complementary for application in horses.     

Transarterial coil embolization of the internal and external carotid and maxillary arteries for prevention of hemorrhage from guttural pouch mycosis in horses

OBJECTIVES: To develop a transarterial coil embolization technique for occlusion of the internal carotid artery (ICA), external carotid artery (ECA), and maxillary arteries (MA) in normal horses and to evaluate this technique for prevention of hemorrhage in horses affected with guttural pouch mycosis. ANIMALS: Ten adult, normal horses and 4 horses with guttural pouch mycosis. METHODS: All horses had transarterial coil embolization of the rostral and caudal ICA, caudal MA, and rostral ECA. In 1 affected horse, an aberrant actively bleeding branch of the ECA was also occluded. Normal horses had a premortem angiogram, and were killed either at 1 or 2 weeks or 1, 2, or 3 months after the procedure. Specimens from the ICA, ECA and MA were evaluated by light microscopy. RESULTS: No surgical complications were observed, except 1 horse that developed laryngeal hemiplegia and 1 pilot horse that had embolization of the cerebral arterial circle. In normal horses, premortem angiography confirmed complete occlusion of all vessels, and coils were positioned as intended. All normal horses had partially maturing to mature, continuous thrombi occluding at the coils. In affected horses, no further episodes of epistaxis were observed. By day 60, all mycotic plaques had resolved without further treatment. Ophthalmic complications were not observed. CONCLUSION: Transarterial embolization provided a safe, rapid, and effective method for ICA, ECA, and MA occlusion in normal and affected horses. In affected horses, the technique was possible despite active bleeding, allowing adequate identification and occlusion of all sources of hemorrhage.     

Sex does not influence serum osteocalcin levels in standardbred horses of different ages.

The influence of sex on serum osteocalcin levels was studied in 99 Standardbred horses, 52 males and 47 females. The age varied between the day of birth and five years old. A significant inverse correlation (r = -0.87, p < 0.01) was observed between serum osteocalcin level and the age of the animal. Sex did not influence serum osteocalcin level (p > 0.05). When the subjects were divided into five age groups of six months or less, 6 to 18, 18 to 24, 24 to 36 and between 36 and 60 months, no significant influence of sex was noted. A predictive model for serum osteocalcin levels (y = 52.19 - 0.026 age in days, r2 = 0.76) for Standardbred horses aged between the day of birth and five years is described.     

Small intestine adenocarcinoma in conjunction with multiple adenomas causing acute colic in a horse.

An 11-year-old Andalusian stallion developed marked signs of colic associated with an acute small intestine obstruction. Exploratory laparotomy revealed a distal jejunum full-thickness wall induration and multiple small adherent intraluminal masses. Fifteen centimeters of jejunum, including the induration, and several intraluminal masses were resected. Histologic examination revealed an adenocarcinoma and multiple polypoid adenomas. The horse was discharged, and no complications were reported 12 months postoperatively. Colic was considered secondary to partial jejunal lumen obstruction by the adenocarcinoma. Adenocarcinoma recurrence or transformation from remaining adenomas into an adenocarcinoma is still a major risk.