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.     

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.     

Levels of supplementation of inorganic selenium and vitamin E for meat quail aged 0 to 14 and 14 to 35 days

Two experiments were carried out to determine the levels of supplementation of inorganic selenium (Se) and vitamin E (VE ) in diets of quails aged 0–14 and 14–35 days old. A completely randomized design was used in a factorial design (Se = 0.1125; 0.2250; 0.3375 and 0.4500 mg kg^?1 diet^?1 × VE  = 10; 23; 36 and 49 IU  kg^?1 diet^?1). In experiment 1, quail (n  = 2,400) were aged 0–14 days and were divided into 16 treatments, with three replicates of 50 birds. In experiment 2, quail (n  = 1,680) were aged 14–35 days and were divided into the same treatments, with three replicates of 35 birds. At age 0–14 days, the levels of VE did not affect performance (p  > .05); however, the feed conversion (FC ) was influenced by a quadratic effect (p  = .0515), according to the level of Se, with a higher level estimated at 0.29 mg Se kg^?1 diet^?1. At age 14–35 days, there was a linear effect with interaction (Se × VE ), for FC (p  = .0150) and weight gain (WG ; p  = .0266). FC (Se, p  = .0048 and VE , p  = .0019) and WG (Se, p  = .0049 and VE , p  = .0068) improved linearly with increasing levels of Se and VE . The feed intake (FI ) decreased linearly (p  = .0582) as a function of VE . The carcass yield showed a quadratic effect (p  = .0056) on the levels of VE , with a higher yield estimation of 27.24 IU VE /kg of diet. It can be concluded that the optimum level of supplementation at age 0–14 days was 0.29 mg Se kg^?1 diet^?1 and 10 IU VE  kg^?1 diet^?1 and at age 14–35 days, it was 0.4500 mg Se kg^?1 diet^?1 and 49 IU of VE  kg^?1 diet^?1.     

Plasma Antimullerian Hormone as a Predictor of Ovarian Antral Follicular Population in Bos indicus (Nelore) and Bos taurus (Holstein) Heifers

In Bos taurus cattle, antimullerian hormone (AMH) has been demonstrated to have a high degree of correlation with ovarian antral follicle count and the number of healthy follicles and oocytes. To document the correlation between the plasma concentration of AMH and follicular number in Bos indicus and Bos taurus heifers, Nelore (Bos indicus, n = 16) and Holstein heifers (Bos taurus, n = 16) had their ovarian follicular waves synchronized. After synchronization, ovarian antral follicular population (AFP) was evaluated three times at 60‐day (d) intervals (T‐120 d, 120 days before plasma AMH determination; T‐60 d, 60 days before; and T0, at the time of plasma AMH determination). The plasma AMH concentration was positively correlated with the number of ovarian follicles on the day of the follicular wave emergence in Bos indicus (Nelore) and Bos taurus (Holstein) heifers at each evaluation time (p < 0.05). The AFP was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.05). Similarly, the AMH concentration was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.0001). When heifers were classified as to present high or low AFP according to the mean of the AFP within each genetic group, high‐AFP heifers presented a greater (p < 0.0001) AMH concentration than low‐AFP heifers, regardless of the genetic group. In conclusion, the AFP is positively correlated with plasma AMH concentration in both Bos indicus (Nelore) and Bos taurus (Holstein) heifers. Furthermore, Bos indicus (Nelore) heifers presented both greater plasma AMH concentrations and AFP than Bos taurus (Holstein) heifers.