Recovery of multireassortant bluetongue virus serotype 6 sequences from a mule deer (Odocoileus hemionus) and Dorset sheep (Ovis aries) in Colorado.

We report the discovery of two bluetongue virus serotype 6 (BTV-6) reassortants recovered from a domestic sheep and a free-ranging mule deer in northern Colorado. At the time of this publication, whole-genome sequencing of BTV-6 isolates in the Western U.S. have not been undertaken. These findings reflect the incursive movement of geographically distinct BTV serotypes into important agricultural areas of the U.S. and demonstrate reassortment with regionally circulating serotypes.

Monitoring longitudinal immunological responses to bluetongue virus 17 in experimentally infected sheep.

Bluetongue virus (BTV) is an economically important pathogen of ruminant species with worldwide prevalence. While many BTV infections are asymptomatic, animals with symptomatic presentation deteriorate quickly with the sickest succumbing to disease within one week. Animals that survive the infection often require months to recover. The immune response to BTV infection is thought to play a central role in controlling the disease. Key to understanding BTV disease is profiling vertebrate host immunological cellular and cytokine responses. Studies to characterize immune responses in ruminants have been limited by a lack of species-specific reagents and assay technology. Here we assess the longitudinal immunological response to experimental BTV-17-California (CA) infection in sheep using the most up to date assays. We infected a cohort of sheep with BTV-17-CA and longitudinally monitored each animal for clinical disease, viremia and specific immunological parameters (B cells, T cells, monocytes) by RT-qPCR, traditional flow cytometry and/or fluorescent based antibody arrays. BTV-inoculated sheep exhibited clinical signs characteristic of bluetongue virus disease. Circulating virus was demonstrated after 8 days post inoculation (DPI) and remained detectable for the remainder of the time course (24 DPI). A distinct lymphopenia was observed between 7 and 14 DPI that rebounded to mock-inoculated control levels at 17 DPI. In addition, we observed increased expression of pro-inflammatory cytokines after 8 DPI. Taken together, we have established a model of BTV infection in sheep and have successfully monitored the longitudinal vertebrate host immunological response and viral infection progression using a combination of traditional methods and cutting-edge technology.

Behavioral and Physiologic Effects of a Single Dose of Oral Gabapentin in Rabbits (Oryctolagus cuniculus).

Stress in rabbits (Oryctolagus cuniculus) may influence veterinarians' ability to assess their health and can lead to complications such as gastrointestinal hypomotility and poor anesthetic outcomes. Gabapentin has been used as an anxiolytic in various species, but little information is available on its use in rabbits. To assess the effect of gabapentin on stress in rabbits, 5 female and 3 male New Zealand white rabbits, aged 8-12 months, weighing 3-4.5 kg, were administered a single dose (25 mg/kg) of oral compounded gabapentin. Effects on individual behaviors and selected physiologic parameters were assessed by a blinded observer using a human intruder test and tractability score (summed total 0-8, most to least tractable). Heart and respiratory rate, rectal temperature, body weight, and fecal output were also recorded. Baseline measurements for each rabbit were assessed immediately prior to gabapentin administration, and at 1, 2, and 4 hours post-administration. With this method rabbits acted as their own concurrent control group. Rabbits were assessed at 7 AM, 11 AM, and 3 PM. Data were analyzed as continuous, binary, and continuous nonparametric (P ≤ .05). No significant differences in physiologic parameters were observed between baseline and the post-administration timepoints. Fecal output was reduced similar to baseline measurements. Behaviors pressing down decreased (at 2 and 4 hours; P = .05 and P = .013, respectively) and approaching human increased (at 2 hours; P = .022) post-gabapentin compared to baseline. Tractability scores were improved at the 2-hour timepoint compared to baseline (Friedman P = .0461; Wilcoxon P = .0413). These results suggest gabapentin 25 mg/kg orally decreased reactivity with a peak effect at 2 hours, without significant effects on measured physiologic parameters. Oral gabapentin in rabbits should be considered to reduce stress in the presence of humans and to facilitate handling.

Bluetongue Research at a Crossroads: Modern Genomics Tools Can Pave the Way to New Insights.

Bluetongue virus (BTV) is an arthropod-borne, segmented double-stranded RNA virus that can cause severe disease in both wild and domestic ruminants. BTV evolves via several key mechanisms, including the accumulation of mutations over time and the reassortment of genome segments.Additionally, BTV must maintain fitness in two disparate hosts, the insect vector and the ruminant. The specific features of viral adaptation in each host that permit host-switching are poorly characterized. Limited field studies and experimental work have alluded to the presence of these phenomena at work, but our understanding of the factors that drive or constrain BTV's genetic diversification remains incomplete. Current research leveraging novel approaches and whole genome sequencing applications promises to improve our understanding of BTV's evolution, ultimately contributing to the development of better predictive models and management strategies to reduce future impacts of bluetongue epizootics.

Pharmacokinetics and Efficacy of a Long-lasting, Highly Concentrated Buprenorphine Solution in Rats.

Buprenorphine (Bup) is an opioid analgesic that is commonly used in laboratory rodents to provide postoperative analgesia. However, dosing every 4 to 6 h is necessary to maintain an analgesic plasma concentration of the drug. A long lasting, highly concentrated veterinary formulation of Bup (LHC-Bup) has been used to provide prolonged analgesia in cats and nonhuman primates. In the current study, we evaluated the duration of efficacy of LHC-Bup to determine if this formulation would provide a similarly prolonged analgesia in rats. Drug concentrations were measured after subcutaneous injection of 0.5 mg/kg LHC-Bup in both male and female rats. Plasma levels were measured at 0.25, 0.5, 1, 2, 4, 8, 12, 24, 36, 48, and 72 h. Male and female rats had peak plasma levels of LHC-Bup at 90 ng/mL and 34 ng/mL, respectively, at 15 min after administration, with a steady decrease by 24 h to 0.7 ng/mL in males and 1.3 ng/mL in females. Mechanical pain tolerance was evaluated after LHC-Bup administration using a Randall-Selitto analgesiometer to assess paw withdrawal. Male rats had a significantly longer paw withdrawal time for up to 12 h after administration, and females had longer paw withdrawal times for up to 24 h. An experimental laparotomy model was then used to assess the clinical efficacy of LHC-Bup at 0.5 mg/kg. LHC-Bup treatment was associated with a greater total distance traveled, reduced time to retrieve a food treat, and reduced grooming from 3 to 12 h after surgery as compared with saline controls. Groups receiving LHC-Bup showed coprophagy whereas other rats did not. These results suggest that administering LHC-Bup at 0.5 mg/kg provides therapeutic plasma concentrations for 12 to 24 h after administration and analgesic efficacy for at least 12 h after dosing. As such, LHC-Bup is a suitable alternative to Bup-HCl.