Laryngeal Thermal Injury Model.

A lack of reliable laryngeal thermal injury models precludes laryngeal burn wound healing studies and investigation of novel therapeutics. We hypothesize that a swine laryngeal burn model can allow for laryngeal burn evaluation over time. Twelve Yorkshire crossbreed swine underwent tracheostomy and endoscopically directed laryngeal burns using heated air (150-160°C). Swine larynges were evaluated and sectioned/stained at 12 hours, 1, 3, 7, 14, and 21 days. A board-certified veterinary pathologist assessed anatomic regions (left and right: epiglottis, true/false vocal folds, and subglottis) using a nine criteria histological injury scoring scale. Six swine were euthanized at scheduled endpoints, three prematurely (airway concerns), and three succumbed to airway complications after 16 to 36 hours. Endoscopic and gross examination from scheduled endpoints revealed massive supraglottic edema and tissue damage, particularly around the arytenoids, extending transglottically. Swine from premature endpoints had comparatively increased edema throughout. Microscopic evaluation documented an inverse relationship between injury severity score and time from injury. Inflammation severity decreased over time, nearly resolving by 14 days. Neutrophils predominated early with histiocytes appearing at 3 days. Granulation tissue appeared at 3 days, and early epiglottic and/or subglottic fibrosis appeared by 7 days and matured by 14 days. Edema, abundant initially, decreased by day 3 and resolved by day 7. This approach is the first to provide longitudinal analysis of laryngeal thermal injuries, reflecting some of the first temporal wound healing characteristic data in laryngeal thermal injuries and providing a platform for future therapeutic studies.

Evaluation of Volume of Intramuscular Injection into the Caudal Thigh Muscles of Female and Male BALB/c Mice (Mus musculus).

This study presents recommendations for intramuscular injection into the caudal thigh muscle of mice according to analysis of in vivo imaging of intramuscularly injected iohexol, a radiocontrast agent commonly used in CT imaging. An experienced laboratory animal technician using a Hamilton syringe intramuscularly injected iohexol into isoflurane-anesthetized female and male BALB/c mice. Injected volumes (25, 50, 100, and 200 µL) underwent CT scanning at 9 time points over a 3-h period. The distribution of the injectate in the muscles of the rear leg was examined over time for each volume group. Results indicated that 25- and 50-µL volumes remain intramuscularly. At 100 µL, mild to moderate leakage into the extramuscular tissues occurred. At 200 µL, leakage into the extramuscular tissues was moderate to severe. Our results suggest volumes of 50 µL or less are recommended for the caudal thigh muscles of mice when intramuscular pharmacokinetics are needed; volumes greater than 50 µL display variable distribution into extramuscular tissues, thus potentially yielding different pharmacokinetic profiles.

Comparison of Saliva Collection Methods for the Determination of Salivary Cortisol Levels in Rhesus Macaques (Macaca mulatta), Cynomolgus Macaques (Macaca fascicularis), and African Green Monkeys (Chlorocebus aethiops).

The ability to quickly and accurately determine cortisol as a biomarker for stress is a valuable tool in assessing the wellbeing of NHP. In this study, 2 methods of collecting saliva (a commercial collection device and passive drool) and the resulting free salivary cortisol levels were compared with total serum cortisol concentration in rhesus macaques (Macaca mulatta), cynomolgus macaques (Macaca fascicularis) and African green monkeys (Chlorocebus aethiops) at 2 collection time points. Serum and salivary cortisol levels were determined using a competitive quantitative ELISA. In addition, both saliva collection methods were evaluated for volume collected and ease of use. Compared with passive drool, the experimental collection device was more reliable in collecting sufficient volumes of saliva, and the resulting salivary cortisol values demonstrated stronger correlation with serum cortisol concentration in all species and collection days except cynomolgus macaques on day 1. This saliva collection device allows quick and reliable sample collection for the determination of salivary cortisol levels. In addition, the results might provide a useful tool for evaluating hypothalamic-pituitary-adrenal axis activity or the physiologic stress reaction in NHP as well as a biomarker of psychologic stress states in a variety of situations.

Pharmacokinetics of 2 Formulations of Transdermal Fentanyl in Cynomolgus Macaques (Macaca fascicularis).

Fentanyl is a µ-opioid agonist that often is used as the analgesic component for balanced anesthesia in both human and veterinary patients. Minimal information has been published regarding appropriate dosing, and the pharmacokinetics of fentanyl are unknown in NHP. The pharmacokinetic properties of 2 transdermal fentanyl delivery methods, a solution (2.6 and 1.95 mg/kg) and a patch (25 µg/h), were determined when applied topically to the dorsal scapular area of cynomolgus macaques (Macaca fascicularis). Serum fentanyl concentrations were analyzed by using liquid chromatography-mass spectrometry. Compared with the patch, the transdermal fentanyl solution generated higher drug concentrations over longer time. Adverse reactions occurred in the macaques that received the transdermal fentanyl solution at 2.6 mg/kg. Both preparations showed significant interanimal variability in the maximal serum drug levels, time to achieve maximal fentanyl levels, elimination half-life, and AUC values. Both the maximal concentration and the time at which this concentration occurred were increased in macaques compared with most other species after application of the transdermal fentanyl patch and compared with dogs after application of the transdermal fentanyl solution. The pharmacokinetic properties of transdermal fentanyl in macaques are markedly different from those in other veterinary species and preclude its use as a long-acting analgesic drug in NHP.

Measurement of fecal corticosterone metabolites as a predictor of the habituation of rhesus macaques (Macaca mulatta) to jacketing.

Jacket use in NHP is a common practice and is often considered a form of refinement during experiments necessitating extended periods of catheterization. An important consideration when using jackets is the physiologic effects that jacketing has on NHP and its potential to confound research. Several studies have evaluated the stress response and habituation of NHP to various forms of restraint, but none have looked directly at the timeframe necessary for the habituation of rhesus macaques (Macaca mulatta) to jackets. We set out to determine whether 3 d was a sufficient timeframe for this species to become habituated to a jacket, with or without an undershirt, by evaluating 2 major physiologic parameters. After jacket placement, we measured food consumption and collected fecal samples to measure fecal corticosterone metabolites (FCM) daily for 2 wk. FCM measurements for NHP without undershirts were significantly increased for days 2 and 3 after jacketing before returning to baseline levels. FCM measurements for NHP with undershirts were significantly increased for only 1 d after jacketing, suggesting that the undershirt has a positive effect on jacket habituation. There were no measurable differences in food consumption during the jacket habituation period. Furthermore, no significant differences were noted between sexes. These findings suggest that FCM levels return to baseline 3 d after jacketing and could be a useful predictor of jacket habituation in rhesus macaques.

Effects of housing density on Long Evans and Fischer 344 rats.

At many breeding facilities, rats are housed at relatively high densities until they are 5 weeks old, at which point they are either shipped for research or rehoused at standard cage densities according to weight. The authors carried out a pilot study in Long Evans and in Fischer 344 rats to investigate whether continuing to house rats at high densities (24 in(2) floor space per rat) past the age of 5 weeks, through puberty and into adulthood would alter behavioral or physiological parameters compared with raising rats at standard densities (about 72 in(2) floor space per rat). After rats reached puberty, the authors rehoused them with unfamiliar cagemates. The researchers evaluated clinical and behavioral signs of stress, weight, blood glucose concentration, white blood cell count and serum corticosterone concentration. Overall, cage density had little effect on the parameters measured, though gender seemed to affect stress in Long Evans rats. The results suggest that rats of these strains can be raised at the higher densities tested until any age and regrouped with unfamiliar cagemates without compromising rats' welfare or subsequent experimental data.

Comparative study of lung cytologic features in normal rhesus (Macaca mulatta), cynomolgus (Macaca fasicularis), and African green (Chlorocebus aethiops) nonhuman primates by use of bronchoscopy.

Invasive bronchoscopy and bronchoaveolar lavage (BAL) fluid collection represents an important tool in studies of the respiratory system of nonhuman primates. Bronchoscopy and BAL fluid collection was performed on groups of rhesus (Macaca mulatta) and cynomolgus (Macaca fasicularis) macaques and African green monkeys (Chlorocebus aethiops), and the resulting comparative lavage cytologic features are described. Analysis of the BAL fluid did not reveal significant differences among species with respect to total cells recovered or differential cellular composition. This description of the method used to lavage the nonhuman primates and the resulting lung cytologic findings provide important comparative data for three species commonly used in biomedical research.