Journal of the American Association for Laboratory Animal Science : JAALAS最新文献

Intrafacility transport of mice is an essential function for both laboratory and husbandry personnel. However, transport may induce a stress response that can alter research findings and negatively impact animal welfare. To determine minimally adverse intrafacility transport methods, in-cage noise and vibration exposure during transport on a variety of transport vehicles (hand carrying, stainless steel rack, flatbed cart, metal teacart, plastic teacart, and a cart with pneumatic wheels) were measured. Under-cage and in-cage padding was tested for its ability to decrease noise and vibration on each vehicle. Behavioral (open field test and elevated plus maze) and corticosterone responses of mice were then measured following transport on the most adverse (metal teacart) and least adverse (pneumatic cart) methods of multicage transport. Behavioral measures showed no difference between transported mice and untransported mice in both single- and group-housed settings. Plasma corticosterone was significantly elevated in mice transported on the metal teacart immediately following transport and continued to have elevated trends in circadian peaks during the 48h of sampling. The cart with pneumatic wheels was most effective at reducing noise and vibration, reflected in posttransport corticosterone readings that remained equivalent to those in untransported mice. This study demonstrates that mitigation of noise and vibration during cart transport may decrease the impact of transport on certain stress parameters in mice.

The ability to apply findings from animal studies efficiently and effectively is predicated on an understanding of biology and pathobiology, how that biology relates to the human systems being modeled, and how the studies are conducted and reported. This overview discusses various factors in research within the animal environment (referred to as extrinsic factors) that the NIH now expects to be documented to foster replicability in science and expand interpretations of study outcomes. Specifically, an important extrinsic factor in research with animals is that of individual personnel who perform handling practices, participate in research interactions, and share an overall presence in the housing facility with animals, all of which can confound reproducibility efforts in biomedical science. An improved understanding of the influences and behaviors of animal research personnel on animal responses is critical with regard to research results and the interpretation of data collected from animal models of biomedical disease.

Nonhuman primates used in biomedical research may experience clinically significant weight loss for a variety of reasons. Episodes of anorexia (complete loss of appetite) or hyporexia (decreased appetite) can result in significant weight loss, potentially altering animal welfare and scientific studies. The FDA has approved several appetite stimulants for use in domestic species, but currently none are approved for use in NHP. Treatment of inappetence and weight loss in NHP often relies on the extralabel use of these compounds. Capromorelin is a ghrelin receptor agonist. As a growth hormone secretagogue, capromorelin increases appetite, leading to weight gain. Studies in several species have shown a positive correlation between capromorelin administration and weight gain; in 2017, an oral solution of capromorelin received FDA approval for use in dogs. We tested this solution in healthy adult rhesus macaques (n = 3 males and 3 females) for its effects on body weight and insulin like growth factor-1 (IGF-1). A control group (n = 2 males and 2 females) was used for comparison. Treated macaques received a 3mg/kg oral dose daily for 7 d. Clinical signs were observed daily. Weights were collected before, during and at the end of treatment. Blood was drawn before, during and after treatment for measurement of IGF-1 levels and standard hematology and biochemistry parameters. Baseline-adjusted mean body weights and IGF-1 levels were significantly higher in treated as compared with control monkeys after 7 d of beginning treatment (body weight of 10.5±0.1kg (mean ± SEM) and 10.1±0.1kg, respectively; IGF-1 of 758±43ng/mL and 639±22ng/mL, respectively). Capromorelin administration was not associated with appreciable changes in hematologic and biochemical values in treated macaques. These findings suggest that capromorelin may be useful for treating inappetence and weight loss in NHP, and based on blood analysis, a 7-d course of treatment does not appear to cause acute toxicity.

Despite the long-term contributions of the spiny mouse (Acomys cahirinus) to research, basic knowledge of appropriate nutrition is lacking for this species. In the wild, spiny mice eat a high-fiber, high-protein food source. In the research setting, spiny mice are prone to obesity that can lead to diabetes mellitus. Common dietary modifications for weight control in humans with diabetes mellitus consist of increased fiber and protein. We hypothesized that increasing the dietary protein or fiber of spiny mice would reduce weight gain and improve their glycemic control, whereas the combination of protein and fiber in the diet would achieve optimal weight management and glycemic control without diet-related pathologic changes. We randomly assigned cages of young adult spiny mice (n = 34) to one of 4 diets: high protein (HP), high fiber (HF), a combination of both high protein and high fiber (HPF), or the base (control) diet (BD). Over the 8-wk study, spiny mice given HF diets maintained baseline weights despite the elevated dietary protein. None of the diets altered blood glucose levels; all diet groups maintained mean blood glucose levels within normal ranges. Spiny mice seem particularly sensitive to changes within their environment, as seen by increased food waste and transient elevated blood glucose levels when the spiny mice were transitioned to novel diets. The short-term elevations in protein and fiber that we tested were well tolerated by spiny mice. Although HF was effective in controlling weight, the ideal percentage of fiber still needs to be determined. The combination diet (HPF) maintained weight and body condition scores and showed a nonsignificant elevation of blood glucose that warrants a longer diet trial before our recommending this specific combination.

Many experiments require the collection of serial blood samples from mice. However, the size of mice limits the volume of blood that can be safely collected as a survival procedure. In IACUC protocols, investigators may report the amount of blood they collect from mice as a number of drops. Many institutions, including ours, use an anecdotal conversion factor (1drop of mouse blood = 25μL) to ensure that blood-collection volumes are compliant with institutional guidelines. To our knowledge, previous work has not experimentally determined the volume of a drop of mouse blood. In this 10-wk crossover experiment, 2 phlebotomists bled 30 C57BL/6J mice from 3 sites (facial, saphenous, and tail) using one or 2 different needle gauge sizes per site. Male and female mice were weighed weekly and divided among 5 groups (n = 6): left and right tail vein, left and right saphenous vein, and facial vein. A single blood drop from each site was weighed, and the volume of each drop was calculated using the average blood density determined from 8 mice terminally bled at the end of the study. Venipuncture site and side significantly influenced blood-drop weight and thus calculated volume. Facial vein puncture produced the largest drop volume (mean: 21.7μL), followed by the saphenous vein (mean: 9.97μL) and tail vein (mean: 4.96μL). Collection from the facial vein was associated with more hemorrhage and morbidity. Left-sided venipuncture was associated with slightly larger-volume blood drops, though the effect size of side was small. The results of this study may be useful in more accurately estimating blood loss via conversion of drops to volume. Our data indicate that blood collection from saphenous and tail veins minimizes blood loss relative to facial vein puncture and may optimize both serial collection of small-volume blood samples and animal welfare.

The New Zealand white rabbit (Oryctolagus cuniculus) is a frequently used surgical model. Pain management after surgery is a critical aspect of animal welfare. Recently, a long-acting buprenorphine formulation (Ethiqa XR; EXR) was approved for use in rats and mice but has not yet been investigated in rabbits. The current study aimed to determine whether a single subcutaneous dose of 0.15mg/kg of EXR could achieve and maintain therapeutic buprenorphine plasma concentrations (0.1ng/mL) for 72h in male and female rabbits. We also evaluated the safety profiles of EXR and the fentanyl patch (FP) by assessing fecal output after surgery, because opioids are known to decrease intestinal motility. Behavior and pain scores were compared for rabbits that received either EXR or the FP after undergoing an annulus puncture procedure to induce osteoarthritis. EXR at 0.15mg/kg SC provided a shorter time to onset and sustained analgesia for 72h in male and female rabbits, whereas the FP provided suboptimal analgesia after 48h. Both EXR and FP reduced fecal output after surgery. Output returned to baseline levels within 72h for the EXR group and remained slightly below baseline at 96h after surgery for the fentanyl group. Grimace pain scores revealed no significant difference between treatment groups. These results suggest that EXR is a safe and effective option for postoperative pain management in rabbits.

Heat supplementation during surgery is a common practice; however, thermal support is not commonly used during anesthesia induction. Mice lose body temperature quickly, and air movement can exacerbate this, potentially putting mice at a thermal deficit before surgery. Whether the method of warming during induction affects overall heat loss during anesthesia is unknown. We hypothesized that the method of heating would affect body temperature (Tb) during anesthesia induction, maintenance, recovery, and once placed back on the rack. Mice (C57BL/6NHsd-6M/6F [C57BL/6]; Hsd:Athymic Nude-Foxn1nu [Nude]; N = 24;12M/12F) were assigned to a treatment in a factorial design: thermal chamber (TC; ambient temperature [Ta] = 28.8°C); heating pad (HP; induction chamber placed on an electric heating pad;Ta = 28.4°C); and control (Ctrl; Ta = 21.6°C). During induction, one mouse at a time was anesthetized with isoflurane over a 3min period and then maintained under anesthesia for 10min on a hot water heating pad (33 °C). Then isoflurane was stopped and time to ambulation was recorded. Tb and activity were tracked in the home cage on the rack before and after anesthesia. During induction, Ctrl mice lost significantly more heat (-2.8 °C) than did TC (+0.2 °C) and HP mice (+0.1 °C) but TC and HP were not different. During anesthesia maintenance, Ctrl mice regained 1 °C, but their Tb was still lower than that of the treated groups. Nude mice consistently had a lower Tb than C57BL/6 mice, regardless of treatment or anesthesia phase. C57BL/6 Ctrl mice took longer to ambulate than either HP or TC mice, but the method of heating did not differentially affect Nude mice. In general, C57BL/6 as compared with Nude and females as compared with males were comparatively more active and had higher Tb during certain times of day, regardless of the heating methods. Overall, our findings support the provision of heat during anesthesia induction, regardless of method, to reduce overall Tb loss during a short anesthesia event.

Overdose of carbon dioxide gas (CO₂) is a common euthanasia method for rodents; however, CO₂ exposure activates nociceptors in rats at concentrations equal to or greater than 37% and is reported to be painful in humans at concentrations equal to or greater than 32.5%. Exposure of rats to CO₂ could cause pain before loss of consciousness. We used 2 standardized loss of righting reflex (LORR) methods to identify CO₂ concentrations associated with unconsciousness in Wistar, Long???Evans, and Sprague???Dawley rats (n = 28 animals per strain). A rotating, motorized cylinder was used to test LORR while the rat was being exposed to increasing concentrations of CO₂. LORR was defined based on a 15-second observation period. The 2 methods were 1) a 1-Paw assessment (the righting reflex was considered to be present if one or more paws contacted the cylinder after the rat was positioned in dorsal recumbency), and 2) a 4-Paw assessment (the righting reflex was considered to be present if all 4 paws contacted the cylinder after the rat was positioned in dorsal recumbency). Data were analyzed with Probit regression, and dose-response curves were plotted. 1-Paw EC₉₅ values (CO₂ concentration at which LORR occurred for 95% of the population) were Wistar, 27.2%; Long???Evans, 29.2%; and Sprague???Dawley, 35.0%. 4-Paw EC₉₅ values were Wistar, 26.2%; Long???Evans, 25.9%, and Sprague???Dawley, 31.1%. Sprague???Dawley EC₉₅ values were significantly higher in both 1- and 4-Paw tests as compared with Wistar and Long???Evans rats. No differences were detected between sexes for any strain. The 1-Paw EC₉₅ was significantly higher than the 4-Paw EC₉₅ only for Sprague-Dawley rats. These results suggest that a low number of individual rats from the strains studied may experience pain during CO₂ euthanasia.

Vehicular whole-body vibration (WBV) can have long-term adverse effects on human quality of life. Animal models can be used to study pathophysiologic effects of vibration. The goal of this study was to assess animal cooperation and well-being to determine the feasibility of a novel seated rat model for investigating the effects of WBV on biologic systems. Twenty-four male Sprague???Dawley rats were used. The experiment consisted of an acclimation phase, 2 training phases (TrP1 and TrP2), and a testing phase (TeP), including weekly radiographic imaging. During acclimation, rats were housed in pairs in standard cages without vibration. First, experimental (EG; n = 18) and control group 1 (C1; n = 3) rats were placed in a vibration apparatus without vibration, with increasing duration over 5 d during TrP1. EG rats were exposed to vertical random WBV that was increased in magnitude over 5 d during TrP2 until reaching the vibration signal used during TeP (15min, 0.7m??s-2 root mean square, unweighted). C1 rats were placed in the vibration apparatus but received no vibration during any phase. Control group 2 (C2; n = 3) rats remained in the home cages. Cooperation was evaluated with regard to rat-apparatus interactions and position compliance. Behavior, weight, and fecal glucocorticoid metabolite concentrations (fGCM) were used to evaluate animal well-being. We observed good cooperation and no behavioral patterns or weight loss between phases, indicating little or no animal stress. The differences in fGCM concentration between groups indicated that the EG rats had lower stress levels than the control rats in all phases except TrP1. Thus, this model elicited little or no stress in the conscious, unrestrained, seated rats.

Blood collection is frequently used for neonatal and juvenile mice in toxicology, developmental, and immunology studies and is often a terminal procedure. However, the use of nonterminal blood collection techniques, including the submandibular and the submental collection techniques described for adult mice, may offer opportunities to reduce animal numbers and refine current methods. The use of the submental technique has not been described for neonatal or juvenile mice. In this study, we compared the submental and submandibular blood collection techniques to determine their suitability for use in neonatal and juvenile mice. Male and female CD1 mice, ages 7, 14, 21, and 28 d, were randomized by sex into submental (n = 16), submandibular (n = 16), or control (n = 8) groups. Each mouse was weighed, bled per its assigned group (or only restrained in the case of control mice), and then decapitated without anesthesia for terminal blood collection. Blood collection volume and corticosterone concentrations were measured. The 2 methods showed significant differences in the volume of blood collected at ages 14 and 28, with the submandibular technique yielding significantly higher volumes. No significant differences were detected in corticosterone levels between the 2 techniques based on age or sex. A subset of mice (n = 8, 2 per age group) were bled via submental or submandibular technique and were evaluated 48 h later for gross and histopathologic evidence of trauma. Seven of the 8 mice showed expected inflammation and healing at the collection sites, with 4 mice having embedded strands of fur in the tissue. These data indicate that the submental blood collection is a viable method for nonterminal blood collection method in neonatal and juvenile mice, especially when smaller amounts of blood are needed.