Animals have been used in research for over 2,000 y. From very crude experiments conducted by ancient scholars, animal research, as a science, was refined over hundreds of years to what we know it as today. However, the housing conditions of animals used for research did not improve significantly until less than 100 years ago when guidelines for housing research animals were first published. In addition, it was not until relatively recently that some extrinsic factors were recognized as a research variable, even when animals were housed under recommended guidelines. For example, temperature, humidity, light, noise, vibration, diet, water, caging, bedding, etc., can all potentially affect research using mice, contributing the inability of others to reproduce published findings. Consequently, these external factors should be carefully considered in the design, planning, and execution of animal experiments. In addition, as recommended by others, the housing and husbandry conditions of the animals should be described in detail in publications resulting from animal research to improve study reproducibility. Here, we briefly review some common, and less common, external factors that affect research in one of the most popular animal models, the mouse.

Decreased appetite is a common clinical problem in captive rhesus macaques (Macaca mulatta). Mirtazapine, a tetracyclic antidepressant originally developed for humans, has shown promise as a safe and effective promoter of weight gain and appetite in several veterinary species including rhesus and cynomolgus macaques. Although mirtazapine is available as oral formulations, transdermal delivery in macaques with reduced appetite would allow quick, painless, topical application. Here we describe the pharmacokinetics of a single application of a widely available veterinary transdermal mirtazapine formulation in 6 rhesus macaques. A dose of 0.5 mg/kg of transdermal mirtazapine ointment that has proven to be effective in rhesus was applied to the caudal pinnae of 3 female and 3 male young adult macaques. Serum was collected at 0, 0.5, 1, 3, 6, 8, 12, 24, 36, 48, and 72 h after administration. Our data indicate transdermal mirtazapine is absorbed at a lower level in rhesus as compared with published values in domestic cats (rhesus peak serum concentration: 1.2 ± 0.3 ng/mL), while drug half-life is longer than that reported in cats (rhesus: 33 ± 7 h). Mirtazapine reaches peak plasma concentrations in rhesus at 16 ± 10 h after administration; our model indicates that up to 5 d of serial dosing may be necessary to reach steady state. Our preliminary data also suggest that sex differences may contribute to efficacy and/or indicate sex-based differences, as male macaques reached T_max more quickly than females (19 ± 2 h in females and 8 ± 3 h in males) and showed higher variation in half-life (33 ± 4 h in females and 34 ± 11 h in males). While previous work indicates clinical efficacy of the 0.5-mg/kg dosage in macaques, further investigation is warranted to determine if rhesus may benefit from higher recommended doses than companion animal species.

Rabbits can develop anemia due to serial phlebotomy or secondary to induced disease states. This study evaluated the effects of a single injection and three consecutive injections of erythropoietin in rabbits at 150 IU/kg and 1,000 IU/kg in order to determine whether these dosages produce a sustained increase in hematocrit. Analysis of CBC and chemistry parameters showed significant elevation in hematocrit one week after administration of 1,000 IU/kg erythropoietin for three consecutive days. These results indicate that this dosage regimen can increase hematocrit in apparently healthy, nonanemic rabbits for one week.

Animal-based research is essential to the study of sepsis pathophysiology, diagnostics, and therapeutics. However, animal models of sepsis are often associated with high mortality because of the difficulty in predicting imminent death based on premortem assessment of the animals. The use of validated visual scoring would allow researchers to systematically identify humane endpoints but visual approaches require high interobserver agreement for accurate results. The objective of this study was to establish a scoring system for mice undergoing cecal ligation and puncture (CLP)-induced sepsis based on 3 visual parameters: respiratory status, activity and response to stimulus (ASR), and eye appearance, with scores ranging from 0 to 3. In the first study, we evaluated interobserver agreement. Veterinary and investigative staff assessed 283 mice with CLP and had substantial to near-perfect agreement for all 3 parameters as evaluated using weighted Cohen κ statistic. The second study assessed the ability of the scoring system and temperature to predict death. The scoring system and subcutaneous transpond- ers were used to monitor C57BL/6J mice (n = 80, male and female) until death or for 7 days after CLP. Results showed that the scoring system discriminates between surviving (n = 26) and nonsurviving (n = 54) septic mice. The scoring system was accurate in predicting death, with an AUC of 0.8997. The sensitivity and specificity of the ASR parameter were 96% and 92%, respectively, and for the eye parameter were 94% and 73%. A sum of the ASR and eye scores that was 5 or more was also predictive of death. Temperature was a quantitative predictor, with sensitivity and specificity of 93% and 92%, respectively. This scoring system refines the CLP model by allowing identification of humane endpoints and avoidance of spontaneous death.

Depending on the strain of immunodeficient mice, Corynebacterium bovis infection can be asymptomatic or cause transient or prolonged skin disease. C. bovis infection of NOD. Cg- Prkdc^scid Il2rg^tm1Wjl /SzJ (NSG) mice results in clinical skin disease that progresses in severity. Amoxicillin metaphylaxic and prophylaxic therapy prevents transmission and infection of mice after exposure to C. bovis and inhibits the growth of C. bovis isolates at therapeutic doses that are clinically achievable in mice. Amoxicillin is not efficacious for treatment of transient clinical skin disease in athymic nude mice, but the efficacy of amoxicillin treatment has not previously been characterized in C. bovis -infected NSG mice. In the current study, NSG mice were treated with amoxicillin beginning at 5 wk after exposure to C. bovis, at which time they had well-established clinical signs of disease. Clinical signs were scored to assess disease progression, regression, and reappearance. Our results showed that amoxicillin treatment for 3 or 6 wk reduced the clinical scores of NSG mice with C. bovis -associated clinical disease. In addition, withdrawal of treatment led to the recurrence of clinical signs. Collectively, our data suggest that amoxicillin treatment is effective in alleviating the clinical signs associated with C. bovis infection for the duration of treatment in NSG mice. Clinical intervention with antibiotics for C. bovis -infected NSG mice can be an option for management of C. bovis -related clinical disease either before or during facility-wide remediation efforts.

Mice are widely used as small animal models for influenza infection and immunization studies because of their susceptibility to many strains of influenza, obvious clinical signs of infection, and ease of handling. Analgesia is rarely used in such studies even if nonstudy effects such as fight wounds, tail injuries, or severe dermatitis would otherwise justify it because of concerns that treatment might have confounding effects on primary study parameters such as the course of infection and/or the serological response to infection. However, analgesia for study-related or -unrelated effects may be desirable for animal welfare purposes. Opioids, such as extended-release buprenorphine, are well-characterized analgesics in mice and may have fewer immune-modulatory effects than other drug classes. In this study, BALB/c and DBA/2 mice were inoculated with influenza virus, and treatment groups received either no analgesics or 2 doses of extended-release buprenorphine 72 h apart. Clinical signs, mortality, and influenza-specific antibody responses were comparable in mice that did or did not receive buprenorphine. We therefore conclude that extended-release buprenorphine can be used to alleviate incidental pain during studies of influenza infection without altering the course of infection or the immune response.

A detailed understanding of mosquito probing and ingestion behaviors is crucial in developing novel interventions to interrupt the transmission of important human and veterinary pathogens, but these behaviors are difficult to observe as the mouthparts are inserted into the skin of the host. Electropenetrography (EPG) allows indirect observation, recording, and quantification of probing and ingestion behaviors of arthropods by visualizing the electrical waveform associated with these behaviors. The study of mosquito probing and ingestion behaviors has been limited to the use of human hands as host, which is not suitable for pathogen transmission studies. Mouse models of mosquito-borne diseases are a widely used and indispensable tool in this research, but previous attempts to use direct current EPG to study mosquito probing behaviors on mice have been unsuccessful. Accordingly, the present study used alternating or direct current (AC-DC) EPG to observe the ingestion behaviors of adult Aedes aegypti mosquitoes on a mouse host in real time under BSL-2 containment conditions with enhanced BSL-3 practices. Our results show that waveform families previously identified during Ae. aegypti probing and ingestion on human hands were observed using 100 mV of AC at an input resistance (Ri) of 10⁷ Ohms (Ω) on CD1 mice. This work is a proof of concept for using mouse models for studying mosquito probing and ingestion behaviors with AC-DC EPG. In addition, these data show that the experimental setup used in these experiments is sufficient for conducting studies on mosquito probing and ingestion behaviors under BSL-2 containment conditions enhanced with BSL-3 practices. This work will serve as a foundation for using EPG to investigate the effects of pathogen infection on mosquito probing behaviors and to understand the real-time dynamics of pathogen transmission.

Southern giant pouched rats (Cricetomys ansorgei) are a small muroid species native to the sub-Saharan Africa. Their exceptionally developed olfactory system, trainability, and relatively small size makes them useful working animals for various applications in humanitarian work. At our institution, a breeding colony of Southern giant pouched rats is maintained to study their physiology and utility as scent detectors. This case report describes the occurrence of spontaneous pituitary neoplasms with distinct clinical presentations in 2 geriatric (approximately 7.5 y old) wild-caught female Southern giant pouched rats. The first pouched rat displayed vestibular deficits, including left-sided head tilt, ataxia, disorientation, and circling. MRI revealed a large, focal heterogeneous mass arising from the pituitary fossa. The second pouched rat presented with polyuria, polydipsia, and hyperglycemia but no neurologic signs. Examination after euthanasia revealed a prolactin (PRL)-expressing pituitary carcinoma and adenoma in the first and second pouched rat, respectively, associated with mammary hyperplasia in both animals. This is the first report of spontaneous PRL-producing pituitary tumors in Southern giant pouched rats.