Molecular diagnostics (PCR and RT-PCR) have become commonplace in laboratory animal research and diagnostics, augmenting or replacing serological and microbiologic methods. This overview will discuss the uses of molecular diagnostics in the diagnosis of pathogenic infections of laboratory animals and in monitoring the microbial status of laboratory animals and their environment. The article will focus primarily on laboratory rodents, although PCR can be used on samples from any laboratory animal species.

Results of animal experiments are used for understanding the pathophysiology of diseases, assessing safety and efficacy of newly developed drugs, and monitoring environmental health hazards among others. Systematic reviews and meta-analyses of animal data are important tools to condense animal evidence and translate the data into practical clinical applications. Such studies are conducted to explore heterogeneity, to generate new hypotheses about pathophysiology and treatment, to design new clinical trial modalities, and to test the efficacy and the safety of the various interventions. Here, we provide an overview regarding the importance of systematic reviews and meta-analyses of animal data and discuss common challenges and their potential solutions. Current evidence highlights various problems and challenges that surround these issues, including lack of generalizability of data obtained from animal models, failure in translating data obtained from animals to humans, poor experimental design and the reporting of the animal studies, heterogeneity of the data collected, and methodologic weaknesses of animal systematic reviews and meta-analyses. Systematic reviews and meta-analyses of animal studies can catalyze translational processes more effectively if they focus on a well-defined hypothesis along with addressing clear inclusion and exclusion criteria, publication bias, heterogeneity of the data, and a coherent and well-balanced assessment of studies' quality.

The porcine ischemia-reperfusion model is one of the most commonly used for cardiology research and for testing interventions for myocardial regeneration. In creating ischemic reperfusion injury, the anesthetic protocol is important for assuring hemodynamic stability of the animal during the induction of the experimental lesion and may affect its postoperative survival. This paper reviews the many drugs and anesthetic protocols used in recent studies involving porcine models of ischemiareperfusion injury. The paper also summarizes the most important characteristics of some commonly used anesthetic drugs. Literature was selected for inclusion in this review if the authors described the anesthetic protocol used and also reported the mortality rate attributed to the creation of the model. This information is an important consideration because the anesthetic protocol can influence hemodynamic stability during the experimental induction of an acute myocardial infarction, thereby impacting the survival rate and affecting the number of animals needed for each study.

The use of a nonsurgical embryo transfer technique in rodents eliminates the potential pain, distress, and health complications that may result from a surgical procedure and as such, represents a refinement in rodent assisted reproductive techniques. A nonsurgical technique has not been previously developed for use with rat embryos. Here we describe an efficient method to deliver either fresh or cultured blastocyst stage embryos to the uterine horn of pseudopregnant female rats using a rat nonsurgical embryo transfer (rNSET) device. The rNSET device is composed of a Teflon catheter and a hub that attaches to a 2 μL pipette. Oxytocin is used to dilate the cervix before the delivery of blastocysts, allowing passage of the rNSET catheter directly into the uterine horn for embryo delivery. The efficiency of recovery of pups after nonsurgical embryo transfer is similar to the efficiency after surgical embryo transfer. Furthermore, the technique is not stressful to the subjects, as demonstrated by the absence of a decrease in weight or increase in fecal corticosterone level in recipients of embryos delivered nonsurgically, without the use of anesthesia or analgesia.

Bedding material is a critical component of the mouse environment and affects animal wellbeing and research integrity. Corn cob (CC) bedding has been a common bedding choice in research despite several potential negative aspects of its use. We investigated the use of compressed paper (CP) bedding as a refinement to CC bedding. CP bedding demonstrated greater total and immediate absorption, compared with CC bedding. CP-bedded cages had a reduced frequency of early cage changing prior to the Guide-recommended 2-wk interval for IVC; this reduction was proportional to room census. Intracage ammonia levels were lower in CP-bedded IVC compared with CC-bedded IVC, independent of the age, sex, and number of mice per cage. By contrast, ammonia levels were similar between CP-bedded and CC-bedded static cages. Collectively, these data support the use of CP bedding as a refinement for CC in ventilated mouse cages, in light of increased husbandry efficiency and its positive effect on the welfare of mice.

Traditional laboratory caging for nonhuman primates is typically configured in a 2-tiered manner, with caging arranged in 2 horizontal rows stacked vertically. Studies of the effects of cage row have yielded inconsistent results with respect to impacts on psychological well-being. This study tests whether rhesus macaques (Macaca mulatta) housed in the bottom tier of caging display poorer responses to attempted positive human interaction than those in the upper tier, suggesting that humans are a greater stressor for animals housed in bottom rows. The attempted positive social interaction took the form of offering a food treat by hand. This study involved 270 male and female singly-housed rhesus macaques, ranging in age from 2.4 to 27.4 y of age. Cage position was characterized not only with respect to tier, but also with respect to proximity of the cage rack to the room door. A single technician recorded whether the animal retrieved the treat within 10 s and also recorded all social behaviors directed toward the technician during the test. No effects of cage tier were detected, nor were effects of proximity to the door found. However, significant contrasts were detected with respect to subjects' sex, age, and lifetime tenure in indoor caging. Females were less likely than males to take treats from a human's hand, and were more likely to show fear. Both increased age and tenure in caging were associated with an increased probability of taking the treat. These findings may have implications for programs aiming to monitor and address fearful behavior.

Rodent vivaria have traditionally used soiled bedding sentinel (SBS) health-monitoring programs to detect and exclude adventitious pathogens that could affect research results. Given the limitations of SBS, a likely reduction in animal usage, and a decrease in animal care staff labor, exhaust air dust (EAD) health monitoring has been evaluated by several groups for its efficacy in detecting pathogens when used as a complete replacement for traditional SBS health-monitoring programs. Compared with SBS, EAD has also been shown to provide increased sensitivity for the detection of multiple pathogens. After implementing EAD at our institution, we conducted an analysis to compare the annual costs of the 2 health-monitoring programs. The EAD program was found to be 26% less expensive than SBS. In addition to these cost savings, EAD decreased the amount of time spent by the staff on heath-monitoring activities. For veterinary technicians, this decrease in time was calculated as a savings of 150 h annually, almost 3 h each week. Finally, the EAD program replaced the use of live sentinel animals, decreasing the associated yearly usage from 1,676 animals to zero.

Recently, the zebrafish has gained in popularity as a vertebrate animal model for biomedical research. Commercial zebrafish housing systems are available and are designed to maximize stocking density of fish for a given space, but these systems are expensive and purchasing them may not be feasible for emerging laboratories with limited funding. In this article, we describe the construction of a simple and affordable recirculating zebrafish housing system. This system can be constructed in 3 working days, with materials readily available in hardware stores. The cost for construction of the system was only 3,000 MYR (750 USD). The system consists of a water reservoir, a supply line that delivers water to the shelves holding the zebrafish tanks, and a drainage line that receives water from both the supply line and the shelves containing the fish tanks and returns this water to the reservoir. This system also has a 3-stage filtration process to ensure that clean water is delivered to the zebrafish tank. The system can house up to 360 zebrafish. This low-cost housing system may make research using zebrafish feasible some laboratories.

Intraperitoneal (IP) injection is a common route of anesthetic administration in mice. Ketamine-xylazine (KX) anesthesia is one of the most widely used IP protocols, but has limitations. Etomidate is an alternative to ketamine that has been used in both human and veterinary medicine yet has not been widely studied in mice. The purpose of this study was to evaluate etomidate-xylazine (EX) anesthesia as an alternative to KX. We hypothesized that EX would be as safe and effective as KX, with both sex- and strain-dependent differences. Male and female Crl:CD1(ICR), C57BL/6NCrl, BALB/cJ and NU/J mice were given a single IP dose of ketamine 100 mg/kg and xylazine 10 mg/kg or etomidate 20 mg/kg and xylazine 10 mg/kg. Sedation times were similar between KX and EX, with CD1 mice exhibiting shorter sedation times. Surgical anesthesia was achieved in 44% of EX mice, compared with 4% of KX mice. C57BL/6NCrl mice were significantly more likely to achieve surgical anesthesia when given EX (94%) or KX (18%) than were other strains. In all strains except C57BL/6NCrl mice, females were more likely to reach surgical anesthesia than males. Several mice experienced an adverse hyperexcitement response during induction, with BALB/cJ (79%) and NU/J (87%) mice given EX significantly more likely than other strains to experience hyperexcitement. EX and KX protocols had no overall differences in lowest respiration rate, lowest systolic blood pressure, lowest rectal temperature, or levels of acidosis, although the lowest heart rates were significantly higher with EX, indicating that EX and KX have similar safety profiles. Thus, EX and KX administration were associated with several significant physiologic differences when comparing sexes or individual strains. Our results indicate that EX is an equally effective sedative and a more effective surgical anesthetic than KX; however, EX is only recommended for invasive procedures in C57BL/6 mice due to the high rate of hyper-excitement and inconsistent surgical depth seen in other strains. Further study is needed to optimize EX for use in multiple mouse strains.

Due to their unpredictability and variable effects, injectable anesthetic regimens in laboratory rodent species warrant refinement. In our study we sought to evaluate alfaxalone, which has gained recent popularity in veterinary medicine, alone and in combination with dexmedetomidine to evaluate their anesthetic ability in Sprague–Dawley rats when administered intraperitoneally. Three doses of alfaxalone only and 4 dose combinations of alfaxalone-dexmedetomidine were tested in males and female rats. The time to induction, anesthetic duration, pulse rate, respiratory rate, temperature, and time to recovery were recorded by a blind observer. The level of anesthesia induced by the various anesthetic protocols was assessed by using pedal withdrawal reflex to a noxious stimulus and scored according to the response. Dependent on the treatment group, atipamezole or saline was administered intraperitoneally once animals reached 60 min of anesthesia. Regardless of the dose, alfaxalone alone achieved only a sedative level of anesthesia, whereas all alfaxalone-dexmedetomidine combinations led to a surgical level of anesthesia in all animals. Anesthesia regimens using alfaxalone alone and in combination with dexmedetomidine demonstrated sex-associated differences, with female rats maintaining longer durations of sedation or anesthesia than their male counterparts. Both male and female rats displayed decreases in physiologic parameters consistent with the effects of dexmedetomidine. Given the results described herein, we recommend 20 mg/kg alfaxalone for sedation and 30 mg/kg alfaxalone combined with 0.05 mg/kg dexmedetomidine for surgical anesthesia in female rats. Appropriate doses of alfaxalone only and alfaxalone-dexmedetomidine for male rats were not determined in this study and need further evaluation.

In this study, adult intact male and female (n = 10) naked mole rats (Heterocephalus glaber) were anesthetized by using a combination of ketamine (20 mg/kg IM), and alfaxalone (4.0 mg/kg IM). Induction and recovery times were recorded. Vital parameters, including heart rate, respiratory rate, and reflexes, were monitored every 5 min during the anesthetic period. Anesthetic induction was smooth and rapid. Induction time was significantly longer in male rats (median, 325 s; range, 180 to 385 s) than in females (median, 145 s; range, 118 to 180 s). In addition, overall duration of loss of righting reflex was shorter in male mole rats (median, 50 min; range, 36 to 65 min) than females (median, 70 min; range, 60 to 85 min). Males largely had intact withdrawal reflexes, whereas females showed variable loss of both forelimb and hindlimb withdrawal reflexes. Neither recovery time (mean ± 1 SD, 16 ± 13 min) nor vital parameters differed between sexes. None of animals showed any anesthesia-related adverse responses. According to these findings, intramuscular AK is a safe and effective protocol that provides brief, light anesthesia in male naked mole rats and deeper anesthesia in females. We recommend adding analgesics when this AK protocol is used for pain-inducing or invasive procedures, and further studies evaluating higher doses and different combinations are indicated.

Pain management in rabbits can be difficult because they are adept at hiding pain and can be stressed by handling and restraint for injection. The use of opioid analgesics with prolonged durations of activity could alleviate pain, but associated adverse effects including gastrointestinal ileus, inappetence, and tissue reactions have been reported. In this study, we compared gross tissue reactions at the site of injection, food consumption, and fecal production after single injections of buprenorphine HCl (Bup; n = 7), sustained-release buprenorphine (BupSR; n = 8), and high-concentration buprenorphine (BupHC; n = 7) during the first 3 d after minor survival surgery. We also measured plasma concentrations of the parent drug, buprenorphine, and 3 metabolites (buprenorphine-3-glucuronide (B3G), norbuprenorphine-3β-glucuronide (N3G), and norbuprenorphine (NB)). Plasma levels of buprenorphine remained above the theoretical minimal analgesic concentration for 4 h for Bup and 42 h for BupHC. For BupSR, plasma levels of buprenorphine remained above the theoretical minimal analgesic concentration for approximately 77 h, starting 15 h after administration. For all 3 formulations, N3G was the most prominent metabolite in the blood. No injection site reactions were visible grossly in any rabbit. Relative to baseline measures and compared with controls (n = 8), food consumption was suppressed on days 1 through 3 in rabbits that received BupSR and on days 2 through 3 in those given BupHC. Feces production on day 3 was reduced to a greater extent in BupSR rabbits than control animals. Two rabbits in the BupHC group exhibited neurologic signs after drug administration. These adverse effects should be considered when choosing a long-lasting buprenorphine formulation to manage pain in rabbits. There is an erratum to this article in Volume 60 Issue 1: https://www.ingentaconnect.com/contentone/aalas/jaalas/2021/00000060/00000001/art00015

The Northern greater galago (Otolemur garnettii) is a prosimian primate most commonly used to study the evolutionary development of vision and somatosensation. This study aimed to investigate the efficacy and cardiopulmonary effects of 3 sedation protocols commonly used in other primate species: 1) alfaxalone (Alf; 8 mg/kg IM) 2) ketamine alone (Ket; 20 mg/kg IM) and 3) ketamine + dexmedetomidine (Ket+Dex; 4 mg/kg + 25 μg/kg IM) with reversal (atipamezole; 250 μg/kg IM). A total of 34 animals were evaluated, including 11 juveniles and 23 adults. Cardiopulmonary parameters such as indirect blood pressure, heart rate, respiratory rate, and SpO₂ were measured, and blood was collected for blood gas analysis and a chemistry panel. To examine the efficacy of each sedation protocol, induction time, immobilization time, and recovery time were recorded. Subjective measures of quality and efficacy included quality of induction, pedal withdrawal reflex, palpebral reflex, muscle tension, rectal temperature, and quality of recovery. All 3 protocols successfully immobilized the animals and all animals recovered from sedation. Heart rates were highest among the Ket group and the lowest for the Ket+Dex group. On average, the Alf group was immobilized for twice as long as either the Ket or Ket+Dex groups. The Ket+Dex group had the fastest average recovery time and subjectively had the best quality of recovery. Based on these results, Ket+Dex is recommended over Alf or Ket alone for brief sedation of healthy galagos.

We studied domestic ferrets (Mustela putorius furo) to evaluate the physiologic effects of routine surgery. Standard plasma biochemistry panels and ¹H-NMR spectroscopy of heparinized whole blood were performed on samples taken 24 h prior to and immediately after surgery from female and male ferrets undergoing routine gonadectomy. Increases in plasma glucose, phosphorus, potassium, and creatine kinase concentrations associated with the duration of surgery were identified on plasma biochemistry panels. Whole-blood NMR spectra allowed us to identify 42 metabolites and one drug residue. Variations between pre- and postoperative metabolite concentrations were most pronounced for female ferrets, which underwent more prolonged surgery than males. Affected metabolites included organic acids and osmolytes (betaine, methylmalonate, <small>D</small>-lactate), fatty acids and lipids (2-hydroxy-3-methylbutyric acid), and amino acid groups (acetylglycine, alloisoleucine, leucine, and isoleucine). These findings indicate that ¹H-NMR spectroscopy of whole blood provides insight into metabolic perturbations in domestic ferrets undergoing surgery that are not detected in routine clinical chemistry panels.

A proposal for the use of porcine pancreatic elastase (PPE) to develop a mouse model of pulmonary emphysema raised concerns about introducing contaminating porcine viruses into our barrier facility. Porcine Circovirus (PCV) is a known contaminant of vaccines and cell cultures that have been exposed to porcine-derived reagents. Endemic infection of PCV3 in laboratory mice has been reported, and some evidence supports natural PCV infection in wild mice. PPE samples from 2 different vendors tested positive for DNA from both PCV2 and 3. To allow model development with these reagents to proceed, we developed a protocol that would meet scientific objectives, minimize exposure of mice, and provide information on the potential for the virus to spread. Five d after BALB/c mice received intralaryngeal administration of PPE, lungs were harvested and analyzed for evidence of disease. Tissues from other major organs were submitted to test for disseminated PCV2 and 3 DNA. Similarly, tissues (including lungs) from direct contact nude sentinel mice were analyzed for the presence of the virus. To evaluate the possibility of endemic PCV2/3 infection, we also surveyed non-porcine reagent exposed mice on other studies. PCV2 and 3 was not detected in any of the tissues submitted. Although this study provided no evidence of infection and transmission of PCV2/3 from the contaminated PPE sample over the 5 d study, further work is needed to understand the risks and impact of introducing PCV contaminated cells or reagents into barrier maintained rodent colonies.