The Association of Primate Veterinarians (APV) recognizes that antimicrobials are commonly prescribed for prophylactic, therapeutic, and experimental management of nonhuman primates (NHP). While clinicians should use antimicrobials to treat documented cases of illness, the decision to prescribe antimicrobials must take into account the increasing resistance of bacterial populations, leading to decreasing efficacy of critical pharmaceuticals in both human and veterinary medicine. The intent of this document is to provide guidance to veterinarians, research staff, and institutional animal care and use committees (IACUCs) on the use of antimicrobials in NHP.

The Association of Primate Veterinarians (APV) supports the responsible use of primates in biomedical research. Honoring the spirit of the 3Rs, these guidelines focus on refinement to explore post research retirement of eligible primates. APV supports the careful evaluation of each animal to determine if retirement is appropriate. Depending on prior use of the animal, animal reuse/reassignment may be an option prior to a decision to retire a primate. The following guidelines aim to provide information to institutions, researchers, animal care-givers, veterinarians, and Institutional Animal Care and Use Committees (IACUC) on the practical aspects of post research retirement of primates.

A point-of-care (POC) device to measure mouse glucose and lipid profiles is an important unmet need for cost-effective, immediate decision making in research. We compared metabolic analyte profiles obtained using a human clinical POC device with those from a veterinary laboratory chemical analyzer (LCA). Unfasted terminal blood samples were obtained by cardiac puncture from C57Bl/6J mice used in a diet-induced obesity model of type 2 diabetes mellitus; age-matched C57Bl/6J controls; a transgenic mouse model of Alzheimer's disease on a C57BL/6J background (16 wk old); and aged C57BL/6J mice (24 to 60 wk old). Aliquots of the blood were immediately assayed onsite using the POC device. Corresponding serum aliquots were sent analyzed by LCA. Measures from the POC and LCA devices were compared by using the Bland–Altman and Passing–Bablok methods. Of a total of 40 aliquots, LCA results were within reported reference ranges for each model. POC results that fell beyond the device range were excluded from the analyses. The coefficient of determination and Passing–Bablok analysis demonstrated that POC glucose and HDL had the best agreement with LCA. The Bland–Altman analysis found no value-dependent bias in glucose and no significant bias in HDL. The remaining lipid analytes (cholesterol and triglyceride) showed significant bias. Until an improved, validated mouse POC device with lipid profile capability is available, the POC device that we tested appears adequate for screening glucose and HDL in mouse blood. Disadvantages of this clinical POC device are the narrow human ranges relative to ranges found in mice and its limited precision as compared with the LCA. This study demonstrates that when the samples are within the device range limits, this human POC device can accurately track metabolic syndrome and be used to compare patterns in glucose and HDL.

Southern giant pouched rats (Cricetomys ansorgei) are muroid rodents native to subSaharan Africa. They are increasingly used as service animals because of their keen sense of smell and are primarily known for clearing minefields in Africa. The objectives of this study were to determine hematologic and biochemical reference intervals from clinically healthy wild-caught captive adult rats, to describe the cytochemical staining reactions of peripheral blood leukocytes, and to document urinalysis findings. Blood samples were collected from the coccygeal artery of 60 isoflurane-anesthetized rats (36 males and 24 females) and analyzed with automated hematologic and biochemical analyzers; manual differential cell counts were performed on modified Wright–stained blood smears. Urine was collected by cystocentesis, and dipsticks were analyzed on a urine analyzer, with visual examination of unstained sediments. Samples from a male rat with chronic renal disease were excluded from analysis. Reference intervals were determined according to guidelines established by the American Society of Veterinary Clinical Pathology. Lymphocytes were the dominant leukocyte in peripheral blood and granular lymphocytes were identified in most animals. Male rats had significantly higher RBC, absolute reticulocyte counts, and MCV than did female rats. Minor sex-associated differences in urea nitrogen concentration and GGT activity were noted. Leukocytes showed unique cytochemical staining characteristics. Small amounts of protein and bilirubin were found in the urine of rats of both sexes and of female rats, respectively, particularly in concentrated urine. These results will provide benchmarks for determining health status and identifying disease in this species of rat.

Fenbendazole is a broad-spectrum benzimidazole commonly used in laboratory animal medicine as an anthelmintic for elimination of pinworms. This drug is generally regarded as safe, with minimal side effects. Some data in rodent species indicate multiple physiologic effects of fenbendazole, including changes in immune parameters and behavior, but no studies to date have evaluated possible effects on reproduction in mice. The purpose of the current study was to determine the effects of several treatment regimens of fenbendazole on reproductive parameters in C57BL/6J mice. Uninfected mice were given fenbendazole-treated feed continuously or every other week until pups were born or weaned. This treatment also was combined with environmental decontamination. No significant differences in litter size, survival rate, or weaning weight were detected between groups. Under the conditions of this study, fenbendazole treatment does not affect reproduction in C57BL/6J mice.

The goals of this study were to examine the effect of stocking density on the stress response and disease susceptibility in juvenile rainbow trout (Oncorhynchus mykiss). Fish were sorted into one of 2 stocking densities (high density "HD", 20-40 kg/m³) or (low density, "LD", 4-8 kg/m³) and 3 stress indices (cortisol levels in serum and water, and neutrophil: lymphocyte (N:L) ratios from blood smears) were measured at multiple time points over 21 d. Serum cortisol was significantly increased at 1 h in LD samples and at 14 d in HD samples. Water cortisol concentrations were significantly higher in LD tanks as compared with HD tanks on day 14. N:L ratios were significantly higher in HD tanks on day 14 as compared with LD tanks and with baseline. The effect of stocking density on mortality after exposure to infectious hematopoietic necrosis virus (IHNV) was compared between fish held in HD or LD conditions, with or without prior acclimation to the different density conditions. No significant differences in survival were found between HD and LD treatments or between acclimated and nonacclimated treatments. Cumulative results indicate that 1) 1 to 4 gram rainbow trout did not generally demonstrate significant differences in stress indices at the density conditions tested over a 21-d period, 2) independent differences were found in 3 stress indices at day 14 after sorting into LD and HD holding conditions; and 3) LD and HD stocking densities did not have a significant effect on mortality due to IHNV.

Compassion Fatigue (CF) is commonly observed in professions associated with human and animal care. The COVID-19 pandemic compelled laboratory animal research institutions to implement new work practices in order to maintain essential animal care operations. These modifications ranged from shift changes to last-resort measures, such as culling animal colonies, to accommodate reduced staffing. Such changes could cause personnel to experience increased stress, isolation, and helplessness—all of which can increase CF risk. In the current study, 200 persons involved with animal research completed an online survey to gauge whether CF among laboratory animal personnel had increased during the pandemic. The survey examined professional quality of life, self-assessed levels of CF, institutional changes, perceived changes in animal welfare, and institutional measures intended to alleviate CF. A total of 86% of participants had experienced CF at some point in their career, with 41% experiencing a CF event (new or worsening symptoms of CF) during the pandemic. In addition, 90% of participants who reported a CF event also reported subsequent effects on their personal or professional lives. Health, employment, and animal-related stress that arose due to the pandemic were all found to influence CF scores significantly. Although 96% of respondents were considered essential workers, 67% did not feel as valued for their work as other essential personnel. Furthermore, 88% of personnel responsible for the euthanasia of healthy animals who experienced a CF event reported that CF also affected their personal life, professional life, or both, and 78% responded that interventions from internal CF programs or leadership did not help to alleviate symptoms of CF. The COVID-19 pandemic and resultant institutional changes will likely have lasting effects on persons and organizations. By determining and subsequently mitigating sources of CF, we can better assist the laboratory animal community during future crises.

The number of zebrafish in biomedical research has increased exponentially over the past decades, leading to pressure on the laboratory animal community to develop and refine techniques to monitor zebrafish health so that suitable stocks can be maintained for research. The water filtration assay is a promising technique in which water from a zebrafish system is filtered, and the filter analyzed by PCR. In the present report, we studied how the volume of water tested and the concentration of bacterial pathogens affected test results. To do so, we used stock solutions of 3 zebrafish pathogens: Edwardsiella ictaluri, Aeromonas hydrophila, and Mycobacterium marinum. We used these stocks to create solutions with known concentrations of each pathogen, ranging between 10² and 10⁷ Colony Forming Units (CFU) per ml. One, 2, and 3 L of each solution was filtered using positive pressure, and the filters were submitted to a commercial lab for PCR testing. Results were fit with a logistic regression model, and the probability of obtaining a positive result were calculated. Test sensitivity varied by organism, but in general, test results were positively correlated with the volume of the water filtered and with the concentration of bacteria in solution. We conclude that a positive result can be expected for E. ictaluri at 10⁵ CFU per mL, A. hydrophila at 10⁶ CFU per ml, and M. marinum at 10⁶ CFU per mL, when 3 L of solution are filtered.

Buprenorphine, an analgesic commonly used in rodent surgery, requires repeated dosing every 4 to 6 h in order to provide adequate analgesia. However, redosing requires repeated handling, which may itself cause stress. Buprenorphine SR-LAB, which reportedly maintains serum levels of buprenorphine greater than 1 ng/mL for 48 to 72 h, is commercially available. However, the viscosity of the product and small dosing volumes make accurate dosing a challenge. Simbadol is a concentrated formulation of buprenorphine hydrochloride labeled for use in cats with recommended dosing frequency of every 24 h. We measured serum concentrations over time after a single injection of this product in C57BL/6NCrl mice and compared it to standard buprenorphine (Buprenex) and Buprenorphine SR-LAB. Male and female mice were injected subcutaneously with one of the 3 buprenorphine formulations at a dose of 1 mg/kg at time 0. Groups of mice (n = 8) were euthanized at 1, 4, 8, 12, 16 h for all groups and 24 h for the Simbadol and the Buprenorphine SR-LAB. Liquid chromatography-mass spectrometry (LC-MS/MS) was used to determine concentrations of buprenorphine in each serum sample. High concentrations were observed in both Simbadol and standard buprenorphine groups one hour after injection (>50 ng/mL). These groups had similar buprenorphine concentration curves, including rates of decline. The standard buprenorphine group had mean concentrations less than 1 ng/mL by 12 h and the Simbadol group by 16 h. In contrast, the Buprenorphine SR-LAB group remained above the 1 ng/mL therapeutic threshold throughout the 24 h. In addition, clinical signs, including increased activity, that lasted for up to an hour after the injection in the Simbadol and standard buprenorphine groups. We conclude that Simbadol does not offer dosing advantages over the standard buprenorphine formulation when given at 1 mg/kg. Buprenorphine SR-LAB maintained a steady concentration of buprenorphine above 1 ng/mL for at least 24 h, and as such is a superior choice for providing long-term analgesia.

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.

This study evaluated the effects of using a heated anesthesia breathing circuit in addition to forced-air warming on body temperature in anesthetized rhesus macaques as compared with forced-air warming alone. Hypothermia is a common perianesthetic and intraoperative complication that can increase the risk of negative outcomes. Body heat is lost through 4 mechanisms during anesthesia: radiation, conduction, convection, and evaporation. Typical warming methods such as forced-air warming devices, conductive heating pads, and heated surgical tables only influence radiative and conductive mechanisms of heat loss. A commercially available heated breathing circuit that delivers gas warmed to 104 °F can easily be integrated into an anesthesia machine. We hypothesized that heating the inspired anesthetic gas to address the evaporative mechanism of heat loss would result in higher body temperature during anesthesia in rhesus macaques. Body temperatures were measured at 5-min intervals in a group of 10 adult male rhesus macaques during 2 anesthetic events: one with a heated anesthesia breathing circuit in addition to forced-air warming, and one with forced-air warming alone. The addition of a heated breathing circuit had a significant positive effect on perianesthetic body temperature, with a faster return to baseline temperature, earlier nadir of initial drop in body temperature, and higher body temperatures during a 2-h anesthetic procedure. Use of a heated anesthesia breathing circuit should be considered as a significant refinement to thermal support during macaque anesthesia, especially for procedures lasting longer than one hour.

Published data are sparse regarding the recognition of clinically relevant pain and appropriate analgesia in amphibians. The amphibian analgesia literature has primarily focused on nociceptive pathways in a single species, the northern leopard frog (Rana pipiens). The objective of the current study was to assess the analgesic efficacy and safety of oral tramadol and subcutaneous morphine in a commonly maintained zoo and pet species, White's tree frog (Litoria caerulea). We hypothesized that tramadol and morphine would provide dose-dependent antinociception, as measured by significant increases in hindlimb withdrawal latency after exposure to a noxious thermal stimulus. Two randomized, placebo-controlled, complete crossover studies were performed, with tramadol (n = 12) administered at 15, 25, and 40 mg/kg PO and morphine (n = 12) administered at 5 and 10 mg/kg SC. Hindlimb withdrawal latency was measured for a maximum of 72 h. No adverse side effects or signs of sedation were observed with any dose or drug evaluated. No significant difference in withdrawal latency was detected between the control and either tramadol or morphine. These negative results were surprising, suggesting that the thermal nociceptive model may not be biologically relevant in amphibian species.

Determining the clinical efficacy of analgesic drugs in amphibians can be particularly challenging. The current study investigated whether a thermal nociceptive stimulus is useful for the evaluation of analgesic drugs in 2 amphibian species. The objectives of this study were 2-fold: 1) compare 2 models of nociception (thermal and mechanical) using 2 frog species; White's Tree Frogs (Litoria caerulea; WTF) and Northern Leopard Frogs (Lithobates pipiens; NLF) after administration of saline or morphine sulfate; and 2) evaluate antinociceptive efficacy of morphine sulfate at 2 doses in a common amphibian research species, the NLF, using a mechanical stimulus. Neither WTF nor NLF displayed consistent drug-dependent changes in withdrawal responses to a noxious thermal stimulus applied using the Hargreaves apparatus, but NLF exposed to the noxious mechanical stimulus demonstrated a significant dose-dependent antinociceptive response to morphine sulfate. These results indicate that morphine is not antinociceptive in WTF, supporting previously reported results, and demonstrate the importance of using an appropriate experimental antinociceptive test in amphibians. Our data suggest that nociception in amphibian species may be best evaluated by using mechanical nociceptive models, although species differences must also be considered.

Rectal prolapse (RP) is a common clinical condition in mice, that does not have a recognized or documented standard of care. At our institution, an average of 240 mice develop RP each year. Our practice has been to recommend euthanasia upon identifying a RP based on its appearance as a painful or distressful condition. This study aimed to assess treatment options that would maintain the RP mucosa and allow mice to reach their study endpoint, and to evaluate the perception of this condition as a painful or distressful event. This study used 120 mice with spontaneous RP, concurrently assigned to ongoing research protocols. Mice were randomly assigned to 1 of 3 treatment groups: petroleum jelly, lidocaine jelly, or no treatment. Fecal samples were collected for pathogen testing, and all mice received an initial base score, followed by weekly blind scores. Upon euthanasia, RP tissue was collected for histopathology. Of the 120 mice identified with RP, 47 mice were breeders; 28% successfully produced 22 additional litters after developing RP. Seventy-three were nonbreeders, with 92% reaching their research study endpoint. No statistically significant differences were detected between the 3 treatment groups based on gross mucosal health, pain and distress, or histopathology. In this study, none of the mice in any group were euthanized based on the RP endpoint scoring criteria. These findings demonstrate that treatment is unnecessary for RP, and mice with RP did not show signs of pain or distress. In adherence to the 3Rs, this study supports animal number reduction and clinical refinement, allowing mice with RPs to reach their intended research study endpoints or produce additional litters.

One strategy commonly employed for rodent surgeries is a "tips-only" surgical technique, which restricts the surgeon to using only the sterile working ends of the surgical instruments to manipulate the surgical field and sterilizes instrument tips with a hot bead sterilizer between consecutive rodents. Despite the common use of the "tips-only" technique, research is lacking on the number of sequential surgeries for which the same set of hot bead-sterilized instruments can be used before introducing bacterial contamination. We performed serial mouse surgeries using the "tips-only" technique under 3 different conditions (aseptic, fur contamination, or cecal contamination) and assessed aerobic bacterial growth before and after each round of hot bead sterilization. Instrument tips showed an increasing probability of contamination of at least one instrument in a series of consecutive surgeries. The probability that all surgical instrument tips in the series were sterile after hot bead sterilization fell by 4% for each surgery involving inadvertent or fur contamination and by 11.5% for each surgery with contamination for all surgical types combined (including entering the gastrointestinal tract). Based on our results, hot bead sterilization is not adequate for surgeries associated with gross contamination. Under our experimental conditions and assuming independence of outcomes between consecutive surgeries, up to 5 surgeries associated with minor or inadvertent contamination could be performed in series with a probability higher than 80% that all instrument tips were sterile for all surgeries. A case-by-case risk assessment should be conducted to derive institutional guidelines for the maximal number of surgeries that can be performed in sequence using the "tips-only" technique with hot bead sterilization of the same set of surgical instruments between surgeries. Full sterilization of instruments after every surgery provides the greatest confidence in maintaining sterility.