Despite the enactment in the early 1990s of regulations requiring social housing of nonhuman primates (NHP), single housing is still prevalent in American research facilities. The publication of the 2011 edition of The Guide for the Care and Use of Laboratory Animals has increased
emphasis on the implementation of social housing as the default housing method for NHP. Overestimation of the risks inherent in social housing coupled with underestimation of both the benefits of social housing and the risks inherent in long-term single housing has prevented large-scale transitions
to social housing. Available caging and housing space often requires research facilities to use isosexual pairs to accomplish social housing. Pair housing presents unique challenges but can be used safely with a thorough understanding of macaque ethology. Here we review literature on the risks
and benefits of pair housing macaques in research facilities and provide a concise best-practice approach to pair housing.
The most widely used technique for determination of fibrinogen concentration is the Clauss fibrinogen (FIBClauss) assay, which measures the clotting time of plasma after addition of excess thrombin. More recently, the PT-derived fibrinogen (FIBPT) assay has been
developed, based on the relationship between fibrinogen concentration and the kinetics of clot formation during the prothrombin time. The objective of this study was to compare the fibrinogen concentration determined by the FIBClauss and FIBPT assays in citrated plasma samples from
healthy dogs (n = 40), monkeys (n = 40), rabbits (n = 26), and rats (n = 58) by using an automated coagulation analyzer. Results of a t test analysis indicated that the mean plasma fibrinogen concentrations measured by the 2 assays for all 4 species were
significantly different. According to Pearson correlation coefficients, the FIBPT assay displayed a high correlation (0.93 to 0.98) with the FIBClauss assay for all species. When the FIBPT and FIBClauss assays were compared by using Deming regression, positive or negative constant and proportional
biases emerged for all species. Intra- and interassay coefficients of variation for the FIBPT and FIBClauss assays were 0.8% to 2.3% and 1.8% to 7.4%, respectively. In conclusion, the FIBPT assay is a rapid and economical method for estimating fibrinogen concentration in plasma samples from
dogs, monkeys, rabbits, and rats. However, it should not be used without restriction. Further studies are required to investigate the performance of this assay in animals with various pathologic states, including coagulopathy, dysfibrinogenemia, and hypo- or hyperfibrinogenemia.
We measured daily intracage ammonia levels and performed weekly assessments of CD1 male, female, and breeder mice housed within disposable, ventilated cages that remained unchanged for 28 d. We tested housing groups comprising 1, 3, or 5 sex-matched mice per cage and breeder pairs with
litters. Mice housed in cages with higher concentrations of ammonia developed degeneration and inflammatory lesions in the nasal passages. Mean ammonia exposure levels that caused rhinitis were 181 ppm for 18 d. Ammonia exposures of 93 ppm for 16 d caused necrosis of the olfactory epithelium,
whereas 52 ppm for 13 d caused epithelial degeneration. Observers could not detect visible signs of rhinitis or identify cages with elevated ammonia levels, nor did they identify any sick or distressed mice. Observers consistently assigned poorer welfare scores as cages became dirtier. We
conclude that we can extend the cage-change interval to at least 28 d for disposable, ventilated caging housing a single CD1 mouse. Cages containing 3 CD1 mice of either sex should be changed biweekly, and cages containing 5 CD1 mice or breeder pairs should be changed at least once weekly.
During the past few decades, the development and use of individually ventilated cages (IVC), which are now commercially available for housing laboratory mice and rats, have increased. Because limited information is available regarding the influence of caging systems on the growth of
rats, the present study assessed body weight and food and water consumption in growing male rats that were housed in IVC and open-top cages (OTC). We allocated 21-d-old male Wistar outbred rats (HsdOla:WI; n = 24) into 2 groups, which then were housed in pairs in IVC (n = 12)
and OTC (n = 12). After an 8-d acclimatization period, body weight and food and water consumption were assessed every 3 d until the rats were 94 d old. There were no significant differences between the body weights of rats housed in IVC compared with OTC over the 65-d observation period.
Food and water consumption were greater in rats housed in OTC compared with IVC, becoming significantly different when the rats were 50 and 53 d old, respectively. In conclusion, IVC and OTC housing conditions influenced food and water intakes but not body weight in growing male rats. Further
research is needed to clarify the exact basis for these changes in food and water consumption.
Wooden objects are often used as nonhuman primate enrichment to provide variety and novelty, promote exploratory behavior, and supply an outlet for curiosity. However, concerns have been raised regarding the ability to sanitize wood by using conventional cage-wash procedures. To address
this concern, we examined sanitation outcomes between soiled plastic toys and manzanita wooden manipulanda immediately after a cage-wash cycle. Both an ATP luminometer device, which is capable of providing an immediate assessment of sanitation levels, and traditional bacterial culture were
used, with the secondary goal of comparing these methods for sanitation monitoring. Results showed that the wooden objects did not differ from plastic toys with respect to the overall efficacy of cage-wash sanitization. Therefore, manzanita wood can be used as nonhuman primate enrichment without
risking pathogen transmission when items are rotated among animals.
Primate facilities provide environmental enrichment to improve animal wellbeing, increase opportunities for expression of species-typical behaviors, and decrease the occurrence of stereotypic behaviors. The current study assessed the efficacy of 3 types of foraging enrichment: puzzle
balls, supertubes, and shakers. We assigned 48 rhesus macaques to 3 experimental groups, each of which received (after a 3-wk baseline observation period) 1 of the 3 enrichment devices intermittently for 3 wk. Observations were collected during 10-min sessions by using 1–0 sampling with
15-s intervals (480 h total). Observations were collected at the same 10 specified time points each week during the baseline period and after enrichment. Data were analyzed by using generalized linear mixed-effects modeling under the assumption that the underlying response followed a Poisson
distribution. Foraging behavior increased significantly in all 3 groups and remained increased in some groups when enrichment was removed after 43 h. The 3 enrichment devices had different effects on individual expression of stereotypy: supertubes decreased it, shakers increased it, and puzzle
balls led to a decrease followed by an increase. We present potential reasons for the changes in stereotypy and postulate a likely balance between the beneficial and negative effects of enrichment in any given environment.
Appropriate cleaning and disinfection procedures in zebrafish (Danio rerio) laboratories are crucial in preventing the spread of aquatic animal pathogens and minimizing the build-up of waste products and biologic matter. The procedures selected should accomplish these goals and
incorporate the individual needs of various laboratories. In this study of a single zebrafish facility, we assessed the efficacy of 2 different cleaning and disinfection procedures for nets, tanks, and lids. ATP levels were used as a surrogate biomarker for microbial burden. We measured the
number of relative light units (RLU), as an expression of the amount of ATP present, on items before and after disinfection and calculated the percentage reduction. We compared daily replacement of a commercial net disinfection product in J lab with weekly replacement in H lab and found a
96.6% reduction in RLU in H lab and a 91.2% reduction in J lab. These results indicate that either replacement schedule is effective. Evaluation of tanks and lids soaked in a bleach disinfection bath for 30 or 60 min revealed a 99.7% reduction in RLU at 30 min compared with 97.1% at 60 min.
Therefore a 30-min soak in a bleach bath achieved a similar level of disinfection as did a 60-min soak. The current results demonstrate that these cleaning and disinfection methods are efficacious.
Most published biomedical research involving animal models is evaluated carefully to ensure that appropriate ethical standards are met. In the current study, 500 journals randomly selected from MedLine were assessed for whether they presented animal research. Of the 138 journals that
did, the instructions to authors of 85 (61.6%) included a requirement for author assurance of adherence to ethical standards during experiments involving animals. In comparison to a wider range of biologic journals, biomedical science journals were more likely to have some sort of ethical
policy concerning the reporting and presentation of animal experiments.
Pinworm detection in laboratory rodents typically is accomplished by using the tape test or various modifications of fecal flotation test to detect eggs. Direct examination of intestinal contents remains the 'gold standard' for pinworm detection, with the limitation of euthanasia of
animals. Here, we compare traditional and real-time PCR methodologies during screening for and confirming the presence of Aspiculuris tetraptera. Two sets of pooled fecal samples collected from each of 521 microisolation cages in a mouse facility suspected to be pinworm-positive were
tested by PCR and fecal flotation methods. The number of PCR-positive cages was 48 (9.2%) compared with 5 (0.96%) by the fecal flotation method. All of the cages determined to be positive by fecal flotation were positive by PCR. We evaluated 8 positive cages containing 26 mice from the screening
group 5 wk later to confirm the initial findings; for 7 of these cages, PCR results from the initial screening were confirmed by fecal centrifugation concentration (FCC) or direct worm detection. Among the 26 mice, 4 were pinworm-positive by FCC, 5 by maceration, and 16 by PCR. All 4 mice
positive by FCC were positive by PCR; PCR was positive for 7 of the 9 mice in which pinworms were detected by FCC or maceration. Our study demonstrates that real-time PCR for survival testing of mice for A. tetraptera effectively augments current detection methods for quarantine and
routine health monitoring.
Treatment of Northern fowl mite (Ornithonyssus sylviarum) infestation on poultry in research facilities can be challenging. The mite has a rapid reproductive cycle (egg to adult in 5 to 7 d), and chemical treatments can be toxic to birds, personnel, and the environment. In addition,
antimite treatment may interfere with experimental research designs. The current study evaluated the efficacy of topical application of an entomopathogenic fungus, Beauveria bassiana, in the treatment of a naturally occurring infestation of Northern fowl mites in pen-housed roosters
(n = 14; age, 18 mo). Two groups of 7 roosters each were used in 2 experiments: Beauveria (30 mL, 2.9 × 1010 spores per bird) compared with water (30 mL, control), and Beauveria compared with the common topical organophosphate agent tetrachlorvinphos–dichlorvos
(30 mL). We also assessed a higher dose of Beauveria (300 mL, 2.9 × 1011 spores per bird) in the 7 birds that were not exposed to tetrachlorvinphos–dichlorvos. Beauveria reduced mite levels relative to the control group but did not outperform tetrachlorvinphos–dichlorvos
when used at an equal volume and frequency. Increasing the volume and frequency of Beauveria application improved outcomes such that visual inspection failed to detect any mites. The results presented here suggest that, when applied in sufficient doses, Beauveria effectively
reduces mites on poultry and can be an important part of an integrated pest management program. Additional research is needed to document the most effective dose, frequency, and location of B. bassiana application to control Northern fowl mites in poultry.
Rodents are often anesthetized by using ketamine and medetomidine, with reversal by atipamezole. Methods vary for times of administration of the atipamezole, and literature is lacking regarding appropriate reversal time. We investigated the recovery of mice reversed with atipamezole
10 min (early) or 40 min (late) after induction of anesthesia. Time to regain pinch-reflex or righting reflex did not differ between the 2 reversal points, but time to walking was significantly greater in mice that underwent early reversal with atipamezole. This delay was not mitigated by
administration of atropine as part of the anesthetic regimen. Inclusion of acetylpromazine in the anesthetic regimen shortened the time needed to reach a surgical plane of anesthesia but also prolonged recovery times as determined by righting reflex and time to walking.
Endometriosis affects a large percentage of the rhesus macaques (Macaca mulatta) at our institution. When the disease is diagnosed in macaques on long-term research protocols, the treatment of choice in our facility is monthly administration of medroxyprogesterone acetate (MPA)
to decrease estrogen release and subsequently diminish clinical signs associated with the disease. Because hormonal fluctuations associated with the normal menstrual cycle are known to affect parameters of glucoregulatory function in rhesus macaques, we evaluated the effect of MPA treatment
on glucoregulatory function cross-sectionally in 6 animals and longitudinally in 4 animals with endometriosis. Our hypothesis was that monthly administration of MPA for the treatment of endometriosis would negatively affect glucoregulatory function in rhesus macaques. We found that adult female
rhesus macaques on MPA therapy for 1.4 to 36.1 mo had lower insulin sensitivity than did age- and weight-matched healthy control animals. In addition, glucoregulatory function was reduced after MPA treatment as compared with pretreatment levels in a group of 4 macaques. These data suggest
that glucoregulatory function should be considered when endometriosis treatment is planned for rhesus macaques.
Alopecia in nonhuman primates in the biomedical research setting is often attributed to compromised psychologic wellbeing. Behavioral causes, mainly hair plucking, have become the unconfirmed and exclusive default diagnosis, and the possibility that alopecia may be secondary to a primary
medical or dermatologic disease is often overlooked. Although nonbehavioral causes of alopecia in nonhuman primates are described in the literature, few prospective hypothesis-based studies have investigated medical and behavioral etiologies concurrently. We therefore undertook such a study
with the aim of designing a clinical diagnostic guide for approaching cases of nonhuman primate alopecia. Because most cases of alopecia in nonhuman primates in the literature and at our facility are not associated with a definitive diagnosis, the hypothesis we tested was that the well-established
diagnostic evaluation for alopecia used for traditional veterinary species is not applicable to nonhuman primates. Discounting differences in histopathology and behavioral assessment, the current study revealed few clinically relevant significant differences between nonhuman primates with
and without alopecia. As a result, our hypothesis was confirmed, and we conclude that the standard dermatologic diagnostic plan typically described for alopecia diagnosis in traditional veterinary species and used as the basis for assessment of alopecia in nonhuman primates should be reassessed.
The chronically catheterized fetal sheep is a widely used model for fetal physiologic and pathophysiologic investigations. Catheterization involves opening the amniochorion to gain access to the fetus. In the current study, we explored the role of the amnion and amniochorion in maintaining
normal amniotic fluid volume (AFV) and composition and fetal blood-gas status after surgery. Fetal sheep were catheterized at 119.6 ± 0.3 (mean ± SE, n = 25) d gestation (term, approximately 147 d). An opening equal to approximately 5% of total membrane surface area was
created by resecting a portion of the amnion or amniochorion during surgery. The uterine wall was closed in all animals. Compared with control sheep (AFV = 992 ± 153 mL, n = 11), resection of the amnion had no significant effect on AFV (745 ± 156 mL, n = 7) measured
5 d after surgery, whereas resection of the amniochorion resulted in extensive loss of amniotic fluid (AFV = 131 ± 38 mL, n = 7). This loss resulted from extensive entry of amniotic fluid into the space between the chorion and uterine wall. Amniotic fluid, fetal plasma, and urinary
solute concentrations; arterial pH; oxygen tension; and carbon dioxide tension were unchanged. A small opening in the amnion has minimal effects on ovine AFV, whereas a small opening in the amniochorion results in oligohydramnios. In addition, the amnion appears to be the primary site that
limits the rate of amniotic fluid absorption by the chorionic vasculature.
Several Pacific 2-spot octopuses (Octopus bimaculoides) shipped from California and held in a recirculating seawater system at Illinois College exhibited an unusual postshipping stress behavior not previously documented in the literature. Ink, normally ejected into the surrounding
seawater, was uncharacteristically retained in the mantle cavity. We describe the resulting behaviors, discuss successful resuscitation efforts, and briefly consider the possible role(s) that ink may have played in the death of one octopus.
A 6-y-old male cynomolgus macaque presented with noticeable swelling of the left forearm and signs of discomfort, as indicated by nonuse of the arm even in a behavioral task that he previously had been well-motivated to perform. Examination under anesthesia revealed lacerations to the
arm. Radiography of the forearm showed no fractures, indicating that the damage was limited to soft tissue. The daily operant behavioral session assessed the amount of force the monkey emitted when touching the screen with the affected arm and how long each touch was sustained. We then used
these parameters (force and duration of touch) as objective measures of putative pain relief and recovery of function to guide the medical treatment. The affected monkey received ketoprofen, buprenorphine, or their combination but continued to perform poorly during daily operant behavioral
sessions. Only after treatment with dexamethasone did performance return to preinjury levels, suggesting inflammation near the radial or ulnar nerve. These findings indicate that performance of a trained operant task performance can be useful in guiding medical treatment, evaluating pain relief,
and objectively monitoring health in laboratory animals.