Bluegill fish (Lepomis macrochirus) are a popular sportfish across North America. Research involving bluegill has focused mainly on locomotion, environmental monitoring, bioaccumulation, and toxicology. With fish becoming more popular research models, bluegill use may increase. Consideration for sedation and anesthesia in bluegill is lacking. MS-222 is a commonly used anesthetic in fish that requires a 21-d washout period before entry into the food chain. Other, safer options for anesthesia should be available. In this study, we first determined a suitable MS-222 dose for general anesthesia, then compared it with 2 different concentrations of alfaxalone (5 and 10 mg/L). Both concentrations of alfaxalone were adequate to reach the desired anesthetic plane, although time to effect was dose-dependent and longer in these groups when compared with MS-222. Time to recovery was also prolonged in both alfaxalone groups compared with the MS-222 group. We also assessed anesthetic degradation in the water bath over time. In this study, we show that sedation with alfaxalone at 5 and 10 mg/L is just as effective as MS-222 with no degradation of either anesthetic over the time measured.

To maintain rodent colonies free from harmful infectious agents, laboratory animal care programs frequently employ the use of sterilized caging and supplies. Sterilized caging is important for preventing the spread of infectious pathogens from contaminated fomites, for biocontainment, and for safety. We designed several experiments to determine the effects of commonly encountered processes on steam penetration and substrate heat exposure. We used biologic indicators as a proxy for steam penetration. We used the time bedding spent above 121 °C as a proxy for substrate cumulative heat exposure. This temperature was measured using a high-temperature data logger. We first examined the effect of stacking cages with various bedding types on steam penetration. We then autoclaved soiled bedding and studied the variables of bedding type, bagging style, presence of diet and water in the cage, and amount of time between cage change out on steam penetration and cumulative heat exposure. For clean bedding, we found adequate steam penetration regardless of bedding type, cycle program, or location of the cage in the bulk autoclave. For the soiled bedding experiments, there were no differences between bedding types noted. Placement of cages inside plastic bagging increased the amount of time the bedding spent above 121 °C on average but not significantly. There was no difference in steam penetration of bedding or time spent above 121 °C for 2- or 4-wk cage change-out schedules. When cages were autoclaved with diet and water, the time the bedding spent above 121 °C was significantly less than when autoclaving bedding alone, but there was adequate steam penetration for all cages. This study demonstrates that common practices in the industry are effective. Based on the results of this study, it is recommended that each institution evaluate their autoclaving practices and confirm that those practices are sufficient and effective.

Abstract

There are few reports describing spontaneous neoplasms in cynomolgus macaques, despite the frequent use of this species in laboratory research. This report describes cytologic, histologic, and immunohistochemical findings of a cutaneous to subcutaneous nerve sheath tumor located within the haired skin of the abdomen of a 2.5-y-old, intact, female, captive Mauritius cynomolgus macaque. The nerve sheath tumor was well demarcated, partially encapsulated, densely cellular, and extended from the subcutis to the most superficial dermis, abutting the epidermis. Neoplastic cells formed intersecting streams and had a high mitotic count (18 per 2.37 mm²). Due to the substantial morphologic overlap of this neoplasm with amelanotic melanoma, particularly the close association with the epidermis, immunohistochemistry was required for definitive diagnosis. Neoplastic cells were immunoreactive to vimentin, S-100, SOX10, laminin, collagen IV, and CD56, and negative for melan-A, tyrosinase, MITF, and HMB45. This immunohistochemical profile is diagnostic for nerve sheath tumor based on human and canine criteria and rules out amelanotic melanoma. Despite incomplete excision, the nerve sheath tumor had not grossly recurred after 1 mo, at which point the animal was euthanized for unrelated reasons. This report underscores the importance of using an immunohistochemical panel in cases of cutaneous and subcutaneous spindle cell neoplasms, as there is substantial morphologic and immunohistochemical overlap between nerve sheath tumors and melanocytic neoplasms due to their shared neuroectodermal origin. To our knowledge, this is the first report of a nerve sheath tumor in a cynomolgus macaque, and one of the few reports of spontaneous neoplasia in this species.

Chlamydia muridarum (Cm) is a moderately prevalent, gram-negative, intracellular bacterium that affects laboratory mice, causing subclinical to severe disease, depending on the host’s immune status. The effectiveness of various antibiotic regimens aimed at eradicating Cm in both immunodeficient and immunocompetent laboratory mice was evaluated. NSG mice were cohoused with Cm-shedding BALB/cJ mice for 14 d to simulate natural exposure. Four groups of 8 infected NSG mice were treated for 7 d with either 0.08% sulfamethoxazole and 0.016% trimethoprim (TMS) in water, 0.0625% doxycycline in feed, 0.124%/0.025% TMS in feed, or 0.12% amoxicillin in feed. A control group was provided standard water and feed. The impact of treatment on gastrointestinal microbiota (GM) was investigated using next-generation shotgun sequencing on the last day of treatment. TMS and amoxicillin had negligible effects on GM, while doxycycline had the largest effect. All antibiotic-treated NSG mice exhibited clinical disease, including dehydration, hunched posture, greater than 20% weight loss, and dyspnea, leading to euthanasia 21 to 40 d posttreatment (32.6 ± 4.2 d; mean ± SD). Untreated controls were euthanized 14 to 33 d postexposure (23.75 ± 5.9 d). All mice were fecal PCR positive for Cm at euthanasia. Histologic evaluation revealed multifocal histiocytic and neutrophilic bronchointerstitial pneumonia and/or bronchiolitis featuring prominent intralesional chlamydial inclusion bodies in all mice. Subsequently, groups of 8 C57BL/6J, BALB/cJ, NOD.SCID, and NSG mice infected with Cm were treated with 0.124%/0.025% TMS in feed for 7 (BALB/cJ and C57BL/6J) or 21 d (NSG and NOD.SCID). All immunocompetent and NOD.SCID mice were negative for Cm by PCR 14 d posttreatment, remained clinically normal, and had no evidence of Cm infection at necropsy, and all NSG mice remained Cm positive and were euthanized. While these findings highlight the difficulties in eradicating Cm from highly immunodeficient mice, eradication of Cm from immunocompetent or moderately immunocompromised mice with antibiotics is feasible.

Abstract

While rodents are used extensively for studying pain, there is a lack of reported direct comparisons of thermal and mechani- cal pain testing methods in rats of different genetic backgrounds. Understanding the range of interindividual variability of withdrawal thresholds and thermal latencies based on these testing methods and/or genetic background is important for appropriate experimental design. Testing was performed in two common rat genetic backgrounds: outbred Sprague–Dawley (SD) and inbred Fischer 344 (F344). Male and female, 10- to 14-wk-old F344 and SD rats were used to assess withdrawal thresholds in 3 different modalities: the Randall-Selitto test (RST), Hargreaves test (HT), and tail flick test (TFT). The RST was performed by using an operator-controlled handheld instrument to generate a noxious pressure stimulus to the left hind paw. The HT and the TFT used an electronically controlled light source to deliver a noxious thermal stimulus to the left hind paw or tail tip, respectively. Rats of each sex and genetic background underwent one type of test on day 0 and day 7. Withdrawal thresholds and thermal latencies were compared among tests. No significant differences were observed. Our findings can serve as a guide for researchers considering these nociceptive tests for their experiments.

Abstract

Soft-pelleted, high-fat diets (HFD) are greasy and crumble easily leading to food wastage and hair coat grease accumulation when mice are fed using commercially available feeders. The ideal HFD feeder design should reduce food wastage, facilitate mouse weight gain, and minimize variables such as hair coat grease accumulation that have the potential to alter scratching behaviors. Our study compared the feeding efficiency of 2 commercially available feeders (feeders A and E) to 4 novel feeder designs (feeders B, C, D, and F). Novel feeders had alterations in feeding aperture size, feeding surface area, feeder configuration, and level of food presentation. Male C57BL/6NCrl mice (n = 120; 4/cage) were randomly assigned to cages containing one of the 6 feeder types and were fed HFD for 12 wk. Feeders and cage bottoms were weighed before use and then weekly at the time of cage change. Mice were weighed before starting the HFD and then biweekly. Scratching behavior was video recorded at 0, 4, 8, and 12 wk. Hair coat grease accumulation was visually scored biweekly. Feeder A use was associated with the highest feed cost due to HFD wastage ($36.98 ± 1.54/cage/wk). Mice fed using Feeder A had the highest average weight gain (23.75 ± 0.8 g, P < 0.005). However, mice also had significantly higher hair coat grease accumulation scores (P < 0.05) and significantly increased scratching frequency at 4 wk (P < 0.05) when compared with mice fed using other feeder types. Novel feeder designs utilized 10 to 21 times less HFD dispensed when compared to feeder A. Mice fed using novel feeders also displayed improved welfare, as evidenced by low hair coat grease accumulation scores, and no significant differences in scratching frequency when compared with baseline behavior.

Environmental enrichment is the provision of different substrates to mimic an animal’s natural environment and encourage natural, species-specific behavior. However, the use of enrichment to improve breeding efficiency in mouse models for neurologic conditions is not well described. There are reports that diminished environmental stimuli and chronic isolation can result in the early expression of the Parkinson phenotype in mice with a genetic predisposition to the disease. In this study, we compared the provision of crinkle paper, DietGel, and their combination on reproductive parameters in B6.Cg-Tg(THY1-SNCA*A53T)M53Sud mice. We found that enhanced enrichment combined with enhanced nutrition increased dam weight and decreased the interlitter intervals. In addition, enhanced enrichment increased the production index, number of pups born, pups weaned, and the percent survival of pups. This study underscores the importance of incorporating enrichment to enhance the reproductive parameters in mice that are models of Parkinson disease.

Abstract

The Whitten effect is a widely used tool for manipulating the mouse estrous cycle and generating reproductively active females within the laboratory setting. Typically, peak numbers of sexually receptive mice occur following exposure to male pheromones, resulting in a higher number of successful copulations on the third day after exposure. Although this method has improved efficiencies, the percentage of females mated and subsequently deemed to be pregnant/pseudopregnant remains relatively low, around 50%. In experiment 1, we aimed to 1) further understand cyclicity; 2) determine whether the initial cycle stage plays an importance on day 3 receptivity; and 3) identify any repetitive patterns/cycle stabilization. Mice (n = 27) were assigned to group cages according to cycle stage (proestrus, estrus, metestrus, diestrus). Experiment 2 was developed to determine an optimum treatment to promote receptivity by exposure to various pheromone stimuli. Mice (n = 45) were randomly assigned to 5 treatment groups (PBS-treated sham soiled bedding, male soiled bedding, live male, pregnant females, and lactating females). In both experiments, daily vaginal cytology was performed for 21 days to determine the cycle stage. Results from experiment 1 indicate that the initial cycle stage did not contribute to day 3 receptivity, although synchronization within several groups/cages was noted, and that the greatest numbers of estrous animals were obtained on days 6 and 7. Experiment 2 revealed that exposure to live males and lactating females both significantly improved receptivity compared with the PBS, male soiled bedding, and pregnant female groups. These results indicate that current strategies used for routine synchronization could be further improved through alternative housing regimens without compromising animal welfare.

Unnecessary and excessive fluid therapy increases the risk of adverse effects such as pulmonary edema. To prevent this, a mini-fluid challenge (MFC) has been utilized to predict whether fluid therapy will improve circulatory dynamics in human intensive care medicine. The study described here investigated whether MFC is also efficacious in pigs. Thirty-two domestic pigs anesthetized and maintained under mechanical ventilation were treated with successive IV fluid administrations of 2, 1, 1, and 2 mL/kg over a 10-min period for a total dose of 6 mL/kg of Ringer lactate. The percentage increase in mean arterial pressure (MAP) at 2, 3, and 4 mL/kg of cumulative fluid administration was examined to determine whether responders could be identified that would benefit hemodynamically from higher doses of fluids. For the purposes of this study, a 10% increase or more in MAP after 6 mL/kg of fluid administration defined responders, and an increase of less than 10% in MAP was used to define nonresponders. The percentage increase in MAP at 2, 3, and 4 mL/kg fluid administration was evaluated to determine whether this could predict responder status. Eleven of the 32 animals were determined to be responders. Responder status was predicted with high accuracy by the administration of 3 mL/kg (AUC = 0.98) and was moderately predicted with administration of 2 mL/kg (AUC = 0.80), as well as pulse pressure variation (AUC = 0.75). Thus, MFC may be helpful to maintain tissue perfusion in pigs through the use of managed fluid therapy.