Effect of Housing Condition and Diet on the Gut Microbiota of Weanling Immunocompromised Mice.

Gastrointestinal microbiota are affected by a wide variety of extrinsic and intrinsic factors. In the husbandry of laboratory mice and design of experiments, controlling these factors where possible provides more reproducible results. However, the microbiome is dynamic, particularly in the weeks immediately after weaning. In this study, we characterized the baseline gastrointestinal microbiota of immunocompromised mice housed under standard conditions for our facility for 6 weeks after weaning, with housing either in an isolator or in individually ventilated cages and a common antibiotic diet (trimethoprim sulfamethoxazole). We compared these conditions to a group fed a standard diet and a group that was weaned to a standard diet then switched to antibiotic diet after 2 weeks. We found no clear effect of diet on richness and α diversity of the gastrointestinal microbiota. However, diet did affect which taxa were enriched at the end of the experiment. The change to antibiotic diet during the experiment did not convert the gastrointestinal microbiome to a state similar to mice consistently fed antibiotic diet, which may highlight the importance of the initial post-weaning period in the establishment of the gastrointestinal microbiome. We also observed a strong effect of housing type (isolator compared with individually ventilated cage) on the richness, α diversity, ß diversity, and taxa enriched over the course of the experiment. Investigating whether the diet or microbiome affects a certain strain's phenotype is warranted in some cases. However, our findings do not suggest that maintaining immunocompromised mice on antibiotic feed has a clinical benefit when potential pathogens are operationally excluded, nor does it result in a more consistent or controlled microbiome in the post-weaning period.

Effect of 3 Euthanasia Methods on Serum Yield and Serum Cortisol Concentration in Zebrafish (Danio rerio).

Zebrafish are an important model in neuroscience and developmental biology and are also an emerging model in hematology and immunology. Little information is available for zebrafish regarding the physiologic impact of different euthanasia methods and whether a chosen method of euthanasia can impact serum yield. These parameters could impact the choice of euthanasia method for a study. To that end, the current study compared 3 methods of adult zebrafish euthanasia and their effects on 3 distinct criteria; time to loss of opercular movement, volume of serum obtained, and serum cortisol concentration. Blood was collected using a postmortem tail amputation and centrifugation blood collection technique. Time to loss of opercular movement differed significantly among euthanasia methods, with animals undergoing rapid chilling displaying the shortest time (mean Rapid Chilling: 40 s; Benzocaine: 86 s; MS222: 96 s). All methods of euthanasia resulted in a comparable average serum yield (Rapid Chilling = 7.5 µL; Benzocaine = 8.5 µL; MS222 = 7.5 µL per fish). None of the euthanasia methods tested resulted in average cortisol concentrations above the reported physiologic range. Although no significant differences were observed in serum yield or serum cortisol concentration, rapid chilling remains the preferred method for painless, humane euthanasia.

Safety Evaluation of a 405-nm LED Device for Direct Antimicrobial Treatment of the Murine Brain.

Antimicrobial resistance is a growing problem in human medicine that extends to biomedical research. Compared with chemical-based therapies, light-based therapies present an alternative to traditional pharmaceuticals and are less vulnerable to acquired bacterial resistance. Due to immunologic privilege and relative tissue sensitivity to topical antibiotics, the brain poses a unique set of difficulties with regard to antimicrobial therapy. This study focused on 405-nm 'true violet' light-which has been shown to kill multiple clinically relevant bacterial species in vitro yet leave mammalian cells unscathed-and its effect on the murine brain. We built a 405-nm LED array, validated its power and efficacy against a clinical bacterial isolate in vitro, and then, at the time of craniotomy, treated mice with various doses of 405-nm light (36, 45, and 54 J/cm²). The selected doses caused no behavioral derangements postoperatively or any observable brain pathology as determined postmortem by histologic evaluation and immunofluorescence staining for caspase 3 and glial fibrillary acidic protein, markers of apoptosis and necrosis. True-violet light devices may present an inexpensive refinement to current practices for maintaining open craniotomy sites or reducing bacterial loads in contaminated surgical sites.