HIV-infected people develop reproducible disruptions in their gastrointestinal microbiota. Despite the suppression of HIV viremia via long-term antiretroviral therapy (ART), alterations still occur in gut microbial diversity and the commensal microbiota. Mounting evidence suggests these microbial changes lead to the development of gut dysbiosis—persistent inflammation that damages the gut mucosa—and correlate with various immune defects. In this study, we examined how early ART intervention influences microbial diversity in SIV-infected rhesus macaques. Using 16S rRNA sequencing, we defined the fecal microbiome in macaques given daily ART beginning on either 3 or 7 d after SIV infection (dpi) and characterized changes in composition, α diversity, and β diversity from before infection through 112 dpi. The dominant phyla in the fecal samples before infection were Bacteroidetes, Firmicutes, Spirochaetes, and Proteobacteria. After SIV infection and ART, the relative abundance of Firmicutes and Bacteroidetes did not change significantly. Significant reductions in α diversity occurred across time when ART was initiated at 3 dpi but not at 7 dpi. Principal coordinate analysis of samples revealed a divergence in β diversity in both treatment groups after SIV infection, with significant differences depending on the timing of ART administration. These results indicate that although administration of ART at 3 or 7 dpi did not substantially alter fecal microbial composition, the timing of early ART measurably altered phylogenetic diversity.

Currently, metabolic cages (MC) are the only way to achieve serial sampling of urine and feces in rodents. However, the use of this caging creates a dramatic change from an animal's usual microenvironment. Here we sought to examine the effect of MC on physiologic parameters that are stress-responsive in rats. We surgically implanted 8 male Wistar rats (weight, 150 to 175 g) with telemetric transmitters and allowed them to recover for at least 2 wk. At the beginning of the study, the rats were moved to conventional open-top cages, and telemetry recording was initiated. After 24 h, the rats were moved to MC or to another conventional cage and the recording continued for another 24 h. Finally, the rats were returned to their home cages, and telemetry recording was performed for a final 24 h. After 10 days, this process was then repeated, with MC and conventional assignments switched. During the 78-h monitoring period, we recorded heart rate, arterial blood pressure, locomotor activity, body weight, and food and water consumption. Heart rate and arterial blood pressure showed transient but significant changes. Locomotor activity during the dark phase was greatly decreased in MC compared with conventional cages, perhaps due to space constraints. In addition, when the rats were housed in MC, they showed a small but significant weight loss. Food consumption did not differ between housing environments, but water consumption was lower when rats were in MC. In conclusion, the housing of rats in MC for 24 h can elicit mild and reversible cardiovascular changes. This finding is consistent with European Directive 2010/63/EU, which considers short-term (less than 24 h) restraint in MC a procedure of mild severity.

Osteosarcoma (OSA) is the most common primary bone tumor in both dogs and humans. The dog is an important research model for OSA, yet dogs have much higher prevalence of bone tumors than do humans, a disparity that has yet to be explained. Neoplastic transformation of cells within or adjacent to bone infarcts into primary bone tumors has been described in humans but only sparsely characterized in the veterinary literature. In this study, 653 cases of canine bone infarcts were received through a referral veterinary osteopathology service over a 14-y period. We identified an idiopathic disorder affecting the nutrient artery, termed canine idiopathic arteriopathy (CIA), which to our knowledge has no direct counterpart in human medicine. This disorder was documented alongside ischemic necrosis of the medullary cavity in 114 bone infarcts in 108 dogs. We hypothesize that CIA precipitated an ischemic environment, resulting in development of a bone infarct down- stream of the abnormal artery. In 52% (59 of 114) of cases, bone infarcts demonstrated evidence of repair (termed reparative bone infarcts [RBI]), while in 48% (55 of 114) of infarcts, a bone tumor was also present, including pleomorphic sarcoma, OSA, fibrosarcoma, and chondrosarcoma. In some cases, a spectrum of tumors was present. We hypothesize that the ischemic infarct environment provoked bone marrow mesenchymal stem cells (MSCs) to attempt repair of the stroma, and in approximately half of cases, MSCs underwent neoplastic transformation (BINT) to produce tumors. The most common sites of bone infarcts were the distal femur, distal radius, proximal humerus, and distal tibia, coinciding with common sites of canine OSA. The authors propose that CIA leading to bone infarcts and infarct-derived tumors, in combination with possible underdiagnosis of canine bone infarcts and misdiagnosis of some RBI as neoplasia, may contribute to the higher reported proportion of bone tumors in dogs compared with humans.

Eliminating unnecessary pain is an important requirement of performing animal experimentation, including reducing and controlling pain of animals used in pain research. The goal of this study was to refine an adjuvant-induced monoarthritis model in rats by providing analgesia with a transdermal fentanyl solution (TFS). Male and female Sprague–Dawley rats, single- or pair-housed, were injected with 20 μL of complete Freund adjuvant (CFA) into the left ankle joint. CFA-injected rats treated with a single dose of transdermal fentanyl solution (0.33 or 1 mg/kg) were compared with an untreated CFA-injected group and sham groups that received either no treatment or TFS treatment (1 mg/kg) during 72 h. At the tested doses, TFS reduced mechanical hyperalgesia and improved the mobility, stance, rearing, and lameness scores at 6 h after CFA injection. Joint circumferences were not reduced by TFS treatment, and no significant differences were detected between the 2 doses of TFS, or between single- and pair-housed rats. Treatment with TFS did not appear to interfere with model development and characteristics. However, overall, the analgesic effect was transient, and several opioid-related side effects were observed.

Multiple animal models have been developed to investigate the pathogenesis of colorectal cancer and to evaluate potential treatments. One model system uses azoxymethane, a metabolite of cycasin, alone and in conjunction with dextran sodium sulfate to induce colon cancer in rodents. Azoxymethane is metabolized by hepatic P450 enzymes and can also be eliminated through the kidneys. In this study, C57BL/6J mice were fed either standard or high-fat diet and then all mice received azoxymethane at 10 mg/kg body weight twice a week for 6 wk. Shortly after the end of treatment, high mortality occurred in mice in the high-fat diet group. Postmortem examination revealed hepatic and renal pathology in mice on both diets. Histologic changes in liver included hepatocytomegaly with nuclear pleomorphism and bile duct hyperplasia accompanied by mixed inflammatory-cell infiltrates. Changes in the kidneys ranged from basophilia of tubular epithelium to tubular atrophy. The results indicate that further optimization of this model is needed when feeding a high-fat diet and giving multiple azoxymethane doses to induce colon cancer in C57BL/6J mice.

Small-animal irradiators are widely used in oncologic research, and many experiments use mice to mimic radiation treatments in humans. To improve fractionated high-precision irradiation in mice with orthotopic pancreatic tumors, we evaluated 3 positioning methods: no positioning aid, skin marker, and immobilization devices (immobilization masks). We retrospectively evaluated the translation vector needed for optimal tumor alignment (by shifting the mouse in left-right, in cranio-caudal, and in anterior-posterior direction) on cone-beam CT from our small-animal radiotherapy system. Of the 3 methods, the skin marker method yielded the smallest mean translation vector (3.8 mm) and was the most precise method overall for most of the mice. In addition, the skin marker method required supplemental rotation (that is, roll, pitch, and yaw) for optimal tumor alignment only half as often as positioning without a positioning aid. Finally, the skin marker method had the highest scores for the quality of the fusion results. Overall, we preferred the skin marker method over the other 2 positioning methods with regard to optimal treatment planning and radiotherapy in an orthotopic mouse model of pancreatic cancer.

The salt calcium chloride (CaCl₂) is widely used in industry as a food additive; levels for human consumption are regulated by international or governmental agencies. Generally, the food industry relies on toxicity studies conducted in mammals such as mice, rats, and rabbits for determining food safety. However, testing in mammals is time-consuming and expensive. Zebrafish have been used in a range of toxicological analyses and offer advantages with regard to sensitivity, time, and cost. However, information in not available with regard to whether the sensitivity of zebrafish to CaCl₂ is comparable to the concentrations of CaCl₂ used as food additives. The aim of this study was to compare the CaCl₂ tolerance of zebrafish embryos and larvae with concentrations currently approved as food additives. Acute toxicity, embryotoxicity, cardiotoxicity, and neurotoxicity assays were used to determine the threshold toxic concentration of CaCl₂ in zebrafish embryos and larvae. The data showed that doses above 0.4% had toxic effects on development and on the activity of the cardiac and neuronal systems. Furthermore, all embryos exposed to 0.8 and 1.6% of CaCl₂ died after 24 hpf. These findings are consistent with the limits of CaCl₂ concentrations approved by Codex Alimentarius. Therefore, zebrafish embryos could be suitable for screening food additives.