Venipuncture Site Influences Blood-drop Volume in C57BL/6 Mice.

Many experiments require the collection of serial blood samples from mice. However, the size of mice limits the volume of blood that can be safely collected as a survival procedure. In IACUC protocols, investigators may report the amount of blood they collect from mice as a number of drops. Many institutions, including ours, use an anecdotal conversion factor (1drop of mouse blood = 25µL) to ensure that blood-collection volumes are compliant with institutional guidelines. To our knowledge, previous work has not experimentally determined the volume of a drop of mouse blood. In this 10-wk crossover experiment, 2 phlebotomists bled 30 C57BL/6J mice from 3 sites (facial, saphenous, and tail) using one or 2 different needle gauge sizes per site. Male and female mice were weighed weekly and divided among 5 groups (n = 6): left and right tail vein, left and right saphenous vein, and facial vein. A single blood drop from each site was weighed, and the volume of each drop was calculated using the average blood density determined from 8 mice terminally bled at the end of the study. Venipuncture site and side significantly influenced blood-drop weight and thus calculated volume. Facial vein puncture produced the largest drop volume (mean: 21.7µL), followed by the saphenous vein (mean: 9.97µL) and tail vein (mean: 4.96µL). Collection from the facial vein was associated with more hemorrhage and morbidity. Left-sided venipuncture was associated with slightly larger-volume blood drops, though the effect size of side was small. The results of this study may be useful in more accurately estimating blood loss via conversion of drops to volume. Our data indicate that blood collection from saphenous and tail veins minimizes blood loss relative to facial vein puncture and may optimize both serial collection of small-volume blood samples and animal welfare.

Assessment of Oral Albendazole and Fumagillin in the Treatment of Pseudoloma neurophilia in Adult Zebrafish.

Pseudoloma neurophilia ( Pn ), the causative agent of the most commonly reported disease of zebrafish, is a microsporidian parasite that confounds research by inducing behavioral and physiologic changes in zebrafish. Although a treatment for P. neurophilia has not been documented in zebrafish, albendazole (ALB) and fumagillin (FUM) have been used to treat microsporidian infections of other fish species. To investigate the efficacy of oral ALB and FUM in the treatment of Pn, we performed a pilot study that demonstrated the safety and palatability of novel gel-based diets containing FUM or ALB in adult AB zebrafish. In a subsequent study, approximately 250 adult AB zebrafish (previously infected with Pn ) were treated with these medicated diets for 4 wk. At 4 different time points (weeks 0, 5, 10, and 16 of the study), fish were euthanized and whole-body qPCR was performed to assess Pn prevalence across treatment and control groups. There was no statistically significant association between treatment group and Pn prevalence at any time point, although potential biologically relevant reductions in Pn prevalence occurred in the combination therapy group at weeks 5 and 16 and in the ALB group at week 5. Based on high-performance liquid chromatography analyses, the medicated diets contained less ALB and more FUM than expected, highlighting the importance of validating medicated feed concentrations to ensure safety, efficacy, and consistency. While Pn remains challenging to eradicate and control, results of this study warrant further investigation into the utility of ALB and FUM as potential treatments for this pathogen.

Spontaneous Pituitary Neoplasm in Two Female Geriatric Southern Giant Pouched Rats (Cricetomys ansorgei).

Southern giant pouched rats (Cricetomys ansorgei) are a small muroid species native to the sub-Saharan Africa. Their exceptionally developed olfactory system, trainability, and relatively small size makes them useful working animals for various applications in humanitarian work. At our institution, a breeding colony of Southern giant pouched rats is maintained to study their physiology and utility as scent detectors. This case report describes the occurrence of spontaneous pituitary neoplasms with distinct clinical presentations in 2 geriatric (approximately 7.5 y old) wild-caught female Southern giant pouched rats. The first pouched rat displayed vestibular deficits, including left-sided head tilt, ataxia, disorientation, and circling. MRI revealed a large, focal heterogeneous mass arising from the pituitary fossa. The second pouched rat presented with polyuria, polydipsia, and hyperglycemia but no neurologic signs. Examination after euthanasia revealed a prolactin (PRL)-expressing pituitary carcinoma and adenoma in the first and second pouched rat, respectively, associated with mammary hyperplasia in both animals. This is the first report of spontaneous PRL-producing pituitary tumors in Southern giant pouched rats.

Comparison of Effects of Capromorelin and Mirtazapine on Appetite in New Zealand White Rabbits (Oryctolagus cuniculus).

Inappetence is a welfare concern in rabbits (Oryctolagus cuniculus), as it can lead to potentially fatal gastrointestinal stasis. In other species, inappetence is commonly treated with appetite stimulants; however, few published studies have evaluated the efficacy of appetite stimulants in rabbits. We performed 2 studies to evaluate the effects of capromorelin and mirtazapine on appetite in New Zealand White (NZW) rabbits. In the first study, healthy rabbits ( n = 9) were evaluated using a randomized crossover design and 9 treatments: capromorelin 4 mg/kg oral (PO) once a day (SID), capromorelin 8 mg/kg PO SID, saline control PO SID, capromorelin 4 mg/kg PO twice a day (BID), capromorelin 8 mg/kg PO BID, saline control PO BID, mirtazapine 0.5 mg/kg transdermal (TD) SID, mirtazapine 1 mg/kg TD SID, and saline control TD SID for 3 d with a 1-wk washout period between treatments. Treatment efficacy was assessed by measuring daily feed intake and fecal output and by weighing rabbits twice a week. Overall, feed intake and fecal output were higher for all treatments as compared with controls, except for fecal output in the capromorelin 4 mg/kg and 8 mg/kg PO SID groups. Feed intake and fecal output were significantly higher with mirtazapine as compared with capromorelin. Body weight and erythema/petechia of the pinnae were greater in the mirtazapine 1 mg/kg TD SID group than in the control group. A second study evaluated rabbits that had undergone surgery (castration, n = 7) and then received one of 3 treatments: capromorelin 8 mg/kg PO BID, mirtazapine 1 mg/kg TD SID, or saline PO BID for 3 d postoperatively. Feed intake and fecal output in the postoperative mirtazapine group were not significantly different from those of the capromorelin and control groups. Due to its superior efficacy as compared with capromorelin in healthy NZW rabbits, we recommend considering mirtazapine as a treatment for inappetence in NZW rabbits.

Hybridization is limited between two lineages of freeze-resistant Trichinella during coinfection in a mouse model.

Hybridization between two closely related but distinct genetic lineages may lead to homogenization of the two lineages with potentially novel phenotypes, or selective pressure to avoid hybridization if the two lineages are truly distinct. Trichinella nativa and Trichinella T6 are zoonotic nematode parasites which can be distinguished genetically despite occasional hybridization. Here, using an experimental murine model, we attempt to determine whether there are barriers to hybridization when sizeable numbers of each lineage are allowed to coinfect a host. Two mice were independently infected with equal numbers of T. nativa and T6. The offspring of these coinfections were genotyped at two microsatellite loci and one mitochondrial locus capable of distinguishing T. nativa from T6 genotypes. Among larvae in the F1 generation, offspring of every possible mating were encountered. Most larvae (63.6%) derived from T. nativa×T. nativa matings, while 21.1% of offspring were the product of T6×T6 matings, and only 15.3% were hybrid offspring of T. nativa×T6 crosses, differing markedly from null expectations. In this experimental model, T. nativa and Trichinella T6 were able to mate, but ratios of offspring indicated pre- or post-zygotic barriers to hybridization that may include assortative mating, genetic incompatibilities, and/or differences in the fitness of offspring. These barriers would limit gene flow between these two lineages in a natural setting, serving as a barrier to their homogenization and promoting their persistence as distinct and separate entities.