Void spot assay procedural optimization and software for rapid and objective quantification of rodent voiding function, including overlapping urine spots

Mouse urinary behavior is quantifiable and is used to pinpoint mechanisms of voiding dysfunction and evaluate potential human therapies. Approaches to evaluate mouse urinary function vary widely among laboratories, however, complicating cross-study comparisons. Here, we describe development and multi-institutional validation of a new tool for objective, consistent, and rapid analysis of mouse void spot assay (VSA) data. Void Whizzard is a freely available software plugin for FIJI (a distribution of ImageJ) that facilitates VSA image batch processing and data extraction. We describe its features, demonstrate them by evaluating how specific VSA method parameters influence voiding behavior, and establish Void Whizzard as an expedited method for VSA analysis. This study includes control and obese diabetic mice as models of urinary dysfunction to increase rigor and ensure relevance across distinct voiding patterns. In particular, we show that Void Whizzard is an effective tool for quantifying nonconcentric overlapping void spots, which commonly confound analyses. We also show that mouse genetics are consistently more influential than assay design parameters when it comes to VSA outcomes. None of the following procedural modifications to reduce overlapping spots masked these genetic-related differences: reduction of VSA testing duration, water access during the assay period, placement of a wire mesh cage bottom on top of or elevated over the filter paper, treatment of mesh with a hydrophobic spray, and size of wire mesh opening. The Void Whizzard software and rigorous validation of VSA methodological parameters described here advance the goal of standardizing mouse urinary phenotyping for comprehensive urinary phenome analyses.

A reappraisal of the role of mosquitoes in the transmission of myxomatosis in Britain

SUMMARYField experiments were made in southern England to re-examine the possibility that mosquitoes in Britain might feed on wild rabbits and hence be vectors of myxomatosis. Mosquitoes of several species were attracted to rabbits enclosed hi cylindrical traps and in a trap in which the animal was placed in a wire mesh cage. Substantial numbers of mosquitoes were also caught biting, or attempting to bite, tethered rabbits which were not in cages or traps. Evidence that mosquitoes fed on wild rabbits under natural conditions was obtained from results of precipitin tests made on blood-smears collected from mosquitoes caught resting amongst vegetation. On a few evenings mosquitoes were seen to be attracted to healthy wild rabbits and apparently attempting to feed on them. Batches of two mosquito species collected from the field were infected with myxoma virus.It was concluded that contrary to previous beliefs mosquitoes in Britain feed to a certain extent on wild rabbits, and therefore are potential vectors of myxomatosis. No attempts were made to assess their relative importance in the transmission of the disease, which in Britain is transmitted mainly by the rabbit flea.

Studies on the ecology of Oncopera intricata Walker (Lepidoptera: Hepialidae). 2. Selection of oviposition sites by the female moths.

The results are reported of studies on the effect of length and density of pasture cover on the selection of oviposition sites by the mated female moths of Oncopera intricata Walker. Moths were placed in a wire-mesh cage, 7 ft in diameter and 2 ft high, in which they were given a choice of pasture cover in which to oviposit. It was found that the length of the grass cover was important in causing settling by the moths since when only short cover was provided, few moths laid their eggs in the plots irrespective of the density of the cover provided. When long cover was provided in some of the plots the preferred sites were where the cover was long and dense but a reasonable number of moths oviposited in short sparse cover. Where long cover alone was provided the dense plots were preferred to the sparse. The implications of these results on the pattern of distribution and survival in natural populations is discussed.