Improved ex vivo method for microbiocidal activity across vertebrate species

The field of ecoimmunology is currently undergoing rapid expansion, whereby biologists from a wide range of ecological disciplines are increasingly interested in assessing immunocompetence in their study organisms. One of the key challenges to researchers is determining what eco-immune measures to use in a given experiment. Moreover, there are limitations depending on study species, requirements for specific antibodies, and relevance of the methodology to the study organism. Here we introduce an improved ex vivo method for microbiocidal activity across vertebrate species. The utility of this assay is that it determines the ability of an organism to remove a pathogen that could be encountered in the wild, lending ecological relevancy to the technique. The applications of this microbiocidal assay are broad, as it is readily adaptable to different types of microbes as well as a wide variety of study species. We describe a method of microbiocidal analysis that will enable researchers across disciplines to effectively employ this method to accurately quantify microbial killing ability, using readily available microplate absorbance readers

Sexual Coloration and Performance Capacity in Male Side-blotched Lizards (Uta stansburiana)

In many species of lizards, males fight over territories that allow them access to females, and during these interactions males will display sexual signals to competing individuals. Social signals such as coloration, are seemingly favored by sexual selection as they are thought to provide information about individual quality, such as performance capacity. Side-blotched Lizards, Uta stansburiana, exhibit throat color polymorphism (i.e. orange, yellow, and blue badges), which signal alternative life history strategies to conspecifics. Although morphological, behavioral and physiological characteristics have been shown to differ across male morphs, color variation within each morph has not been compared to aspects of individual quality. In wildcaught adult male U. stansburiana, we tested for associations between coloration (colorimetrics and size of throat badge and side-blotch), a range of morphological traits (snout–vent length, mass, limb and head measures), and performance capacity (sprint speed). Across morphs, sprint speed is positively correlated with aspects of coloration including throat badge and side-blotch size. Sprint speed is also positively correlated with mass, but not snout-vent length and hind limb length. Within morphs, blue hue, yellow brightness, and orange saturation are positively correlated with sprint speed, while other colorimetric relationships are not significant. Our results reveal sexual coloration in male U. stansburiana to be indicative of performance capacity, which may be used as competitive signals under sexual selection

Variations, validations, degradations, and noninvasive determination of pregnancy using fecal steroid metabolites in free-ranging pronghorn

Pregnancy status is a key parameter used to assess reproductive performance of a species as it represents a starting point for measuring vital rates. Vital rates allow managers to determine trends in populations such as neonate survival and recruitment; two important factors in ungulate population growth rates. Techniques to determine pregnancy have generally involved capture and restraint of the animal to obtain blood samples for determining serum hormone levels. Non-invasive pregnancy assessment, via feces, eliminates any hazards between handler and animal, as well as removes handling-induced physiological biases. Using noninvasive fecal sampling, we conducted hormone validations, investigated pregnancy rates, and determined hormone degradation rates across five subpopulations of pronghorn (Antilocapra americana) in Idaho. Samples were collected during April-May of 2018 and 2019 from adult pronghorn of known sex and age class. Metabolites of testosterone, cortisol, 17β-estradiol, and progesterone were measured in fecal samples, and concentrations of estradiol and progesterone were examined for pregnancy determination. Average fecal progesterone metabolite (FPM) levels of pregnant females were more than double compared to levels of nonpregnant females. Fecal estrogen metabolite (FEM) levels did not differ during concurrent sampling. The largest difference in FPM levels between pregnant and nonpregnant females began on 28 April. Pregnancy determination sampling showed average FPM levels for all five subpopulations were significantly different than the nonpregnant female validation group. Nonetheless, pregnancy rates for some subpopulations lacked conclusive estimates due to early fecal sampling. Fecal glucocorticoid metabolites (FGM) levels significantly differed between pregnant females and male pronghorn, but did not differ from nonpregnant females. Degradation rates of FPM and FGM differed across days, with values for FPM from Day 1 being significantly different from all subsequent days, and after Day 9 for FGM, demonstrating the requirement of fresh samples to accurately measure hormone concentrations. We concluded that a noninvasive method to diagnosis pregnancy is possible in pronghorn via progesterone metabolites if fresh samples are collected during late gestation

Demographic Assessment of the Triploid Parthenogenetic Lizard Aspidoscelis Neotesselatus at the Northern Edge of its Range

Aspidoscelis neotesselatus (Colorado Checkered Whiptail) is a hybrid-derived triploid parthenogenetic lizard with a natural range overlapping with six counties in southeastern Colorado, USA. It has also become established by anthropogenic causation in Grant County, Washington State, approximately 1,600 km northwest of its range in Colorado. Large parts of its natural range are within military reservations. Reduced genetic variation in all-female species makes them especially susceptible to environmental disturbances, such as military activities. At Fort Carson (FC), we estimated an abundance index via a catch-per-unit estimator, weekly survival using Cormack-Jolly-Seber models, and body condition and clutch size as indicators of population health across three low-impact training areas (TA; 45, 48, and 55). Abundance estimates varied across TAs from a low of 0.99 to a high of 6.12 females per hectare. Body condition only marginally varied by age class and TAs. Apparent monthly survival was relatively low in all areas and even lower at TA 55 than at TA 48 (0.638 versus 0.771); however, the uncertainty around those estimates was large. Results suggest that TA 48 supported a large fraction of reproductive females that were successful in producing eggs, providing further insight into where monitoring and conservation efforts should be concentrated within FC

Interacting stressors and the potential for adaptation in a changing world: responses of populations and individuals

To accurately predict the impact of environmental change, it is necessary to assay effects of key interacting stressors on vulnerable organisms, and the potential resiliency of their populations. Yet, for the most part, these critical data are missing. We examined the effects of two common abiotic stressors predicted to interact with climate change, salinity and temperature, on the embryonic survival and development of a model freshwater vertebrate, the rough-skinned newt (Taricha granulosa) from different populations. We found that salinity and temperature significantly interacted to affect newt embryonic survival and development, with the negative effects of salinity most pronounced at temperature extremes. We also found significant variation among, and especially within, populations, with different females varying in the performance of their eggs at different salinity–temperature combinations, possibly providing the raw material for future natural selection. Our results highlight the complex nature of predicting responses to climate change in space and time, and provide critical data towards that aim

Sickness Behaviors Across Vertebrate Taxa: Proximate and Ultimate Mechanisms

There is nothing like a pandemic to get the world thinking about how infectious diseases affect individual behavior. In this respect, sick animals can behave in ways that are dramatically different from healthy animals: altered social interactions and changes to patterns of eating and drinking are all hallmarks of sickness. As a result, behavioral changes associated with inflammatory responses (i.e. sickness behaviors) have important implications for disease spread by affecting contacts with others and with common resources, including water and/or sleeping sites. In this Review, we summarize the behavioral modifications, including changes to thermoregulatory behaviors, known to occur in vertebrates during infection, with an emphasis on non-mammalian taxa, which have historically received less attention. We then outline and discuss our current understanding of the changes in physiology associated with the production of these behaviors and highlight areas where more research is needed, including an exploration of individual and sex differences in the acute phase response and a greater understanding of the ecophysiological implications of sickness behaviors for disease at the population level

Stoichiometric and Stable Isotope Ratios of Wild Lizards in an Urban Landscape Vary with Reproduction, Physiology, Space and Time

Spatial and temporal variation in stoichiometric and stable isotope ratios of animals contains ecological information that we are just beginning to understand. In both field and lab studies, stoichiometric or isotopic ratios are related to physiological mechanisms underlying nutrition or stress. Conservation and ecosystem ecology may be informed by isotopic data that can be rapidly and non-lethally collected from wild animals, especially where human activity leaves an isotopic signature (e.g. via introduction of chemical fertilizers, ornamental or other non-native plants or organic detritus). We examined spatial and temporal variation in stoichiometric and stable isotope ratios of the toes of Uta stansburiana (side-blotched lizards) living in urban and rural areas in and around St. George, Utah. We found substantial spatial and temporal variation as well as context-dependent co-variation with reproductive physiological parameters, although certain key predictions such as the relationship between δ15N and body condition were not supported. We suggest that landscape change through urbanization can have profound effects on wild animal physiology and that stoichiometric and stable isotope ratios can provide unique insights into the mechanisms underlying these processes