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Introduction: Laboratory rat welfare is critically influenced by laboratory animal personnel through their implementation, or lack of implementation, of various enrichment techniques. One such promising technique is heterospecific play, or “rat tickling”, which mimics aspects of rat rough-and-tumble play and can contribute to improving welfare, but may be infrequently implemented. The theory of planned behavior can be used to study implementation by measuring intentions and beliefs about rat tickling, including behavioral attitudes (whether it is good or bad), subjective norms (whether there is social/professional pressure to provide it), and control beliefs (whether they feel in control of providing it). Therefore, the objective of this study was to identify current rat tickling prevalence and predictors among laboratory animal personnel in the United States and Canada. Our hypothesis was that rat tickling prevalence would be low and associated with beliefs about the practice, enrichment, and laboratory animals in general. Methods: Laboratory animal personnel were recruited from widespread online promotion. A total of 794 personnel (mean = 40±11 years, 80% white, 80% female) completed at least 50% of the mixed methods online survey and met inclusion criteria of currently working with laboratory rats in the USA or Canada. The survey included questions about demographics, enrichment practices and beliefs, attitudes towards rats, general positive behaviors (e.g. talking to laboratory animals), and both practices and beliefs about rat tickling. Qualitative data were coded using thematic analysis. Quantitative data were analyzed using general linear models. Results: Laboratory personnel reported low levels of rat tickling implementation, with 89% of participants reporting using it never or rarely. Laboratory personnel reported 2 key benefits (handling: 61%, welfare: 55%) and 3 key barriers (time: 59%, personnel: 22%, and research: 22%) to rat tickling using qualitative analysis. Current and planned rat tickling were positively associated with more positive beliefs (social/professional pressure p<0.0001, control of providing tickling p<0.0001) and familiarity with tickling (p<0.0001). Current rat tickling was also positively associated with more positive general behaviors towards laboratory animals, such as naming animals (p<0.0001). Future rat tickling was positively associated with more positive attitudes about it (p<0.0001) and a desire to implement more enrichment (p<0.01). Conclusion: Our findings show that even though rat tickling implementation is currently low, it is positively associated with personnel beliefs, familiarity, general attitudes, and a desire for more enrichment. That is, laboratory animal personnel were more likely to provide rat tickling if they were more familiar with it, thought providing it was both good and under their control, and felt subject to social/professional pressure, as well as if they wanted to provide more enrichment and generally had more positive behaviors towards laboratory animals. There is potential to increase rat tickling by increasing personnel familiarity with the procedure through training, decreasing the time required, and changing personnel beliefs–thereby improving rat welfare. [ABSTRACT FROM AUTHOR]

Social learning has important ecological and evolutionary consequences but the role of certain factors, such as social rank, neophobia (i.e., avoidance of novel stimuli), persistence, and task-reward association, remain less understood. We examined the role of these factors in social learning by captive coyotes (Canis latrans) via three studies. Study 1 involved individual animals and eliminated object neophobia by familiarizing the subjects to the testing apparatus prior to testing. Studies 2 and 3 used mated pairs to assess social rank, and included object neophobia, but differed in that study 3 decoupled the food reward from the testing apparatus (i.e., altered task-reward association). For all three studies, we compared performance between coyotes that received a demonstration from a conspecific to control animals with no demonstration prior to testing. Coyotes displayed social learning during study 1; coyotes with a demonstrator were faster and more successful at solving the puzzle box but did not necessarily use the same modality as that observed to be successful. In study 2, there was no difference in success between treatment groups but this is likely because only one coyote within each pair was successful so successful coyote results were masked by their unsuccessful mate. In study 3, there was no difference in success between treatment groups; only two coyotes, both dominant, hand-reared males with demonstrators were able to perform the task. However, coyotes with a demonstrator were less neophobic, measured as latency to approach the object, and more persistent, measured as time spent working on the apparatus. Social rank was the best predictor of neophobia and persistence and was also retained in the best model for time to eat inside the apparatus, a post-trial measurement of object neophobia. These results suggest coyotes are capable of social learning for novel tasks but social rank, neophobia, and persistence influence their social-learning capabilities. This study contributes to understanding the mechanisms underlying how animals gain information about their environment. [ABSTRACT FROM AUTHOR]

An appreciation of body size allometry is central for understanding insect pollination ecology. A recent model utilises allometric coefficients for five of the seven extant bee families (Apoidea: Anthophila) to include crucial but difficult-to-measure traits, such as proboscis length, in ecological and evolutionary studies. Melittidae were not included although they are important pollinators in South Africa where they comprise an especially rich and morphologically diverse fauna. We measured intertegular distance (correlated with body size) and proboscis length of 179 specimens of 11 species from three genera of Melittidae. With the inclusion of Melittidae, we tested the between family differences in the allometric scaling coefficients. AIC model selection was used to establish which factors provide the best estimate of proboscis length. We explored a hypothesis that has been proposed in the literature, but which has not been tested, whereby body and range sizes of bees are correlated with rainfall regions. We tested this by using body size measurements of 2109 museum specimens from 56 species of Melittidae and applied the model coefficients to estimate proboscis length and foraging distance. Our results from testing differences across bee families show that with the addition of Melittidae, we retained the overall pattern of significant differences in the scaling coefficient among Apoidea, with our model explaining 98% of the variance in species-level means for proboscis length. When testing the relationship between body size and rainfall region we found no relationship for South African Melittidae. Overall, this study has added allometric scaling coefficients for an important bee family and shown the applicability of using these coefficients when linked with museum specimens to test ecological hypothesis. [ABSTRACT FROM AUTHOR]

Cryptic species are a challenge for systematics, but their elucidation also may leave critical information gaps about the distribution, conservation status, and behavior of affected species. We use the leopard frogs of the eastern U.S. as a case study of this issue. We refined the known range of the recently described Rana kauffeldi, the Atlantic Coast Leopard Frog, relative to the region’s two other leopard frog species, conducted assessments of conservation status, and improved methods for separating the three species using morphological field characters. We conducted over 2,000 call and visual surveys and took photographs of and tissue samples from hundreds of frogs. Genetic analysis supported a three-species taxonomy and provided determinations for 220 individual photographed frogs. Rana kauffeldi was confirmed in eight U.S. states, from North Carolina to southern Connecticut, hewing closely to the Atlantic Coastal Plain. It can be reliably differentiated in life from R. pipiens, and from R. sphenocephala 90% of the time, based on such characters as the femoral reticulum patterning, dorsal spot size and number, and presence of a snout spot. However, the only diagnostic character separating R. kauffeldi from R. sphenocephala remains the breeding call described in 2014. Based on our field study, museum specimens, and prior survey data, we suggest that R. kauffeldi has declined substantially in the northern part of its range, but is more secure in the core of its range. We also report, for the first time, apparent extirpations of R. pipiens from the southeastern portion of its range, previously overlooked because of confusion with R. kauffeldi. We conclude with a generalized ecological research agenda for cryptic species. For R. kauffeldi, needs include descriptions of earlier life stages, studies of niche partitioning with sympatric congeners and the potential for hybridization, and identification of conservation actions to prevent further declines. [ABSTRACT FROM AUTHOR]

Indo-Pacific red lionfish (Pterois volitans) have invaded the western Atlantic, and most recently the northern Gulf of Mexico (nGOM), at a rapid pace. Given their generalist habitat affinities and diet, and strong ecological overlap with members of the commercially valuable snapper-grouper complex, increased density and abundance of lionfish could result in significant competitive interactions with nGOM commercially important species. We experimentally investigated the intensity of behavioral interactions between lionfish and indigenous, abundant and economically important juvenile nGOM red snapper (Lutjanus campechanus), and other increasingly abundant juvenile tropical snapper species (gray snapper—L. griseus and lane snapper—L. synagris) in large outdoor mesocosms to examine snapper vulnerabilities to lionfish competition. When paired with lionfish, red snapper swimming activity (i.e., time swimming and roving around experimental tank or at structure habitat during experiments) was significantly lower than in intraspecific control trials, but gray and lane snapper swimming activities in the presence of lionfish did not significantly differ from their intraspecific controls. Additionally in paired trials, red and lane snapper swimming activities were significantly lower than those of lionfish, while no significant difference in swimming activities was observed between lionfish and gray snapper. We found that red snapper prey consumption rates in the presence of lionfish were significantly lower than in their intraspecific 3-individual control trials, but when paired together no significant differences in prey consumption rates between red snapper and lionfish were observed. When paired with lane or gray snapper, lionfish were observed having comparatively higher prey consumption than snappers, or as observed in lionfish intraspecific 1-individual controls. However, lane and gray snapper consumption rates in the presence of lionfish did not significantly differ from those in intraspecific controls. These findings suggest that competition between juvenile snappers and invasive lionfish may be variable, with lionfish exhibiting differing degrees of competitive dominance and snappers exhibiting partial competitive vulnerability and resistance to lionfish. While the degree of intensity at which these interactions may occur in nGOM reefs may differ from those observed in our findings, this study enables greater understanding of the potential ecological effects of red lionfish on native reef fishes. [ABSTRACT FROM AUTHOR]

In Chesapeake Bay, the soft-shell clam Mya arenaria (thin-shelled, deep-burrowing) exhibits population declines when predators are active, and it persists at low densities. In contrast, the hard clam Mercenaria mercenaria (thick-shelled, shallow-burrowing) has a stable population and age distribution. We examined the potential for habitat and predators to control densities and distributions of bivalves in a field caging experiment (Mya only) and laboratory mesocosm experiments (both species). In the field, clams exposed to predators experienced 76.3% greater mortality as compared to caged individuals, and blue crabs were likely responsible for most of the mortality of juvenile Mya. In mesocosm experiments, Mya had lower survival in sand and seagrass than in shell hash or oyster shell habitats. However, crabs often missed one or more prey items in seagrass, shell, and oyster shell habitats. Predator search times and encounter rates declined when prey were at low densities, likely due to the added cost of inefficient foraging; however, this effect was more pronounced for Mya than for Mercenaria. Mercenaria had higher survival than Mya in mesocosm experiments, likely because predators feeding on Mercenaria spent less time foraging than those feeding on Mya. Mya may retain a low-density refuge from predation even with the loss of structurally complex habitats, though a loss of habitat refuge may result in clam densities that are not sustainable. A better understanding of density-dependent predator-prey interactions is necessary to prevent loss of food-web integrity and to conserve marine resources. [ABSTRACT FROM AUTHOR]

The ability to recognize individuals and track growth over time is crucial to population dynamics research as well as studies of animal behavior. Invertebrates are particularly difficult to track as they often molt, have regenerative capabilities, or lack hard parts to attach markers. We tested, in laboratory and field studies, a new way of marking sea anemones (order Actiniaria) by injection of three vital stains (i.e., neutral red, methylene blue, and fluorescein). Neutral red and methylene blue did not affect growth or survival, but fluorescein was lethal at high concentrations. Marked individuals could be identified up to seven months after injection with neutral red, six weeks with methylene blue, and three days with low concentrations of fluorescein. Neutral red could be used for long-term monitoring of growth and survival in the field, and in combination with methylene blue could be used to mark individuals in distinguishable patterns for short-term studies such as examining predator-prey interactions, movement of individuals, and recruitment survival. [ABSTRACT FROM AUTHOR]

Ecomorphology links microhabitat and morphology. By comparing ecomorphological associations across clades, we can investigate the extent to which evolution can produce similar solutions in response to similar challenges. While Anolis lizards represent a well-studied example of repeated convergent evolution, very few studies have investigated the ecomorphology of geckos. Similar to anoles, gekkonid lizards have independently evolved adhesive toe pads and many species are scansorial. We quantified gecko and anole limb length and microhabitat use, finding that geckos tend to have shorter limbs than anoles. Combining these measurements with microhabitat observations of geckos in Queensland, Australia, we observed geckos using similar microhabitats as reported for anoles, but geckos with relatively longer limbs were using narrower perches, differing from patterns observed in anoles and other lizards. We also observed arboreal geckos with relatively shorter proximal limb segments as compared to rock-dwelling and terrestrial geckos, similar to patterns observed for other lizards. We conclude that although both geckos and anoles have adhesive pads and use similar microhabitats, their locomotor systems likely complement their adhesive pads in unique ways and result in different ecomorphological patterns, reinforcing the idea that species with convergent morphologies still have idiosyncratic characteristics due to their own separate evolutionary histories. [ABSTRACT FROM AUTHOR]

When laboratory dogs are rehomed into private households, they experience an extreme change in their life situation. They leave their familiar, limited environment in the research facility and encounter a multitude of animate and inanimate stimuli in their new home. Although literature reports have described the experiences with rehoming as being mostly positive, scientific observations of the dogs in everyday situations have not been done. Hence, we conducted an observational test with 74 laboratory beagles 6 weeks after adoption in their new homes. This test included standardized tasks and elements; the dogs were observed during specific interactions with their new owners and during a walk. Furthermore, the owners of these 74 and of 71 additional dogs participated in standardized phone interviews 1 and 12 weeks after adoption, during which they answered questions about the dogs’ behavior in everyday situations. In the observational test, the dogs behaved mostly friendly towards humans and dogs, were tolerant during manipulations by the owner and were relaxed during the walk, even in traffic. Eighty percent (of n = 71) of the dogs walked well behaved on the leash without pulling. According to the interviews, the majority of the dogs showed desired, friendly and relaxed behavior, and the survey results reflected the bonding between dog and owner. The analysis of a possible influence of various factors (age, sex, origin, etc.) using mixed regression models confirmed the results from two previous behavior tests and interviews. Specifically, dogs that had been bred in the research facility scored significantly better than dogs that the research facility had purchased from commercial laboratory dog breeders (p = 0.0113). The results of this study demonstrate a successful adaptation of the rehomed beagles to their new life situation. [ABSTRACT FROM AUTHOR]

Most statistical and mechanistic models used to predict mosquito borne disease transmission incorporate climate drivers of disease transmission by utilizing environmental data collected at geographic scales that are potentially coarser than what mosquito populations may actually experience. Temperature and relative humidity can vary greatly between indoor and outdoor environments, and can be influenced strongly by variation in landscape features. In the Aedes albopictus system, we conducted a proof-of-concept study in the vicinity of the University of Georgia to explore the effects of fine-scale microclimate variation on mosquito life history and vectorial capacity (VC). We placed Ae. albopictus larvae in artificial pots distributed across three replicate sites within three different land uses–urban, suburban, and rural, which were characterized by high, intermediate, and low proportions of impervious surfaces. Data loggers were placed into each larval environment and in nearby vegetation to record daily variation in water and ambient temperature and relative humidity. The number of adults emerging from each pot and their body size and sex were recorded daily. We found mosquito microclimate to significantly vary across the season as well as with land use. Urban sites were in general warmer and less humid than suburban and rural sites, translating into decreased larval survival, smaller body sizes, and lower per capita growth rates of mosquitoes on urban sites. Dengue transmission potential was predicted to be higher in the summer than the fall. Additionally, the effects of land use on dengue transmission potential varied by season. Warm summers resulted in a higher predicted VC on the cooler, rural sites, while warmer, urban sites had a higher predicted VC during the cooler fall season. [ABSTRACT FROM AUTHOR]

Timing of reproduction in birds is important for reproductive success and is known to depend on environmental cues such as day length and food availability. However, in equatorial regions, where day length is nearly constant, other factors such as rainfall and temperature are thought to determine timing of reproduction. Rainfall can vary at small spatial and temporal scales, providing a highly fluctuating and unpredictable environmental cue. In this study we investigated the extent to which spatio-temporal variation in environmental conditions can explain the timing of breeding of Red-capped Lark, Calandrella cinerea, a species that is capable of reproducing during every month of the year in our equatorial east African study locations. For 39 months in three climatically-distinct locations, we monitored nesting activities, sampled ground and flying invertebrates, and quantified rainfall, maximum (Tmax) and minimum (Tmin) temperatures. Among locations we found that lower rainfall and higher temperatures did not coincide with lower invertebrate biomasses and decreased nesting activities, as predicted. Within locations, we found that rainfall, Tmax, and Tmin varied unpredictably among months and years. The only consistent annually recurring observations in all locations were that January and February had low rainfall, high Tmax, and low Tmin. Ground and flying invertebrate biomasses varied unpredictably among months and years, but invertebrates were captured in all months in all locations. Red-capped Larks bred in all calendar months overall but not in every month in every year in every location. Using model selection, we found no clear support for any relationship between the environmental variables and breeding in any of the three locations. Contrary to popular understanding, this study suggests that rainfall and invertebrate biomass as proxy for food do not influence breeding in equatorial Larks. Instead, we propose that factors such as nest predation, female protein reserves, and competition are more important in environments where weather and food meet minimum requirements for breeding during most of the year. [ABSTRACT FROM AUTHOR]

The effects of climate change on animal populations may be shaped by habitat characteristics at both micro- and macro-habitat level, however, empirical studies integrating these two scales of observation are lacking. As analyses of the effects of climate change commonly rely on data from a much larger scale than the microhabitat level organisms are affected at, this mismatch risks hampering progress in developing understanding of the details of the ecological and evolutionary responses of organisms and, ultimately, effective actions to preserve their populations. Cavity nesters, often with a conservation status of concern, are an ideal model because the cavity is a microenvironment potentially different from the macroenvironment but nonetheless inevitably interacting with it. The lesser kestrel (Falco naumanni) is a cavity nester which was until recently classified by as Vulnerable species. Since 2004, for nine years, we collected detailed biotic and abiotic data at both micro- and macro-scales of observation in a kestrel population breeding in the Gela Plain (Italy), a Mediterranean area where high temperatures may reach lethal values for the nest content. We show that macroclimatic features needed to be integrated with both abiotic and biotic factors recorded at a microscale before reliably predicting nest temperatures. Among the nest types used by lesser kestrels, we detected a preferential occupation of the cooler nest types, roof tiles, by early breeders whereas, paradoxically, late breeders nesting with hotter temperatures occupied the overheated nest holes. Not consistent with such a suggested nest selection, the coolest nest type did not host a higher reproductive success than the overheated nests. We discussed our findings in the light of cavity temperatures and nest types deployed within conservation actions assessed by integrating selected factors at different observation scales. [ABSTRACT FROM AUTHOR]

Ocean acidification is considered as a crucial stressor for marine communities. In this study, we tested the effects of the IPCC RPC6.0 end-of-century acidification scenario on a natural plankton community in the Gullmar Fjord, Sweden, during a long-term mesocosm experiment from a spring bloom to a mid-summer situation. The focus of this study was on microzooplankton and its interactions with phytoplankton and mesozooplankton. The microzooplankton community was dominated by ciliates, especially small Strombidium sp., with the exception of the last days when heterotrophic dinoflagellates increased in abundance. We did not observe any effects of high CO2 on the community composition and diversity of microzooplankton. While ciliate abundance, biomass and growth rate were not affected by elevated CO2, we observed a positive effect of elevated CO2 on dinoflagellate abundances. Additionally, growth rates of dinoflagellates were significantly higher in the high CO2 treatments. Given the higher Chlorophyll a content measured under high CO2, our results point at mainly indirect effects of CO2 on microzooplankton caused by changes in phytoplankton standing stocks, in this case most likely an increase in small-sized phytoplankton of <8 μm. Overall, the results from the present study covering the most important part of the growing season indicate that coastal microzooplankton communities are rather robust towards realistic acidification scenarios. [ABSTRACT FROM AUTHOR]

The laboratory rat is emerging as an attractive preclinical animal model of autism spectrum disorder (ASD), allowing investigators to explore genetic, environmental and pharmacological manipulations in a species exhibiting complex, reciprocal social behavior. The present study was carried out to compare two commonly used strains of laboratory rats, Sprague-Dawley (SD) and Long-Evans (LE), between the ages of postnatal day (PND) 26–56 using high-throughput behavioral phenotyping tools commonly used in mouse models of ASD that we have adapted for use in rats. We detected few differences between young SD and LE strains on standard assays of exploration, sensorimotor gating, anxiety, repetitive behaviors, and learning. Both SD and LE strains also demonstrated sociability in the 3-chamber social approach test as indexed by spending more time in the social chamber with a constrained age/strain/sex matched novel partner than in an identical chamber without a partner. Pronounced differences between the two strains were, however, detected when the rats were allowed to freely interact with a novel partner in the social dyad paradigm. The SD rats in this particular testing paradigm engaged in play more frequently and for longer durations than the LE rats at both juvenile and young adult developmental time points. Results from this study that are particularly relevant for developing preclinical ASD models in rats are threefold: (i) commonly utilized strains exhibit unique patterns of social interactions, including strain-specific play behaviors, (ii) the testing environment may profoundly influence the expression of strain-specific social behavior and (iii) simple, automated measures of sociability may not capture the complexities of rat social interactions. [ABSTRACT FROM AUTHOR]

Food neophobia is a reaction to novel food observed in many animal species, particularly omnivores, including Rattus norvegicus. A neophobic reaction is typically characterised by avoidance of novel food and the necessity to assess both its potential value and toxicity by the animal. It has been hypothesised that this reaction is not observed in rats inhabiting a changeable environment with a high level of variability with regard to food and food sources. This study was conducted in such changeable conditions and it aims to demonstrate the behaviour of wild rats R. norvegicus in their natural habitat. The rats were studied in a farm setting, and the experimental arena was demarcated by a specially constructed pen which was freely accessible to the rats. At regular intervals, the rats were given new flavour- and smell-altered foods, while their behaviour was video-recorded. The results obtained in the study seem to confirm the hypothesis that rats inhabiting a highly changeable environment do not exhibit food neophobia. The observed reaction to novel food may be connected with a reaction to a novel object to a larger extent than to food neophobia. The value of the results obtained lies primarily in the fact that the study was conducted in the animals’ natural habitat, and that it investigated their spontaneous behaviours. [ABSTRACT FROM AUTHOR]

This Formal Comment responds to Jordan et al., and stresses that if scientific findings are to be robust, training in experimental design and statistics is critical to ensure that research questions, design considerations, and analyses are aligned. [ABSTRACT FROM AUTHOR]