Efficient coding of natural images in the mouse visual cortex.

How the activity of neurons gives rise to natural vision remains a matter of intense investigation. The mid-level visual areas along the ventral stream are selective to a common class of natural images-textures-but a circuit-level understanding of this selectivity and its link to perception remains unclear. We addressed these questions in mice, first showing that they can perceptually discriminate between textures and statistically simpler spectrally matched stimuli, and between texture types. Then, at the neural level, we found that the secondary visual area (LM) exhibited a higher degree of selectivity for textures compared to the primary visual area (V1). Furthermore, textures were represented in distinct neural activity subspaces whose relative distances were found to correlate with the statistical similarity of the images and the mice's ability to discriminate between them. Notably, these dependencies were more pronounced in LM, where the texture-related subspaces were smaller than in V1, resulting in superior stimulus decoding capabilities. Together, our results demonstrate texture vision in mice, finding a linking framework between stimulus statistics, neural representations, and perceptual sensitivity-a distinct hallmark of efficient coding computations.

Meso-Py: Dual Brain Cortical Calcium Imaging in Mice during Head-Fixed Social Stimulus Presentation.

We present a cost-effective, compact foot-print, and open-source Raspberry Pi-based widefield imaging system. The compact nature allows the system to be used for close-proximity dual-brain cortical mesoscale functional-imaging to simultaneously observe activity in two head-fixed animals in a staged social touch-like interaction. We provide all schematics, code, and protocols for a rail system where head-fixed mice are brought together to a distance where the macrovibrissae of each mouse make contact. Cortical neuronal functional signals (GCaMP6s; genetically encoded Ca2+ sensor) were recorded from both mice simultaneously before, during, and after the social contact period. When the mice were together, we observed bouts of mutual whisking and cross-mouse correlated cortical activity across the cortex. Correlations were not observed in trial-shuffled mouse pairs, suggesting that correlated activity was specific to individual interactions. Whisking-related cortical signals were observed during the period where mice were together (closest contact). The effects of social stimulus presentation extend outside of regions associated with mutual touch and have global synchronizing effects on cortical activity.

Ongoing declines for the world's amphibians in the face of emerging threats.

Systematic assessments of species extinction risk at regular intervals are necessary for informing conservation action1,2. Ongoing developments in taxonomy, threatening processes and research further underscore the need for reassessment3,4. Here we report the findings of the second Global Amphibian Assessment, evaluating 8,011 species for the International Union for Conservation of Nature Red List of Threatened Species. We find that amphibians are the most threatened vertebrate class (40.7% of species are globally threatened). The updated Red List Index shows that the status of amphibians is deteriorating globally, particularly for salamanders and in the Neotropics. Disease and habitat loss drove 91% of status deteriorations between 1980 and 2004. Ongoing and projected climate change effects are now of increasing concern, driving 39% of status deteriorations since 2004, followed by habitat loss (37%). Although signs of species recoveries incentivize immediate conservation action, scaled-up investment is urgently needed to reverse the current trends.

Two new species of semiaquatic Anolis (Squamata: Dactyloidae) from Costa Rica.

We describe two new species of Draconura-clade semiaquatic anoles from the central Pacific versant of Costa Rica. The two new species are similar to Anolis aquaticus in external appearance and ecology but differ from this species in male dewlap coloration and scalation. Anolis robinsoni sp. nov. and A. riparius sp. nov. differ from each other mainly in male dewlap color. All three species are distinct according to diagnostic morphological traits and a phylogenetic analysis of mitochondrial DNA sequences (669 bases of COI gene). We discuss the distribution and ecology of Anolis aquaticus and the new species.

Uncovering the effect of different brain regions on behavioral classification using recurrent neural networks.

As our ability to record neural activity from a larger number of brain areas increases, we need to develop tools to understand how this activity is related to ongoing behavior. Recurrent neural networks (RNNs) have been shown to perform successful classification for sequence data. However, they are black box models: once trained, it is difficult to uncover the mechanisms that they are using to classify. In this study, we analyze the effect of RNNs on classifying behavior using a simulated dataset and a widefield neural activity dataset as mice perform a self-initiated behavior. We show that RNNs are comparable to, or outperform, traditional classification methods such as Support Vector Machine (SVM), and can also lead to accurate prediction of behavior. Using dimensionality reduction, we visualize the activity of the RNNs to better understand the classification mechanisms of the RNNs. Finally, we are able to accurately pinpoint the effect of different regions on behavioral classification. This study highlights the utility and interpretability of RNNs while classifying behavior using neural activity from different regions.

Recent findings of Isthmohyla pictipes (Anura: Hylidae) in Costa Rica: variation and implications for conservation.

We report recent findings of Isthmohyla pictipes (Cope, 1875) in the Cordillera de Talamanca, Costa Rica, roughly two decades after it was last registered. We provide notes on microhabitat use, color variation, external morphology of adults and larvae, and geographic variation, and discuss some taxonomic characters employed to differentiate I. pictipes from I. tica (Starrett, 1966) and I. xanthosticta (Duellman, 1968). We also report fluorescence on the ventral surfaces of I. pictipes. Our findings are expected to shed light on the taxonomy of this species and should be useful in further population assessments and conservation plans.

Automated task training and longitudinal monitoring of mouse mesoscale cortical circuits using home cages.

We report improved automated open-source methodology for head-fixed mesoscale cortical imaging and/or behavioral training of home cage mice using Raspberry Pi-based hardware. Staged partial and probabilistic restraint allows mice to adjust to self-initiated headfixation over 3 weeks' time with ~50% participation rate. We support a cue-based behavioral licking task monitored by a capacitive touch-sensor water spout. While automatically head-fixed, we acquire spontaneous, movement-triggered, or licking task-evoked GCaMP6 cortical signals. An analysis pipeline marked both behavioral events, as well as analyzed brain fluorescence signals as they relate to spontaneous and/or task-evoked behavioral activity. Mice were trained to suppress licking and wait for cues that marked the delivery of water. Correct rewarded go-trials were associated with widespread activation of midline and lateral barrel cortex areas following a vibration cue and delayed frontal and lateral motor cortex activation. Cortical GCaMP signals predicted trial success and correlated strongly with trial-outcome dependent body movements.

Toxicity and Alkaloid Profiling of the Skin of the Golfo Dulcean Poison Frog Phyllobates vittatus (Dendrobatidae).

Frogs in the genus Phyllobates are known for the presence of batrachotoxin, a highly toxic alkaloid, in their skin. Nevertheless, Phyllobates frogs from Costa Rica and Panama (P. lugubris and P. vittatus) are considered non-toxic, as they have been reported to harbor low concentrations of this alkaloid. However, the potential toxicity of Central American Phyllobates has not been assessed experimentally. Our goal was to determine the toxicity of the whole skin of P. vittatus, an endemic species from the Southeastern Pacific region of Costa Rica. We performed median lethal dose (LD50) tests in mice to determine general toxicity, and an irritant assay based on the behavioral responses of mice to subcutaneous injection, to determine differences in irritability, as a measure of toxicity, among three study localities. Using UPLC-ESI-QTOF, we obtained chemical profiles of the methanolic extract of frog skins. Due to the absence of mortality at the studied doses, we were unable to estimate LD50. However, we recorded a list of toxicity symptoms in mice that are consistent with cardiotoxic effects, and found that mice presented more symptoms at higher concentrations of skin extracts during the first hour of the LD50 assays, recovering completely at all doses by the end of the assay. On the other hand, we did not detect differences in irritability among studied localities. Additionally, we putatively identified three toxic alkaloids (Batrachotoxinin A, DHQ 251A and Lehm 275A). This study provides the first experimental data on the toxicity and associated symptoms in mice, as well as the chemical profile of the skin of P. vittatus. We suggest that the skin alkaloids of P. vitattus may confer a chemical defense towards predators.

Physiological Signature of Memory Age in the Prefrontal-Hippocampal Circuit.

The long-term storage of episodic memory requires communication between prefrontal cortex and hippocampus. However, how consolidation alters dynamic interactions between these regions during subsequent recall remains unexplored. Here we perform simultaneous electrophysiological recordings from anterior cingulate cortex (ACC) and hippocampal CA1 in mice during recall of recent and remote contextual fear memory. We find that, in contrast to recent memory, remote memory recall is accompanied by increased ACC-CA1 synchronization at multiple frequency bands. The augmented ACC-CA1 interaction is associated with strengthened coupling among distally spaced CA1 neurons, suggesting an ACC-driven organization of a sparse code. This robust shift in physiology permits a support vector machine classifier to accurately determine memory age on the basis of the ACC-CA1 synchronization pattern. Our findings reveal that memory consolidation alters the dynamic coupling of the prefrontal-hippocampal circuit and results in a physiological signature of memory age.

Batrachochytrium dendrobatidis infection in amphibians predates first known epizootic in Costa Rica.

Emerging infectious diseases are a growing threat to biodiversity worldwide. Outbreaks of the infectious disease chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd), are implicated in the decline and extinction of numerous amphibian species. In Costa Rica, a major decline event occurred in 1987, more than two decades before this pathogen was discovered. The loss of many species in Costa Rica is assumed to be due to Bd-epizootics, but there are few studies that provide data from amphibians in the time leading up to the proposed epizootics. In this study, we provide new data on Bd infection rates of amphibians collected throughout Costa Rica, in the decades prior to the epizootics. We used a quantitative PCR assay to test for Bd presence in 1016 anuran museum specimens collected throughout Costa Rica. The earliest specimen that tested positive for Bd was collected in 1964. Across all time periods, we found an overall infection rate (defined as the proportion of Bd-positive individuals) of 4%. The number of infected individuals remained relatively low across all species tested and the range of Bd-positive specimens was shown to be geographically constrained up until the 1980s; when epizootics are hypothesized to have occurred. After that time, infection rate increased three-fold, and the range of specimens tested positive for Bd increased, with Bd-positive specimens collected across the entire country. Our results suggest that Bd dynamics in Costa Rica are more complicated than previously thought. The discovery of Bd's presence in the country preceding massive declines leads to a number of different hypotheses: 1) Bd invaded Costa Rica earlier than previously known, and spread more slowly than previously reported; 2) Bd invaded multiple times and faded out; 3) an endemic Bd lineage existed; 4) an earlier Bd lineage evolved into the current Bd lineage or hybridized with an invasive lineage; or 5) an earlier Bd lineage went extinct and a new invasion event occurred causing epizootics. To help visualize areas where future studies should take place, we provide a Bd habitat suitability model trained with local data. Studies that provide information on genetic lineages of Bd are needed to determine the most plausible spatial-temporal, host-pathogen dynamics that could best explain the epizootics resulting in amphibian declines in Costa Rica and throughout Central America.

The skipper butterflies (Lepidoptera: Hesperiidae) of the Reserva Ecológica Leonelo Oviedo, San José, Costa Rica

Hesperiidae is one of the most diverse families of butterflies in Costa Rica, with approximately 486 species. Even so, there are few butterfly lists where this group has been included. In this paper, we present information on seasonality, abundance and natural history features of this family for the Leonelo Oviedo Ecological Reserve (RELO), a 2 ha forest embedded in an urban matrix. Over the course of two years, a monthly sampling was carried out on a 270 m trail across the Reserve from 08:00 to 12:00, collecting all the individuals located within 5 m on each side of the trail. To better represent the richness, individuals were also randomly collected for more than ten years, but the butterflies collected in this way were not included in the statistical analysis. Photographs were taken of all the species in order to provide an identification guide. For the cryptic species, drawings and dissections of the genitalia were made. For the community indexes we used Microsoft Excel and the Shannon index with base two logarithm. For the summary of the monthly data analysis were done according to dry and wet season. For a comparison of richness and abundance we did a g-test to evaluate if there are differences between seasons; however, with the use of the R package vegan a hierarchical cluster analysis was done using the Jaccard index with Wards minimum variance agglomerative method. With R package pvclust the uncertainty of the clusters based on a bootstrap with 10 000 iterations. 423 individuals of 49 species were included in the statistical analysis, from a total of 435 individuals of 58 species. A tendency to greater richness and abundance of skippers was found during the dry season. Through the cluster analysis, it was possible to determine that in relation to the diversity of skippers, both wet seasons are grouped significantly (P = 0.05). The dry seasons are also grouped significantly (P = 0.05). The reserve has connectivity with other green areas via a stream. During the wet season, plant growth increases connectivity, which could lead to the entry of new individuals of different species that are not permanent residents of RELO and establish small populations, increasing the richness and abundance of species. This added to the variation in the occurrence of some species of butterflies in response to seasonal variations and differences in the availability of resources in different seasons explains the grouping of species between seasons.

Con aproximadamente 486 especies, Hesperiidae es una de las familias más diversas de mariposas de Costa Rica. A pesar de eso, en pocas listas de especies locales son incluidas. En este trabajo, presentamos información sobre la estacionalidad, abundancia e historia natural de los hespéridos de la Reserva Ecológica Leonelo Oviedo. Ésta reserva se encuentra dentro de una matriz urbanizada y por un período de dos años se realizaron muestreos mensuales a lo largo de un sendero de 270 m que la atraviesa. Las recolectas se llevaron a cabo desde las 08:00 hasta las 12:00, atrapando todos los individuos encontrados a cada lado del sendero a una distancia de hasta 5 m. Para representar mejor la riqueza de especies, se siguió recolectando individuos de manera aleatoria por más de diez años posteriores al muestreo inicial. Consecuentemente, estos individuos no fueron incluidos en el análisis estadístico realizado. Se tomaron fotografías de todas las especies obtenidas en total, para proporcionar una guía de identificación de especies. Ademas, para las especies crípticas se realizaron disecciones e ilustraciones de los genitales masculinos. Para el análisis de diversidad se utilizó el índice de Shannon con logaritmo en base dos y analizando los datos por separado de las estación lluviosa y seca. Para la comparación de riqueza y abundancia se realizó una prueba G y se obtuvo que hay mayor diversidad y abundancia durante la estación seca. Se utilizó el paquete vegan R para un análisis de conglomerados de Jaccard con un indice Ward de mínima varianza. Con el Paquete R pvclust la incertidumbre de los conglomerados se realizó con un bootstrap de 10 000 interacciones. 423 individuos de 49 especies fueron incluidas en el análisis estadístico, de un total de 435 individuos de 58 especies tratadas en este trabajo. Con el análisis de conglomerados fue posible determinar que la diversidad de Hesperiidae en las dos estaciones lluviosas analizadas se agrupan significativamente (P = 0.05). Lo mismo que sucede con las estaciones secas (P = 0.05). La Reserva presenta conectividad con otras áreas boscosas a través de una quebrada. Durante la estación lluviosa, el crecimiento vegetativo aumenta el paso de individuos lo que puede propiciar la llegada de nuevas especies que no necesariamente son residentes permanentes de la Reserva y establecerse en pequeñas poblaciones que incrementan la diversidad y abundancia temporalmente, Lo anterior, mas las variaciones estacionales en la disponibilidad de recursos explican la similitud de la composición de especies entre cada una de las estaciones.

Shifts in the diversity of an amphibian community from a premontane forest of San Ramón, Costa Rica

Biological communities are experiencing rapid shifts of composition in Neotropical ecosystems due to several factors causing population declines. However, emerging evidence has provided insights on the adaptive potential of multiple species to respond to illnesses and environmental pressures. In Costa Rica, the decline of amphibian populations is a remarkable example of these changes. Here we provide evidence of variation in the amphibian richness of a premontane forest of San Ramón (Costa Rica) across a ~30 year period. We also quantified changes in the composition and abundance of the leaf-litter frog community occurring in the same premontane forest, by comparing diversity data with a difference of ~18 years. We evaluated the similarity of species richness from 1980s to 2010s based on several sources, and the dissimilarity of species diversity in the site comparing 28 standardized surveys from 1994-1995 and 2011-2012. We compared the relative abundance of some frogs that inhabit the leaf-litter layer between these same periods. Our results show that there is more similarity in amphibian richness between 1980s and 2010s (~ 52 %) than between 1980s and 1990s (~ 40 %). The richness of leaf-litter anurans was ~ 65 % similar between 1990s and 2010s. The diversity of leaf-litter anuran was clearly different between 1994-1995 and 2011-2012, and it was clustered among those periods. We determined that the amphibian community in this premontane forest drastically changed: many species have disappeared, or gradually declined through the decades (e.g. Pristimantis ridens, P. cruentus, Craugastor bransfordii) as in other well studied localities of Costa Rica, while some few species flourished after being almost absent from the site in the 1990s (e.g. Craugastor crassidigitus, Lithobates warszewistchii). Currently dominant species such as C. crassidigitus would be using developed resistance against Bd-fungus as an advantage (apparent competition) in the premontane forest where the disease is more virulent than in lowlands. Our analysis supports the hypothesis of individualized responses of anuran populations under distinct site and elevations. We suggest to continue monitoring the amphibian communities of premontane tropical forests to understand how this ecosystem gradually resist and adapts to this catastrophic time of biodiversity loss.

Las comunidades biológicas están experimentando rápidos cambios de composición en los ecosistemas neotropicales debido a varios factores que causan disminuciones de las poblaciones. Sin embargo, la evidencia emergente ha proporcionado información sobre el potencial de adaptación de múltiples especies para responder a enfermedades y presiones ambientales. En Costa Rica, el declive de las poblaciones de anfibios es un ejemplo notable de estos cambios. Aquí proporcionamos evidencia de variación en la riqueza de anfibios de un bosque premontano de San Ramón (Costa Rica) en un período de ~ 30 años. También cuantificamos los cambios en el ensamblaje, y las oscilaciones poblacionales, de la comunidad de ranas de hojarasca que ocurren en el mismo bosque premontano, tras comparar datos de diversidad con una diferencia de ~ 18 años. Evaluamos la similitud de la riqueza de especies desde la década de 1980 hasta la de 2010 basados en varias fuentes, así como la disimilitud de la diversidad de especies en el sitio comparando 28 muestreos estandarizados entre 1994-1995 y 2011-2012. Comparamos la abundancia relativa de algunas ranas que habitan la hojarasca entre 1994-1995 y 2011-2012. Nuestros resultados muestran que existe una similitud un poco mayor en la riqueza de anfibios entre los años 1980 y 2010 que entre los 1980 y 1990 (~ 40 %). La riqueza de los anuros de la hojarasca fue aproximadamente un 65 % similar entre la década de 1990 y 2010. La diversidad de anuros de la hojarasca fue claramente diferente entre 1994-1995 y 2011-2012, y se agrupó entre esos períodos. Determinamos que la comunidad de anfibios en este bosque premontano cambió drásticamente: muchas especies han desaparecido o han disminuido gradualmente a lo largo de las décadas como en otras localidades bien estudiadas de Costa Rica (por ejemplo, Pristimantis ridens, P. cruentus, Craugastor bransfordii), mientras algunas pocas especies florecieron tras casi desaparecer de nuestro sitio en la década de 1990 (por ejemplo, Craugastor crassidigitus, Lithobates warszewistchii). Actualmente, las especies dominantes como C. crassidigitus podrían tener resistencia contra el hongo quitridio, desarrollada como una ventaja (competición aparente) en el bosque premontano donde la enfermedad es más virulenta que en las tierras bajas. Nuestro análisis apoya la hipótesis de las respuestas individualizadas de las poblaciones de anuros en distintos sitios y elevaciones. Sugerimos continuar con el monitoreo de las comunidades de anfibios de los bosques neotropicales premontanos para comprender cómo este ecosistema resiste gradualmente y se adapta a este momento catastrófico de pérdida de biodiversidad.

Individualized tracking of self-directed motor learning in group-housed mice performing a skilled lever positioning task in the home cage.

Skilled forelimb function in mice is traditionally studied through behavioral paradigms that require extensive training by investigators and are limited by the number of trials individual animals are able to perform within a supervised session. We developed a skilled lever positioning task that mice can perform within their home cage. The task requires mice to use their forelimb to precisely hold a lever mounted on a rotary encoder within a rewarded position to dispense a water reward. A Raspberry Pi microcomputer is used to record lever position during trials and to control task parameters, thus making this low-footprint apparatus ideal for use within animal housing facilities. Custom Python software automatically increments task difficulty by requiring a longer hold duration, or a more accurate hold position, to dispense a reward. The performance of individual animals within group-housed mice is tracked through radio-frequency identification implants, and data stored on the microcomputer may be accessed remotely through an active internet connection. Mice continuously engage in the task for over 2.5 mo and perform ~500 trials/24 h. Mice required ~15,000 trials to learn to hold the lever within a 10° range for 1.5 s and were able to further refine movement accuracy by limiting their error to a 5° range within each trial. These results demonstrate the feasibility of autonomously training group-housed mice on a forelimb motor task. This paradigm may be used in the future to assess functional recovery after injury or cortical reorganization induced by self-directed motor learning. NEW & NOTEWORTHY We developed a low-cost system for fully autonomous training of group-housed mice on a forelimb motor task. We demonstrate the feasibility of tracking both end-point, as well as kinematic performance of individual mice, with each performing thousands of trials over 2.5 mo. The task is run and controlled by a Raspberry Pi microcomputer, which allows for cages to be monitored remotely through an active internet connection.

An Automated Home-Cage System to Assess Learning and Performance of a Skilled Motor Task in a Mouse Model of Huntington's Disease.

Behavioral testing is a critical step in assessing the validity of rodent models of neurodegenerative disease, as well as evaluating the efficacy of pharmacological interventions. In models of Huntington's disease (HD), a gradual progression of impairments is observed across ages, increasing the need for sensitive, high-throughput and longitudinal assessments. Recently, a number of automated systems have been developed to perform behavioral profiling of animals within their own home-cage, allowing for 24-h monitoring and minimizing experimenter interaction. However, as of yet, few of these have had functionality for the assessment of skilled motor learning, a relevant behavior for movement disorders such as HD. To address this, we assess a lever positioning task within the mouse home-cage. Animals first acquire a simple operant response, before moving to a second phase where they must learn to hold the lever for progressively longer in a rewarded position range. Testing with this paradigm has revealed the presence of distinct phenotypes in the YAC128 mouse model of HD at three early symptomatic time points. YAC128 mice at two months old, but not older, had a motor learning deficit when required to adapt their response to changes in task requirements. In contrast, six-month-old YAC128 mice had disruptions of normal circadian activity and displayed kinematic abnormalities during performance of the task, suggesting an impairment in motor control. This system holds promise for facilitating high throughput behavioral assessment of HD mouse models for preclinical therapeutic screening.

Mesoscale brain explorer, a flexible python-based image analysis and visualization tool.

Imaging of mesoscale brain activity is used to map interactions between brain regions. This work has benefited from the pioneering studies of Grinvald et al., who employed optical methods to image brain function by exploiting the properties of intrinsic optical signals and small molecule voltage-sensitive dyes. Mesoscale interareal brain imaging techniques have been advanced by cell targeted and selective recombinant indicators of neuronal activity. Spontaneous resting state activity is often collected during mesoscale imaging to provide the basis for mapping of connectivity relationships using correlation. However, the information content of mesoscale datasets is vast and is only superficially presented in manuscripts given the need to constrain measurements to a fixed set of frequencies, regions of interest, and other parameters. We describe a new open source tool written in python, termed mesoscale brain explorer (MBE), which provides an interface to process and explore these large datasets. The platform supports automated image processing pipelines with the ability to assess multiple trials and combine data from different animals. The tool provides functions for temporal filtering, averaging, and visualization of functional connectivity relations using time-dependent correlation. Here, we describe the tool and show applications, where previously published datasets were reanalyzed using MBE.

Shared Epizoic Taxa and Differences in Diatom Community Structure Between Green Turtles (Chelonia mydas) from Distant Habitats.

The first reports of diatoms growing on marine mammals date back to the early 1900s. However, only recently has direct evidence been provided for similar associations between diatoms and sea turtles. We present a comparison of diatom communities inhabiting carapaces of green turtles Chelonia mydas sampled at two remote sites located within the Indian (Iran) and Atlantic (Costa Rica) Ocean basins. Diatom observations and counts were carried out using scanning electron microscopy. Techniques involving critical point drying enabled observations of diatoms and other microepibionts still attached to sea turtle carapace and revealed specific aspects of the epizoic community structure. Species-poor, well-developed diatom communities were found on all examined sea turtles. Significant differences between the two host sea turtle populations were observed in terms of diatom abundance and their community structure (including growth form structure). A total of 12 and 22 diatom taxa were found from sea turtles in Iran and Costa Rica, respectively, and eight of these species belonging to Amphora, Chelonicola, Cocconeis, Navicula, Nitzschia and Poulinea genera were observed in samples from both locations. Potential mechanisms of diatom dispersal and the influence of the external environment, sea turtle behaviour, its life stage, and foraging and breeding habitats, as well as epibiotic bacterial flora on epizoic communities, are discussed.

Cost effective raspberry pi-based radio frequency identification tagging of mice suitable for automated in vivo imaging.

BACKGROUND: Automation of animal experimentation improves consistency, reduces potential for error while decreasing animal stress and increasing well-being. Radio frequency identification (RFID) tagging can identify individual mice in group housing environments enabling animal-specific tracking of physiological parameters. NEW METHOD: We describe a simple protocol to radio frequency identification (RFID) tag and detect mice. RFID tags were injected sub-cutaneously after brief isoflurane anesthesia and do not require surgical steps such as suturing or incisions. We employ glass-encapsulated 125kHz tags that can be read within 30.2±2.4mm of the antenna. A raspberry pi single board computer and tag reader enable automated logging and cross platform support is possible through Python. RESULTS: We provide sample software written in Python to provide a flexible and cost effective system for logging the weights of multiple mice in relation to pre-defined targets. COMPARISON WITH EXISTING METHODS: The sample software can serve as the basis of any behavioral or physiological task where users will need to identify and track specific animals. Recently, we have applied this system of tagging to automated mouse brain imaging within home-cages. CONCLUSIONS: We provide a cost effective solution employing open source software to facilitate adoption in applications such as automated imaging or tracking individual animal weights during tasks where food or water restriction is employed as motivation for a specific behavior.

High-throughput automated home-cage mesoscopic functional imaging of mouse cortex.

Mouse head-fixed behaviour coupled with functional imaging has become a powerful technique in rodent systems neuroscience. However, training mice can be time consuming and is potentially stressful for animals. Here we report a fully automated, open source, self-initiated head-fixation system for mesoscopic functional imaging in mice. The system supports five mice at a time and requires minimal investigator intervention. Using genetically encoded calcium indicator transgenic mice, we longitudinally monitor cortical functional connectivity up to 24 h per day in >7,000 self-initiated and unsupervised imaging sessions up to 90 days. The procedure provides robust assessment of functional cortical maps on the basis of both spontaneous activity and brief sensory stimuli such as light flashes. The approach is scalable to a number of remotely controlled cages that can be assessed within the controlled conditions of dedicated animal facilities. We anticipate that home-cage brain imaging will permit flexible and chronic assessment of mesoscale cortical function.

Diatoms and Other Epibionts Associated with Olive Ridley (Lepidochelys olivacea) Sea Turtles from the Pacific Coast of Costa Rica.

Although the sea turtles have long been familiar and even iconic to marine biologists, many aspects of their ecology remain unaddressed. The present study is the first of the epizoic diatom community covering the olive ridley turtle's (Lepidochelys olivacea) carapace and the first describing diatoms living on sea turtles in general, with the primary objective of providing detailed information on turtle epibiotic associations. Samples of turtle carapace including the associated diatom biofilm and epizoic macro-fauna were collected from Ostional beach (9° 59´ 23.7´´ N 85° 41´ 52.6´´ W), Costa Rica, during the arribada event in October 2013. A complex diatom community was present in every sample. In total, 11 macro-faunal and 21 diatom taxa were recorded. Amongst diatoms, the most numerous were erect (Achnanthes spp., Tripterion spp.) and motile (Haslea sp., Navicula spp., Nitzschia spp., Proschkinia sp.) forms, followed by adnate Amphora spp., while the most common macro-faunal species was Stomatolepas elegans (Cirripedia). Diatom densities ranged from 8179 ± 750 to 27685 ± 4885 cells mm-2. Epizoic microalgae were either partly immersed or entirely encapsulated within an exopolymeric coat. The relatively low diatom species number, stable species composition and low inter-sample dissimilarities (14.4% on average) may indicate a mutualistic relationship between the epibiont and the basibiont. Dispersal of sea turtle diatoms is probably highly restricted and similar studies will help to understand both diatom diversity, evolution and biogeography, and sea turtle ecology and foraging strategies.