Trachymyrmex septentrionalis ants promote fungus garden hygiene using Trichoderma-derived metabolite cues.

Fungus-growing ants depend on a fungal mutualist that can fall prey to fungal pathogens. This mutualist is cultivated by these ants in structures called fungus gardens. Ants exhibit weeding behaviors that keep their fungus gardens healthy by physically removing compromised pieces. However, how ants detect diseases of their fungus gardens is unknown. Here, we applied the logic of Koch's postulates using environmental fungal community gene sequencing, fungal isolation, and laboratory infection experiments to establish that Trichoderma spp. can act as previously unrecognized pathogens of Trachymyrmex septentrionalis fungus gardens. Our environmental data showed that Trichoderma are the most abundant noncultivar fungi in wild T. septentrionalis fungus gardens. We further determined that metabolites produced by Trichoderma induce an ant weeding response that mirrors their response to live Trichoderma. Combining ant behavioral experiments with bioactivity-guided fractionation and statistical prioritization of metabolites in Trichoderma extracts demonstrated that T. septentrionalis ants weed in response to peptaibols, a specific class of secondary metabolites known to be produced by Trichoderma fungi. Similar assays conducted using purified peptaibols, including the two previously undescribed peptaibols trichokindins VIII and IX, suggested that weeding is likely induced by peptaibols as a class rather than by a single peptaibol metabolite. In addition to their presence in laboratory experiments, we detected peptaibols in wild fungus gardens. Our combination of environmental data and laboratory infection experiments strongly support that peptaibols act as chemical cues of Trichoderma pathogenesis in T. septentrionalis fungus gardens.

Combining surface and soil environmental DNA with artificial cover objects to improve terrestrial reptile survey detection.

Reptiles are increasingly of conservation concern due to their susceptibility to habitat loss, emerging disease, and harvest in the wildlife trade. However, reptile populations are often difficult to monitor given the frequency of crypsis in their life history. This difficulty has left uncertain the conservation status of many species and the efficacy of conservation actions unknown. Environmental DNA (eDNA) surveys consistently elevate the detection rate of species they are designed to monitor, and while their use is promising for terrestrial reptile conservation, successes in developing such surveys have been sparse. We tested the degree to which inclusion of surface and soil eDNA sampling into conventional artificial-cover methods elevates the detection probability of a small, cryptic terrestrial lizard, Scincella lateralis. The eDNA sampling of cover object surfaces with paint rollers elevated per sample detection probabilities for this species 4-16 times compared with visual surveys alone. We readily detected S. lateralis eDNA under cover objects up to 2 weeks after the last visual detection, and at some cover objects where no S. lateralis were visually observed in prior months. With sufficient sampling intensity, eDNA testing of soil under cover objects produced comparable per sample detection probabilities as roller surface methods. Our results suggest that combining eDNA and cover object methods can considerably increase the detection power of reptile monitoring programs, allowing more accurate estimates of population size, detection of temporal and spatial changes in habitat use, and tracking success of restoration efforts. Further research into the deposition and decay rates of reptile eDNA under cover objects, as well as tailored protocols for different species and habitats, is needed to bring the technique into widespread use.

El interés por la conservación de los reptiles es cada vez mayor debido a su susceptibilidad ante la pérdida del hábitat, enfermedades emergentes y la captura para el mercado de fauna. Sin embargo, las poblaciones de reptiles son difíciles de monitorear por lo frecuente que es la cripsis en sus historias de vida. Esta dificultad deja incierto el estado de conservación de muchas especies y desconocida la eficacia de las acciones de conservación. Los censos de ADN ambiental (DNAa) elevan sistemáticamente la tasa de detección de las especies que monitorean, y aunque su uso es prometedor para la conservación de los reptiles terrestres, han sido escasos los éxitos en el desarrollo de dichos censos. Analizamos el grado al que la inclusión del muestreo de DNAa superficial y del suelo a los métodos convencionales de cobertura artificial eleva la probabilidad de detección de una pequeña lagartija terrestre críptica: Scincella lateralis. El muestreo de DNAa de las superficies con cobertura de objetos con rodillos de pintura elevó las probabilidades de detección por muestra para esta especie 4-16 veces más que los censos visuales. Detectamos fácilmente el DNAa de S. lateralis bajo los objetos de cubierta hasta dos semanas después de la última detección visual y en algunos objetos de cubierta en donde no se había observado en los meses previos a S. lateralis. Con suficiente intensidad de muestreo, el análisis de DNAa del suelo bajo objetos de cubierta produjo probabilidades de detección por muestra comparables como métodos de rodillo superficial. Nuestros resultados sugieren que la combinación del DNAa y los métodos de objetos de cobertura puede incrementar considerablemente el poder de detección de los programas de monitoreo de reptiles, lo que permite estimaciones más precisas del tamaño poblacional, detección de los cambios espaciales y temporales en el uso de hábitat y el éxito de rastreo de los esfuerzos de restauración. Además, se necesita la investigación sobre las tasas de depósito y descomposición del DNAa de reptiles bajo objetos de cubierta, así como los protocolos hechos para diferentes especies y hábitats, para que la técnica entre al uso difundido.

Chemical Gradients of Plant Substrates in an Atta texana Fungus Garden.

Many ant species grow fungus gardens that predigest food as an essential step of the ants' nutrient uptake. These symbiotic fungus gardens have long been studied and feature a gradient of increasing substrate degradation from top to bottom. To further facilitate the study of fungus gardens and enable the understanding of the predigestion process in more detail than currently known, we applied recent mass spectrometry-based approaches and generated a three-dimensional (3D) molecular map of an Atta texana fungus garden to reveal chemical modifications as plant substrates pass through it. The metabolomics approach presented in this study can be applied to study similar processes in natural environments to compare with lab-maintained ecosystems. IMPORTANCE The study of complex ecosystems requires an understanding of the chemical processes involving molecules from several sources. Some of the molecules present in fungus-growing ants' symbiotic system originate from plants. To facilitate the study of fungus gardens from a chemical perspective, we provide a molecular map of an Atta texana fungus garden to reveal chemical modifications as plant substrates pass through it. The metabolomics approach presented in this study can be applied to study similar processes in natural environments.

Moving eDNA surveys onto land: Strategies for active eDNA aggregation to detect invasive forest insects.

The use of environmental DNA (eDNA) surveys to monitor terrestrial species has been relatively limited, with successful implementations still confined to sampling DNA from natural or artificial water bodies and soil. Sampling water for eDNA depends on proximity to or availability of water, whereas eDNA from soil is limited in its spatial scale due to the large quantities necessary for processing and difficulty in doing so. These challenges limit the widespread use of eDNA in several systems, such as surveying forests for invasive insects. We developed two new eDNA aggregation approaches that overcome the challenges of above-ground terrestrial sampling and eliminate the dependency on creating or utilizing pre-existing water bodies to conduct eDNA sampling. The first, "spray aggregation," uses spray action to remove eDNA from surface substrates and was developed for shrubs and other understorey vegetation, while the second, "tree rolling," uses physical transfer via a roller to remove eDNA from the surface of tree trunks and large branches. We tested these approaches by surveying for spotted lanternfly, Lycorma delicatula, a recent invasive pest of northeastern USA that is considered a significant ecological and economic threat to forests and agriculture. We found that our terrestrial eDNA surveys matched visual surveys, but also detected L. delicatula presence ahead of visual surveys, indicating increased sensitivity of terrestrial eDNA surveys over currently used methodology. The terrestrial eDNA approaches we describe can be adapted for use in surveying a variety of forest insects and represent a novel strategy for surveying terrestrial biodiversity.

An hPSC-Derived Tissue-Resident Macrophage Model Reveals Differential Responses of Macrophages to ZIKV and DENV Infection.

Zika virus (ZIKV) and dengue virus (DENV) are two closely related flaviviruses that lead to different clinical outcomes. The mechanism for the distinct pathogenesis of ZIKV and DENV is poorly understood. Here, we investigate ZIKV and DENV infection of macrophages using a human pluripotent stem cell (hPSC)-derived macrophage model and discover key virus-specific responses. ZIKV and DENV productively infect hPSC-derived macrophages. DENV, but not ZIKV, infection of macrophages strongly activates macrophage migration inhibitory factor (MIF) secretion and decreases macrophage migration. Neutralization of MIF leads to improved migratory ability of DENV-infected macrophages. In contrast, ZIKV-infected macrophages exhibit prolonged migration and express low levels of pro-inflammatory cytokines and chemokines. Mechanistically, ZIKV disrupts the nuclear factor κB (NF-κB)-MIF positive feedback loop by inhibiting the NF-κB signaling pathway. Our results demonstrate the utility of hPSC-derived macrophages in infectious disease modeling and suggest that the distinct impact of ZIKV and DENV on macrophage immune response may underlie different pathogenesis of Zika and dengue diseases.

Stability of Wake-Sleep Cycles Requires Robust Degradation of the PERIOD Protein.

Robustness in biology is the stability of phenotype under diverse genetic and/or environmental perturbations. The circadian clock has remarkable stability of period and phase that-unlike other biological oscillators-is maintained over a wide range of conditions. Here, we show that the high fidelity of the circadian system stems from robust degradation of the clock protein PERIOD. We show that PERIOD degradation is regulated by a balance between ubiquitination and deubiquitination, and that disruption of this balance can destabilize the clock. In mice with a loss-of-function mutation of the E3 ligase gene ß-Trcp2, the balance of PERIOD degradation is perturbed and the clock becomes dramatically unstable, presenting a unique behavioral phenotype unlike other circadian mutant animal models. We believe that our data provide a molecular explanation for how circadian phases, such as wake-sleep onset times, can become unstable in humans, and we present a unique mouse model to study human circadian disorders with unstable circadian rhythm phases.

Left inferior parietal representations for skilled hand-object interactions: evidence from stroke and corticobasal degeneration.

Patients with ideomotor apraxia (IM) are frequently more impaired in the production and imitation of object-related (transitive) than non-object-related, symbolic (intransitive) gestures, but reasons for this dissociation, and its anatomical underpinnings, remain unclear. Our theoretical model of praxis (Buxbaum, 2001) postulates that left inferior parietal lobe (IPL) gesture representations store information about postures and movements of the body and hand for skillful manipulation of familiar objects; in contrast, bilateral fronto-parietal dynamic calculations provide constantly-updated information about the current position and movement of the body and hand for both familiar and novel, transitive and intransitive movements. This account predicts distinct patterns of IM in patients with left IPL damage versus bilateral fronto-parietal involvement. Consistent with predictions, 16 stroke patients with left IPL damage were more impaired with transitive than intransitive gestures, whereas 4 patients with bilateral fronto-parietal damage due to corticobasal degeneration (CBD) were not [F (1, 18) = 8.5 p < .01]. Additionally, the hand posture component of transitive gestures was the most impaired aspect of gesture in CVA, but tended to be the least impaired aspect of gesture in CBD [F (3, 54) = 5.1, p < .005]. Finally, CVA patients were more impaired with transitive hand postures than meaningless or intransitive hand postures, whereas CBD patients showed the opposite pattern. These data indicate that the left IPL mediates representations of skilled hand-object interactions, as distinct from dynamic coding of the body in space, and suggest that the IPL maps between representations of object identity in the ventral stream and spatial body representations mediated by the dorsal system.

Neural substrates of knowledge of hand postures for object grasping and functional object use: evidence from fMRI.

A number of lines of evidence suggest that computation of hand posture differs for object grasping as compared to functional object use. Hand shaping for grasping appears to rely strongly upon calculations of current object location and volume, whereas hand shaping for object use additionally requires access to stored knowledge about the skilled manipulation specific to a given object. In addition, the particular hand postures employed for functional object use may be either prehensile (clenching, pinching) or non-prehensile (e.g., palming, poking), in contrast to the prehensile postures that are obligatory for grasping. In this fMRI study, we assessed the hypothesis that a left-hemisphere-lateralized system including the inferior parietal lobe is specifically recruited for the computation and recognition of hand postures for functional object use. Fifteen subjects viewed pictures of manipulable objects and determined whether they would be grasped with a pinch or clench (Grasp condition), functionally used with a pinch or clench (Prehensile Use condition), or functionally used with a palm or poke hand posture (Non-prehensile Use condition). Despite the fact that the conditions were equated for behavioral difficulty, significantly greater activations were observed in the left inferior frontal gyrus (IFG), posterior superior temporal gyrus (STG), and inferior parietal lobule (IPL) in Non-prehensile Use trials as compared to Grasp trials. Comparison of Non-prehensile Use and Prehensile Use activations revealed significant differences only in the left IPL. These data confirm the importance of the left IPL in storing knowledge of hand postures for functional object use, and have implications for understanding the interaction of dorsal and ventral visual processing systems.

On beyond mirror neurons: internal representations subserving imitation and recognition of skilled object-related actions in humans.

A considerable recent literature argues that the same representations, encoded by inferior prefrontal and parietal cells known as "mirror neurons", may be activated in both production and recognition of object-related actions. Here, we test several predictions derived from the contemporary literature on the parity between production and recognition and the putative emergence of the mirror neuron system from a system coding hand-object interactions. Forty-four patients with left-hemisphere stroke, 21 of whom exhibited ideomotor apraxia, performed a number of pantomime imitation and recognition tasks, and performance was scored with respect to hand posture, arm posture, amplitude, and timing. Consistent with predictions, there were strong relationships between object-related pantomime imitation and object-related pantomime recognition, and between imitation and recognition of the hand posture component of object-related actions. Skilled object-related gesture representations are likely to be closely tied to evolutionarily more primitive systems controlling object grasping, to emerge from a mapping between object and action information coded by ventral and dorsal streams, and to be lateralized to the left hemisphere in humans.