{"title":"A versatile recording device for the analysis of continuous daily external activity in colonies of highly eusocial bees.","authors":"Arthur Roque Justino, Klaus Hartfelder","doi":"10.1007/s00359-024-01709-2","DOIUrl":"10.1007/s00359-024-01709-2","url":null,"abstract":"<p><p>As pollinators, bees are key to maintaining the biodiversity of angiosperm plants, and for agriculture they provide a billion-dollar ecosystem service. But they also compete for resources (primarily nectar and pollen), especially the highly social bees that live in perennial colonies. So, how do they organize their daily temporal activities? Here, we present a versatile, low-cost device for the continuous, automatic recording and data analysis of the locomotor activity in the colony-entrance tube of highly eusocial bees. Consisting of an in-house built block containing an infrared detector, the passage of bees in the colony entrance tunnel is registered and automatically recorded in an Arduino environment, together with concomitant recordings of temperature and relative humidity. With a focus on the highly diverse Neotropical stingless bees (Meliponini), we obtained 10-day consecutive recordings for two colonies each of the species Melipona quadrifasciata and Frieseomelitta varia, and also for the honey bee. The Lomb-Scargle periodogram analysis identified a predominant circadian rhythmicity for all three species, but also indications of ultradian rhythms. For M. quadrifasciata, which is comparable in size to the honey bee, we found evidence for a possibly anticipatory activity already before sunrise. As all three species also presented activity at night in the colony entrance tube, this also raises questions about sleep organization in social insects. The cost and versatility of the device and the open-source options for data analysis make this an attractive system for conducting studies on circadian rhythms in social bees under natural conditions, complementing studies on flower visits by these important pollinators.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"885-900"},"PeriodicalIF":1.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141428354","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Koustubh M Vaze, Giulia Manoli, Charlotte Helfrich-Förster
{"title":"Characterization of pre-diapause phase in the northern Drosophila species D. ezoana.","authors":"Koustubh M Vaze, Giulia Manoli, Charlotte Helfrich-Förster","doi":"10.1007/s00359-024-01707-4","DOIUrl":"10.1007/s00359-024-01707-4","url":null,"abstract":"<p><p>Drosophila ezoana is a virilis group Drosophila species inhabiting northern latitudes. The flies enter adult reproductive diapause to survive winter upon exposure to short photoperiod conditions (short-day) over several consecutive days. Insect pre-diapause phase - the duration between the beginning of exposure to short days and expression of diapause is thought to be comprised of two distinct phases - (a) photoperiodic time measurement that detects short-days, followed by (b) physiological events leading to the expression of diapause phenotype. A short-day dependent segment of the pre-diapause phase thus approximates the process of photoperiodic time measurement. Continuous darkness has been found to be a neutral condition with respect to diapause regulation in many insect species. The effect of variable number of short-days followed by continuous darkness on diapause incidence thus allows identification of short-day dependent segment of pre-diapause phase thereby mapping the process of photo-periodic time measurement. Although, few weeks of exposure to short-days in adult stage is known to be sufficient for the expression of diapause in D. ezoana, the number of short days required for the completion of photo-periodic time measurement has never been systematically analysed. Our experiments show that continuous darkness is a neutral condition for diapause regulation also in D. ezoana. We utilized the neutral nature of continuous darkness to map the process of photoperiodic time measurement in the D. ezoana strain 124OJ8 which showed that integration of short-day photic cues over the first 10 days of pre-diapause phase is essential for diapause induction.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"901-908"},"PeriodicalIF":1.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141447644","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Elizabeth R Elliott, Kaitlyn E Brock, Rachael M Vacassenno, Douglas A Harrison, Robin L Cooper
{"title":"The effects of doxapram and its potential interactions with K2P channels in experimental model preparations.","authors":"Elizabeth R Elliott, Kaitlyn E Brock, Rachael M Vacassenno, Douglas A Harrison, Robin L Cooper","doi":"10.1007/s00359-024-01705-6","DOIUrl":"10.1007/s00359-024-01705-6","url":null,"abstract":"<p><p>The channels commonly responsible for maintaining cell resting membrane potentials are referred to as K2P (two-P-domain K<sup>+</sup> subunit) channels. These K<sup>+</sup> ion channels generally remain open but can be modulated by their local environment. These channels are classified based on pharmacology, pH sensitivity, mechanical stretch, and ionic permeability. Little is known about the physiological nature of these K2P channels in invertebrates. Acidic conditions depolarize neurons and muscle fibers, which may be caused by K2P channels given that one subtype can be blocked by acidic conditions. Doxapram is used clinically as a respiratory aid known to block acid-sensitive K2P channels; thus, the effects of doxapram on the muscle fibers and synaptic transmission in larval Drosophila and crawfish were monitored. A dose-dependent response was observed via depolarization of the larval Drosophila muscle and an increase in evoked synaptic transmission, but doxapram blocked the production of action potentials in the crawfish motor neuron and had a minor effect on the resting membrane potential of the crawfish muscle. This indicates that the nerve and muscle tissues in larval Drosophila and crawfish likely express different K2P channel subtypes. Since these organisms serve as physiological models for neurobiology and physiology, it would be of interest to further investigate what types of K2P channel are expressed in these tissues. (212 words).</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"869-884"},"PeriodicalIF":1.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141159270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Perceptually salient differences in a species recognition cue do not promote auditory streaming in eastern grey treefrogs (Hyla versicolor).","authors":"Lata Kalra, Shoshana Altman, Mark A Bee","doi":"10.1007/s00359-024-01702-9","DOIUrl":"10.1007/s00359-024-01702-9","url":null,"abstract":"<p><p>Auditory streaming underlies a receiver's ability to organize complex mixtures of auditory input into distinct perceptual \"streams\" that represent different sound sources in the environment. During auditory streaming, sounds produced by the same source are integrated through time into a single, coherent auditory stream that is perceptually segregated from other concurrent sounds. Based on human psychoacoustic studies, one hypothesis regarding auditory streaming is that any sufficiently salient perceptual difference may lead to stream segregation. Here, we used the eastern grey treefrog, Hyla versicolor, to test this hypothesis in the context of vocal communication in a non-human animal. In this system, females choose their mate based on perceiving species-specific features of a male's pulsatile advertisement calls in social environments (choruses) characterized by mixtures of overlapping vocalizations. We employed an experimental paradigm from human psychoacoustics to design interleaved pulsatile sequences (ABAB…) that mimicked key features of the species' advertisement call, and in which alternating pulses differed in pulse rise time, which is a robust species recognition cue in eastern grey treefrogs. Using phonotaxis assays, we found no evidence that perceptually salient differences in pulse rise time promoted the segregation of interleaved pulse sequences into distinct auditory streams. These results do not support the hypothesis that any perceptually salient acoustic difference can be exploited as a cue for stream segregation in all species. We discuss these findings in the context of cues used for species recognition and auditory streaming.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"853-867"},"PeriodicalIF":1.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140908973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Zsolt Pirger, Péter Urbán, Bence Gálik, Bence Kiss, Antal Tapodi, János Schmidt, Gábor K Tóth, Joris M Koene, György Kemenes, Dóra Reglődi, Tibor Kiss, István Fodor
{"title":"Same same, but different: exploring the enigmatic role of the pituitary adenylate cyclase-activating polypeptide (PACAP) in invertebrate physiology.","authors":"Zsolt Pirger, Péter Urbán, Bence Gálik, Bence Kiss, Antal Tapodi, János Schmidt, Gábor K Tóth, Joris M Koene, György Kemenes, Dóra Reglődi, Tibor Kiss, István Fodor","doi":"10.1007/s00359-024-01706-5","DOIUrl":"10.1007/s00359-024-01706-5","url":null,"abstract":"<p><p>Evidence has been accumulating that elements of the vertebrate pituitary adenylate cyclase-activating polypeptide (PACAP) system are missing in non-chordate genomes, which is at odds with the partial sequence-, immunohistochemical-, and physiological data in the literature. Multilevel experiments were performed on the great pond snail (Lymnaea stagnalis) to explore the role of PACAP in invertebrates. Screening of neuronal transcriptome and genome data did not reveal homologs to the elements of vertebrate PACAP system. Despite this, immunohistochemical investigations with an anti-human PAC<sub>1</sub> receptor antibody yielded a positive signal in the neuronal elements in the heart. Although Western blotting of proteins extracted from the nervous system found a relevant band for PACAP-38, immunoprecipitation and mass spectrometric analyses revealed no corresponding peptide fragments. Similarly to the effects reported in vertebrates, PACAP-38 significantly increased cAMP synthesis in the heart and had a positive ionotropic effect on heart preparations. Moreover, it significantly modulated the effects of serotonin and acetylcholine. Homologs to members of Cluster B receptors, which have shared common evolutionary origin with the vertebrate PACAP receptors, PTHRs, and GCGRs, were identified and shown not to be expressed in the heart, which does not support a potential role in the mediation of PACAP-induced effects. Our findings support the notion that the PACAP system emerged after the protostome-deuterostome divergence. Using antibodies against vertebrate proteins is again highlighted to have little/no value in invertebrate studies. The physiological effects of vertebrate PACAP peptides in protostomes, no matter how similar they are to those in vertebrates, should be considered non-specific.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"909-925"},"PeriodicalIF":1.9,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11551080/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141472748","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nuria Saiz, Ángel L Alonso-Gómez, Sergio Bustamante-Martínez, Nuria de Pedro, María J Delgado, Esther Isorna
{"title":"Is there direct photoentrainment in the goldfish liver? Wavelength-dependent regulation of clock genes and investigation of the opsin 7 family.","authors":"Nuria Saiz, Ángel L Alonso-Gómez, Sergio Bustamante-Martínez, Nuria de Pedro, María J Delgado, Esther Isorna","doi":"10.1007/s00359-024-01722-5","DOIUrl":"https://doi.org/10.1007/s00359-024-01722-5","url":null,"abstract":"<p><p>Widespread direct photoentrainment in zebrafish peripheral tissues is linked to diverse non-visual opsins. To explore whether this broadly distributed photosensitivity is specific to zebrafish or is a general teleost feature, we investigated hepatic photosynchronization in goldfish. First, we focused on the opsin 7 family (OPN7, a key peripheral novel opsin in zebrafish), investigating its presence in the goldfish liver. Subsequently, we studied whether light can directly entrain the goldfish liver and retina clocks. Silico analysis revealed seven OPN7 paralogs from four gene families, suggesting expansion through whole-genome and tandem duplications. The paralogs of families OPN7a, OPN7b, and OPN7d were mainly localized in neural tissues, while OPN7c paralogs were more abundant in peripheral tissues-including the liver-suggesting divergent roles. Light (independently of the wavelength employed) directly induced the per2a clock gene in the retina both in vivo and in vitro, confirming expected photoentrainment. However, in the liver, photoinduction of per1a and cry1a only occurred in vivo, not in vitro. These results suggest an indirect light-entrainment mechanism of the goldfish hepatic clock, possibly mediated by other oscillators or photosensitive organs. Our findings challenge the assumption of widespread direct photosensitivity in the peripheral tissues of teleosts. Further research is needed to understand the role of tissue-specific photoentrainment and non-visual opsins in diverse teleost species.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":""},"PeriodicalIF":1.9,"publicationDate":"2024-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142523719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Facundo Fainstein, Franz Goller, Gabriel B Mindlin
{"title":"Song-like activation of syringeal and respiratory muscles during sleep in canaries.","authors":"Facundo Fainstein, Franz Goller, Gabriel B Mindlin","doi":"10.1007/s00359-024-01720-7","DOIUrl":"https://doi.org/10.1007/s00359-024-01720-7","url":null,"abstract":"<p><p>Sleep replay activity involves the reactivation of brain structures with patterns similar to those observed during waking behavior. In this study, we demonstrate that adult male canaries exhibit spontaneous, song-like peripheral reactivation during night sleep. Our findings include: (1) the presence of activity in respiratory muscles, leading to song-like air sac pressure patterns of low amplitude, (2) the simultaneous occurrence of respiratory replay events and reactivation of syringeal muscles, and (3) the reactivation of syringeal muscles without concurrent respiratory system activity. This song-like reactivation of peripheral motor systems enables the identification of specific motor patterns, with replay events preserving individual morphological and temporal properties. The activation of peripheral motor systems in songbirds and the differences in activation patterns between species give unique insights into the fictive behavioral output of activation of a complex learned motor behavior during sleep, shedding light on the neural control mechanisms and potential functions.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":""},"PeriodicalIF":1.9,"publicationDate":"2024-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142481299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Evangeline M Rose, Chelsea M Haakenson, Aliyah Patel, Shivika Gaind, Benjamin D Shank, Gregory F Ball
{"title":"Song system neuroanatomy, and immediate early gene expression in a finch species with extensive male and female song.","authors":"Evangeline M Rose, Chelsea M Haakenson, Aliyah Patel, Shivika Gaind, Benjamin D Shank, Gregory F Ball","doi":"10.1007/s00359-023-01651-9","DOIUrl":"10.1007/s00359-023-01651-9","url":null,"abstract":"<p><p>Birdsong is a relatively well-studied behavior, both due to its importance as a model for vocal production learning and as an intriguing complex social behavior. Until the last few decades, work on birdsong focused almost exclusively on males. However, it is now widely accepted that female song not only exists, but is fairly common throughout the oscine passerines. Despite this, and the large number of researchers who have begun exploring female song in the field, researchers in the lab have been slow to adopt model species with female song. Studying female song in the lab is critical for our understanding of sex-specific factors in the physiology controlling this fascinating behavior. Additionally, as a model for vocal production learning in humans, understanding the mechanistic and neuroendocrine control of female song is clearly important. In this study, we examined the red-cheeked cordon bleu (RCCB), an Estrildid finch species with extensive female song. Specifically, we found that there were no significant sex differences in circulating levels of testosterone and progesterone, nor in song production rate. There were no significant differences in cell densities in the three nuclei of the song control system we examined. Additionally, the volume of the robust nucleus of the arcopallium was not significantly different and we report the smallest sex difference in HVC yet published in a songbird. Finally, we demonstrated similar levels of motor driven immediate early gene expression in both males and females after song production.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"735-749"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9761542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Kendra C Buresch, Noelle D Huget, William C Brister, Elaine Y Zhou, Abraham S Lineaweaver, Chloe Rifai, Jinyang Hu, Zoe E Stevenson, Jean G Boal, Roger T Hanlon
{"title":"Evidence for tactile 3D shape discrimination by octopus.","authors":"Kendra C Buresch, Noelle D Huget, William C Brister, Elaine Y Zhou, Abraham S Lineaweaver, Chloe Rifai, Jinyang Hu, Zoe E Stevenson, Jean G Boal, Roger T Hanlon","doi":"10.1007/s00359-024-01696-4","DOIUrl":"10.1007/s00359-024-01696-4","url":null,"abstract":"<p><p>Octopuses integrate visual, chemical and tactile sensory information while foraging and feeding in complex marine habitats. The respective roles of these modes are of interest ecologically, neurobiologically, and for development of engineered soft robotic arms. While vision guides their foraging path, benthic octopuses primarily search \"blindly\" with their arms to find visually hidden prey amidst rocks, crevices and coral heads. Each octopus arm is lined with hundreds of suckers that possess a combination of chemo- and mechanoreceptors to distinguish prey. Contact chemoreception has been demonstrated in lab tests, but mechanotactile sensing is less well characterized. We designed a non-invasive live animal behavioral assay that isolated mechanosensory capabilities of Octopus bimaculoides arms and suckers to discriminate among five resin 3D-printed prey and non-prey shapes (all with identical chemical signatures). Each shape was introduced inside a rock dome and was only accessible to the octopus' arms. Octopuses' responses were variable. Young octopuses discriminated the crab prey shape from the control, whereas older octopuses did not. These experiments suggest that mechanotactile sensing of 3D shapes may aid in prey discrimination; however, (i) chemo-tactile information may be prioritized over mechanotactile information in prey discrimination, and (ii) mechanosensory capability may decline with age.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"815-823"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140112162","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Vladimir V Popov, Dmitry I Nechaev, Alexander Ya Supin, Evgeniya V Sysueva
{"title":"Monaural and dichotic forward masking in the dolphin's auditory system.","authors":"Vladimir V Popov, Dmitry I Nechaev, Alexander Ya Supin, Evgeniya V Sysueva","doi":"10.1007/s00359-023-01675-1","DOIUrl":"10.1007/s00359-023-01675-1","url":null,"abstract":"<p><p>Short-latency auditory-evoked potentials (AEPs) were recorded non-invasively in the bottlenose dolphin Tursiops truncatus. The stimuli were two sound clicks that were played either monaurally (both clicks to one and the same acoustic window) or dichotically (the leading stimulus (masker) to one acoustic window and the delayed stimulus (test) to the other window). The ratio of the levels of the two stimuli was 0, 10, or 20 dB (at 10 and 20 dB, the leading stimulus was of a higher level). The inter-stimulus intervals (ISIs) varied from 0.15 to 10 ms. The test response magnitude was assessed by correlation analysis as a percentage of the control (non-masked) response. At monaural stimulation, the test response was of a constant magnitude (5-6% of the control) at ISIs of 0.15-0.3 ms and recovered at longer ISIs. At dichotic stimulation, the deepest suppression of the test response occurred at ISIs of 0.5-0.7 ms. The response was slightly suppressed at short ISIs (0.15-0.3 ms) and recovered at ISIs longer than 0.5-0.7 ms. The relation of parameters of the forward masking to echolocation in dolphins is discussed.</p>","PeriodicalId":54862,"journal":{"name":"Journal of Comparative Physiology A-Neuroethology Sensory Neural and Behavioral Physiology","volume":" ","pages":"751-759"},"PeriodicalIF":1.9,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41160301","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"心理学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}