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AbstractUnifying models have shown that the amount of space used by animals (e.g., activity space, home range) scales allometrically with body mass for terrestrial taxa; however, such relationships are far less clear for marine species. We compiled movement data from 1,596 individuals across 79 taxa collected using a continental passive acoustic telemetry network of acoustic receivers to assess allometric scaling of activity space. We found that ectothermic marine taxa do exhibit allometric scaling for activity space, with an overall scaling exponent of 0.64. However, body mass alone explained only 35% of the variation, with the remaining variation best explained by trophic position for teleosts and latitude for sharks, rays, and marine reptiles. Taxon-specific allometric relationships highlighted weaker scaling exponents among teleost fish species (0.07) than sharks (0.96), rays (0.55), and marine reptiles (0.57). The allometric scaling relationship and scaling exponents for the marine taxonomic groups examined were lower than those reported from studies that had collated both marine and terrestrial species data derived using various tracking methods. We propose that these disparities arise because previous work integrated summarized data across many studies that used differing methods for collecting and quantifying activity space, introducing considerable uncertainty into slope estimates. Our findings highlight the benefit of using large-scale, coordinated animal biotelemetry networks to address cross-taxa evolutionary and ecological questions.
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Organismos Acuáticos , Peces , Animales , Fenómenos de Retorno al Lugar HabitualRESUMEN
Cell fate determination depends in part on the establishment of specific transcriptional programs of gene expression. These programs result from the interpretation of the genomic cis-regulatory information by sequence-specific factors. Decoding this information in sequenced genomes is an important issue. Here, we developed statistical analysis tools to computationally identify the cis-regulatory elements that control gene expression in a set of coregulated genes. Starting with a small number of validated and/or predicted cis-regulatory modules (CRMs) in a reference species as a training set, but with no a priori knowledge of the factors acting in trans, we computationally predicted transcription factor binding sites (TFBSs) and genomic CRMs underlying coregulation. This method was applied to the gene expression program active in Drosophila melanogaster sensory organ precursor cells (SOPs), a specific type of neural progenitor cells. Mutational analysis showed that four, including one newly characterized, out of the five top-ranked families of predicted TFBSs were required for SOP-specific gene expression. Additionaly, 19 out of the 29 top-ranked predicted CRMs directed gene expression in neural progenitor cells, i.e., SOPs or larval brain neuroblasts, with a notable fraction active in SOPs (11/29). We further identified the lola gene as the target of two SOP-specific CRMs and found that the lola gene contributed to SOP specification. The statistics and phylogeny-based tools described here can be more generally applied to identify the cis-regulatory elements of specific gene regulatory networks in any family of related species with sequenced genomes.
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Drosophila melanogaster/genética , Redes Reguladoras de Genes , Genoma/genética , Elementos Reguladores de la Transcripción/genética , Algoritmos , Animales , Secuencia de Bases , Sitios de Unión/genética , Biología Computacional/métodos , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/clasificación , Drosophila melanogaster/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Inmunohistoquímica , Hibridación in Situ , Larva/genética , Larva/crecimiento & desarrollo , Filogenia , Interferencia de ARN , Reproducibilidad de los Resultados , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Lipid and fatty acid datasets are commonly used to assess the nutritional composition of organisms, trophic ecology, and ecosystem dynamics. Lipids and their fatty acid constituents are essential nutrients to all forms of life because they contribute to biological processes such as energy flow and metabolism. Assessment of total lipids in tissues of organisms provides information on energy allocation and life-history strategies and can be an indicator of nutritional condition. The analysis of an organism's fatty acids is a widely used technique for assessing nutrient and energy transfer, and dietary interactions in food webs. Although there have been many published regional studies that assessed lipid and fatty acid compositions, many only report the mean values of the most abundant fatty acids. There are limited individual records available for wider use in intercomparison or macro-scale studies. This dataset consists of 4856 records of individual and pooled samples of at least 470 different marine consumer species sampled from tropical, temperate, and polar regions around Australia and in the Southern, Indian, and Pacific Oceans from 1989 to 2018. This includes data for a diverse range of taxa (zooplankton, fish, cephalopods, chondrichthyans, and marine mammals), size ranges (0.02 cm to ~13 m), and that cover a broad range of trophic positions (2.0-4.6). When known, we provide a record of species name, date of sampling, sampling location, body size, relative (%) measurements of tissue-specific total lipid content and abundant fatty acids, and absolute content (mg 100 g-1 tissue) of eicosapentaenoic acid (EPA, 20:5n3) and docosahexaenoic acid (DHA, 22:6n3) as important long-chain (≥C20 ) polyunsaturated omega-3 fatty acids. These records form a solid basis for comparative studies that will facilitate a broad understanding of the spatial and temporal distribution of marine lipids globally. The dataset also provides reference data for future dietary assessments of marine predators and model assessments of potential impacts of climate change on the availability of marine lipids and fatty acids. There are 480 data records within our data file for which the providers have requested that permission for reuse be granted, with the likely condition that they are included as a coauthor on the reporting of the dataset. Records with this condition are indicated by a "yes" under "Conditions_of_data_use" in Data S1: Marineconsumer_FAdata.csv (see Table 2 in Metadata S1 for more details). For all other data records marked as "No" under "Conditions_of_data_use," there are no copyright restrictions for research and/or teaching purposes. We request that users acknowledge use of the data in publications, research proposals, websites, and other outlets via formal citation of this work and original data sources as applicable.
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Ecosistema , Ácidos Grasos , Animales , Ácidos Grasos/análisis , Ácidos Grasos/metabolismo , Cadena Alimentaria , Peces , Zooplancton , MamíferosRESUMEN
Multiscale analysis of morphogenesis requires to follow and measure in real-time the in vivo behaviour of large numbers of individual cells over long period of time. Despite recent progress, the large-scale automated tracking of cells in developing embryos and tissues remains a challenge. Here we describe a genetic tool for the random and sparse labelling of individual cells in developing Drosophila tissues. This tool is based on the conditional expression of a nuclear HaloTag protein that can be fluorescently labelled upon the irreversible binding of a cell permeable synthetic ligand. While the slow maturation of genetically encoded fluorescent renders the tracking of individual cells difficult in rapidly dividing tissues, nuclear HaloTag proteins allowed for rapid labelling of individual cells in cultured imaginal discs. To study cell shape changes, we also produced an HaloTag version of the actin-bound protein LifeAct. Since sparse labelling facilitates cell tracking, nuclear HaloTag reporters will be useful for the single-cell analysis of fate dynamics in Drosophila tissues cultured ex vivo.
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Rastreo Celular , Análisis de la Célula Individual , Animales , DrosophilaRESUMEN
Understanding the relationship between mercury in seafood and the distribution of oceanic methylmercury is key to understand human mercury exposure. Here, we determined mercury concentrations in muscle and blood of bigeye and yellowfin tunas from the Western and Central Pacific. Results showed similar latitudinal patterns in tuna blood and muscle, indicating that both tissues are good candidates for mercury monitoring. Complementary tuna species analyses indicated species- and tissue- specific mercury patterns, highlighting differences in physiologic processes of mercury uptake and accumulation associated with tuna vertical habitat. Tuna mercury content was correlated to ambient seawater methylmercury concentrations, with blood being enriched at a higher rate than muscle with increasing habitat depth. The consideration of a significant uptake of dissolved methylmercury from seawater in tuna, in addition to assimilation from food, might be interesting to test in models to represent the spatiotemporal evolutions of mercury in tuna under different mercury emission scenarios.
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Mercurio , Compuestos de Metilmercurio , Animales , Humanos , Mercurio/análisis , Compuestos de Metilmercurio/análisis , Músculos/química , Océano Pacífico , Agua de Mar , AtúnRESUMEN
Knowledge of the three-dimensional movement patterns of elasmobranchs is vital to understand their ecological roles and exposure to anthropogenic pressures. To date, comparative studies among species at global scales have mostly focused on horizontal movements. Our study addresses the knowledge gap of vertical movements by compiling the first global synthesis of vertical habitat use by elasmobranchs from data obtained by deployment of 989 biotelemetry tags on 38 elasmobranch species. Elasmobranchs displayed high intra- and interspecific variability in vertical movement patterns. Substantial vertical overlap was observed for many epipelagic elasmobranchs, indicating an increased likelihood to display spatial overlap, biologically interact, and share similar risk to anthropogenic threats that vary on a vertical gradient. We highlight the critical next steps toward incorporating vertical movement into global management and monitoring strategies for elasmobranchs, emphasizing the need to address geographic and taxonomic biases in deployments and to concurrently consider both horizontal and vertical movements.
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The stereotyped arrangement of sensory bristles on the adult fly thorax arises from a self-organized process, in which inhibitory Notch signaling both delimits proneural stripes and singles out sensory organ precursor cells (SOPs). A dynamic balance between proneural factors and Enhancer of split-HLH (E(spl)-HLH) Notch targets underlies patterning, but how this is regulated is unclear. Here, were identify two classes of E(spl)-HLH factors, whose expression both precedes and delimits proneural activity, and is dependent on proneural activity and required for proper SOP spacing within the stripes, respectively. These two classes are partially redundant, since a member of the second class, that is normally cross-repressed by members of the first class, can functionally compensate for their absence. The regulation of specific E(spl)-HLH genes by proneural factors amplifies the response to Notch as SOPs are being selected, contributing to patterning dynamics in the notum, and likely operates in other developmental contexts.
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Traditionally, large planktivorous elasmobranchs have been thought to predominantly feed on surface zooplankton during daytime hours. However, the recent application of molecular methods to examine long-term assimilated diets, has revealed that these species likely gain the majority from deeper or demersal sources. Signature fatty acid analysis (FA) of muscle tissue was used to examine the assimilated diet of the giant manta ray Mobula birostris, and then compared with surface zooplankton that was collected during feeding and non-feeding events at two aggregation sites off mainland Ecuador. The FA profiles of M. birostris and surface zooplankton were markedly different apart from similar proportions of arachidonic acid, which suggests daytime surface zooplankton may comprise a small amount of dietary intake for M. birostris. The FA profile of M. birostris muscle was found to be depleted in polyunsaturated fatty acids, and instead comprised high proportions of 18:1ω9 isomers. While 18:1ω9 isomers are not explicitly considered dietary FAs, they are commonly found in high proportions in deep-sea organisms, including elasmobranch species. Overall, the FA profile of M. birostris suggests a diet that is mesopelagic in origin, but many mesopelagic zooplankton species also vertically migrate, staying deep during the day and moving to shallower waters at night. Here, signature FA analysis is unable to resolve the depth at which these putative dietary items were consumed and how availability of this prey may drive distribution and movements of this large filter-feeder.
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Ácidos Grasos Insaturados/metabolismo , Ácidos Grasos/metabolismo , Conducta Alimentaria/fisiología , Rajidae/metabolismo , Animales , Ecuador , Músculos/metabolismo , Rajidae/fisiología , Zooplancton/metabolismoRESUMEN
In epithelia, mitotic cells round up and push against their neighbors to divide. Mitotic rounding results from increased assembly of F-actin and cortical recruitment of Myosin II, leading to increased cortical stability. Whether this process is developmentally regulated is not well known. Here, we examined the regulation of cortical stability in Sensory Organ Precursor cells (SOPs) in the Drosophila pupal notum. SOPs differed in apical shape and actomyosin dynamics from their epidermal neighbors prior to division, and appeared to have a more rigid cortex at mitosis. We identified RhoGEF3 as an actin regulator expressed at higher levels in SOPs, and showed that RhoGEF3 had in vitro GTPase Exchange Factor (GEF) activity for Cdc42. Additionally, RhoGEF3 genetically interacted with both Cdc42 and Rac1 when overexpressed in the fly eye. Using a null RhoGEF3 mutation generated by CRISPR-mediated homologous recombination, we showed using live imaging that the RhoGEF3 gene, despite being dispensable for normal development, contributed to cortical stability in dividing SOPs. We therefore suggest that cortical stability is developmentally regulated in dividing SOPs of the fly notum.
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The emergence of spatial patterns in developing multicellular organisms relies on positional cues and cell-cell communication. Drosophila sensory organs have informed a paradigm in which these operate in two distinct steps: Prepattern factors drive localized proneural activity, then Notch-mediated lateral inhibition singles out neural precursors. Here we show that self-organization through Notch signaling also establishes the proneural stripes that resolve into rows of sensory bristles on the fly thorax. Patterning, initiated by a gradient of Delta ligand expression, progresses through inhibitory signaling between and within stripes. Thus, Notch signaling can support self-organized tissue patterning as a prepattern is transduced by cell-cell interactions into a refined arrangement of cellular fates.
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Tipificación del Cuerpo/fisiología , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriología , Receptores Notch/metabolismo , Órganos de los Sentidos/embriología , Animales , Tipificación del Cuerpo/genética , Comunicación Celular , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Modelos Teóricos , Receptores Notch/genética , Órganos de los Sentidos/citología , Transducción de Señal , Células Madre/metabolismo , Tórax/inervaciónRESUMEN
The characterization of diet for the giant manta ray Manta birostris has been problematic given their large-scale movement patterns and the difficulty in obtaining stomach contents from this species. The large majority of existing information is based on observational data limited to feeding events at the sea surface during daylight. Recently discovered aggregation sites for the giant manta ray off mainland Ecuador are some of the most accessible to date and provide a unique opportunity for researchers to gather much needed information on this elusive species. To assess how important surface zooplankton is to giant manta ray diet, we conducted stable isotope analysis (15N and 13C) on M. birostris muscle and surface zooplankton. Trophic position estimates placed M. birostris overall at a secondary consumer level of approximately 3.4 but there was large variation in δ15N and δ13C values among individuals. Manta birostris muscle tissue δ13C values were also not consistent with this species feeding predominantly on surface zooplankton and suggest that the majority of dietary intake is of mesopelagic origin. Given the conservative life history and fisheries pressure on large planktivores, knowledge of their trophic role and foraging strategies is essential to better understand their ecology and develop effective conservation measures.
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Large tropical and sub-tropical marine animals must meet their energetic requirements in a largely oligotrophic environment. Many planktivorous elasmobranchs, whose thermal ecologies prevent foraging in nutrient-rich polar waters, aggregate seasonally at predictable locations throughout tropical oceans where they are observed feeding. Here we investigate the foraging and oceanographic environment around Lady Elliot Island, a known aggregation site for reef manta rays Manta alfredi in the southern Great Barrier Reef. The foraging behaviour of reef manta rays was analysed in relation to zooplankton populations and local oceanography, and compared to long-term sighting records of reef manta rays from the dive operator on the island. Reef manta rays fed at Lady Elliot Island when zooplankton biomass and abundance were significantly higher than other times. The critical prey density threshold that triggered feeding was 11.2 mg m-3 while zooplankton size had no significant effect on feeding. The community composition and size structure of the zooplankton was similar when reef manta rays were feeding or not, with only the density of zooplankton changing. Higher zooplankton biomass was observed prior to low tide, and long-term (~5 years) sighting data confirmed that more reef manta rays are also observed feeding during this tidal phase than other times. This is the first study to examine prey availability at an aggregation site for reef manta rays and it indicates that they feed in locations and at times of higher zooplankton biomass.
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Arrecifes de Coral , Conducta Alimentaria , Conducta Predatoria , Rajidae/fisiología , Movimientos del Agua , Animales , Australia , ZooplanctonRESUMEN
We present the first photographic evidence of the presence of the giant manta ray Manta birostris in east Australian waters. Two individuals were photographed off Montague Island in New South Wales and off the north east coast of Tasmania, during summer 2012 and 2014, respectively. These sightings confirm previous unverified reports on the species occurrence and extend the known distribution range of M. birostris to 40°S. We discuss these findings in the context of the species' migratory behaviour, the regional oceanography along the south east Australian coastline and local productivity events.
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Notch signaling depends on regulated intracellular trafficking of the receptor and its ligands (Kopan and Ilagan, Cell 137:216-233, 2009; Le Borgne et al., Development 132:1751-1762, 2005). Here we describe two methods to study the intracellular trafficking of Notch and Delta in Drosophila. First, an ex vivo antibody uptake assay is used to monitor endocytosis of Notch and Delta by living cells in dissected explants (Le Borgne and Schweisguth, Dev Cell 5:139-148, 2003). Second, real-time imaging of fluorescent proteins that are expressed at physiological levels is used to study trafficking of Notch in living flies (Venken et al., Science 314:1747-1751, 2006; Couturier et al., Nat Cell Biol 14, 131-139, 2012).
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Anticuerpos/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila/citología , Drosophila/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Proteínas de la Membrana/metabolismo , Receptores Notch/metabolismo , Animales , Anticuerpos/análisis , Disección/métodos , Proteínas de Drosophila/análisis , Endocitosis , Proteínas Fluorescentes Verdes/análisis , Proteínas Fluorescentes Verdes/metabolismo , Inmunohistoquímica/métodos , Péptidos y Proteínas de Señalización Intracelular/análisis , Sustancias Luminiscentes/análisis , Sustancias Luminiscentes/metabolismo , Proteínas de la Membrana/análisis , Microscopía/métodos , Imagen Óptica/métodos , Transporte de Proteínas , Receptores Notch/análisisRESUMEN
Signaling and endocytosis are highly integrated processes that regulate cell fate. In the Drosophila melanogaster sensory bristle lineages, Numb inhibits the recycling of Notch and its trafficking partner Sanpodo (Spdo) to regulate cell fate after asymmetric cell division. In this paper, we have used a dual GFP/Cherry tagging approach to study the distribution and endosomal sorting of Notch and Spdo in living pupae. The specific properties of GFP, i.e., quenching at low pH, and Cherry, i.e., slow maturation time, revealed distinct pools of Notch and Spdo: cargoes exhibiting high GFP/low Cherry fluorescence intensities localized mostly at the plasma membrane and early/sorting endosomes, whereas low GFP/high Cherry cargoes accumulated in late acidic endosomes. These properties were used to show that Spdo is sorted toward late endosomes in a Numb-dependent manner. This dual-tagging approach should be generally applicable to study the trafficking dynamics of membrane proteins in living cells and tissues.
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Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Endosomas/metabolismo , Hormonas Juveniles/metabolismo , Receptores Notch/metabolismo , Animales , Cadherinas/metabolismo , División Celular , Drosophila melanogaster/citología , Células Epiteliales/metabolismo , Proteínas Fluorescentes Verdes/metabolismo , Concentración de Iones de Hidrógeno , Proteínas de Microfilamentos/metabolismo , Microscopía Fluorescente , Transporte de Proteínas , Proteínas Recombinantes de Fusión/metabolismo , Imagen de Lapso de TiempoRESUMEN
Numb acts as a cell-fate determinant during asymmetric and stem cell divisions in both vertebrates and invertebrates [1, 2]. In Drosophila, Numb is unequally segregated in asymmetrically dividing sensory organ precursor cells (SOPs). Numb is inherited by the pIIb cell (Notch OFF) and is absent from the pIIa cell (Notch ON) [3, 4]. Numb is required to establish directional Notch signaling during cytokinesis [3, 5-7]. Using real-time imaging of a functional GFP-tagged Numb, we show that Numb relocalizes during cytokinesis from the basal cortex of pIIb to subapical endosomes. This relocalization appeared to depend on its interaction with the α-adaptin [8, 9]. Live imaging of Sanpodo (Spdo), a membrane protein interacting with Numb and regulating the trafficking of Notch [6, 7, 10-15], revealed that Spdo is internalized during cytokinesis and coaccumulates with Numb in pIIb endosomes. Using a GFP-tagged Notch [6], we found that Notch coaccumulates with Spdo in a Numb-dependent manner in these pIIb endosomes. Numb was, however, dispensable for the internalization of Notch and Spdo. We propose that Numb interacts with internalized Spdo-Notch oligomers at sorting endosomes and inhibits the recycling of Notch, thereby creating an asymmetry in Notch distribution along the pIIa-pIIb interface and regulating binary fate choice.
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División Celular Asimétrica/fisiología , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiología , Drosophila/fisiología , Endosomas/metabolismo , Hormonas Juveniles/fisiología , Receptores Notch/metabolismo , Secuencia de Aminoácidos , Animales , Cromosomas Artificiales Bacterianos , Proteínas de Drosophila/genética , Hormonas Juveniles/genética , Microscopía Fluorescente , Modelos Biológicos , Datos de Secuencia Molecular , Células-Madre Neurales/fisiología , Transporte de Proteínas/fisiología , Células Receptoras Sensoriales/fisiologíaRESUMEN
Assessing the trophic role and interaction of an animal is key to understanding its general ecology and dynamics. Conventional techniques used to elucidate diet, such as stomach content analysis, are not suitable for large threatened marine species. Non-lethal sampling combined with biochemical methods provides a practical alternative for investigating the feeding ecology of these species. Stable isotope and signature fatty acid analyses of muscle tissue were used for the first time to examine assimilated diet of the reef manta ray Manta alfredi, and were compared with different zooplankton functional groups (i.e. near-surface zooplankton collected during manta ray feeding events and non-feeding periods, epipelagic zooplankton, demersal zooplankton and several different zooplankton taxa). Stable isotope δ(15)N values confirmed that the reef manta ray is a secondary consumer. This species had relatively high levels of docosahexaenoic acid (DHA) indicating a flagellate-based food source in the diet, which likely reflects feeding on DHA-rich near-surface and epipelagic zooplankton. However, high levels of ω6 polyunsaturated fatty acids and slightly enriched δ(13)C values in reef manta ray tissue suggest that they do not feed solely on pelagic zooplankton, but rather obtain part of their diet from another origin. The closest match was with demersal zooplankton, suggesting it is an important component of the reef manta ray diet. The ability to feed on demersal zooplankton is likely linked to the horizontal and vertical movement patterns of this giant planktivore. These new insights into the habitat use and feeding ecology of the reef manta ray will assist in the effective evaluation of its conservation needs.
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Elasmobranquios/fisiología , Ácidos Grasos Omega-6/metabolismo , Conducta Alimentaria/fisiología , Zooplancton , Animales , Isótopos de Carbono/análisis , Ácidos Grasos Omega-6/análisis , Isótopos de Nitrógeno/análisisRESUMEN
Cell-fate diversity can be generated by the unequal segregation of the Notch regulator Numb at mitosis in both vertebrates and invertebrates. Whereas the mechanisms underlying unequal inheritance of Numb are understood, how Numb antagonizes Notch has remained unsolved. Live imaging of Notch in sensory organ precursor cells revealed that nuclear Notch is detected at cytokinesis in the daughter cell that does not inherit Numb. Numb and Sanpodo act together to regulate Notch trafficking and establish directional Notch signalling at cytokinesis. We propose that unequal segregation of Numb results in increased endocytosis in one daughter cell, hence asymmetry of Notch at the cytokinetic furrow, directional signalling and binary fate choice.
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Citocinesis , Proteínas de Drosophila/metabolismo , Endocitosis , Hormonas Juveniles/metabolismo , Receptores Notch/metabolismo , Animales , Animales Modificados Genéticamente , Núcleo Celular/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/citología , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Dinaminas/genética , Dinaminas/metabolismo , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Hormonas Juveniles/genética , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Microscopía Fluorescente , Mutación , Unión Proteica , Interferencia de ARN , Receptores Notch/genética , Imagen de Lapso de TiempoRESUMEN
Manta rays Manta alfredi are present all year round at Lady Elliot Island (LEI) in the southern Great Barrier Reef, Australia, with peaks in abundance during autumn and winter. Drivers influencing these fluctuations in abundance of M. alfredi at the site remain uncertain. Based on daily count, behavioural, weather and oceanographic data collected over a three-year period, this study examined the link between the relative number of sightings of manta rays at LEI, the biophysical environment, and the habitat use of individuals around the LEI reef using generalised additive models. The response variable in each of the three generalised additive models was number of sightings (per trip at sea) of cruising, cleaning or foraging M. alfredi. We used a set of eleven temporal, meteorological, biological, oceanographic and lunar predictor variables. Results for cruising, cleaning and foraging M. alfredi explained 27.5%, 32.8% and 36.3% of the deviance observed in the respective models and highlighted five predictors (year, day of year, wind speed, chlorophyll-a concentration and fraction of moon illuminated) as common influences to the three models. There were more manta rays at LEI in autumn and winter, slower wind speeds, higher productivity, and around the new and full moon. The winter peak in sightings of foraging M. alfredi was found to precede peaks in cleaning and cruising activity around the LEI reef, which suggests that enhanced food availability may be a principal driver for this seasonal aggregation. A spatial analysis of behavioural observations highlighted several sites around the LEI reef as 'multi-purpose' areas where cleaning and foraging activities commonly occur, while the southern end of the reef is primarily a foraging area. The use of extensive citizen science datasets, such as those collected by dive operators in this study, is encouraged as they can provide valuable insights into a species' ecology.