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  • Population structure of short-finned pilot whales in the oceanic archipelago of Madeira based on photo-identification and genetic analyses: implications for conservation
    Publication . Alves, Filipe; Quérouil, Sophie; Dinis, Ana; Nicolau, Cátia; Ribeiro, Cláudia; Freitas, Luís; Kaufmann, Manfred; Fortuna, Caterina
    1. Pilot whales Globicephala spp. are known to display a hierarchical social pattern, but longitudinal data to infer population structure of short-finned pilot whales Globicephala macrorhynchus are rare. 2. Using data collected between 2003-2011 in the oceanic archipelago of Madeira, the grouping structure of short-finned pilot whales was studied using photo-identification methods and mtDNA sequences and microsatellite markers to test the hypotheses that (1) there is at least one pelagic and one or more island-associated communities, and (2) groups are made of related individuals, with a matrilineal social structure. 3. Pilot whales demonstrated a large degree of variability in site fidelity, including residents (up to 14-year interval), regular visitors and transients. The social and temporal analyses revealed a well-differentiated society with long-lasting relationships (of years). The genetic analyses suggested that individuals of the three residency patterns may not be genetically isolated, and that small groups are made up of related individuals, suggesting some degree of social philopatry, while large groups are probably temporary associations of smaller groups. 4. It is proposed that the pilot whales encountered in Madeira belong to a single population encompassing several clans, possibly three clans of island-associated whales and others of transients, each containing two to three matrilineal pods, each with a mean of 15 individuals (SD=9, range: 4-29). We suggest that the clans interact for mating purposes when they meet. 5. For management decisions, it is considered that the island-associated whales should not be regarded as demographically independent populations, but instead as stable social entities to be included in governmental management plans and requiring periodic evaluation of their status. The high proportion of marked individuals and low rate of mark change encourages further research in this species.
  • Bottlenose dolphin Tursiops truncatus group dynamics, site fidelity, residency and movement patterns in the Madeira Archipelago (North-East Atlantic)
    Publication . Dinis, A.; Alves, F.; Nicolau, C.; Ribeiro, C.; Kaufmann, M.; Cañadas, A.; Freitas, L
    The bottlenose dolphin Tursiops truncatus is one of the most frequently sighted cetacean species in the Madeira Archipelago (North-East Atlantic); however, little is known about its population ecology in these waters. Photo-identification undertaken during systematic, non-systematic and opportunistic surveys conducted between 2001 and 2012 was used to assess group dynamics, site fidelity, residency and movement patterns of bottlenose dolphins in the archipelago. Three different patterns of residency were assigned, based on individual sighting histories. Group size ranged between 2 and 90 individuals (median 12), with no significant monthly difference in group size. All resident dolphins formed a core, complex network with migrant and transient dolphins. In contrast, satellite clusters were formed exclusively by transient individuals. The lagged identification rate (LIR) model indicated that a dolphin remained in the area for an average of 90 days, whereas the average time an individual spent outside the study area was approximately 313 days. The Madeira Archipelago appears to be only a section of a much larger home range for this population, with the dolphins exhibiting three different patterns of occurrence that associate with each other. The information gained about this poorly studied population contributes to the overall knowledge of bottlenose dolphins in insular oceanic habitats.
  • Determination of phthalic acid esters and di(2-ethylhexyl) adipate in fish and squid using the ammonium formate version of the QuEChERS method combined with gas chromatography mass spectrometry
    Publication . Sambolino, Annalisa; Ortega-Zamora, Cecilia; González-Sálamo, Javier; Dinis, Ana; Cordeiro, Nereida; Canning-Clode, João; Hernández-Borges, Javier
    In the present study, the ammonium formate version of the QuEChERS method, considered highly advantageous in relation to instrument maintenance and other issues, was applied for the first time to extract a group of twelve phthalic acid esters (PAEs, i.e. dipropyl phthalate, DPP; diisobutyl phthalate, DIBP; dibutyl phthalate, DBP; diisopentyl phthalate, DIPP; di-n-pentyl phthalate, DNPP; dihexyl phthalate, DHP; butyl benzyl phthalate, BBP; dicyclohexyl phthalate, DCHP; di(2-ethylhexyl) phthalate, DEHP; di-n-octyl phthalate, DNOP; diisononyl phthalate, DINP; and diisodecyl phthalate, DIDP) and one adipate (di(2-ethylhexyl) adipate, DEHA) from two species of fish (Scomber colias and Katsuwonus pelamis) and one of squid (Loligo gahi). The method was validated in terms of linearity, trueness and matrix effects. Determination coefficients (R2 ) for matrix-matched calibration curves were higher than 0.99 in all cases, being the lowest calibration levels in the range 0.5–10 ng/g. Mean recovery values were between 70 and 117% with relative standard deviation values ≤20%. Matrix effects were soft (between − 20 and +20%) for most analytes and matrices, except in squid samples, which was mostly medium with a moderate ion suppression. The analysis of 10 samples of each type showed the presence of DIBP, DBP and DEHP at concentrations up to 44.2 ± 2.1 ng/g of wet weight in some of the samples and species, still not representing concerning values when considering the daily intake of such species of seafood in the human diet (tolerable daily intake -TDI- values were not exceeded). Results demonstrated that the ammonium formate version of the QuEChERS method can be applied with success for the extraction and determination of the selected PAEs and DEHA in fish and squid samples.
  • Daytime dive characteristics from six short-finned pilot whales Globicephala macrorhynchus off Madeira Island
    Publication . Alves, Filipe; Dinis, A.; Ribeiro, C.; Nicolau, C.; Kaufmann, M.; Fortuna, C. M.; Freitas, L.
  • Social structure of a population of bottlenose dolphins (Tursiops truncatus) in the oceanic archipelago of Madeira, Portugal
    Publication . Dinis, Ana; Alves, Filipe; Nicolau, Cátia; Ribeiro, Cláudia; Kaufmann, Manfred; Cañadas, Ana; Freitas, Luís
    In order to investigate social structure, 11 years of individual photo-identification data of bottlenose dolphin were analysed. We examined the type of association indices between pairs of identified individuals; the patterns of affiliation between indi vidual dolphins and the probabilities of association between individuals over time. Between 2001 and 2012, there were 272 encounters which resulted in the identification of 501 individuals. The discovery curve resulting from the photo-identification analysis indicated an open population with regular recruitment of new individuals. All individuals were found to be asso ciated at an association index of ,0.05. A total of 291 individuals recorded from 2004 to 2012 were used to assess the tem poral pattern of the social structure. The model fit to the Standardized Lagged Association Rate (SLAR) that best described the studied bottlenose dolphin population was ‘casual acquaintances’, and the analysis of associations over time showed a decreasing SLAR curve that falls until reaching the null rate, confirming random associations. The decline of the SLAR curve after 500 days (1.4 years) suggests disassociation over that time period which can be explained by demographic events such as mortality or emigration. In an open ocean habitat like Madeira this is not unexpected, as there are neither geographic boundaries nor enclosed environments. This population presented a dynamic and fluctuating social structure, where groups change in size and composition. In future conservation efforts this population should be considered as one large community, where individuals associate, disassociate and reassociate with each other over time.
  • Ecology and conservation of bottlenose dolphins in Madeira Archipelago, Portugal
    Publication . Dinis, Ana Margarida Brites Caetano; Kaufmann, Manfred Josef; Cañadas, Ana
    This study aimed to provide an insight on the ecology of the bottlenose dolphin population in Madeira archipelago. To achieve this, population structure; group dynamics, site-fidelity, residency and movement patterns within and out of the study area; survival and abundance estimates and spatial and temporal distribution and habitat preferences related to physiographic parameters using data collected between 2001-2011, were investigated. Photo-identification data analysis revealed strong evidences that bottlenose dolphins seen in the archipelago of Madeira belong to an open population with regular recruitment of new animals to the area. This population exhibited a typical fission-fusion society, in which short-term acquaintances prevail, with only a few long-lasting associations. Photo-identification methods demonstrated that there is a large variability in residency pattern, with resident, transient and migrant individuals. Only a small number of dolphins were found to be resident (4.3%). Social network diagram as well as SLAR analysis supported the existence of a mixed population of residents, migrants and transients. Mark-recapture methods estimated a high survival rate, within the range of other long-lived cetacean species. The resident community is composed of app. 180 individuals. In addition, around 400 dolphins of different residency patterns were found to use the south area of Madeira Island. Spatial distribution indicated that bottlenose dolphins were regularly found in shallow and closer to shore areas, suggesting the existence of biological processes influenced by bathymetry. Moreover, temporal patterns revealed no strong seasonal fluctuation in the presence of bottlenose dolphins in Madeira archipelago waters. Bottlenose dolphins are listed under the Annex II of the EU habitats Directive that requires the designation of Special Areas of Conservation (SACs) for this species; as such, the knowledge gained through this work can be used by governmental authorities to the establishment and management of areas for the conservation of bottlenose dolphin in Madeira archipelago.
  • Optimization and validation of a micro–QuEChERS method for phthalates detection in small samples of cetacean blubber
    Publication . Sambolino, Annalisa; Rodriguez, Marta; De la Fuente, Jesus; Arbelo, Manuel; Fernández, Antonio; Kaufmann, Manfred; Cordeiro, Nereida; Dinis, Ana
    In this study, an innovative method was developed to detect and quantify phthalates in fresh cetacean blubber. An adaptation of the ammonium formate QuEChERS method was used and adapted as a micro-extraction for small quantities of samples. Significantly, this technique utilized minimal quantities of reagents and salts, with the additional implementation of rigorous Quality Assurance/Quality Control protocols to further reduce background contamination. To ensure the reliability of this method, comprehensive validation procedures were conducted, with a specific focus on two widely studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus). Determination coefficients (R2) for matrix-matched calibration were >0.93 with limits of quantifications (LOQ) of the method in the range of 5–10 ng/g. Mean recovery values were between 40 and 100 %. This novel methodology holds particular relevance for environmental research studies, offering the capability to detect emerging contaminants with minimal sample requirements. This aspect is particularly valuable in investigations that involve free-ranging animals and rely on biopsy sampling. It allows for the assessment of contaminant levels in healthy individuals within wild populations, enhancing our understanding of ecological impacts and potential conservation measures. • A micro-extraction adaptation of the ammonium formate QuEChERS method was developed and applied to a small quantity of fresh cetacean blubber to detect phthalates. • Small quantities of reagents and salts were used, and additional Quality Assurance/ Quality Control procedures were taken to further minimize background contamination. • Method validation was carried out for two cosmopolitan and extensively studied cetacean species: the common bottlenose dolphin (Tursiops truncatus) and the short-finned pilot whale (Globicephala macrorhynchus).
  • Seasonal variation in microplastics and zooplankton abundances and characteristics: the ecological vulnerability of an oceanic island system
    Publication . Sambolino, Annalisa; Herrera, Inma; Álvarez, Soledad; Rosa, Alexandra; Alves, Filipe; Canning-Clode, João; Cordeiro, Nereida; Dinis, Ana; Kaufmann, Manfred
    The ingestion of microplastics (MPs - plastic particles <5 mm) by planktivorous organisms represents a signif icant threat to marine food webs. To investigate how seasonality might affect plastic intake in oceanic islands' ecosystems, relative abundances and composition of MPs and mesozooplankton samples collected off Madeira Island (NE Atlantic) between February 2019 and January 2020 were analysed. MPs were found in all samples, with fibres accounting for 89 % of the particles. MPs and zooplankton mean abundance was 0.262 items/m3 and 18.137 individuals/m3 , respectively. Their monthly variations follow the seasonal fluctuation of environmental parameters, such as currents, chlorophyll-a concentration, sea surface temperature and precipitation intensity. A higher MPs/zooplankton ratio was recorded in the warm season (May-Oct), reaching 0.068 items/individual when considering large-sized particles (1000–5000 μm). This is the first study to assess the seasonal variability of MPs in an oceanic island system providing essential information respecting its ecological impact in pelagic environments.
  • Survival and abundance of short-finned pilot whales in the archipelago of Madeira, NE Atlantic
    Publication . Alves, Filipe; Dinis, Ana; Nicolau, Cátia; Ribeiro, Cláudia; Kaufmann, Manfred; Fortuna, Caterina; Freitas, Luís
    Estimates of population parameters for the short-finned pilot whale, Globicephala macrorhynchus, are scarce in literature, contributing to an International Union for Conservation of Nature (IUCN) status of Data Deficient. In this study, photo-identi fication data collected over 7 yr from Madeira were used to estimate for the first time survivorship, capture probability, and abundance in this species using mark-recap ture methodology. The Cormack-Jolly-Seber model estimated that the adult island associated (i.e., resident and regular visitor) whales had a constant survival rate of 0.960 (95% CI: 0.853–0.990) and an annual capture probability varying between 0.372 (CI: 0.178–0.619) and 0.843 (CI: 0.619–0.947). A parameterization of the Jolly-Seber model estimated that 140 island-associated whales (CI: 131–151) used the area throughout the course of the study. Based on a closed population model, the most precise (lower CV) annual estimate of the total number of pilot whales using the southern and eastern waters of Madeira (~900 km2 ) in a 3 mo period covering summer/autumn was 334 animals (CI: 260–437). No trend was observed. Despite including biases, the approach used in this study provided plausible estimates of population parameters, which can contribute to the regional conservation strategies.
  • Spatial and temporal distribution of bottlenose dolphins, Tursiops truncatus, in the Madeira archipelago, NE Atlantic
    Publication . Dinis, Ana; Carvalho, A.; Alves, F.; Nicolau, C.; Ribeiro, C.; Kaufmann, M.; Cañadas, A.; Freitas, L.
    One of the first steps in understanding the relationships between populations and their habitats is to determine which areas they use with higher frequency. This study used systematic and non-systematic survey data from 2001-2002 and 2004-2012 to determine encounter rates and investigate temporal and spatial distribution of bottlenose dolphins around Madeira, Desertas and Porto Santo islands. A total 24,914 km of search effort was carried out and 199 sightings were recorded. Highest encounter rates were found off the east coast of Madeira and off Porto Santo. Moreover, higher encounter rates occurred over bathymetries ranging between 500-1,000 m during systematic surveys whereas in non systematic surveys relative high encounter rates were found in depths of 2,000-2,500m. Most dolphins were found to be distributed in depths <1,000m and at no more than 10 km offshore indicating a preference for shallower waters. Dolphins were sighted during the whole year and there were no significant differences in encounter rate between months. These results suggest the existence of preferential areas for this species based on static bathymetric features. The fact that the dolphins prefer inshore areas that are more exposed to anthropogenic activities should be taken into account when discussing bottlenose dolphin conservation measures in the Madeira archipelago.