Population structure, habitat use and conservation of short-finned pilot whales Globicephala macrorhynchus in the Archipelago of Madeira

This thesis provides information on the grouping structure, survival, abundance, dive characteristics and habitat preferences of short-finned pilot whales occurring in the oceanic archipelago of Madeira (Portugal, NE Atlantic), based on data collected between 2001-2011, and contributes for its conservation. Photo-identification methods and genetic analyses demonstrated that there is a large degree of variability in site fidelity, including resident, regular visitor and transient whales, and that they may not be genetically isolated. It is proposed that the pilot whales encountered in Madeira belong to a single population encompassing several clans, possibly three clans of island-associated (i.e. resident and regular visitor) whales and others of transients, each containing two to three matrilineal pods. Mark-recapture methods estimated that the island-associated community is composed of less than 150 individuals and that their survival rate is within the range of other long-lived cetacean species, and that around 300 whales of different residency patterns uses the southern area of the island of Madeira from mid-summer to mid-autumn. No significant trend was observed between years. Time-depth recorders deployed in adult whales during daytime revealed that they spend over ¾ of their time at the surface, that they have a low diving rate, and that transient whales also forage during their passage. The analyses of visual data collected from nautical and aerial line-transect surveys indicate a core/preferred habitat area in the south-east of the island of Madeira. That area is used for resting, socializing, foraging, breeding, calving and birthing. Thus, that area should be considered as an important habitat for this species, at least seasonally (during autumn) when the species is more abundant, and included in conservation plans. No direct threat needing urgent measures was identified, although the impact of some activities like whale-watching or marine traffic should be assessed.

Ecology and conservation of bottlenose dolphins in Madeira Archipelago, Portugal

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.

Gonad development and hormone titres in Loggerhead Sea Turtles (Caretta caretta) in the NE Atlantic

The study proposed to describe sexual development in pelagic stage loggerhead sea turtles Caretta caretta and compare this to hatchlings and adults. It is meant as an ontogenic approach, in order to understand reproductive development and population composition and their dynamics in the pelagic environment. The study focused on the pelagic loggerheads that are found in the waters offshore Madeira Island (Portugal) in the North-eastern Atlantic and use it as a developmental habitat. The innovating character of this work relied on the lack of any description regarding the gonad ontogenesis and reproductive development for the pelagic stage in any of the 7 existing sea turtle species, all of them in danger of extinction. Three methods were used to diagnose the sex of each juvenile individual and asses the level of reproductive development: (1) laparoscopy, (2) gonad biopsy and (3) the assessment of two sex steroids circulating levels, namely testosterone and estradiol. In order to cover all life stages and compare data obtained for the juvenile stage, hatchlings and nesting female adults were sampled at the nearest nesting rookery at Boa Vista Island in the Cape Verde Archipelago. Gonads from dead hatchlings were collected for gonad histology and blood was collected from nesting females for sex steroids assessment. Laparoscopies revealed to be a valid sexing method for the juvenile stage, since gonads are morphologically differentiated at these size classes. Moreover, laparoscopy was validated using gonad histology. Gonad histology of juveniles showed that gonads are already completely differentiated into ovaries or testes at the size classes examined, but development seems to be quiescent. Males present already developed seminiferous tubules with spermatogonia lining the interior of the seminiferous tubule. Female gonads present oocytes at different development stages, but only oocytes up to stage III were observed. The maximum oocyte diameter in each individual correlated with body size, suggesting that reproductive development is an on-going process in juvenile females. The circulating levels of both testosterone and estradiol in juveniles of both sexes were very low and consistently lower than the ones observed in the nesting females from Boa Vista Island. No bimodal distribution was found for any of the sex steroids analysed and thus circulating hormone levels were not a reliable tool for sexing juvenile individuals with a non-invasive technique. The ratio testosterone:estradiol did not show a bimodal distribution either. The levels of testosterone correlated with sea surface temperature. The fact that temperatures observed during this study were below 24ºC might have hindered a differential testosterone pattern between juvenile males and females. Sex ratios for this population were generated according to laparoscopy results and compared among years and size classes. An overall sex ratio of 2 females for each male was found, but they varied among size classes but not among years. Possible causes for the sex ratios observed are discussed. This study is a contribution to our knowledge on the pelagic stage of loggerhead turtles, namely on the population structure regarding sex ratio, which is a vital tool for implementing conservation strategies.

Argilas modificadas como catalisadores ácidos sólidos

Important bentonitic deposits are present in Porto Santo Island, part of the Madeira Archipelago. Several locations were selected and samples were collected and characterised. The bentonite obtained at Serra de Dentro (SD) was selected for further laboratorial work. The fine fraction of SD bentonite was purified using several methods and the sodium homoionic form was prepared. This was the starting material used in the three generic types of modifications: metal exchange, acid activation and pillaring. These modifications produce materials with markedly different acidic (e.g. Brönsted and/or Lewis acidity), textural (e.g. increase of the surface area and active site accessibility) or structural (e.g. creation of permanent porous structures) properties. The wide range of materials obtained (including reference clays counterparts) was characterised in terms of chemical, structural, textural and catalytic properties. Limonene is an important raw material produced in Portugal, and its aromatisation reaction was chosen for the catalytic characterisation of the clay catalysts prepared.

Contribution to the chemoreception capacity of juvenile Loggerhead sea turtles (Caretta caretta, L.)

Loggerhead sea turtle juveniles (Caretta caretta), pelagic stage, are found in waters of Madeira archipelago. Pelagic turtles are in the main growth phase of their life cycle and consequently higher energy needs. However, knowledge about the ecology of pelagic loggerhead sea turtles is still quite rudimentary, mainly about the mechanisms that lead them to find food in the vast ocean. Studies with other pelagic species, such as procellariiform birds, revealed that the olfactory system play an important role for the detection of feeding areas, through the detection of concentration peaks of DMS (dimethylsulfide), a scent compound that naturally exists in the marine environment and it is related to areas of high productivity. Based on the assumption that loggerhead sea turtles use a similar mechanism, behavioural experiments were conducted in order to analyze the chemoreception capacity to DMS (airborne chemoreception - theoretically responsible for the long distance detection of areas with food patches; and aquatic chemoreception - theoretically responsible for the short distance detection of preys). The first step was to observe if pelagic loggerheads demonstrate sensitivity to DMS and the second was to verify if they really use the DMS, in natural conditions, as an airborne cue to find areas where food patches might be available. Four juveniles of loggerhead sea turtles were tested in captivity and three wild turtles in the open ocean. The results of airborne chemoreception experiments in captivity revealed that one turtle clearly demonstrated sensitivity to DMS and the sea experiments confirmed this result. However, the experiments were not conclusive on the question whether the pelagic turtles actually use the DMS as an airborne cue to detect long distance food patches. In aquatic chemoreception experiments was not observed sensitivity to DMS by the three sea turtles tested. In the classical conditioning experiment, where DMS and food were given nearly at the same time revealed that after a certain period of time, the sea turtle tested did not associated the DMS stimulus with a possible food reward. The main cause of mortality of loggerhead sea turtles in Madeira waters is due to the accidental capture (bycatch) by deep pelagic longlines fishery which the target species is the black-scabbard (Aphanopus carbo) fish. Chub mackerel (Scomber japonicus) is one of the baits used in this fishery. Aquatic chemoreception experiments were conducted in order to evaluate the attractiveness of the chub mackerel for sea turtles. For the three sea turtles tested, the results showed that in 90% of the cases the sea turtles were extremely attracted by the underwater smell of this fish.

Content in metallic ions of wines from the Madeira and Azores archipelagos

This study determines for the first time Na, K, Ca, Mg, Fe, Cu, Zn, Mn, Sr, Li and Rb contents in wines from the archipelagos of Madeira and Azores (Portugal). The greater part of the mean content for the different parameters fell within the ranges described in the literature, except for sodium whose higher content may be due to the effect of marine spray. ANOVA was used to establish the metals with significant differences in mean content between the wines from both archipelagos, between table and liquor wines of Madeira, and between wines of Pico and Terceira Islands from the Azores archipelago. Principal component analysis shows differences in the wines according to the wine-making process and/or the equipment employed. Stepwise linear discriminant analysis achieves a good classification and validation of wines according to the archipelago of origin, and the island in the case of Azores wines.

New invertebrate vectors of Okadaic acid from the North Atlantic Waters - Portugal (Azores and Madeira) and Morocco

Okadaic acid and its analogues are potent phosphatase inhibitors that cause Diarrheic Shellfish Poisoning (DSP) through the ingestion of contaminated shellfish by humans. This group of toxins is transmitted worldwide but the number of poisoning incidents has declined over the last 20 years due to legislation and monitoring programs that were implemented for bivalves. In the summer of 2012 and 2013, we collected a total of 101 samples of 22 different species that were made up of benthic and subtidal organisms such echinoderms, crustaceans, bivalves and gastropods from Madeira, São Miguel Island (Azores archipelago) and the northwestern coast of Morocco. The samples were analyzed by UPLC-MS/MS. Our main objective was to detect new vectors for these biotoxins. We can report nine new vectors for these toxins in the North Atlantic: Astropecten aranciacus, Arbacia lixula, Echinaster sepositus, Holothuria sanctori, Ophidiaster ophidianus, Onchidella celtica, Aplysia depilans, Patella spp., and Stramonita haemostoma. Differences in toxin contents among the species were found. Even though low concentrations were detected, the levels of toxins that were present, especially in edible species, indicate the importance of these types of studies. Routine monitoring should be extended to comprise a wider number of vectors other than for bivalves of okadaic acid and its analogues.

Aluminium tolerance in bean traditional cultivars from Madeira

Common bean (Phaseolus vulgaris L.) is the most important legume crop in the world, providing low-cost, high quality protein, minerals and dietary fiber for human nutrition. The crop was originated from diversity centers in America and exhibits adaptation abilities to different environmental conditions, including soil with low pH. Acid soils occupy 30% of the agro ecosystem areas in the world. In Madeira, acid Andosols and unsatured Cambisols are the dominant groups of soils. Generally, under acidic and infertile conditions, besides of H+ toxicity, soluble aluminium (Al) is the most important abiotic factor limiting plant development and crop productivity. In the field, the hidden roots are also affected and the reduction of root growth under Al stress can be clearly observed in early stages. Seedlings of fifty bean accessions from the Archipelago of Madeira were tested under controlled conditions in the presence of 50 mM Al at pH 4.4. In general, the tested germplasm appeared to be sensitive or very sensitive to Al toxicity. However, fifteen traditional cultivars clearly exhibited elevated Al-tolerance, with an average root relative elongation (RRE) exceeding 50%, while top six accessions surpassed the 60% RRE mark. The Madeira bean germplasm is a valuable resource for sustainable crop production in acid soils and it could be used as parental lines in breeding programs aimed for Al tolerance in common beans.

Discrepancies in HLA Typing by PCR-SSOP and SBT Techniques: a case study

Six hundred twenty-one samples from Portugal, the Cabo Verde archipelago, and Guinea-Bissau were typed for HLA-A, HLA-B, and HLADRB1usingthepolymerasechainreaction–sequence-specificoligonucleotide probe (PCR-SSOP) method and the sequence-based typing (SBT) method to characterizeandcomparediscrepanciesbetweenthetwomethods.Fifty-three alleles (4.27% of 1,242 chromosomes typed) identified by the PCR-SSOP method were not concordant with the results obtained using the SBT method. Thirty-four (2.74% of total chromosomes typed) PCR-SSOP mistyping results were discrepancies inside the same allele group and 19 others (1.53% of total chromosomes typed) were relative to nonconcordant results between different groups. PCR-SSOP allele mistyping is the result of interpretation difficulties resulting from less intense, absent, or dubious hybridization patterns. Noncommercial PCR-SSOP procedures are highly exigent on the technicians’ experience and the availability of properly calibrated high-precision equipment.

Fatores significativos na saúde mental das pessoas idosas

A atenção à saúde mental (SM) das pessoas idosas é prioritária. Os problemas de SM neste grupo populacional têm aumentado sendo importante investir na prevenção e rastreio dos mesmos. Este estudo teve como objetivos caraterizar do ponto de vista da SM a população idosa da Região Autónoma da Madeira (RAM); determinar as prevalências das situações de SM positiva e negativa e avaliar a influência positiva (protetora) ou negativa (de risco) de certos fatores pessoais e do meio na SM. Foi um estudo transversal no qual participaram 342 pessoas com 65 e mais anos, dos dois géneros, residentes na comunidade. Destes 67,0 % apresentaram SM positiva. A probabilidade da SM ser mais positiva era 2,5 vezes superior quando possuíam 1 a 11 anos de escolaridade (OR = 2,5 IC 95% 1,3 – 4,8); 0,3 vezes inferior nas mulheres (OR = 0,3 IC 95% 0,1 - 0,6), nos idosos com redes sociais muito limitadas (OR = 0,3 IC 95% 0,1 - 0,9) e nos que percecionavam a saúde como razoável ou pior (OR = 0,3 IC 95% 0,1 - 0,9). Era menor 0,5 vezes quando percecionavam a saúde como pior comparativamente aos pares (OR = 0,5 IC 95% 0,3 - 0,9), e 0,3 vezes comparativamente à detida um ano antes (OR = 0,3 IC 95% 0,2 - 0,6). Era 0,1 vez inferior (OR = 0,1 IC 95% 0,1-0,7) nos idosos com limitações físicas para satisfazerem necessidades próprias. Estes fatores devem considerar-se na promoção da SM e na prevenção de perturbações da mesma.