In vitro spore germination of Polystichum drepanum, a threatened fern from Madeira Island

Polystichum drepanum (Sw) C. Presl is a threatened fern endemic to a few forest areas in the north-west of Madeira Island. The aims of this work were to establish suitable culture conditions for in vitro germination of spores, and to evaluate short-term storage conditions for P drepanum spores. The highest frequency of germination was obtained in Murishage and Skoog (MS) liquid medium, without agitation. However, gametophytes maintained in MS liquid medium did not grow and, after 4 weeks, became anoxic and died. Thus, after germination in liquid medium, gametophytes were transferred to an MS double-phase culture system for further growth. The effects of storage period, temperature, and relative humidity during storage on in vitro spore germination were studied. Spore viability was assessed after 2, 4 and 6 months, and high viability (> 94%) was observed in all the assays. However, germination capability decreased with increased storage periods. The number of sporophytes obtained also decreased with prolonged storage periods. The results indicate that spores of R drepanum stored for 4 months at 21 degrees C maintain high viability and high germination frequency. ne sporophytes obtained were acclimatised in a mixture of peat and vermiculite [2:1 (v/v)] under high relative humidity (90-95%). Seventy-five sporophytes were successfully acclimatised to ex vitro conditions and showed active growth in the glasshouse.

Temporal distribution of the short-beaked common dolphin (Delphinus delphis) in the south of Madeira Island (Portugal) and relationship with oceanographic variables

Dissertação de mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015

Predicting the impacts of climate change on the distribution and conservation of endemic forest land snails of Madeira Island

Climate change is emerging as one of the major threats to natural communities of the world’s ecosystems; and biodiversity hotspots, such as Madeira Island, might face a challenging future in the conservation of endangered land snails’ species. With this thesis, progresses have been made in order to properly understand the impact of climate on these vulnerable taxa; and species distribution models coupled with GIS and climate change scenarios have become crucial to understand the relations between species distribution and environmental conditions, identifying threats and determining biodiversity vulnerability. With the use of MaxEnt, important changes in the species suitable areas were obtained. Laurel forest species, highly dependent on precipitation and relative humidity, may face major losses on their future suitable areas, leading to the possible extinction of several endangered species, such as Leiostyla heterodon. Despite the complexity of the biological systems, the intrinsic uncertainty of species distribution models and the lack of information about land snails’ functional traits, this analysis contributed to a pioneer study on the impacts of climate change on endemic species of Madeira Island. The future inclusion of predictions of the effect of climate change on species distribution as part of IUCN assessments could contribute to species prioritizing, promoting specific management actions and maximizing conservation investment.

Susceptibility of mosquito vectors to Dirofilaria immitis on Madeira Island, Portugal

Dirofilaria immitis (Leidy, 1856), an agent of heartworm disease, is an important parasite from both the veterinary standpoint and as a model to study human filariasis. It is a mosquito-borne filarial nematode which inhabits the right ventricle and pulmonary arteries of dogs. D. immitis is an important disease agent on Madeira Island with about 30% of dogs testing positive for this worm. Nevertheless, the vectors of this parasite in Madeira have never been studied, nor has the interaction between pathogen and vector, or the environmental variables that might influence heartworm transmission. Innate susceptibility to infection is only one component of vector competence, and field isolation of naturally infected mosquitoes has shown the capability of D. immitis to exploit a great diversity of vector species under natural conditions. The purpose of this work was to determine which mosquitoes are vectors of heartworm disease, the relation between population density and environment, and the association between immune response of the vector to the filarial parasite. Seasonal abundance of Culex theileri and Culex pipiens molestus was studied. Correlation and canonical correspondence analysis were performed using abundance data of these two species with selected weather variables, including mean temperature, relative humidity and accumulated precipitation. The most important factor determining Cx. theileri abundance was accumulated precipitation, while Cx. pipiens molestus abundance did not have any relationship with weather variables. Field studies were performed to verify whether Cx. theileri Theobald functions as a natural vector of D. immitis on Madeira Island, Portugal. Cx. theileri tested positive for D. immitis for the first time. The same study was made regarding Cx. p. molestus. Two abnormal L2 stage filarial worms were found in Malpighian tubules in field caught Cx. p. molestus. In the laboratory, two strains of Cx. p. molestus were studied for their susceptibility to D. immitis. None presented infective-stage larvae. Finally, because Cx. p. molestus is an autogenous mosquito, we evaluated the reproductive costs when this mosquito mounts an immune response against D. immitis in the absence of a blood meal. This mosquito showed an active immune response when inoculated intrathoracically with microfilariae (mf) of the heartworm. The ovaries from mosquitoes undergoing melanotic encapsulation developed more eggs than those which could not melanize the mf. This fact is contradictory with some previous studies of reproductive costs in Armigeres subalbatus and Ochlerotatus trivittatus, and it was the first time that an autogenous mosquito was used to study this subject.

Statistical modelling of extreme rainfall in Madeira Island

Extreme rainfall events have triggered a significant number of flash floods in Madeira Island along its past and recent history. Madeira is a volcanic island where the spatial rainfall distribution is strongly affected by its rugged topography. In this thesis, annual maximum of daily rainfall data from 25 rain gauge stations located in Madeira Island were modelled by the generalised extreme value distribution. Also, the hypothesis of a Gumbel distribution was tested by two methods and the existence of a linear trend in both distributions parameters was analysed. Estimates for the 50– and 100–year return levels were also obtained. Still in an univariate context, the assumption that a distribution function belongs to the domain of attraction of an extreme value distribution for monthly maximum rainfall data was tested for the rainy season. The available data was then analysed in order to find the most suitable domain of attraction for the sampled distribution. In a different approach, a search for thresholds was also performed for daily rainfall values through a graphical analysis. In a multivariate context, a study was made on the dependence between extreme rainfall values from the considered stations based on Kendall’s τ measure. This study suggests the influence of factors such as altitude, slope orientation, distance between stations and their proximity of the sea on the spatial distribution of extreme rainfall. Groups of three pairwise associated stations were also obtained and an adjustment was made to a family of extreme value copulas involving the Marshall–Olkin family, whose parameters can be written as a function of Kendall’s τ association measures of the obtained pairs.

Effectiveness of different solid-phase microextraction fibres for differentiation of selected Madeira island fruits based on their volatile metabolite profile: identification of novel compounds

A headspace solid-phase microextraction (HS-SPME) procedure based on five commercialised fibres (85 μm polyacrylate – PA, 100 μm polydimethylsiloxane – PDMS, 65 μm polydimethylsiloxane/divinylbenzene – PDMS/DVB, 70 μm carbowax/divinylbenzene – CW/DVB and 85 μm carboxen/polydimethylsiloxane – CAR/PDMS) is presented for the characterization of the volatile metabolite profile of four selected Madeira island fruit species, lemon (Citrus limon), kiwi (Actinidia deliciosa), papaya (Carica papaya L.) and Chickasaw plum (Prunus angustifolia). The isolation of metabolites was followed by thermal desorption gas chromatography–quadrupole mass spectrometry (GC–qMS) methodology. The performance of the target fibres was evaluated and compared. The SPME fibre coated with CW/DVB afforded the highest extraction efficiency in kiwi and papaya pulps, while in lemon and plum the same was achieved with PMDS/DVB fibre. This procedure allowed for the identification of 80 compounds, 41 in kiwi, 24 in plums, 23 in papaya and 20 in lemon. Considering the best extraction conditions, the most abundant volatiles identified in kiwi were the intense aldehydes and ethyl esters such as (E)-2-hexenal and ethyl butyrate, while in Chicasaw plum predominate 2-hexenal, 2-methyl-4-pentenal, hexanal, (Z)-3-hexenol and cyclohexylene oxide. The major compounds identified in the papaya pulp were benzyl isothiocyanate, linalool oxide, furfural, hydroxypropanone, linalool and acetic acid. Finally, lemon was shown to be the most divergent of the four fruits, being its aroma profile composed almost exclusively by terpens, namely limonene, γ-terpinene, o-cymene and α-terpinolene. Thirty two volatiles were identified for the first time in the fruit or close related species analysed and 14 volatiles are reported as novel volatile metabolites in fruits. This includes 5 new compounds in kiwi (2-cyclohexene-1,4-dione, furyl hydroxymethyl ketone, 4-hydroxydihydro-2(3H)-furanone, 5-acetoxymethyl-2-furaldehyde and ethanedioic acid), 4 in plum (4-hydroxydihydro-2(3H)-furanone, 5-methyl-2-pyrazinylmethanol, cyclohexylene oxide and 1-methylcyclohexene), 4 in papaya (octaethyleneglycol, 1,2-cyclopentanedione, 3-methyl-1,2-cyclopentanedione and 2-furyl methyl ketone) and 2 in lemon (geranyl farnesate and safranal). It is noteworthy that among the 15 volatile metabolites identified in papaya, 3-methyl-1,2-cyclopentanedione was previously described as a novel PPARγ (peroxisome proliferator-activated receptor γ) agonist, having a potential to minimize inflammation.

Cozinha da Madeira website: the online touchpoint for a sustainability oriented service of Madeira Island

Over the past several years technology has been evolving in a way that it has become crucial for most businesses and companies to have interactive technology enabled touchpoints available online. Such interactive touchpoints can be developed as mobile application, webpages, or even through social networks. In the end such touchpoints will most surely represent the most easily reachable and marketable side of the business. Today selling a product alone is no longer enough to make consumers satisfied and complete, businesses and business models are changing. Increasingly, companies are choosing to not just sell products but to combine both sale and service. These service-based approaches will provide the client with a unique and personalized experience of what the company is selling. By selling a service the company transmits values that are more complex than the simple selling of a product. A service is something immaterial, happens over time and exists in the moment of the delivery. When conceiving and designing services, the use of the new technologies becomes a crucial step in order to craft touchpoints that facilitate the whole experience cycle of the service, from attracting, orienting, interacting and retaining the client, as well as providing later support to the consumer to advocate for the service itself. This thesis reports on the design and implementation of the online touchpoint of Cozinha da Madeira, which is a service designed to support tourism, specifically promoting the discovery of tradition and landscapes in the island of Madeira. Such touchpoint developed in the form of a website, embodies completely or partially various stages of the Service Experience cycle, from attracting and connecting, orienting, interacting as well as retaining and advocating. Through this thesis we will describe the design and implementation of such touchpoint as well as the evaluation and possible future implications.

Evolution of the landscape of Madeira Island: long-term vegetation dynamics

The aim of this thesis was to evaluate historical change of the landscape of Madeira Island and to assess spatial and temporal vegetation dynamics. In current research diverse “retrospective techniques”, such as landscape repeat photography, dendrochronology, and research of historical records were used. These, combined with vegetation relevés, aimed to gather information about landscape change, disturbance history, and vegetation successional patterns. It was found that landscape change, throughout 125 years, was higher in the last five decades manly driven by farming abandonment, building growth and exotic vegetation coverage increase. Pristine vegetation was greatly destroyed since early settlement and by the end of the nineteenth century native vegetation was highly devastated due to recurrent antropogenic disturbances. These actions also helped to block plant succession and to modify floristical assemblages, affecting as well as species richness. In places with less hemeroby, although significant growth of vegetation of lower seral stages was detected, the vegetation of most mature stages headed towards unbalance between recovery and loss, being also very vulnerable to exotic species encroachment. Recovery by native vegetation also occurred in areas formerly occupied by exotic plants and agriculture but it was almost negligible. Vegetation recovery followed the successional model currently proposed, attesting the model itself. Yet, succession was slower than espected, due to lack of favourable conditions and to recurrent disturbances. Probable tempus of each seral stage was obtained by growth rates of woody taxa estimated through dendrochronology. The exotic trees which were the dominant trees in the past (Castanea sativa and Pinus pinaster) almost vanished. Eucalyptus globulus, the current main tree of the exotic forest is being replaced by other cover types as Acacia mearnsii. The latter, along with Arundo donax, Cytisus scoparius and Pittosporum undulatum are currently the exotic species with higher invasive behaviour. However, many other exotic species have also proved to be highly pervasive and came together with the ones referred above to prevent native vegetation regeneration, to diminish biological diversity, and to block early successional phases delaying native forest recovery.

Aedes aegypti on Madeira Island (Portugal): genetic variation of a recently introduced dengue vector

The increasing population of Aedes aegypti mosquitoes on Madeira Island (Portugal) resulted in the first autochthonous dengue outbreak, which occurred in October 2012. Our study establishes the first genetic evaluation based on the mitochondrial DNA (mtDNA) genes [cytochrome oxidase subunit I (COI) and NADH dehydrogenase subunit 4 (ND4)] and knockdown resistance ( kdr ) mutations exploring the colonisation history and the genetic diversity of this insular vector population. We included mosquito populations from Brazil and Venezuela in the analysis as putative geographic sources. The Ae. aegyptipopulation from Madeira showed extremely low mtDNA genetic variability, with a single haplotype for COI and ND4. We also detected the presence of two important kdr mutations and the quasi-fixation of one of these mutations (F1534C). These results are consistent with a unique recent founder event that occurred on the island of Ae. aegyptimosquitoes that carry kdr mutations associated with insecticide resistance. Finally, we also report the presence of the F1534C kdr mutation in the Brazil and Venezuela populations. To our knowledge, this is the first time this mutation has been found in South American Ae. aegypti mosquitoes. Given the present risk of Ae. aegypti re-invading continental Europe from Madeira and the recent dengue outbreaks on the island, this information is important to plan surveillance and control measures.

Evolution of the landscape of Madeira Island: long-term vegetation dynamics

The aim of this thesis was to evaluate historical change of the landscape of Madeira Island and to assess spatial and temporal vegetation dynamics. In current research diverse “retrospective techniques”, such as landscape repeat photography, dendrochronology, and research of historical records were used. These, combined with vegetation relevés, aimed to gather information about landscape change, disturbance history, and vegetation successional patterns. It was found that landscape change, throughout 125 years, was higher in the last five decades manly driven by farming abandonment, building growth and exotic vegetation coverage increase. Pristine vegetation was greatly destroyed since early settlement and by the end of the nineteenth century native vegetation was highly devastated due to recurrent antropogenic disturbances. These actions also helped to block plant succession and to modify floristical assemblages, affecting as well as species richness. In places with less hemeroby, although significant growth of vegetation of lower seral stages was detected, the vegetation of most mature stages headed towards unbalance between recovery and loss, being also very vulnerable to exotic species encroachment. Recovery by native vegetation also occurred in areas formerly occupied by exotic plants and agriculture but it was almost negligible. Vegetation recovery followed the successional model currently proposed, attesting the model itself. Yet, succession was slower than espected, due to lack of favourable conditions and to recurrent disturbances. Probable tempus of each seral stage was obtained by growth rates of woody taxa estimated through dendrochronology. The exotic trees which were the dominant trees in the past (Castanea sativa and Pinus pinaster) almost vanished. Eucalyptus globulus, the current main tree of the exotic forest is being replaced by other cover types as Acacia mearnsii. The latter, along with Arundo donax, Cytisus scoparius and Pittosporum undulatum are currently the exotic species with higher invasive behaviour. However, many other exotic species have also proved to be highly pervasive and came together with the ones referred above to prevent native vegetation regeneration, to diminish biological diversity, and to block early successional phases delaying native forest recovery.

Upper crustal structure of Madeira Island revealed from ambient noise tomography

We present the first image of the Madeira upper crustal structure, using ambient seismic noise tomography. 16 months of ambient noise, recorded in a dense network of 26 seismometers deployed across Madeira, allowed reconstructing Rayleigh wave Green's functions between receivers. Dispersion analysis was performed in the short period band from 1.0 to 4.0 s. Group velocity measurements were regionalized to obtain 2D tomographic images, with a lateral resolution of 2.0 km in central Madeira. Afterwards, the dispersion curves, extracted from each cell of the 2D group velocity maps, were inverted as a function of depth to obtain a 3D shear wave velocity model of the upper crust, from the surface to a depth of 2.0 km. The obtained 3D velocity model reveals features throughout the island that correlates well with surface geology and island evolution.

Determination of a Mean Solar Radiation Year and of a Typical Meteorological Year for the region of Funchal in the Madeira Island

This paper presents the determination of a mean solar radiation year and of a typical meteorological year for the region of Funchal in the Madeira Island, Portugal. The data set includes hourly mean and extreme values for air temperature, relative humidity and wind speed and hourly mean values for solar global and diffuse radiation for the period 2004-2014, with maximum data coverage of 99.7%. The determination of the mean solar radiation year consisted, in a first step, in the average of all values for each pair hour/day and, in a second step, in the application of a five days centred moving average of hourly values. The determination of the typical meteorological year was based on Finkelstein-Schafer statistics, which allows to obtain a complete year of real measurements through the selection and combination of typical months, preserving the long term averages while still allowing the analysis of short term events. The typical meteorological year validation was carried out through the comparison of the monthly averages for the typical year with the long term monthly averages. The values obtained were very close, so that the typical meteorological year can accurately represent the long term data series. The typical meteorological year can be used in the simulation of renewable energy systems, namely solar energy systems, and for predicting the energy performance of buildings.

Study of different atmospheric environments associated to storms development in the Madeira Island

The study aims to improve the understanding about different atmospheric environments leading to the development of storms associated with heavy precipitation in Madeira Island. For this purpose, four main goals have been considered: 1) To document the synoptic and mesoscale environments associated with heavy precipitation. 2) To characterize surface precipitation patterns that affected the island during some periods of significant accumulated precipitation using numerical modelling. 3) To study the relationship between surface precipitation patterns and mesoscale environments. 4) To highlight how the PhD findings obtained in the first three goals can be translated into an operational forecast context. Concerning the large scale environment, precipitation over the island was favoured by weather systems (e.g, mesoscale convective systems and low pressure systems), as well as by the meridional transport of high amount of moisture from a structure denominated as “Atmospheric River”. The tropical origin of this moisture is underscored, however, their impact on the precipitation in Madeira was not so high during the 10 winter seasons [2002 – 2012] studied. The main factor triggering heavy precipitation events over the island is related to the local orography. The steep terrain favours orographically-induced stationary precipitation over the highlands, although maximum of precipitation at coastal region may be produced by localized blocking effect. These orographic precipitating systems presented different structures, associated with shallow and deep convection. Essentially, the study shows that the combination of airflow dynamics, moist content, and orography is the major mechanism that produces precipitation over the island. These factors together with the event duration act to define the regions of excessive precipitation. Finally, the study highlights two useful points for the operational sector, regarding the meridional water vapour transport and local effects causing significant precipitation over the Island; RESUMO: O estudo procura melhorar o entendimento sobre os diferentes ambientes atmosféricos que favorecem o desenvolvimento de tempestades associadas com precipitação intensa na ilha da Madeira. Nesse sentido foram definidos quatro objetivos: 1) Documentar os ambientes sinópticos e de mesoescala associados com precipitação intensa; 2) Caracterizar padrões de precipitação na superfície, em eventos de elevada precipitação acumulada, utilizando modelação numérica; 3) Estudar as relações entre os padrões de precipitação e ambientes de mesoescala; 4) Mostrar como tais resultados podem ser utilizados num contexto operacional de previsão do tempo. Em relação a ambientes de larga escala, verificou-se que a ocorrência de eventos de precipitação intensa sobre a ilha foi favorecida por sistemas meteorológicos, assim como pelo transporte meridional de humidade por meio de estruturas atualmente denominadas Rios atmosféricos. Neste último caso é de destacar a origem tropical de humidade, no entanto, o seu impacto na precipitação sobre a Madeira durante os 10 invernos estudados [2002-2012] não foi tão elevada. O principal fator que favorece os eventos de precipitação intensa está relacionado com a orografia local. O terreno complexo da ilha favorece a ocorrência de precipitação estacionária induzida orograficamente sobre as terras mais altas, embora a precipitação nas zonas costeiras possa ser produzida por um efeito localizado de bloqueio. Estes sistemas orográficos precipitantes apresentaram diferentes estruturas, associados a convecção pouco profunda e profunda. O estudo mostra que a combinação entre as características do escoamento, a quantidade de humidade, e a orografia são os condimentos essenciais para o desenvolvimento da precipitação sobre a ilha, atuando de maneira a definir as regiões de precipitação excessiva. Por fim, o estudo destaca dois pontos que podem ser úteis na previsão do tempo operacional, ligados a larga escala e aos efeitos locais, os quais podem levar ao desenvolvimento de tempestades e precipitação intensa sobre a ilha.

Screening of wheat Landraces from Madeira Island for iron tolerance

Since the fifteen century, the rainfed-cultivation of wheat for grain is traditionally performed on the Island of Madeira. Under several microclimatic conditions and along very sloppy mountains, the landraces are grown on isolated terraces of Andosols with high amounts of iron. Iron oxides are the main inorganic binding agent contributing to the stability of aggregates and to soil fertility in long-term sustainable agriculture in acid and iron-rich soils. After a two day period of seedling initial growth, a screening test of sixty traditional wheat (Triticum spp.) landraces from the ISOPlexis Genebank at the University of Madeira, Funchal, was performed using nutrient solutions containing 10 or 600 mM Fe, during five days, under controlled laboratory conditions. The elongation of the longest primary root was measured for each genotype and the mean root increment relative to control (as, % relative root increment or RRI; n=28) calculated. This parameter appeared to be a sensitive indicator of Fe tolerance in wheat. Over 85% of wheat germplasm showed the RRI higher than 50%, while the RRI of seven accessions exceeded 70%. This indicates that those landraces are Fe tolerant and might be of particular interest for cultivation under acid rich iron soils of tropical and subtropical areas.