Diel variation in movement patterns and habitat use by the near-threatened cabrera vole in mediterranean farmland

As variações circadianas no comportamento animal e o seu impacto nas populações constituem desafios importantes em ecologia e conservação. Nesta tese documentam-se as variações circadianas no uso do habitat e padrões de movimento pelo rato de Cabrera, em habitats Mediterrânicos fragmentados. O estudo baseou-se no radio-seguimento de indivíduos em habitats dominados por herbáceas e arbustos. Os resultados indicaram que a proporção de tempo despendido em deslocações, a distância percorrida, e a selecção do tipo de vegetação, estão fortemente interrelacionados, variando consideravelmente ao longo de diferentes períodos do dia. Os ratos movimentaram-se mais frequentemente e maiores distâncias nos períodos diurnos, durante os quais as áreas dominadas por herbáceas foram usadas mais intensivamente. Durante a estação seca houve alguma tendência para a diminuição dos movimentos durante as horas mais quentes. Estes resultados são discutidos no sentido de mostrar como indicadores comportamentais podem contribuir para melhorar a gestão e conservação da espécie; ABSTRACT: Understanding the circadian variations in species behaviour and its impacts on population is a challenging topic in ecology and conservation. This thesis documents the circadian variations in habitat use and movement patterns by Cabrera voles in fragmented Mediterranean farmland. The study was based on radiotracking data of individuals living in habitat patches dominated by wet grasses and shrubs. Results indicated that the proportion of time animals spent moving, the distance moved and the selection strength of vegetation were closely linked behavioural traits, which varied considerably across the 24 hour cycle. Voles moved more frequently and over larger distances during daytime, which was when wet grasses were used more intensively. During the dry season there was some tendency for a decrease in movement activity during the hottest hours of the day. These results are used to discuss how behavioural indicators may be useful to improve conservation management of the species.

Monitoring pasture variability: optical OptRx® crop sensor versus Grassmaster II capacitance probe

Estimation of pasture productivity is an important step for the farmer in terms of planning animal stocking, organizing animal lots, and determining supplementary feeding needs throughout the year. The main objective of this work was to evaluate technologies which have potential for monitoring aspects related to spatial and temporal variability of pasture green and dry matter yield (respectively, GM and DM, in kg/ha) and support to decision making for the farmer. Two types of sensors were evaluated: an active optical sensor(OptRx®, which measures the NDVI, Normalized Difference Vegetation Index) and a capacitance probe (GrassMaster II which estimates plant mass). The results showed the potential of NDVI for monitoring the evolution of spatial and temporal patterns of vegetative growth of biodiverse pasture. Higher NDVI values were registered as pasture approached its greatest vegetative vigor, with a significant fall in the measured NDVI at the end of Spring, when the pasture began to dry due to the combination of higher temperatures and lower soil moisture content. This index was also effective for identifying different plant species (grasses/legumes) and variability in pasture yield. Furthermore, it was possible to develop calibration equations between the capacitance and the NDVI (R2 = 0.757; p < 0.01), between capacitance and GM (R2 = 0.799; p<0.01), between capacitance and DM (R2 = 0.630; p<0.01), between NDVI and GM (R2=0.745; p < 0.01), and between capacitance and DM (R2=0.524; p<0.01). Finally, a direct relationship was obtained between NDVI and pasture moisture content (PMC, in %) and between capacitance and PMC (respectively, R2 = 0.615; p<0.01 and R2=0.561; p <0.01) in Alentejo dryland farming systems.

Calibration of Grassmaster II to estimate green and dry matter yield in Mediterranean pastures: effects of pasture moisture content

Accurate assessment of standing pasture biomass in livestock production systems is a major factor for improving feed planning. Several tools are available to achieve this, including the GrassMaster II capacitance meter. This tool relies on an electrical signal, which is modified by the surrounding pasture. There is limited knowledge on how this capacitance meter performs in Mediterranean pastures. Therefore, we evaluated the GrassMaster II under Mediterranean conditions to determine (i) the effect of pasture moisture content (PMC) on the meter’s ability to estimate pasture green matter (GM) and dry matter (DM) yields, and (ii) the spatial variability and temporal stability of corrected meter readings (CMR) and DM in a bio-diverse pasture. Field tests were carried out with typical pastures of the southern region of Portugal (grasses, legumes, mixture and volunteer annual species) and at different phenological stages (and different PMC). There were significant positive linear relations between CMR and GM (r2 = 0.60, P < 0.01) and CMR and DM (r2 = 0.35, P < 0.05) for all locations (n = 347). Weak relationships were found for PMC (%) v. slope and coefficient of determination for both GM and DM. A significant linear relation existed for CMR v. GM and DM for PMC >80% (r2= 0.57, P < 0.01, RMSE = 2856.7 kg ha–1, CVRMSE=17.1% to GM; and r2= 0.51, P < 0.01,RMSE = 353.7 kg ha–1, CVRMSE = 14.3% to DM). Therefore, under the conditions of this current study there exists an optimum PMC (%) for estimating both GM and DM with the GrassMaster II. Repeated-measurements taken at the same location on different dates and conditions in a bio-diverse pasture showed similar and stable patterns between CMR and DM (r2= 0.67, P < 0.01, RMSE = 136.1 kg ha–1, CVRMSE = 6.5%). The results indicate that the GrassMaster II in-situ technique could play a crucial role in assessing pasture mass to improve feed planning under Mediterranean conditions.

Phytosociological survey on the Lusitan-Extremadurean grasslands of Dactylis hispanica subsp. lusitanica (Stebbins & D.Zohary) Rivas-Mart. & Izco

Deeply connected with forest serial dynamics, and present in almost semi-natural landscapes of Iberian Peninsula, Dactylis hispanica subsp. lusitanica perennial grasslands antalis a major issue on biogeographical analysis, once it lacks on syntaxonomical solutions for its affiliation and respective positioning on serial dynamics of distinct territories, which edaphoclimatic envelopes suggest different floristic assemblages. Mainly part of Agrostion castellanae-Stipion giganteae alliance, the Lusitan-Extremadurean grasslands were studied, with the objective of updating that biogeographic Province, and several phytosociological relevés were made, allowing the syntaxonomical update as well to resolve the serial-dynamics arrangement where this peculiar grasslands makes part of. At last this communities belong to priority 6220 [*Pseudo-stepe with grasses and annuals (Thero-Brachypodietea)], habitat that interests as well for conservational matters, due to its peculiar flora.

Study of the allergenic profile of pollen from Platanus hybrida in the city of Evora, Portugal

Background and Aim: Although grasses and olive are the most relevant allergenic species in the Alentejo region, aggravation of allergic symptoms in the early spring, unrelated with those species pollen seasons, has been reported, particularly in urban environment. Plane trees, hence pollen, are highly abundant in the city of Évora, nonetheless allergen pollen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Platanus hybrida, one of the most representative species in Evora showing pollination prior to the main pollen season in Alentejo. Methods: Pollen from Platanus hybrida and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Sensitization and cross-reactivity was assessed by solid phase immunoblot. Results: Half of the patient exhibited sensitization to pollen extracts of P. Hybrida. Western blot have shown several immunoreactive bands in the Mr 10-90 kDa range. Immunoreactive bands were also observed in the protein profile according to the pI in the pI range 4.0-6.1. Cross-reactivity of P. hybrida with D. glomerata was found. Although several bands are common to D. glomerata, a band with ~50kDa was observed in P. hybrida but not in D. glometata. Conclusion: These results evidenced allergens found in P. hybrida pollen. Moreover, cross–reactivity between P. hybrida and highly allergenic species such as D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by FEDER through the “Programa Operacional Fatores de Competitividade – COMPETE” (Strategic projects of ICAAM and ICT 2013-2015).

Allergenic profile of Quercus rotundifolia pollen in Alentejo, Portugal

Background and Aim: Grasses and olive are the most relevant allergenic species in the Alentejo region. However, aggravation of allergic symptoms has been reported in the early spring, before grass and olive pollen seasons. Quercus pollen is the most abundant pollen type in the early spring in Alentejo, nonetheless its allergen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Quercus rotundifolia among the most representative species showing pollination in April, prior to the main pollen season in Alentejo. Methods: Pollen from Quercus rotundifolia, Olea europaea and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Extract from Quercus ilex pollen was kindly offered by Bial. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Sensitization and cross-reactivity was assessed by solid phase immunoblot. Results: Most of the patient evidenced sensitization to pollen extracts of Q. rotundifolia. Protein profile of Q. rotundifolia has shown several bands in the Mr 10-90 kDa, mostly overlapping with Q. ilex. Western blot have shown several immunoreactive bands. Immunoreactive bands were also observed in the protein profile according to the pI in the range 4.0-6.1. Cross-reactivity between Q. rotundifolia with O. europaea and D. glomerata was found. Conclusion: These results evidenced allergens found in Q. rotundifolia pollen. It also shows that protein profile of Q. rotundifolia and Q. ilex are mostly alike suggesting that similarities in allergen profile are expected. Moreover, cross–reactivity between Q. rotundifolia and highly allergenic species such as O. europaea and D. glomerata was found which probably contributes to the aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by FEDER through the “Programa Operacional Fatores de Competitividade – COMPETE” (Strategic projects of ICAAM and ICT 2013-2015). We also aknowledge Bial-Aristegui for supplying pollen and extract samples of Q. ilex.

Perfil em alergénios do pólen de Platanus hybrida

Objetivos: O plátano (Platanus hybrida) é uma árvore frequentemente utilizada em ambiente urbano, com fins ornamentais. Sendo uma árvore de grande porte, produz pólen em grande quantidade. Embora seja responsável por níveis de exposição a pólen elevados no início da primavera, que são coincidentes com queixas da população, o seu potencial alergénico está pouco caracterizado. Este trabalho teve, assim, como objetivo caracterizar o perfil em alergénios do pólen de plátano na cidade de Évora, Alentejo. Métodos: Prepararam-se extratos de amostras de pólen de Platanus hybrida ou Dactylis glomerata utilizando tampão bicarbonato. Os extratos foram liofilizados e conservados a -80ºC. O conteúdo em proteínas foi determinado pelo método de Bradford. O perfil em alergénios foi avaliado por western blot utilizando soros humanos (obtidos mediante consentimento informado de doentes do Hospital do Espírito Santo de Évora – HESE). Resultados: Observou-se teste positivo a P. hybrida em metade dos soros testados. O perfil em proteínas de P. hybrida exibiu diversas bandas imunorreativas com massas moleculares compreendidas entre 10-90 kDa e com pI no intervalo 4,4-7,0. Foram encontradas imunorreativas comuns a Q. rotundifólia e/ou a D. glomerata. Duas bandas identificadas na gama de 50kDa e 60 kDa parecem específicas de P. hybrida. Também se registou reatividade cruzada com D. glomerata. Conclusões: Este trabalho evidencia alguns alergénios encontrados em pólen de P. hybrida. Para além disso mostra ainda a existência de reatividade cruzada com pólen de gramíneas. Estes resultados sugerem que o pólen de plátano, dada a sua grande abundância na cidade de Évora, poderá contribuir para o agravamento a sintomatologia da população que sofre de polinose, em particular no início da primavera. Agradecimentos: Este trabalho foi financiado por fundos do FEDER através do Programa Operacional Fatores de Competitividade – COMPETE”. Um agradecimento especial ao nosso colega, já falecido, Prof. Rui Brandão, pelo estímulo que deu a este trabalho e pela sua dedicação para a implementação e desenvolvimento da Aerobiologia na Universidade de Évora. Temos a honra de dedicar este trabalho à sua memória. Background and Aim: Although grasses and olive are the most relevant allergenic species in the Alentejo region, aggravation of allergic symptoms in the early spring, unrelated with those species pollen seasons, has been reported, particularly in urban environment. Plane trees, hence pollen, are highly abundant in the city of Évora, nonetheless allergen pollen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Platanus hybrida, one of the most representative species in Evora showing pollination prior to the main pollen season in Alentejo. Methods: Pollen from Platanus hybrida, Quercus rotundifolia or Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Results: Protein profile of P. Hybrida has shown several bands in the Mr 10-90 kDa. Western blot have shown several immunoreactive bands. Protein profile according to the pI showed immunoreactive bands in the pI range 4.0-6.1. Cross-reactivity of P. hybrida with Q. rotundifolia and D. glomerata was found. Conclusion: These results evidenced allergens found in P. hybrida pollen. Moreover, cross–reactivity between P. hybrida and highly allergenic species such as D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by “FEDER - Programa Operacional Factores de Competitividade – COMPETE”. A special acknowledgment to our colleague Prof. Rui Brandão, deceased, for his dedication to the present work, to the implantation and development of Aerobiology in the University of Évora. We have the honour of dedicating this work to the memory of Prof. Rui Brandão.

Perfil em alergénios do pólen de Quercus rotundifólia

Background and Aim: Grasses and olive are the most relevant allergenic species in the Alentejo region. However, aggravation of allergic symptoms has been reported in the early spring, before grass and olive pollen seasons. Quercus pollen is the most abundant pollen type in the early spring in Alentejo, nonetheless its allergen profile has not yet been evaluated. The aim of this work was to characterize the allergen profile of pollen from Quercus rotundifolia the most representative species showing pollination in April, prior to the main pollen season in Alentejo. Methods: Pollen from Quercus rotundifolia, Olea europaea and Dactylis glomerata was extracted with ammonium bicarbonate buffer, lyophilized and stored at -80ºC until analysis. Extract from Quercus ilex pollen was kindly offered by Bial. Protein content was determined by the Bradford method. SDS-PAGE followed by western blot, using allergic patient sera (obtained from the Hospital do Espírito Santo de Évora – HESE), were performed to evaluate the allergen profile of the pollen. Results: Protein profile of Q. rotundifolia has shown several bands in the Mr 10-90 kDa, mostly overlapping with Q. ilex. Western blot have shown several immunoreactive bands. Protein profile according to the pI showed immunoreactive bands in the pI range 4.0-6.1. Cross-reactivity of Q. rotundifolia with O. europaea and D. glomerata was found. Conclusion: These results evidenced allergens found in Q. rotundifolia pollen. It also shows that protein profile of Q. rotundifolia and Q. ilex are mostly alike suggesting that similarities in allergen profile are expected. Moreover, cross–reactivity between Q. rotundifolia and highly allergenic species such as O. europaea and D. glomerata was found which probably contributes for aggravation of pollinosis in the early spring. Acknowledgments: This work was supported by “FEDER - Programa Operacional Factores de Competitividade – COMPETE”. A special acknowledgment to our colleague Prof. Rui Brandão, deceased, for his dedication to the present work, to the implantation and development of Aerobiology in the University of Évora. We have the honour of dedicating this work to the memory of Prof. Rui Brandão.