III Encontro de Estudantes de Doutoramento

em Ambiente e Agricultura

III PhD Students Meeting in Environmental

and Agriculture
29th and 30th November 2018

Book of Abstracts
Title: III PhD Students Meeting in Environmental and Agriculture

Editors:
Marta Laranjo
Ana Alexandre
Teresa Pinto Correia
Cláudia Marques

Address:
Universidade de Évora,
Largo dos Colegiais, 2
7004-516 Évora

Published:
UE – Universidade de Évora
Copyright © 2018, all rights reserved
ISBN: 978-972-778-139-3
Dear participants,

It is our great pleasure to welcome you to the III Encontro de Estudantes de Doutoramento
em Ambiente e Agricultura / PhD Students Meeting in Environmental and Agriculture, held in
Évora on the 29th and 30th November 2018 and organised by ICAAM and IIFA from the
Universidade de Évora. We have put together a two-day program combining communications
by both well-established and junior scientists, organised in a way to encourage discussion and
exchange of ideas and to explore new challenges in research regarding Environmental and
Agricultural Sciences.

The meeting program comprises five sessions, namely Different Approaches Applied to
Environment and to Agriculture, Olive Grove, Vineyard and Other Cultures, Mediterranean
Agro-Silvo-Pastoral Systems, Biodiversity and Ecosystems Functioning and Animal
Production and Health. Each session begins with two plenary lectures followed by several
short talks selected from the abstracts submitted by the PhD students. Furthermore, several
poster presentations will be displayed throughout the meeting. The program also includes a
field trip entitled “As aves do montado e ribeira da Herdade da Mitra”.

This meeting intends to stimulate the interaction between PhD students, to streamline
scientific discussion and highlight the ones who will become the researchers of the future.

Finally, we wish to thank the authors who have contributed to the program of this meeting and
hope you will enjoy the Meeting and enjoy the beautiful city of Évora, an UNESCO World
Heritage. You should find all detailed information in the meeting book, including the scientific
program, abstracts and a list of participants.

Welcome to Évora!

The Organising Committee,

Marta Laranjo, ICAAM

Ana Alexandre, ICAAM
Teresa Pinto Correia, ICAAM
Cláudia Marques, IIFA
 COMMITTEES

Organising Committee

Marta Laranjo, Universidade de Évora - ICAAM

Ana Alexandre, Universidade de Évora - ICAAM

Teresa Pinto Correia, Universidade de Évora - ICAAM

Cláudia Marques, Universidade de Évora – IIFA

Scientific Committee

Ana Teresa Caldeira, Universidade de Évora – HERCULES

Elsa Leclerc Duarte, Universidade de Évora – ICAAM

Luís Leopoldo Silva, Universidade de Évora – ICAAM

Luiz Gazarini, Universidade de Évora – ICAAM

Maria João Cabrita, Universidade de Évora – ICAAM

Teresa Pinto Correia, Universidade de Évora – ICAAM

Organised by

Sponsors
Program

Tuesday, 29th November 2018

09:00 Registration

09:45 Opening Session

Teresa Pinto Correia
Diretora do Instituto de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM)
António Candeias
Diretor do Instituto de Investigação e Formação Avançada (IIFA)

Different Approaches Applied to Environment and to Agriculture

Chair: Manuel Mota

Universidade de Évora – ICAAM, IHC
10:15 Alergia e Ambiente: uma abordagem multi-disciplinar no estudo da doença
alérgica respiratória sazonal
Célia Antunes
Universidade de Évora - ICAAM, ICT
10:55 O potencial desconhecido da cultura do tremoço. Blad e deflamina
Ricardo Boavida Ferreira
Instituto Superior de Agronomia - LEAF

11:35 Coffee break and poster session

Presentations selected form the submitted abstracts

12:00 Atmospheric pollutants NO2 and O3 contribute to augmented allergenic activity
of grass pollen
Ana Galveias
12:15 Time series analysis as a tool to study the behavior of airborne Platanus pollen
during a 11-year period
Beatriz Lara
12:30 Establishment and standardization of soil enzymatic activity assays at Soil
Microbiology Laboratory
Taiana Conceição
12:45 Effect of biological enrichment of compost and its storage period on the
biological control of Sclerotium rolfsii in turfgrass (Agrotis stolonifera L.)
Luísa Coelho
13:00 Lunch
13:45 Field trip (organized by LabOr)
As aves do montado e ribeira da Herdade da Mitra
14:30 Microbiological evaluation and physico-chemical and sensory characterization of
cheese produced in Cape Verde
Elyane Borges Dias
14:45 Effect of species, seasonal variation, and addition of transglutaminase and
glucomannan fiber on the physical properties and sensory evaluation of fish
hams prepared with gilthead sea bream, sea bass, and meagre
Ana Teresa Ribeiro
14:30

Olive Grove, Vineyard and Other Cultures

Chair: Augusto Peixe

Universidade de Évora - ICAAM
15:05 Vírus de plantas: de agentes causadores de doença a protetores de plantas
Maria do Rosário Félix
Universidade de Évora - ICAAM
15:45 Cromatografia de alta resolução: Métodos analíticos para análise vestigial
Marco Richter Gomes da Silva
Universidade Nova de Lisboa - LAQV-requimte
16:25 Coffee break and poster session

Presentations selected form the submitted abstracts

16:45 Study of Portuguese extra virgin olive oils according to the producing agricultural
method: A qualitative and ecological approach
Miguel Ferro
17:00 Evaluation of anthocyanin profile during ripening in Vitis vinifera red wine grape
varieties grown in Alentejo
Maria Inês Rouxinol
17:15 Mycorrhization of Cistus spp. with Terfezia arenaria (Moris) Trappe and its
application in desert truffle cultivation
Rogério Louro

17:30 Sustentabilidade caminho sem volta

André Ramalho Flores
Consultor técnico na área da Sementeira Direta
20:30 Meeting dinner at the restaurant “Cozinha do Cardeal”
Friday, 30th November de 2018

Mediterranean Agro-Silvo-Pastoral Systems

Chair: João Rabaça

Universidade de Évora, ICAAM
09:00 Tecnologias na monitorização do montado: uma abordagem global sobre o ECO-
SPAA
João Serrano
Universidade de Évora - ICAAM
09:40 Agro-Silvo-Pastorícia: Inovação com Tradição
José Mira Potes
Instituto Politécnico de Santarém – Escola Superior Agrária
10:20 Coffee break and poster session

Presentations selected form the submitted abstracts

10:50 Visions and strategies for water management in agriculture: a transdisciplinary
experience in the Alentejo
Taiana Homobono
11:05 Climate change in the development of a model of occurrence of forest fires:
determination of trend series and critical periods
Leonel Nunes
11:20 Evaluation of the interactions soil - pasture - trees and animals in Montado
ecosystem
Emanuel Carreira

Biodiversity and Ecosystems Functioning

Chair: Anabela Belo

Universidade de Évora - ICAAM
11:40 Estradas e biodiversidade. Coexistir ou não coexistir, eis a questão
António Mira
Universidade de Évora – CIBIO-InBIO, ICAAM
12:20 Bem-me-quer, mal-me-quer: uma história sobre cegonhas e linhas eléctricas
Francisco Moreira
Universidade do Porto– CIBIO-InBIO
13:00 Lunch
 Presentations selected form the submitted abstracts
14:05 Floristic analysis of four communities of Prunus lusitanica L. from Europe and
North Africa
Mauro Raposo
14:20 Effect of Alqueva Dam on terrestrial invertebrates communities: threat or
opportunity
Rui Raimundo
14:35 Accounting for connectivity uncertainties in predicting roadkills: a comparative
approach between path selection functions and habitat suitability models
Francesco Valerio
14:50 Regime of fire in Portugal: assumptions and facts
Nuno Guiomar

Animal Production and Health

Chair: Catarina Lavrador

Universidade de Évora - ICAAM
15:10 O papel do útero na sobrevivência e desenvolvimento do concepto
Rita Payan Carreira
Universidade de Évora, CECAV
15:50 Coffee break and poster session
16:05 Doença hemorrágica viral em coelhos bravos
Conceição Peleteiro
Universidade de Lisboa-FMV, CIISA

Presentations selected form the submitted abstracts

16:45 Characterization of the Mertolenga Breed phenotypes under the influence of the
CAP in the period between 1986 and 2012
Luís Santa Maria
17:00 Evaluation of elastase and cathepsin G inhibition on collagen transcription and
prostaglandin secretion in mare endometrium
Ana Amaral
17:15 Assessing pubertal age of male Bísaro pigs through testicular and epididymal
morphometric parameters - preliminary results
Gustavo Paixão
17:30 Environmental conditions in pigs facilities located in Southern Europe
Teresa Morgado
17:45 Closing Session
Including Best Poster Prize
Different Approaches
Applied to Environment and to Agriculture
 Invited Speaker 1

Allergy and Environment: a multidisciplinary approach in the study of seasonal

respiratory allergic diseases

Célia M. Antunes ([email protected])

Department of Chemistry, School of Sciences and Technology, University of Évora

Institute of Earth Sciences, University of Évora

The burden of pollen allergies is increasing worldwide. Due to its environmental etiologic nature,
the disease management in a changing environment has proven to be challenging. The
Laboratory of Applied Biochemistry and Aerobiology is devoted to a multidisciplinary approach
for the study of environmental factors involved in pollen allergy elicitation and aggravation. Our
main goal is to contribute to a better understanding of this complex problem in the pursuing of
the development of mitigation strategies.

In this context, a wide range of methodologies for the study of airborne pollen and allergens,
the effect of air pollutants on pollen physiology and allergenicity, as well as the consequent
impacts for human health are undertaken.

So far, our results have contributed to: the clarification of airborne pollen and allergen loads,
distribution and their transport in the atmosphere in our region; the identification of new
allergenic species and cross-reactivity patterns among allergens and its relevance for
symptomatology; and, finally, to the development of a PolenAlert system aiming to contribute
to a better management of seasonal respiratory allergic disease.
 Invited Speaker 2

O potencial desconhecido da cultura do tremoço. Blad e deflamina

Ricardo Boavida Ferreira
Instituto Superior de Agronomia – LEAF, Universidade de Lisboa
No que diz respeito à sua utilização na nutrição humana, a cultura do tremoço ‘atrasou-se’
relativamente à da soja porque se encontra naturalmente adaptada a condições
Mediterrânicas, caracterizadas por um clima quase semi-desértico. Nestas condições, em que
alimento e hospedeiros escasseiam, as plantas têm de investir muito potencial metabólico em
defesa para conseguir sobreviver. A presença de alcaloides tóxicos foi durante muito tempo
um grande entrave à utilização generalizada dos tremoços na alimentação do homem e dos
animais, embora estas sementes contenham também algumas proteínas com interesse
terapêutico para o homem.

Utilizando técnicas de melhoramento clássicas, os investigadores alemães foram os primeiros a

produzir, há já várias décadas, semente doces (i.e. com teores residuais de alcaloides) de
tremoço, o que possibilitou a introdução desta leguminosa em todos os setores da
alimentação humana e animal. Há mesmo alguns países que utilizam o tremoço numa ampla
gama de produtos alimentares, desde comida para bebés até à piscicultura.

Fatores de natureza política e económica têm condicionado e mesmo restringido a cultura de

tremoço na bacia Mediterrânica, onde os tremoceiros crescem espontaneamente e se
encontram, por isso, muito bem adaptados. É, pois, estranho é que a proteína animal ingerida
pelos Europeus seja essencialmente conseguida à custa da soja, na sua grande maioria
transgénica e importada do continente Americano.

Ao reduzir as importações massivas de soja do continente Americano, o cultivo generalizado

do tremoceiro na bacia Mediterrânica contribuiria para

-Uma balança comercial mais favorável na UE;

-Uma fonte de rendimento para os agricultores das regiões mais pobres da UE;

-Uma agricultura sustentável na bacia Mediterrânica;

-Uma fonte de proteínas edíveis bioativas para a saúde do homem e das plantas.

Por outro lado, a grande euforia que existe a nível mundial relativamente aos compostos
naturais bioativos, assenta basicamente em metabolitos secundários que, se ingeridos fora de
uma alimentação equilibrada e variada, exibem efeitos muito tóxicos, de que são exemplos os
óleos essenciais, polifenóis antioxidantes, alcaloides e muitos péptidos.

Contrariamente à maioria das outras espécies, o tremoço é fonte de três proteínas edíveis
bioativas, cuja ação não se torna tóxica para o homem se ingerido em grande quantidade. São
todas relacionadas com as principais proteínas de reserva da semente, as α-, β-, γ- e δ-
conglutinas. A própria γ-conglutina tem sido estudada por investigadores Italianos pela sua
bioatividade a nível da diabetes. O BCO e a deflamina foram descobertas no nosso grupo.

O BCO (do Inglês Blad-Containing Oligomer) é um oligómero de polipéptidos derivados da β-

conglutina, comestível, de 210 kDa, que se acumula nos cotilédones de plântulas de Lupinus
entre os dias 4 e 14 após o início da germinação. Descobrimos, em 1991, o seu principal
polipéptido, a Blad (20,4 kDa), com a qual temos trabalhado desde então e continuamos a
descobrir coisas novas.

A Blad tem uma atividade antifúngica muito potente contra todos os fungos testados nos
últimos 20 anos (incluindo fungos patogénicos de plantas e do homem, food spoiling e food
poisoning), atividade bactericida contra bactérias Gram- e Gram+ e atividade bioestimulante
potente nas plantas. A sua atividade é igual ou superior à dos melhores fungicidas químicos
atualmente disponíveis. A Blad está já à venda nos E.U.A. (sob o nome comercial FractureTM,
encontra-se incluída na FRAC Code List© de 2018 com o código BM01 e foi recentemente
aprovada para utilização em agricultura biológica pela OMRI) e Canadá e, durante 2019 e 2020,
ficará comercialmente disponível num número de outros países, incluindo a Europa, Austrália,
China, Coreia do Norte, México, Japão, etc. Temos presentemente mais de 40 patentes
internacionais sobre a Blad.

A deflamina é um oligómero de polipéptidos derivados da β- e da δ-conglutinas, comestível,

presente nas sementes secas de sementes de Lupinus (e, em menor grau, também nas
sementes de algumas outras leguminosas e não só), que apresenta potentes atividades anti-
inflamatórias e anticancerígenas. A sua bioatividade resiste à ação de proteases, a baixos
valores de pH, a temperaturas elevadas (pelo menos 180 OC, o que significa que resiste à
cozedura e ao fabrico de biscoitos) e ao processo digestivo. Foi testada em modelos de doença
(ratos), bem como em coelhos. Estamos presentemente a realizar experiências com tumores
humanos em peixes-zebra (como modelos de doença) e estudos de nutrição em leitões
desmamados (como modelos animais de digestão). Temos em preparação testes clínicos. Foi
recentemente submetido um pedido de patente internacional.
 Oral Communication 1

Atmospheric Pollutants NO2 And O3 Contribute To Augmented Allergenic

Activity Of Grass Pollen

Ana Galveias1, Sara Mendes1, Rute Arriegas1, Helena Ribeiro2, Ana R Costa1, Célia M Antunes1,
Ilda Noronha2

1ICT (Institute of Earth Sciences), IIFA & Dep. Chemistry, School of Sciences and Technology, University of Evora, Evora,

Portugal

2ICT (Institute of Earth Sciences) & Faculty of Sciences, University of Porto, Porto, Portugal

e-mail: [email protected]

Superoxide dismutase (SOD), a known enzyme in several species and recently described as a
grass allergen, that catalyzes the dismutation of toxic superoxide radicals to molecular oxygen
and hydrogen peroxide, working against oxidative stress in plants. Airborne pollen during its
transport become in contact with a variety of atmospheric chemicals, including common air
pollutants O3 and NO2. Besides, their direct deleterious action in respiratory air ways, exposure
to these pollutants, both independently or in combination, may change the pollen biochemical
properties thus its allergenicity. Nonetheless, species-specific action of these pollutants or their
synergic effects in pollen are essentially unknown.
The aim of this research was to study the effects of O3, NO2 and the mixture of both pollutants
on SOD activity of Dactylis glomerata pollen, a proposed grass allergen.
Pollen samples were in vitro exposed to O3, NO2 and O3+NO2 in an environmental chamber
system during 6hrs and concentrations around the ambient levels. A blank pollen sample,
subjected to the same procedure but not exposed to the gases was used as control.
Pollen extracts from unexposed (Control) or exposed to pollutants were prepared in phosphate
buffered saline (PBS) and frozen until analyses. Total SOD activity was measured following
riboflavin-induced reduction of nitro blue tetrazolium (NBT) for 40 min, and the absorbance (A)
was measured at 560 nm every 5 min, triggered with cool white fluorescent light and enzymatic
activity (U/mg protein) was determined.
Pollen exposure to O3 alone did not significantly affect SOD activity while exposure to NO2
induced a 50% SOD activity increase (150±25%). When mixed together (O3+NO2) the SOD activity
was higher (200±40%) compared to control and slightly but not significantly higher when
compared to NO2 alone. These results show that exposure to NO2 induced a significant increase
on SOD activity and suggest that, although O3 alone did not induce any change, it may exert a
synergic action on the effect of NO2, contributing to amplify the response.
These results suggest that NO2 stimulates pollen antioxidant defenses while O3 does not,
characterized at least by an augmented SOD activity, an allergen.
Consequently, pollen exposure to NO2 can induce an increase in the allergenicity and directly
correlated with the enhancement of respiratory diseases.

The work is co-funded by the European Union through the European Regional Development Fund, included in the
COMPETE 2020 (Operational Program Competitiveness and Internationalization) through the ICT project (UID/GEO/
04683/2013) with the reference POCI-01-0145-FEDER-007690 and by National Funds through FCT for the project
PTDC/ATP-EAM/0817/2014.
 Oral Communication 2

Time series analysis as a tool to study the behavior of airborne Platanus pollen
during a 11-year period
B. Lara1, J. Rojo1, R. Pérez-Badia1
1ICAM (Institute of Environmental Science), University of Castilla-La Mancha, Toledo, Spain.
Email: [email protected]

Platanus pollen is an important cause of hay fever in many Spanish cities where plane trees
(Platanus orientalis L. var. acerifolia Dryand in Aiton) are widely used as an ornamental species
in parks, gardens and urban green areas. The aim of this study is to analyze the Platanus pollen
time series by a commonly used method for time series analysis as a seasonal-trend
decomposition procedure based on LOESS smoothing which aids to isolate the three different
components (seasonal, trend and residual) to be studied in an independent way. In this study
airborne Platanus pollen was monitored throughout the period 2005-2015 using a Hirst
volumetric spore trap located on the roof of a building belonging to the University of Castilla-
La Mancha (Spain), on the Science & Technology campus in Toledo.

The Platanus pollen season in the city of Toledo started in the latter half of March and is
characterized by an intense and explosive pollination, leading to high airborne pollen
concentrations over a short period of time, exactly most of the pollen season occur during a
40-day period (on average 95% of the annual pollen is recorded over that period). The daily
maximum concentration tended to be recorded in late March or early April and a trend toward
rising the pollen concentration on the peak days has been observed in our data, thus
increasing the number of days with allergy risk in central Spain.

Also, time series clearly showed a 3-year cyclical pattern, in which two years of high pollen
intensity were followed by a third year of lower pollen intensity likely linked to management
techniques such as periodical pruning of plane trees in urban green areas. This information is
crucial to generate prediction models as a tool for evaluating allergy risk to Platanus pollen.
Models for forecasting were constructed combining the cyclical behavior of the pollen
(seasonality of the time series) with the modelling of short-term fluctuations in airborne pollen
concentrations prompted by daily changes in meteorological variables and pollen
concentrations over the previous days. A model based on the 3-year cyclical pattern and
predictive variables over a prior period 7-9 days accounted for 70% of the variance of the daily
pollen concentration. In this case, calibration models based on a 3-year period are more stable
and enable the use of longer prior time periods than those obtained with a 1-year period.
 Oral Communication 3

Establishment and standardization of soil enzymatic activity assays at Soil

Microbiology Laboratory.
1

Taiana Conceição1, Isabel Brito2

2
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e Formação
Avançada), Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Biologia, Universidade de

Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.
Email: [email protected]

By catalyzing many vital reactions necessary for the life processes of soil microorganisms, soil
enzymes regulate the functioning of the ecosystem and play key biochemical functions in the
overall process of organic matter transformation and nutrient cycling in the soil system. Soil
enzymes have been extensively studied and a variety of methods were developed to measure
their activity over the years, as they provide important information about the ecological and
functional status of the soil microorganisms acting as "biological fingerprints" of soil
microorganisms. The adjustment of techniques for the quantification of dehydrogenase (EC
1.1.1.), arylsulfatase (EC 3.1.6.1), β-glucosidase (EC 3.2.1.21), acid phosphatase (EC3.1.3.2), L-
glutaminase (EC 3.5.1.2) and L-asparaginase (EC 3.5.1.1) activities have been performed at Soil
Microbiology Laboratory, to establish a routine with standard procedures that can be applied
in several contexts. Different scale protocols are being tested, the traditional bench scale and
the microplate scale. To test the bench scale protocols under the laboratory conditions a soil
with intentionally induced different microbial activities was studied. One treatment consisted
of soil supplemented with a solution of sucrose, ammonia sulphate and potassium phosphate
(10:1:1) and incubated for 48h/37ºC, to increase biological activity. A second treatment
consisted of a three series of soil sterilization (24h each) to completely inactivate biological
activity, and a third treatment consisted in the soil as it was after field collection. All
treatments were moisture to 70% of water holding capacity either with the nutrient solution
or with sterile water for the two last treatments. The enzymatic assays in doped soil showed a
significantly higher values than fresh soil, and for sterilized soil the values were near zero. So
far, the results have shown that the bench scale protocols used reflect the differences imposed
in the tested soil treatments and can reliably be used in laboratory routine for dehydrogenase,
arylsulfatase, β-glucosidase and acid phosphatase activity evaluation. Though amidohydrolases
(L-glutaminase and L-asparaginase) need more tests to achieve a standard procedure, because
the two different bench scale protocols used have shown discrepancies of pattern recovery.
The microplate format will also be tested as it allows the simultaneous analysis of multiple
enzymes in microliter volumes, with reduce reagent costs and assay time.
 Oral Communication 4

Effect of biological enrichment of compost and its storage period on the

biological control of Sclerotium rolfsii in turfgrass (Agrotis stolonifera L.)
1

2 L. Coelho1,2,3, L. Dionísio1,2, C. Guerrero1,2, M. Reis1,2

Universidade do Algarve, Campus de Gambelas 8005-139 Faro, Portugal.

3

MeditBio (Centre for Mediterranean Bioresources and Food), Universidade do Algarve. Campus de Gambelas,
8005-139 Faro, Portugal.

ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Biologia, Universidade de

Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

Email: [email protected]

The suppressive capacity of composts against soil diseases may be caused by their microbiota
activity. Compost enrichment with antagonistic organisms is a strategy to increase its
effectiveness, allowing a more effective biological control of soil diseases. However, compost
storage may affect its suppressive capacity, depending on compost and the soil disease.
Turfagrass (Agrostis stolonifera L.) are susceptible to a variety of fungal diseases, being
essential to find ecofriendly alternatives for their control. Organic composts can be an efficient
alternative to control plant diseases, reducing pesticide use. In this study two composts from
agroindustrial residues were tested, P1 and P2. From the end of the thermophilic phase until
the end of the process, fungi from both composts were isolated and identified. In in vitro
experiments, Trichoderma atroviride showed the best results on Sclerotium rolfsii control and,
for this reason, it was used in compost enrichment. Samples of both composts and peat
(control) were enriched with T. atroviride suspension with 1.33x106 conidia mL-1, in order to
obtain 9.0x107 conidia L-1 of the substrates. Samples of peat and composts (enriched and not
enriched) were stored, and their suppressive capacity was tested after two weeks and after six
months of storage. Turfgrass was sown in 100 mL pots, with the substrates: peat (T), composts
P1 and P2, and the same substrates submitted to a thermic treatment of 60 ᵒC (t) during seven
days: Tt, P1t and P2t, respectively. The same substrates were enriched (E) with T. atroviride:
TE, P1E, P2E, TTE, P1TE and P2TE, making a total of 12 treatments, with five pots and four
replicates each. When turfgrass covered the surface of the pots, each pot was inoculated with
S. rolfsii, by placing a PDA disk with the pathogenic.
In substrates stored for two weeks, it was verified that T. atroviride increased composts
suppressive capacity. After six months of storage, P1, P2, P1E and P2E maintained their
suppressive capacity, but the severity of the disease increased in P1TE and P2TE.
Considering these previous results, it will be important to evaluate composts suppressive
capacity, with and without antagonist enrichment, after 1 year of storage.

L.C. acknowledges FCT (SFRH/BD/ 109218/2015).

 Oral Communication 5

Microbiological evaluation and physico-chemical and sensory characterization

of cheese produced in Cape Verde

E. Borges Dias1

1I doutoranda em Ciências Agrárias e Ambientais, Universidade de Évora, Pólo da Mitra, Évora, Portugal.

Email: [email protected] / [email protected]

The objective of this study is to establish the microbiological, chemical and sensorial
characterization of the cheese of Cape Verde in order to commercialize them and implement
the protected designation of origin (PDO) system for this excellent food product. The
genuineness and typicality of the product, ensures its protection, enable fraud detection
(through possible forgery) and also contributes to increasing the competitiveness of small-
scale artisanal cheese production units. In this way it is intended to associate this product with
the development and recognition of the region. There are no studies related to the
microbiological, chemical and sensorial characterization of these cheese, so they do not have
certification of Protected Designation of Origin (PDO) and Protected Geographical Indication
(PGI), negatively affecting their economic value. Thus, the exponential growth of the annual
production, the total lack of knowledge of its properties and characteristics and the chemical
composition, justify that studies are directed towards contributing to a more in-depth
knowledge that allows us to make the production of the cheese profitable and the PDO
certification and IGP. The project aims to provide the Cape Verde Region with powerful tools
to progressively achieve a unique position in the export market under competitive conditions
and on the other hand to provide Cape Verde's cheese with structures to implement the
protected designation of origin system (PDO). This competition requires product quality,
supported by modern technologies and high sensitivity, evaluated according to adequate
indexes and compatible with the practices and techniques used. On the other hand, to
guarantee the genuineness of the cheese produced in Cape Verde, it is necessary to know the
product scientifically through microbiological, chemical and sensorial evaluation, allowing a
more efficient control and the detection of possible adulterations.
To this end, the objectives of the proposed work are as follows: To evaluate the
microbiological quality of cheese produced in Cape Verde; Determine the physical-chemical
parameters that define the quality of the cheese; Proceed to the sensorial analysis as an
efficient approach in the identification and establishment of sensory markers of cheese;
Establish the genetic profile of cheese as a way to guarantee its authenticity, typicality and
protection of the product.
 Oral Communication 6

Effect of species, seasonal variation, and addition of transglutaminase and

glucomannan fiber on the physical properties and sensory evaluation of fish hams
prepared with gilthead sea bream, sea bass, and meagre

Ribeiro, A.T.123; Elias, M.34; Teixeira, B.2; Vieira, H.2; Mendes, R.2

1 Polytechnic Institute of Santarém, Superior School of Agriculture, Department of Food Technology, Biotechnology and
Nutrition. Quinta do Galinheiro - S. Pedro. 2001-904 Santarém, Portugal.

2 Portuguese Institute for the Sea and Atmosphere, Department for the Sea and Marine Resources, Rua Alfredo Magalhães
Ramalho, 6, 1495-006 Lisboa, Portugal.

3 ICAAM–Instituto de Ciências Agrarias e Ambientais Mediterrânicas, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554
Évora, Portugal.

4 Departamento de Fitotecnia, Escola de Ciências e Tecnologia, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora,
Portugal.

Email: [email protected]

The increasing consumer demand for healthier products is driving the development of novel fish-based
products, due to the well-known benefits of fish consumption. Within the variety of traditional meat
products in market places, cooked ham is particularly appropriated, on account of its broad public
acceptance.

This work aimed to study the influence of fish species, capture season, and addition of microbial
transglutaminase (MTGase) and glucomannan dietary fiber from konjac (Amorphophallus konjac; KGM)
in the preparation of fish hams and its effects on the physical properties and sensory evaluation.

Sea bass (Dicentrarchus labrax), gilthead sea bream (Sparus aurata), and meagre (Argyrosomus regius),
captured in the summer and winter seasons, were used individually to prepare hams. Different
formulations varying the amount of MTGase (0-0.5 % MTGase) and KGM (0-1% KGM) were done. Color,
water holding capacity, pH, texture (puncture, compression, and compression relaxation tests), and
sensory analysis were determined in the different fish hams.

Results showed that in terms of species used for the preparation of fish hams, hams prepared with
meagre had higher water holding capacity and the textural properties were closer to those of traditional
hams prepared with pork. The capture season also affected several properties of fish hams, with a minor
importance, and it was dependent on the species. The addition of MTGase and KGM in the preparation
of fish hams resulted in hams with textural properties closer to those of pork hams. However, the use of
KGM compromised the visual appearance of fish hams. In general, fish hams were well accepted by the
sensory panelists.

This study showed that the fish hams produced have potential as novel fish-based products. In particular
for each species, fish hams prepared with gilthead sea bream captured during winter or with sea bass
captured in the summer, both prepared with 0.5 % MTGase and 1% KGM, and those with meagre
captured in the winter with 0.5 % MTGase (without KGM) exhibited the most comparable physical
properties to a pork ham.
 Poster 1

Characterization of compounds with biological activity of Opuntia ficus indica

l. Miller

G.C. Murta1, M.R. Martins3 , A.E. Rato2,

1ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e Formação
Avançada), Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

2Laboratório HERCULES - Departamento de Química e Laboratório HERCULES da Universidade de Évora

Universidade de Évora, Polo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

3ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Fitotecnia, Universidade de

Évora, Polo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

Email: [email protected]

The species Opuntia ficus indica (L.) Miller is native to Mexico, found widely distributed
throughout Central America, South America, Australia, South Africa and in Mediterranean
countries. The taxonomic classification of the genus Opuntia is complex, since many of the
classification systems present errors in the concepts of genus and species, which ends up
contributing to the high synonymy. This is an active genus of evolution and characterization,
contributing to erroneous classifications, variation among species induced by genetic
differences resulting from the influence of environmental factors, is perceptible for example in
differences in cladodes size, flower color and length of leaf thorns. The Opuntia species has
high importance in the agricultural economy, since it is a valuable resource, especially in
periods of drought. In Portugal it is sub spontaneous in hedges and weavings, being cultivated
for the formation of artificial hedges. From the morphological point of view, this can be divided
into root, vegetative part, fruit and flower. The vegetative part corresponds to the cladodes,
which consist of modified stalks, which replace the leaves in their photosynthetic function.
These have an ovoid shape about 18-25 cm in length, and in addition to their photosynthetic
function, also serve as storage water. However, the fruit has gained notoriety due to its
nutritional benefits and different pharmacological actions, being rich in bioactive antioxidant
compounds such as betalaínas, polyphenols and ascorbic acid. Its extracts have demonstrated
different activities, such as ant ulcerative, antioxidant, anticancer, neuroprotective,
hepatoprotective and antiproliferative activities, thus justifying the theme of the following
dissertation, since its purpose is to characterize the chemical composition and evaluate some
nutritional and biological properties of the fruits (orange, red and green), cladodes, seed oil
and "pie" (seed residue after oil extraction) of fig, aiming to contribute to the transfer of
knowledge about its nutraceutical properties and, consequent appreciation and proper use of
figs and its derivatives, and also contributing to the establishment of new criteria for the
certification of marketed products.
 Poster 2

COMPARATIVE STUDY OF ATMOSPHERIC CONCENTRATIONS OF POLLEN GRAINS OF

AMARANTACEAE AND SAL K1 PROTEIN IN ALCÁZAR DE SAN JUAN AREA (CIUDAD REAL, SPAIN)

Ana María Burgos-Montero1, Rosa Pérez-Badia2, Francisco Feo Brito3, Carmen Arilla Rodríguez4,
Juan Asturias Ortega4 , Borja Bartolomé Zavala4

1
Allergy Department of Mancha Centro Hospital, Ap 13600 Alcázar de San Juan, Ciudad Real,
Spain.

2
ICAM (Institute of Environmental Sciences), Botanical Area. University of Castilla-La Mancha,
Ap. 45071 Toledo, Spain.

3
Allergy Department of General University Hospital . Ap 13005 Ciudad Real, Spain.
4
I+ D Department, Roxall. Ap. 48170 Zamudio, Bilbao, Spain.

Email: [email protected]

Introduction: Previous studies have shown discrepancies between the pollination period and
the allergic symptoms of patients by inhalation of Amaranthaceae pollen. This is attributed to
the presence of non-aerovant grains, which are not detectable with pollen traps. Chenopodium
Album and Salsola kali are the most representative species of this family from the
allergological point of view and Sal k1 is the most representative allergen from Salsola kali
pollen. The objective of this work was to analyze the correlation between the atmospheric
concentrations of Amaranthaceae pollen grains and the allergen Sal k1 during 2015, 2016 and
2017.

Material and Methods: Pollen samples were taken with a volumetric collector type Hirst
(Burkard®) and bioaerosols with a Burkard type Cyclon® sensor. The levels of Sal k1 were
determined from the samples of bioaerosols with the ELISA-sandwich immunoassay method.
Results: The sum of the daily concentrations of pollen grains was 371 g/m3 in 2015, 291 g/m3 in
2016 and 369 g/m3 in 2017. The highest concentrations of pollen grains were in August-
September, with a peak day on September 7 in 2015 (35 grains/m3), on August 20 in 2016 (30
g/m3) and on August 19 in 2017 (19 g/m3). The highest concentration of Sal k1 was on August
18 (17.5 ng/ml) in 2015, on August 14 (20.1 ng/ml) in 2016 and on August 12 (16.8 ng/ml) in
2017. The relationship between the daily pollen concentrations of Amaranthaceae and the
concentration of the Sal k1 allergen (Spearman statistical test) has resulted significant and with
a positive correlation, with a coefficient of determination of 0.581 for 2015, 0.691 for 2016 and
0.397 for 2017.

Conclusions: The main period of pollination of Amaranthaceae comprises from May to

October, with the existence of two pollen peaks: one in May-June and another in August-
September. However, the peak of pollen in atmospheric air detected in spring does not
correspond to high levels of Sal k1. Probably, in this area, the flowering of the species of
Amaranthaceae in these months correspond mostly to other species such as Chenopodium
album and the full flowering of Salsola kali occurs at the end of summer.
 Poster 3

The adherence of North Alentejo population to Mediterranean Patterns: focus

on the consumption of polyphenol-rich products and its relationship with
saliva composition
T. Louro1, H. Luís2, P. Moreira3 , E. Lamy1,
1
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e Formação
Avançada), Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

2
Faculdade de Medicina Dentária – Universidade de Lisboa / Escola Superior de Saúde – Instituto Politécnico de
Portalegre. Avenida de Santo António, 23, 7300-047 Portalegre, Portugal

3
FCNAUP (Faculdade de Ciências da Nutrição e Alimentação, Universidade do Porto), Rua Dr. Roberto Frias, 4200-
465 Porto, Portugal.

Email: [email protected]

Adherence to Mediterranean Diet (MD) is being lost in most of the countries of the
Mediterranean, namely Spain, Italy or Greece. Although Portugal being geographically not in
the Mediterranean basin, an ancestral influence from the neighbor countries resulted in MD as
culturally settled. Similar to other countries, also in Portugal, a shift in dietary habits has also
been observed in the last decades. One of the characteristics of Mediterranean dietary
patterns is the consumption of non-processed vegetable based products, and consequently to
the consumption of high amounts of antioxidants, among which polyphenols. Despite of what
has been referred above, in nowadays, a characterization of MD adherence in Portuguese
population is missing. Moreover, even when dietary habits are characterized, the consumption
of antioxidants is missing, what represents a limitation, since these compounds have
particularly important health effects.

The major aim of the present study was to characterize the dietary patterns of the population
of North Alentejo, searching for the level of adherence of this population to MD and for the
levels of polyphenols intake. Moreover, and since the consumption of particular types of
polyphenols are considerably linked to salivary proteome, an intervention study was
performed, where the effect of polyphenols consumption, in saliva protein profile, was
assessed.

In terms of MD adherence, it was possible to observe that only a small percentage of North
Alentejo population (around 19%) presents a high score of adherence to MD. Even so, by
comparing with reports from other populations, it is not a very low rate. Concerning the
intervention study, it was possible to observe changes in salivary protein profile after the
consumption of extra dose of polyphenol-rich foods. These changes were in the levels of
salivary proteins that may also be related with oral food perception, suggesting that
polyphenol consumption habits may influence further acceptance of foods.

These results will be detailed and discussed in the proposed presentation.

 Poster 4

Unveiling food consumption patterns of rural dwellers in Europe – a critical approach to

social innovation

Paola A. Hernández1
1
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), , Universidade de Évora, Pólo da Mitra, Ap. 94,
7002-554 Évora, Portugal.

Email: [email protected]

Abstract: The concept of social innovation has increased its popularity in current
developmental research in rural studies in Europe, as it appears to respond to the various
social and economic disparities faced by a fragile rural population and the confronting
dynamics in the way we produce and consume food. Social innovation is a contested concept
among policymakers, advocates and critics; however, it is a useful approach to the set of new
ideas (products, services and models) aimed at meeting social needs and creating new
relationships and collaborations to tackle rural issues. The objective of this thesis paper is to
develop a suitable theoretical framework to sketch a methodology for mixed methods data
collection to unveil the food consumption patterns by rural dwellers in selected regions across
Europe. Assuming new forms of reconnecting producers and consumers are inspired from
innovative food system models, the paper elaborates on the existing literature about social
innovation and alternative food networks, as well as on debates on food security. This PhD
chapter paper carries out a literature review on the topic from academic research and
empirical work and identifies eight variables to be included in the following stage of the PhD
work in the data collection phase. Main results from the research highlight the importance of
alternative local food systems in shortening the linkages between producers and consumers,
as strategies to address the environmental, structural and food quality issues derived from the
current global food regime. However, a holistic understanding of complex local food systems
must move beyond the ‘productivist’ trap of alternative food systems and the focus on market
differentiation, plus acknowledge innovations’ multifunctional nature and reflect on the
degree of embeddedness of actors in these new designed food systems.

Keywords: social innovation, alternative food networks, food relocalization, food access, food
availability

This work is funded by National Funds through FCT - Foundation for Science and Technology under the Project UID/
AGR/ 00115/2013/
 Poster 5
Texture of Cachena meat.

S. Ricardo-Rodrigues1; A.C. Agulheiro-Santos1, M. Laranjo1,2, E. Potes1, M. Elias1

1 Instituto
de Ciências Agrárias e Ambientais Mediterrânicas (ICAAM), Universidade de Évora, Pólo da Mitra, Ap. 94,
7006-554 Évora, Portugal.

2 IIFA-Instituto de Investigação e Formação Avançada, Universidade de Évora, Ap. 94, 7006-554 Évora, Portugal

The meat quality, mainly tenderness, was affected by complex interactions of multiple
antemortem and postmortem factors such as animal genetics, feeding, handling, slaughter
process. Genetics determines an animal's potential for producing tender meat, and its
interaction with ante- and postmortem environment and also management will determine the
ultimate tenderness of the meat from each animal.

Cachena is a cattle breeds, part of the Portuguese genetic heritage, with high interest for the
south Alentejo region due to the high rusticity of these animals. Animals are small and the
meat is known by its excellent characteristics of texture and flavour. The determination of the
factors that affect meat tenderness of Cachena is of extreme importance for the producers
and for the industry. The goal of GOCACHENA project is understanding the relation among
genetics, feeding, age and weight of animals, with the quality, mainly tenderness, of this
exquisite meat. This research work is part of the refereed project and obtained on preliminary
results useful for the persecution of the main goals.

The modalities of this trial are different due to the preparation of the meat of Long dorsal:
sliced (aging 2, 5, 8 days) and entire pieces (aging 2, 8, 10 days). The animals were males, with
slaughter weight from 165 to 225 kg, and age between 6 and 13 months. The texture was
analyzed trough Texture Profile Analyze (TPA) using a Texture Analyzer TA.HD.Plus (©Stable
MycroSystem). From each animal two parts of the Long dorsal muscles were separated and
packed in bags each one with two samples and the measures were done in triplicate in each
slice of each sample. The samples were prepared according to Fabre et al (2018) and Veiseth-
Kent et (2018).

The results shown that there aren’t significant differences for Tenderness, Springiness and
Chewiness for sliced meat, however a slight decrease was noticeable. Considering the entire
pieces, the only significant difference was found in the values of Springiness. The values of
tenderness unexpectedly increase slightly.

The dramatic heterogeneity in the meat of animals may be caused by genetic factors. This
statement confirms the necessity of deep studies like that of GOCACHENA

Acknowledgements: This work was supported by Project GO-301 PDR2020-1.0.1-FEADER 030803. This research was
partially supported by National Funds through the FCT – Fundação para a Ciência e Tecnologia, project
UID/AGR/00115/2013. M. Laranjo acknowledges a Post-Doc research grant from FCT (SFRH/BPD/108802/2015).
 Poster 6

Starch based edible coatings effectiveness in preventing fresh fruit phenolics

degradation
Serranito, Agostinho J. 1*, Rouxinol, M. Inês1**, Rato, A. E.1, Agulheiro-Santos, Ana C.1

1 Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, apartado 94 7002-554, Évora,
Portugal
*Aluno do Curso de Agronomia; ** Aluno de Doutoramento em Bioquímica

Email: [email protected]

Starch-based edible coatings can enhance food quality, safety and stability. Unique
advantages of edible coatings can lead to the development of new products, such as individual
packaging for foods, carriers for various food additives, and nutrient supplements (M.A. García
et al 2009)1.
The extension of post-harvest life on fruits should consider challenges like extending
senescence periods, reducing dehydration, the onset and microbial growth. Traditional
preservation methods are focused on some of these challenges, like creating synthetic film
packaging or antimicrobial treatments (A. El Gaouth, et al 1991)2.
Phenolic compounds are responsible by the fruit’s organoleptic properties, however
despite their numerous beneficial proprieties, they are extremely sensitive and susceptible to
degradation. Edible coatings have a high potential to extend the shelf life of fresh fruits.
The use of coatings made from natural products can be beneficial to prevent the
degradation of phenolics and increase the fruits shelf life.
A sample of strawberries was tested with a modality with edible coating and one
without , the study went for 8 days and samples from all days were analysed by an improved
Folin- Ciocalteau method (data to be published).
The extraction for analysis was made by means of an ethanolic solution. And the
results were expressed in
The results shown that, the fruits without coating there is a degradation on the
phenolic content after the2nd day of conservation. The fruits with coatings kept the phenolic
content concentration stable. The same behaviour can also be observed in weight loss, in
which fruits lost more weight, respiration rate was higher, and pH variation over time. The
berries tested with coatings lost less weight, maintained pH level’s thru time and decreased
the respiration rate.
In sum, the use of edible coatings derived from potato starch can improve and enlarge
the post-harvest life of fruits.
Future studies should have in mind new formulations and ways of application.

1
María A. García , Adriana Pinotti , Miriam N. Martino , and Noemí E. Zaritzky (2009)
Characterization of Starch and Composite Edible Films and Coatings. In : M.E. Embuscado and
K.C. Huber (eds.), Edible Films and Coatings for Food Applications, 169–209
2
El Gaouth A , Arul J , Ponnampalam R , Boulet M (1991) Chitosan coating effect on storability
and quality of fresh strawberry . J. Food Sci. 56 , 6 , 1618 – 1620
Olive Grove, Vineyard and Other Cultures
 Invited Speaker 3
Plant viruses: from causal agents of disease to plant protectors

Maria do Rosário Félix1, Carla Varanda2, Patrick Materatski2, Maria Doroteia Campos2 and
Maria Ivone Clara1

1ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Fitotecnia, Universidade de

Évora, Pólo da Mitra, 7002-554 Évora, Portugal.
1ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e Formação
Avançada), Universidade de Évora, Pólo da Mitra, 7002-554 Évora, Portugal.

Email: [email protected]

Traditionally, plant viruses are viewed as harmful, undesirable pathogens and those that affect
olive trees are not an exception. The symptoms associated to this kind of agents in olive trees
are, among others, chlorotic lesions, defoliation, deformation in fruits and leaves, and death of
stems and trees. Viral diseases are not treatable and as such it is necessary to increase the
knowledge on their epidemiology to improve sanitary rules and prevent infections in the field.
In Portugal, high levels of viruses were detected, reaching up to 100%, in dozens of olive
orchards sampled, being the most prevalent the necroviruses, from Alpha- and Beta-necrovirus
genera. The molecular characterization of viral genomes and their replication strategies have
been studied, and this has allowed the design and the improvement of molecular tools for olive
viruses diagnosis. Apart from the harmful effects, viruses can provide several useful 'designer
functions' or 'sequence modules' with which future vectors for general biotechnology may be
developed. The study of viral genomes, particularly in the context of their pathogenic
interactions with host plants, led us to the design of two types of vectors for plant protection
use. One of those is based on virus-induced gene silencing (VIGS), using the natural plant gene
silencing strategy to protect plants against viruses; and the other is based on the expression of
antimicrobial peptides (AMPs), using the ability of virus genome replication to produce
antibiotics and antifungal molecules to prevent diseases caused by bacteria and/or fungi. In
both, a partial or complete gene is introduced in the vector and then the virus-related
'interfering' molecule is stably introduced into plants via the DNA-transfer mechanism of
Agrobacterium tumefaciens or mechanical viral inoculation. The presence of these viral-vectors
in plants confers them protection against specific diseases. The possibility to manipulate the
plant-virus relationship is a powerful tool to increase and improve strategies for unconventional
crop protection techniques.
 Invited Speaker 4

High Resolution Chromatography. Method for target analysis

Marco Gomes da Silva

LAQV-REQUIMTE, FCT-NOVA, Campus de Caparica, 2829-516, Caparica

Gas chromatography (GC) is a powerful analytical tool that can be applied to a variety of
matrices in order to separate, detect and identify their constituents. Capillary gas
chromatography was in the six decades used for complex matrices characterization as well as
target analysis. New stationary phases, with broader application range, more robust and thus
more versatile, together with developed column manufacturing procedures and
instrumentation allow to enhance the resolution of the chromatographic systems, defining
new productivity and throughput standards. Presently GC/MS is the most frequently used
hyphenate technique for the characterization and identification of many volatile and
semivolatile compounds. However, in spite of the continuous development of equipment,
techniques and analytical methodologies, a total separation of all the sample components is
still unachievable, due to the complexity of the samples and of the analysis (high number of
components, structure similarities, isomers, and wide range of concentrations). High
similarities are thus expectable between the retention times of several analytes,
independently of the stationary phase used, that will result in coelutions. These coelutions are
often impossible to detect and identify with some GC/MS instruments, in spite of the use of
selective single ion monitoring or in tandem (SIM and MS/MS) mode or of complex
deconvolution processes. An additional problem results from the wide range of analyte
concentrations in their matrices. Consequently, the trace level analytes, that sometimes are
the biologically active components in the matrix under study, may never be detected, if they
are coeluting with high concentration compounds. Future developments are thus considered
in order to mitigate these constraints.
 Oral Communication 7

Study of Portuguese extra virgin olive oils according to the producing

agricultural method: A qualitative and ecological approach

M. Ferro1,2,*, M. Cabrita2, J.M. Herrera3, M.F. Duarte1,2

1
Centro de Biotecnologia Agrícola e Agro-Alimentar do Alentejo (CEBAL)/Instituto Politécnico de Beja (IPBeja), 7801-
908 Beja, Portugal.
2
Instituto de Ciências Agrárias e Ambientais Mediterrânicas – ICAAM, Universidade de Évora, Instituto Pólo da
Mitra, Apartado 94, 7006-554, Évora, Portugal.
3
Centro de Investigação em Biodiversidade e Recursos Genéticos – CIBIO-InBIO, Universidade de Évora, Casa
Cordovil 2a Andar, Rua Dom Augusto Eduardo Nunes, 7000-651 Évora, Portugal

*Email: [email protected]

During the past decade, Portugal has significantly increased its olive production, competing for
its place as a leading producer at international markets. Alentejo region, with a total exploited
agricultural area of 2.100.762 ha (58% of the total national explored area), is by far the most
dedicated Portuguese province for olive oil production. A total of 180.607 ha are exclusive
devoted to olive production, with an input of 74% of the whole national olive oil production.
The replacement of traditional orchards by intensive and super-intensive ones represents a
notorious developments in the olive growing sector, focused on increasing productivity to its
maximum. Opposing to the traditional distribution, these intensive and super-intensive
orchards are characterized by making the most out of the cultivated area, in general with
densities as high as 450 trees/ha and 1500 trees/ha, respectively. From the productivity
perspective, super-intensive systems represent a breakthrough in extra virgin olive oil (EVOO)
production, with extremely premature and productive trees aligned in high density systems.
Nevertheless, not only productivity should be considered. The ecological EVOO´s footprint is
quite distinct according to the applied agricultural practices. Soil and water contaminations are
among the most concerning factors when considering these new approaches, and they are
mainly due to excessive application of agrochemicals.
Regarding the EVOO quality, to the best of our knowledge, no study has been reported
associating agricultural practices to EVOO quality, either by fatty acid, phenolic or volatile
characterization.
With this work, we intend to cross several information-related sources, such as the EVOO
chemical characterization (fatty acid, hydrophilic phenolic and volatile profiles), the presence
of chemical pollutants and pesticides in both EVOO and soil/water and its ecological impact,
with distinct agricultural systems (organic, traditional, intensive and super-intensive). With this
information, chemometric studies will be applied in order to create a tool to access the
nutritional value and environmental impact of EVOOs production systems.
M.F. acknowledges FCT (SFRH/BD/140083/2018). Authors are grateful to FCT for the financial support to ICAAM
Research Unit (FCT UID/AGR/00115/2013) and would also like to acknowledge the Program Alentejo 2020, through
the European Fund for Regional Development (FEDER) under the scope of OleaValor – Valorização das Variedades
de Oliveira Portuguesas (ALT20-03-0145-FEDER-000014). J.M.H. is supported by FCT and the European program
Alentejo 2020 through the contract IF/00001/2015 and the project PTDC/AAG-REC/6480/2014.
 Oral Communication 8

Evaluation of anthocyanin profile during ripening in Vitis vinifera red wine

grape varieties grown in Alentejo
Mª Inês Rouxinol*1, Mª Rosário Martins2, João Mota Barroso1, Ana Elisa Rato1
1
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e Formação
Avançada), Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

2
Laboratório HERCULES (Herança Cultural, Estudos e Salvaguarda), Universidade de Évora, 7000-809 Évora

* Biochemistry PhD Student

Email: [email protected]

Anthocyanins are the natural compounds responsible for the red color in wines. They are the
final products of a specific branch on the flavonoid pathway, meaning that they belong to the
phenolics class. Their distinct color makes them extremely important for the fruit and its
subproducts quality (namely red wine), enhancing the sensorial qualities. Also, it has been
reported that anthocyanins (along with other phenolic compounds) have high antioxidant
activity. Wine is widely consumed around the globe, and there is an increasing interest in its
consumption due to its reported health benefits when consumed in moderation. In this study,
four varieties (Syrah, Tempranillo, Touriga Nacional and Trincadeira) produced in Herdade da
Mitra were monitored during version to understand the development of some phytonutrients
and their respective antioxidant potential.

To allow these quantifications, phenolic compounds were extracted and evaluated. Total
phenolic compounds content1 and total flavonoids1 were evaluated by UV/Vis spectroscopy.
Anthocyanins separation, identification and quantification was determined by HPLC-DAD2.
Antioxidant activity of grape extracts was quantified using three different methods3 (DPPH
method, total reducing power and β-carotene/linoleic acid system).

During ripening, Syrah, Touriga Nacional and Trincadeira presented an increasing of phenolic
content, flavonoids and anthocyanins, on the other hand, Tempranillo presented a decrease in
these compounds during verásion. Monoglucoside-anthocyanins have a higher content in
grapes during all maturation process and Malvidin-3-O-glucoside is the most prevalent
anthocyanin. Grape extracts have showed high antioxidant activity by all methods tested,
highlighting the high ability of scavenging free radicals. Results showed a high correlation
between total phenolic content and the antioxidant ability to scavenge free radicals.

References
1. Hosu, A., Cristea, V.-M. & Cimpoiu, C. (2014) Analysis of total phenolic, flavonoids, anthocyanins and
tannins content in Romanian red wines: Prediction of antioxidant activities and classification of wines using artificial
neural networks. Food Chem. 150, 113–118.

2. Antoniolli, A., Fontana, A. R., Piccoli, P. & Bottini, R. Characterization of polyphenols and evaluation of
antioxidant capacity in grape pomace of the cv. Malbec. Food Chem. 178, 172–178

3. Arantes S., Piçarra A., Candeias F., Teixeira D., Caldeira A.T., Martins M.R. (2017) Antioxidant activity and
cholinesterase inhibition studies of four flavouring herbs from Alentejo, Natural Product Research, 31 (18): 2183-
2187.
 Oral Communication 9

Mycorrhization of Cistus spp. with Terfezia arenaria (Moris) Trappe and its application
in desert truffle cultivation

Louro, R.1, Natário, B.1, Santos-Silva, C.1

1ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Biologia, Universidade de Évora, Pólo da
Mitra, Ap. 94, 7002-554 Évora, Portugal.

Desert truffles are of considerable interest for ecological, agroforestry and commercial purposes. They
represent a key component of the mycological flora in arid and semi-arid regions due to their important role
as symbiotic partners of diverse host plants. Plus, their fruitbodies constitutes a potentially important food
source, rich in proteins and poor in carbohydrates and lipids. Desert truffle increasing demand has recently
boosted more research, aiming to achieve new strategies to enable their medium-large-scale cultivation,
but so far only Terfezia boudieri and Terfezia claveryi were successfully cultivated together with
Helianthemum spp. in basic soils.
T. arenaria is the most commercially valued desert truffle specie in central and southern Portugal. Usually,
it is found in acid soils, establishing mycorrhizal associations with several Cistaceae plants. Since, Cistus
species are the most important elements of Mediterranean scrublands in Alentejo (southern Portugal), the
objectives of the proposed research were to expand the current knowledge about the mycorrhizal
associations of Cistus spp. with Terfezia arenaria and develop strategies to allow its application towards
desert truffle cultivation and the production of Cistus mycorrhized plants.
While working toward the proposed goals, we have repeatedly sampled numerous specimens of Terfezia
throughout the Alentejo region and through molecular characterization of the collected specimens we were
able to describe a new taxa, Terfezia lusitanica sp. nov. We also successfully developed novel
micropropagation protocols for Cistus salviifolius and C. ladanifer, the two most common Cistus species in
the region, which enabled us to effectively infect, ex-vitro, the root system of the abovementioned Cistus
species and to obtain high mycorrhization rates in both cases. Recently, we were able to surpass the major
challenge of the proposed research, the isolation of Terfezia spp in pure culture, and now we have
collections of T. arenaria and T. fanfani mycelia growing in agar plates. In the next steps, we aim to develop
an efficient in-vitro mycorrhization protocol using Terfezia mycelium, in order to, characterize the
mycorrhizae formed between these two desert truffles and the abovementioned Cistus species and to
evaluate which are the best plant-symbiont to enable Terfezia medium-large-scale cultivation in acidic
situations.

This work was supported by the project “Cistus spp. mycorrhization with Terfezia arenaria (Moris) Trappe – application in desert
truffles production (ALT20-03-0145-FEDER-000006).
 Poster 7

Olive flowering phenology variation of three traditional Portuguese olive

varieties in Alentejo region

Elsa Lopes1,4, Francisco Mondragão-Rodrigues1,2, Paulo Vivas1, Graça Pacheco de Carvalho1,

Luís Alcino Conceição1,2, Augusto Peixe2, María Vitória Alarcón3 & Julio Salguero4
1Instituto Politécnico de Portalegre, Escola Superior Agrária de Elvas, Portugal, Email: [email protected]
2 ICAAM, (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Portugal.
3Área de Hortofruticultura. Centro de Investigaciones Científicas y Tecnológicas de Extremadura (CICYTEX). Instituto
de Investigaciones Agrarias Finca La Orden-Valdesequer, Badajoz.

4Fisiología Vegetal. Escuela de Ingenierías Agrarias. Universidad de Extremadura, Badajoz.

In Olea europaea L. the beginning, the duration and the end of flowering are determined by
genetic characteristics inherent to each variety and by the physiological response to
meteorological factors of the year, mainly temperature.

During two consecutive campaigns (2017 and 2018), a study was carried out to collect
information on the flowering phenology of the varieties ‘Galega vulgar’, ‘Azeiteira’ and
‘Cobrançosa’, in intensive olive grove on a farm located in a Portuguese town of Monforte.
For the campaign of 2018, the beginning of flowering (marked by stage BBCH 60) took place in
a range of 3 days, between May 21th ('Galega', ‘Azeiteira’) and May 24st ('Cobrançosa'). The
duration of the flowering period varied, depending on the variety, between 11 days
(‘Cobrançosa), 14 days (‘Galega’) and 17 days (‘Azeiteira’).
Compared to 2017, in this campaign all varieties were in bloom much later (approximately 3
weeks for ‘Cobrançosa’ and ‘Galega’ and 4 weeks for ‘Azeiteira’), and the flowering period was
shorter (between 16 and 21 days in 2017).
The obtained results clearly demonstrated an inter-annual variability in the varieties under
study, which is very important to cope with climate changing, presenting highly significant
differences for all of the parameters when comparing the results of both campaigns. These
results are crucial to the phenological characterization of these Portuguese olive varieties under
study, however, to confirm its differentiated behaviour, it is necessary to repeat this
phenological evaluation in further campaigns.

The experimental trial was financed by the EAFRD and the National Funds through the Regional Operational Program
ALENTEJO 2020, Operation ALT20-03- 0145-FEDER- 000014 - "Valuation of Portuguese Olive Varieties (Oleavalor)".
 Poster 8

Endophytic fungal communities of grapevine roots in Italian and

Portuguese commercial vineyards afflicted by esca

J. Concas1,2, S. Murolo1, G. Romanazzi1, M.R. Félix2

1
Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Via Brecce
Bianche, 60131 Ancona, Italy.
2
Departamento de Fitotecnia, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Instituto de Investigação
e Formação Avançada, Universidade de Évora, Núcleo da Mitra, Ap. 94, 7006-554 Évora, Portugal.

Email: [email protected]

Amongst several pathogens reported in grapevine, the agents of esca represent one of the
most destructive, causing serious economic problems in worldwide vineyards. The aim of this
study was to detect difference in fungal endophytic population (species and quantity) with and
without presence of esca pathogens. The survey was carried out from roots of 4 grapevine
varieties: Verdicchio, Chardonnay, Trincaderia and Alicante Bouchet. The sampling was
performed during summer 2017 from 6 commercial vineyards located in Alentejo (Portugal)
and Marche region (Italy), selecting a total of 25 plants with and 25 plants with no symptoms
of esca. A suitable method for external disinfection of vine roots not unsettling endopyhtes
was developed and used successfully. Portions of disinfected roots were putted in growth in
Petri dishes with Potato Dextrose Agar, and each single grown fungus was isolated. Isolates
were grouped according with morphological characteristics, and species were identified by
morphological characterization. The work allowed to isolate in vitro more than 160 fungal
endophytes. The first results showed a sensitive difference (higher than 50%) of isolates
between Verdicchio-Chardonnay and Trincaderia-Alicante Bouchet cvs. In Verdicchio and
Chardonnay cvs, 9 species mainly identified as belonging to Fusarium spp. and Trichoderma
spp. have been found as occurring in more than 26% of plants, and 5 species have been
identified in more than 38% of plants. In Trincaderia and Alicante Bouchet cvs, 3 species
(Epicoccum sp. and 2 unidentified spp.) have been detected only in asymptomatic plants.
Amongst all the not found isolates in plants with symptoms, 13 of them occur as common in
no symptomatic plants of same cv (eg.: Phoma sp., Fusarium sp., Epicoccum sp., Chaetomium
sp., Trichoderma sp.). A specie, not identified as a common agent of esca, occurred only in
plants that showed symptoms, deserving attention. A molecular characterization by ITS
primers is currently underway. This first result allowed already to give a first contribute to
further studies for biological control of esca.
Mediterranean Agro-Silvo-Pastoral Systems
 Invited Speaker 5

Technologies for monitoring montado ecosystem (ECO-SPAA): a global

approach
João Serrano
Prof. Auxiliar com Agregação
Departamento de Engenharia Rural
ICAAM
Universidade de Évora

Abstract

The main objective of this work was to present several proximal and remote sensors
with potential to monitor relevant variables in the complex Mediterranean silvo-pastoral
montado ecosystem (ECO-SPAA). The interest in using these two techniques in a
complementary way can make an important contribution to:
(i) Understanding the interaction between various factors and components that
interrelate in this ecosystem (soil, pasture, trees and animals);
(ii) The determination of key components of pasture productivity and quality;
(iii) Support grazing management decisions; and
(iv) Implementation of site-specific pasture management.
 Invited Speaker 6

Agro-silvo–pastoral Systems: Inovation with Tradition

José Mira Potes

Instituto Politécnico de Santarém – Escola Superior Agrária

A full characterisation of the Mediterranean environment is fundamental, in order to adequately

frame agro-silvo-pastoral systems. Through the disaggregation of the agricultural, forestry and
animal components, the respective productive subsystems are analysed. The integration needed
for the good management of the overall system is performed through the Montado rotation and
extensive livestock feeding scheme, finally focusing on the importance of the available
environmental services. The Dynamics of the Montado will also be discussed.
 Oral Communication 10

Visions and strategies for water management in agriculture: a transdisciplinary experience in

the Alentejo
Homobono, T1.; Guimarães, M.H.2; Madureira, L.1
1
Universidade Trás-os-Montes e Alto Douro, Departamento Economia, de Sociologia e Gestão, Quinta de Prados, 5000-554, Vila
Real.
2
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Biologia, Universidade de Évora, Pólo
da Mitra, Ap. 94, 7002-554 Évora, Portugal.

Email: [email protected]

Agriculture faces the challenge of adapting to climate change and the context of natural resource crises
while continuing to respond to global food needs. Thus, high ecological value agriculture, agro-ecological
approaches and agro-silvo-pastoral systems can help to avoid damages to the ecosystems goods and
services where they are developed. The complexity of these challenges requires the development of
scientific approaches that promote the analysis of the agricultural sector in a holistic and inclusive way.
The project proposes the implementation of a reflective and adaptive scientific production process,
based on the principles of transdisciplinarity. The participatory approaches adopted put local actors as
an active part in exploring the diversity of perspectives in understanding the water management role in
agricultural sector decision making. This project is based on a case study in the municipality of Serpa, in
the region of Alentejo (Portugal) and is supported by the international project DIVERCROP
(https://divercropblog.wordpress.com/), which aims to analyze and understand productive systems in
the wester Mediterranean basin, at different scales. Through the transdisciplinary premises the projects
aim to contribute to the construction of a systemic view of the relationship between water resources
management and territorial development. The case study contextualization will be carried out through
the socio-ecological systems framework proposed by Elinor Ostrom, which promotes a holistic
understanding of complex issues such as the development of agricultural production in the current
context. This characterization is based on the results of semi-structured interviews with key actors of the
studied region, and will be complemented by a literature review. The results obtained in this stage will
allow the design of the participatory approach stage, in which the Territory Game. This technique will be
applied to identify the main perceptions about water changes and their effects on agricultural and land
use activities, as well as, to the creation of a vision for the territory and the pathways needed to achieve
the desirable future. To evaluate the transdisciplinary approach used, questionnaires will be applied to
detect factors that can influence the actor’s participation in the knowledge sharing. Two main results
are expected at the end of the project: identification of measures adopted and challenges encountered
regarding water management in the municipality studied, and to contribute to the promotion of
transdisciplinarity in the management of natural resources.

KEY-WORDS: water management; agriculture; transdisciplinarity; SES framework; Alentejo.

 Oral Communication 11

Climate change in the development of a model of occurrence of forest fires:

determination of trend series and critical periods

Nunes, L.J.R.1,2, Meireles, C.I.R.1,2, Gomes C.J.P.1,2,3, Almeida Ribeiro N.M.C.1,2,4

1
IIFA - Instituto de Investigação e Formação Avançada), Universidade de Évora, Pólo da Mitra, Évora, Portugal.

2
ICAAM - Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Évora, Portugal

3
Departamento da Paisagem, Ambiente e Ordenamento, Universidade de Évora, Évora, Portugal

4
Departamento de Fitotecnia, Universidade de Évora, Évora, Portugal

Email: [email protected]

Climate change is a phenomenon that could significantly affect the day-to-day activities of the
rural communities, with particular emphasis on those directly dependent on the agricultural
and forestry sector. In this way, the present work intends to base the impact that climate
change has on forest risk assessment, namely on how the occurrence of rural fires is affecting
the management of the forest areas and how the occurrence of these fires has evolved in the
near past. Thus, a comparative analysis of the data provided by IPMA - Portuguese Institute of
the Sea and the Atmosphere was carried out for the period from 2001 to 2017, with the
climatic normal for the period between 1971 and 2000, for the variables of Average Air
Temperature and for Precipitation. In this comparative study, the mean monthly values were
considered and the months in which anomalies occurred were determined. Anomalies were
considered in the months in which the Average Air Temperature varied more or less 1 degree
than the value corresponding to the climatic normal, in at least 50% of the national territory.
The same procedure was repeated for the variable Precipitation, counting as anomaly the
occurrence of a variation in precipitation greater or less than 50%, also in 50% of the national
territory. Then the calculation of the moving averages for cycles of 3, 5 and 7 periods was
applied and the trend lines were projected. Subsequently, the relationship between the results
obtained and the occurrence of rural fires was carried out, the spatial distribution of forest
area, species and stand structure, was not analyzed in the present work.
From the obtained results it was possible to confirm the existence of a tendency for the
occurrence of climatic anomalies, highlighting the occurrence of an increasing number of
months with temperatures higher by at least 1 degree. Even without It was also possible to
foresee the co-relation between the occurrence of rural fires and the periods of anomaly and
absence of precipitation. From the results obtained it is also possible to infer that, and
analyzing the tendency for these phenomena to occur, it is necessary to change the "Critical
Period of Rural Fires", since it is statistically verified that what is currently in force does not
covers the entire period where anomalies occur and where large-scale rural fires can
potentially happen.
 Oral Communication 12

Evaluation of the interactions soil - pasture - trees and

animals in Montado ecosystem
E. Carreira1, A. Pereira2, J. Serrano3, M. Carvalho4
1Aluno de Doutoramento em Ciências Agrárias e Ambientais, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-
554 Évora, Portugal.

2ProfessorAuxiliar com agregação do Departamento de Zootecnia, Universidade de Évora, Pólo da Mitra, Ap. 94,
7002-554 Évora, Portugal.

3ProfessorAuxiliar com agregação do Departamento de Engenharia Rural, Universidade de Évora, Pólo da Mitra, Ap.
94, 7002-554 Évora, Portugal.

4Professor Catedrático do Departamento de Fitotecnia, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554
Évora, Portugal

Email: [email protected]

The work associated with ECO-SPAA, a project to be held at Herdade da Mitra (Évora
University), have as main objective the monitoring of the Montado ecosystem (ECO)
through monitoring and study of interactions between soil, pasture, trees and animals
(SPAA). After the third year, and after soil correction and fertilization (2ha), the
component related to the dynamic management of grazing will be implemented
between November 2018 and June 2020. Thus, in two plots with about 2ha each, 12
trees (holm oaks) 6 were selected in a low zone and 6 in a high zone, with 4 exclusion
boxes, 2 under the canopy and 2 outside the canopy . The following measurements are
taken every month (between September and June): 1) NDVI, which is correlated with
the amount of chlorophyll and with the vegetative vigor of the pasture; 2) Capacitance,
which is correlated with pasture production (Kg MS / ha); 3) Soil moisture; 4)
Thermographic photographs, which measure the temperature of the pasture; 5)
Pasture height; 6) Pasture crops, for percentage of dry matter, crude protein and NDF.

Following this, a doctoral student will be held, with the following questions: 1) why are
there differences in pasture growth under and outside the tree canopy?; 2) After
correction of soil pH are there differences in productivity under and outside the tree
canopy?; 3) What is the productivity of animals in this system?; 4) What is the effect of
grazing on pasture productivity?. We will have 4 plots, in 2 we will have a fixed header,
and in the other 2 there will be variable biotic loads depending on the availability of
pasture. To study the subject and try to answer the questions the methodology is as
follows: 1) monthly performance of the measurements described above; 2) weighing
and body condition of the sheep; 3) laboratory analysis of pasture. On the one hand,
the soil-pasture interaction will be evaluated, on the other the pasture-animal
interaction.
Biodiversity and Ecosystems Functioning
 Invited Speaker 7

Roads and biodiversity. To coexist or not to coexist, that is the question.

António Mira

Universidade de Évora. Departamento de Biologia. Pólo da Mitra, 7002-554 Évora

([email protected])

Roads are the most extended infrastructure on Earth and are present in most modern landscapes.
They are critical for connecting human communities, but at the same time they pose a threat to
biodiversity. There are several negative effects of the roads. However, the most relevant are
habitat fragmentation and the associated barrier effect on the movement of animals; and road kill
mortality, which at present is already higher at world level than kills from hunting. There are
currently 64 million km of roads on Earth and an additional 25 million km are expected to be built
by 2050, 90% of which in developing countries in areas where biodiversity is high. Thus, it is
urgent to act for the promotion of more sustainable roads reducing their impact on wildlife.
However, there is still a lack of information and prior to any action it is important to clarify several
issues in order to promote the success of the interventions. This presentation focuses on an
assessment of the main effects of roads on biodiversity and the main issues still open for debate,
for which a research agenda will be needed to find efficient solutions that will reverse the global
decline in biodiversity worldwide.
 Invited Speaker 8
Love me, love me not: a story of storks and power lines

Francisco Moreira

Universidade do Porto– CIBIO-InBIO

After a strong decline since the 1950-60’s, the breeding white stork (Ciconia ciconia) population
has been increasing steeply since the 1980’s. Along this recovery period, a curious behavioural
change has occurred: storks have been “abandoning” their traditional nest sites in trees and
buildings to be increasingly use electricity pylons to build nests. The result is that white storks
are “wired”, so that no less than 25% (!) of the nests in the country are built on pylons, mainly
in very high-tension power lines. These nests can cause management problems to electricity
companies, as they are the cause of power outages. On the other hand, this preference by storks
has a drawback: it increases the risk of mortality for birds, through collision with power line
cables and electrocution. This presentation will report this “love and hate” story, trying to
address the pros and cons for storks nesting on power lines, as well as the drivers of pylon
occupation.
 Oral Communication 13

Floristic analysis of four communities of Prunus lusitanica L. from Europe

and North Africa

M. Raposo¹, C. Meireles¹, S. Del Río², F. Vázquez-Pardo³ & C. Pinto-Gomes¹

¹Departamento de Paisagem, Ambiente e Ordenamento, Instituto de Ciências Agrárias e Ambientais Mediterrâneas

(ICAAM), Escola de Ciência e Tecnologia, Universidade de Évora. Rua Romão Ramalho, n.º 59, 7000-671 Évora,
Portugal.
²Departamento de Biodiversidad y Gestión Ambiental (Área de Botánica), Instituto de Ganadería de Montaña (CSIC-
ULE), Facultad de Ciencias Biológicas y Ambientales, Universidad de León. Campus de Vegazana s/n, E-24071, León,
España.
³Departamento de Producción Florestal y Pastos. Centro de Investigación Finca La Orden-Valdesequera (CICYTEX),
06187 Guadajira (Badajoz), España.
Email: [email protected]

In the scope of a PhD thesis in Agrarian and Environmental Sciences, we aimed to understand
the conservation state of Prunus lusitanica L. communities in its native territory, Europe and
North Africa. To achieve this, a set of field trips to Portugal, Spain, France and Morocco were
performed for data collection. These communities are true paleotropical relics of the Ibero-
Maghreb territories, being recognized as a priority habitat for conservation in Europe
(5230pt2), present in Annex I of the Habitats Directive (92/43/CEE). Also, this taxon is part of
the IUCN Red List, presenting a Vulnerable status at a global level, according to the latest
evaluation carried out using the IUCN criteria. Thus, this work presents a comparative analysis
between four distinct Prunus lusitanica communities, located mainly in the Rife Mountains
(Morocco), the Serra da Estrela (Portugal), the Leon Mountains (Spain) and the Western
Pyrenees (France). In this analysis we compared the floristic originalities of each territory, as
well as their similarities and differences in plant communities, highlighting its biogeographical
and bioclimatic aspects. As a result, some management measures are proposed in order to
improve its conservation status. Through fieldwork, it was also possible to identify new
occurrence sites for this species, which are published for the first time in this work.

Thanks for the cooperation of the teachers Mouhaydine Tlemcani, Sara Del Río, Angel Penas, Jean Jacques Lazare
and the project LIFE-RELICT – Preserving Continental Laurissilva Relics (LIFE16 NAT/PT/000754).
 Oral Communication 14

Effect of Alqueva Dam on terrestrial invertebrates communities: threat or

opportunity
R. Raimundo1,2 , D. Figueiredo3
1
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), IIFA (Instituto de Investigação e
Formação Avançada), Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.
2
CIBIO-InBIO, Universidade de Évora, Casa Cordovil 2ª Andar, Rua Dr. Joaquim Henrique da Fonseca,
7000-890 Évora, Portugal
3
Departamento de Biologia, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.

E-mail: [email protected]
The implementation of a dam has serious impacts in the biodiversity on terrestrial
invertebrates. As consequence, some islands that did not exist before are created. In the
Alqueva (South of Portugal), the effect of the implementation of a large dam raises the main
question: is it a threat or an opportunity for the biological communities? Thus, we focus our
objectives to answer this question and others related with terrestrial invertebrate
communities. The main goal of this Thesis is to measure the long-term influence of the
Alqueva dam on the ecological communities of the terrestrial invertebrates in 30
permanent islands of the Alqueva reservoir archipelago generated by this dam, and
understand the gains and losses of these communities in this enormous transformation of
the landscape. The terrestrial invertebrates target species are spiders, carabid beetles and
butterflies. The data will be analyzing to quantify the effect of the construction of the dam,
comparing two scenarios: before and after the flooding. To achieve this goal, we proposed
four specific objectives: 1. Determine which spiders and carabid beetles communities
presented major differences between the islands in the different time periods; 2. Explore the
relationships between spiders, carabid beetles and butterflies communities and
environmental variables; 3. Explore relationships between parameters of biological diversity
and environmental variables, using multiple regression models; correlations and predicting
distribution of these species in the future; 4. Evaluate the composition and evolution of the
analysed terrestrial invertebrate communities under the theory of island biogeography. To
accomplish the specific objectives, multivariate methods will be used to explore the data to
explain the distribution and changes of terrestrial invertebrate species and communities
present in Alqueva islands.
 Oral Communication 15
Accounting for connectivity uncertainties in predicting roadkills: a comparative

approach between path selection functions and habitat suitability models

1,* 2,3 4 1 1
Francesco Valerio , Filipe Carvalho , A. Márcia Barbosa , António Mira , Sara M. Santos
1
UBC – Conservation Biology Lab; Department of Biology, University of Évora, Évora, Portugal

2
Research Center in Biodiversity and Genetic Resources (CIBIO/InBIO), University of Porto, Campus Agrário de

Vairão, 4485-661 Vairão, Portugal

3
Department of Zoology and Entomology, School of Biological and Environmental Sciences, University of Fort Hare,

Private Bag X1314, Alice 5700, South Africa

4
Department of Biology, University of Évora, Évora - Portugal

E-mail address: [email protected]; (F. Valerio).

Functional connectivity modelling is increasingly being used to predict the best spatial location

for animal road-crossing structures and to mitigate wildlife roadkills. This tool requires

estimation of resistance surfaces, ideally modelled with movement data which are costly to

obtain. An alternative is to use occurrence data within species distribution models to infer

movement resistance, although this remains a controversial issue.

The aim of this study was: (i) to compare the performance of resistance surfaces derived from

path versus occurrence data in identifying road crossing locations of a forest carnivore; (ii) to

assess the influence of movement type (daily vs. dispersal) on this performance.

Resistance surfaces were built for genet (Genetta genetta) in South Portugal using 1) path

selection functions with telemetry data, and 2) species distribution models with occurrence

data. An independent roadkill dataset was used to evaluate the performance of each

connectivity model in predicting roadkill locations.

The results show that resistance surfaces derived from occurrence data are as suitable in

predicting roadkills as are path data for daily movements. When dispersal was simulated, the

performance of both resistance surfaces was equally good at predicting roadkills. Moreover,
contrary to our expectations, we found no significant differences in locations of roadkill

predictions between models based on daily movements and models based on dispersal. Our

results suggest that species distribution models are a cost-effective tool to build functional

connectivity models for road mitigation plans when movement data are not available.

Keywords: movement models; species distribution models; ecological process; mitigation;

roadkill; dispersal period.

FV was supported by a PhD fellowship, while SMS, AMB and FC by postdoctoral contracts, which were all funded by
Fundação para a Ciência e a Tecnologia (SFRH/BD/122854/2016, SFRH/BPD/70124/2010,
IF/00266/2013/CP1168/CT0001 and SFRH/BPD/115228/2016, respectively). We also received support from the
projects LIFE LINES (LIFE14 NAT/PT/00108) and POPCONNECT (PTDC/AAG-MAA/0372/2014).
 Oral Communication 16

Regime of fire in Portugal: assumptions and facts

Nuno Guiomar (ICAAM-Universidade de Évora)

Abstract
The definition of effective strategies for wildfire prevention, pre-suppression and suppression
(both initial and extended attack) requires the strict knowledge of the fire regime. Due to the
impacts that large fires have on landscape, economy and society in general, they foster
reactions aimed at finding quick and easy solutions to a problem that is extremely complex.
However, many of these options are supported by wrong assumptions about fire regime that,
when evolving into axiomatic approaches and from those to dogmatism, not only limit the
implementation of the necessary measures towards fire management, as they tend to increase
the complexity of the problem over time. The claim of a "Portugal without Fires" results from
the widespread incomprehension of the fire regime, while it is believed that strengthening the
heavy aerial support, controlling the ignitions, or increasing the criminal framework for
arsonists will result in a significant decrease of the annual burned area.
The number of fires and the burned area are the basic data used in the majority of approaches
to the fire regime, but few studies have attempted to describe, simultaneously, large sets of
characteristics that may derive therefrom. Since these two components of the fire regime are
driven by different factors, and given the asymmetry in the distribution of the number of fires
by burned area classes, it is critical that the analysis on the indicators derived from these two
basic variables take into account, not only these evidences, but the whole spectrum of
relations between them. Moreover, and since fire regime results from an accumulation of
events that occur in time and overlap in space, the temporal and spatial aggregation of the
indicators must also be conducted. In fact, the analysis of several fire regime indicators at
different temporal and spatial scales allows, not only to detect hardly discernible patterns in
studies focused on individual fires or developed in specific locations, but also to identify
deviations or disruptions in their distribution and, consequently, particular aspects of each fire
regime.
It will be presented the fundamental characteristics of the fire regime in Portugal mainland,
considering not only different spatial and temporal scales, but also the previously described
context that supported the choice of indicators and analytical processes.
 Poster 9

Naturally-produced compounds for the control of two major plant parasitic

nematodes

P. Barbosa1, J. Branco1,2, C. Vicente1, A. Cristina Figueiredo2, M. Mota1,3

1
NemaLab, Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra,
Apartado 94, 7002-554 Évora, Portugal.

2
Centro de Estudos do Ambiente e do Mar (CESAM Lisboa), Faculdade de Ciências da Universidade de Lisboa, Centro
de Biotecnologia Vegetal (CBV), DBV, C2, Piso 1, Campo Grande, 1749-016 Lisboa, Portugal.

3
Departamento Ciências da Vida, Universidade Lusófona de Humanidades e Tecnologias, EPCV, Campo Grande 376,
1749-024 Lisboa, Portugal.

Email: [email protected]

The pine wood nematode, Bursaphelenchus xylophilus, and the root lesion nematode,
Pratylenchus penetrans are two of the main plant parasitic nematodes responsible for
productivity losses in a significant number of plant species. Despite their significant differences
in terms of life cycle, dissemination, plant host range or feeding strategies, both nematodes
are particularly difficult to control. The presently existing control methods are costly and
hazardous to the environment and to humans, and in today´s context of sustainable
development, the selection of environmentally safe and effective ways to control these
nematodes, become imperative. There is thus a need to search for sustainable alternatives,
and plant natural products may play an important role in multi-pest pathogen control. Oxygen-
containing- and monoterpene hydrocarbons were bioassayed separately in the two nematodes
following standard methodologies. Preliminary results indicate that at 2 mg/mL after 24h-
exposure, oxygen-containing terpene molecules (citral, carvacrol, geraniol, etc.) showed a
higher mortality than the monoterpene hydrocarbon molecules (limonene, α-pinene, γ-
terpinene, etc.). For the first group of molecules, corrected mortality was higher in B.
xylophilus (75-100%) when compared with P. penetrans (60-100%). For the second group,
corrected mortality values were slightly higher in P. penetrans (5-10%) in contrast to B.
xylophilus (<5%). Compounds with a nematotoxic activity higher than 80% are being further
tested at 1, 0.5, 0.25 and 0.125 mg/mL.

This work is funded by National Funds through FCT under the PhD grant SFRH/BD/134201/2017, PTDC/AGR-
PRO/2589/2014 and UID/AGR/00115/2013, UID/AMB/50017/2013, FEDER PT2020-Compete 2020.
Animal Production and Health
 Invited Speaker 9

Uterine contributions for conceptus survival and development

R. Payan-Carreira1,2

1
Dept. Medicina Veterinária, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-554 Évora, Portugal.
2
CECAV – Centro de Ciência Animal e Veterinária, UTAD, Vila Real, Portugal.
Email: [email protected]

Contrary to most common assumptions, the uterus is not a friendly place for the embryo, unless
adequately primed by the local interplay of sex steroid receptors. In fact, the interaction
between the mother and embryonic structures is only possible for a short time frame that in
humans is named as the window of implantation. Then, a multitude of structural and molecular
changes occurs in the superficial endometrium that allows the embryo to attach to maternal
structures to form the placenta. In dogs and cats, species presenting decidual placenta, invasion
of maternal endometrium starts at the window of pregnancy. Even is non-decidual species, the
endometrium must be sensitised to allow the attachment of the trophoblast to the epithelial
lining of the maternal uterus.

Implantation and the start of pregnancy approaches a finely tuned inflammatory reaction,
where the maternal uterine defence system tightly controls the trophoblast invading
competence, in a species-specific pattern, that simultaneously ensure the connection between
the two different structures (uterine lining and the trophoblast) while guarantees the embryo
not to be attacked as foreign tissue. In all eutherian (placental) mammals, establishment of
successful pregnancy requires synchronous interactions of the conceptus with the endometrium
of the uterus.

From its role in accepting implantation, as well as for controlling the conceptus development in
early implantation, the uterus is an essential driver of female fertility. Based on studies
developed in dogs, we will discuss some of the changes occurring in normal and pregnant canine
endometrium around the window of implantation.
 Invited Speaker 10
Rabbit haemorrhagic disease virus infection in wild rabbits in Portugal. How to protect the
species? Recent research and future developments

Fábio Abade dos Santos 1, Margarida Duarte 1,2, Maria da Conceição Peleteiro 1

1 Centre for Interdisciplinary Research in Animal Health (CIISA), Faculdade de Medicina Veterinária,
Universidade de Lisboa. Av. da Universidade Técnica, 1300-477 Lisboa, Portugal;

2 Instituto Nacional de Investigação Agrária (INIAV), Quinta do Marquês, 2780-157 Oeiras, Portugal

The European wild rabbit, Oryctolagus cuniculus (Linnaeus, 1758) is a key species of Iberia
ecosystems. Subspecies Oryctolagus cuniculus algirus is only found in the Iberia, in all Portugal
territory and central and southern (?) Spain1. After 2005, the wild rabbit populations declined
abruptly in the Iberian Peninsula, strongly due to Viral Haemorrhagic Disease which emerged in
Europe in 1989 (with the etiological agent rabbit haemorragic disease virus). In Portugal, rabbit
haemorrhagic disease virus 2 (RHDV2 or GI.2) was detected for the first time in 2012 and is presently
disseminated both in the continent and in the islands2,3,4.

Many studies have established the lesional patterns of the disease in non-vaccinated domestic
rabbits to better understand its pathogenesis5. The main goal of this study was to evaluate the
impact of commercial vaccine administration in the wild rabbits’ response to infection at the tissue
level, and to understand how protective the commercial RHDV2 inactivated vaccines developed for
the rabbit industry are in wild rabbits.

Non-vaccinated and vaccinated naturally dead wild rabbits were collected from hunting reserves and
reproduction centres, respectively. Viral loads6 were evaluated and the lesional patterns established.
The fact that lesions were systematically more severe in the non-vaccinated rabbits suggests that the
disease progressed more rapidly in this group that also showed the higher viral loads. However,
vaccinated rabbits were also very susceptible to infection, with equal high mortalities, showing that
the vaccine was not as effective in the wild rabbit as in the domestic rabbit industry.

An effective vaccine for wild rabbits should be delivered easily, not requiring handling of the animals
since the stress involved often causes death and affects immune response.

Recently, the project FIGHT-TWO was approved for financial support by FCT aiming to produce an
oral vaccine. This multi-institution research project is presently at its first steps, and intends to
develop and provide a safe oral vaccine to be distributed in the field as bait aiming the
immunological full protection of wild rabbits against RHDV2 infection. Also, this non-invasive
immunization technology will be used to protect a broad proportion of the populations in affected
areas, crucial to reduce virus transmission and control the infection without capture and
manipulation.

At the same time, because of a strategic partnership established between FMV-UL and FF-UL, an
innovative study is already on-going to produce an automatic applicator of a non-toxic, natural-
repellent of ticks, fleas and flying insects. The repellent will be applied to wild rabbits in field
conditions reducing the interaction with arthropods. By decreasing indirect contact with the virus,
this temporary shield will simultaneously tackle down the main wild-rabbit diseases - RHD and
Myxomatosis.

This project was supported by SFRH/BD/137067/2018 and FCT project UID/CVT/276/2013. Molecular
diagnostic was funded by FUNDO FLORESTAL PERMANENTE (Projeto Avaliação Ecossanitária das Populações
Naturais de Coelho-Bravo Visando o Controlo da Doença Hemorrágica Viral, Dispach 4547/2017, May 31).

References
1. Ferrand, N., 2008. Inferring the evolutionary history of the European rabbit (Oryctolagus cuniculus) from
molecular marker. In: Alves, P., Ferrand, N., Hackl ̈ander, K. (Eds.), Lagomorph Biology: Evolution, Ecology and
Conservation. Springer, Berlin, Germany, pp. 47–63.
2.Abrantes J, Lopes AM, Dalton KP, Melo P, Correia JJ, Ramada M, et al. New variant of rabbit hemorrhagic
disease virus, Portugal, 2012-2013. Emerg Infect Dis. 2013;19(11):1900
3. Duarte M, Carvalho C, Bernardo S, Barros SV, Benevides S, Flor L, et al. Rabbit haemorrhagic disease virus 2
(RHDV2) outbreak in Azores: Disclosure of common genetic markers and phylogenetic segregation within the
European strains. Infect Genet Evol [Internet]. Elsevier B.V.; 2015;35:163–71. Available from:
http://dx.doi.org/10.1016/j.meegid.2015.08.005
4. Abade dos Santos, F.A., Carvalho, C., Nuno, O., Correia, J.J., Henriques, M., Peleteiro, M.C.,
Fevereiro, M., Duarte, M.D., 2017. Detection of rabbit Haemorrhagic disease virus 2 during the wild rabbit
(Oryctolagus cuniculus) eradication from the Berlengas archipelago, Portugal. BMC Vet. Res. 13, 336.
5. Carvalho, C.L., Leclerc Duarte, E., Monteiro, J.M., Afonso, C., Pacheco, J., Carvalho, P., Mendonça, P., Botelho,
A., Albuquerque, T., Themudo, P., Fevereiro, M., Henriques, A.M., Santos Barros, S., Dias Duarte, M., 2017.
Progression of rabbit haemorrhagic disease virus 2 upon vaccination in an industrial rabbitry: A laboratorial
approach. World Rabbit Sci. 25, 73–85.
6.Duarte, M.D., Carvalho, C.L., Barros, S.C., Henriques, A.M., Ramos, F., Fagulha, T., Luís, T., Duarte, E.L.,
Fevereiro, M., 2015. A real time Taqman RT-PCR for the detection of rabbit hemorrhagic disease virus 2
(RHDV2). J. Virol. Methods 219, 90–95.
 Oral Communication 17

Characterization of the Mertolenga Breed phenotypes under the influence of

the CAP in the period between 1986 and 2012

L. Santa-Maria1; C. Roquete2
1
IPBeja (Instituto Politécnico de Beja) Escola Superior Agrária, Departamento de Biociências, Rua Pedro Soares,
7800-295 Beja, Portugal.

2
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Departamento de Zootecnia, Universidade de
Évora, Pólo da Mitra, 7002-554 Évora, Portugal.

Email: [email protected]

Natural conditions in most of the Portuguese territory (marginal soils and severe water
deficit in the summer) do not favor intensive meat production, which in turn determines
a low stocking rate unless the pastures are well managed and the animals adequately
supplemented whenever necessary. These improvements are costly and pointless in
most cases explaining why breeders do not adopt this strategy. As grassland and
forage production are in most cases not suitable for efficient fattening, the conversion
of land use has led to an increase in the number of suckler cows in the period studied
(1986 to 2012), since these animals are less dependent on forage quality and have
lower feed costs under these conditions than in other farming systems. It is under this
complexity of factors, strongly conditioned by the difficult conditions to which they are
generally subjected, that the three phenotypes of the mertolenga breed stand out for
their characteristics of rusticity, ease of delivery and low food requirements. Political
conditions are added to the natural conditions and therefore the question arises as to
the extent to which the productive efficiency of this breed and its phenotypes has been
altered throughout the stages of the CAP, in what parameters and how can it be in the
future ensure the economic viability of its staff regardless of Community support.
 Oral Communication 18
Evaluation of elastase and cathepsin G inhibition on collagen transcription and
prostaglandin secretion in mare endometrium

A. Amaral1,2, C. Fernandes1, K. Lukasik2, A. Szóstek-Mioduchowska2, A. Baclawska2,

S. Morazzo1, MR. Rebordão1,3, P. Pinto-Bravo1,3, D. J. Skarzynski2, G. Ferreira-Dias1
1
CIISA - Centre for Interdisciplinary Research in Animal Health, Faculty of Veterinary
Medicine, University of Lisbon, Portugal;
2
Institute of Animal Reproduction and Food Research, Polish Academy of Science,
Olsztyn, Poland;
3
Coimbra College of Agriculture, Polytechnic Institute of Coimbra, Coimbra, Portugal.

Email: [email protected]

Mare endometrium fibrosis – endometrosis - is a major cause of subfertility/infertility in

mares. Neutrophil extracellular traps (NETs), besides killing pathogens in uterus, may
also contribute to endometrial fibrosis. We have shown that elastase (ELA) and
cathepsin G (CAT) are NETs components that may stimulate in vitro endometrial
collagen deposition. Thus, the aim of this study was to evaluate the effect of sivelestat
(SIV) – an ELA inhibitor - and cathepsin G inhibitor (INH) – a CAT inhibitor - on
collagen transcription and prostaglandin secretion, in different estrous cycle phases.
Follicular-FP (n=8) and mid luteal–MLP (n=7) phase explants were cultured for 24 or
48h with ELA (0.5, 1µg/mL), SIV (10µg/mL), ELA (0.5,1µg/mL) + SIV (10µg/mL), CAT
(0.1, 1µg/mL); INH (1µg/mL) or CAT (0.1, 1µg/mL) + INH (1µg/mL). Collagen 1 (COL1)
gene transcription was determined by qPCR and prostaglandins (PGE2; PGF2α)
secretion in culture medium was assessed by EIA. In FP, at 24h, ELA0.5 and CAT1
increased COL1 transcription (P<0.005; P<0.05 respectively) and PGF2α production
(P<0.05; P<0.01), but its inhibition - ELA0.5+SIV (P<0.05) and CAT1+INH (P<0.05) -
decreased them. Also, ELA0.5+SIV or ELA1+SIV raised PGE2 production (P<0.01). At
48h, ELA1 increased COL1 transcription (P<0.01) and PGF2α production (P<0.001), but
its inhibition (ELA1+SIV) decreased them (P<0.01; P<0.05, respectively). CAT0.1+INH
incubation was also able to reduce COL1 transcripts (P<0.05). PGF2α augmented with
ELA0.5 (P<0.001) but lowered with ELA0.5+SIV (P<0.01) and also PGF2α secretion
increased with CAT0.1 and CAT1 (P<0.05). Besides, ELA1+SIV incubation increased
PGE2 (P<0.05). In MLP, ELA0.5 (24h, P<0.01; 48h, P<0.001) and CAT1 (24h,
P<0.001; 48h, P<0.05) up-regulated COL1 transcription, but ELA0.5+SIV (24h, P<0.05;
48h, P<0.001) and CAT1+INH (24h, P<0.001; 48h, P<0.05) decreased it. At 48h,
incubation with ELA1 also up-regulated COL1 transcription and PGF2α production
(P<0.05), but PGF2α production lowered with ELA1+SIV and also with CAT0.1+INH
incubation (P<0.05). PGE2 production was higher in ELA1+SIV incubation (48h,
P<0.05) and CAT1+INH (24h, P<0.001). Inhibition of ELA and CAT pro-fibrotic action
by SIV and INH, may reduce the establishment of mare endometrial fibrosis by
stimulating the production of anti-fibrotic PGE2 and inhibiting pro-fibrotic PGF2α.

Grants: UID/CVT/00276/2013 (CIISA, Portugal); PTDC-CVT-REP-4202-2014 (FCT,

Portugal); “Iuventus plus” Polish Ministry of Science and Higher Education
(0463/IP1/2015/73); SFRH/BD/101058/2014 (PhD scholarship).
 Oral Communication 19
Assessing pubertal age of male Bísaro pigs through testicular and epididymal
morphometric parameters - preliminary results
G. Paixão1, A. Esteves1, N. Carolino2, M. Pires1, R. Payan-Carreira1,3
1
Animal and Veterinary Research Centre (CECAV), Universidade de Trás-os-Montes e Alto Douro, 5000-801 Vila
Real, Portugal.

2
Instituto Nacional de Investigação Agrária e Veterinária, Unidade Estratégica de Investigação e Serviços de
Biotecnologia e Recursos Genéticos, 2005-048 Vale de Santarém, Portugal (Instituto de Ciências Agrárias e
Ambientais Mediterrânicas)

3
Dept. Medicina Veterinária, Escola de Ciências e Tecnologia, Universidade de Évora, Pólo da Mitra, Ap. 94, 7002-
554 Évora, Portugal.

Email: [email protected]

Bísaro pig (BP) had grown in numbers in the last decade, representing one of the most
important Portuguese livestock breeds. As per farmer’s observations, BP are sexually
precocious, reaching puberty at around five-month-old. On average, boars are bred for the
first time just after completing one year of age, but in the herdbook, breeding records start as
early as four-month-old. Despite the farmers’ random observations and breeding records, little
is known about the pubertal age of BP. This study aims to estimate the age of puberty in male
Bísaro pigs through testicular and epididymal morphometry. Fifty-six pairs of testis and
epididymis were collected from male Bísaro pigs ranging in age from 1 to 8 months of age.
Samples were collected post-mortem (n=26) or from surgical castration (n=30), from May 2017
to April 2018, sourced from six different farms. After collection, testis and epididymis were
trimmed, weighed and measured. Testicles were then cut longitudinally to obtain three tissue
samples of <1 cm deep and fixed in 10% buffered formalin solution. Tissue samples were
processed for routine haematoxylin–eosin staining. Microscopic parameters used for the
morphometric studies included the spermatogenesis scoring, the number of Sertoli and Leydig
cells, the diameter of seminiferous tubule, and the presence of spermatozoa in the epididymis
and vas deferens. Measurements were evaluated using an image processing software
(ImageJ®). Correlations between testicular and epididymal length, width, depth, weight and
volume, were highly positive (r: 0.866-0.997; p<0.001; n= 56). Positive correlation was also
found between Leydig cell and nucleus diameter (r: 0.732; p<0.001; n=52), but no significant
correlations were found between the diameter of seminiferous tubule and the Leydig cell and
nucleus. Differently, tubular diameter increased proportionally to the age (R2: 0.69; p<0.001;
n=52). A GLM model was used to predict BP testis mean±SEM length, width and depth and
animal’s age when spermatozoa were found in the epididymis (6.98±0.87, 4.50±0.51 and
3.77±0.13 cm, at 117.50±6.36 days), and in the vas deferens (7.34±0.79, 4.75±0.49 and
3.98±0.47 cm, at 145.71±3.79 days). The farm had significant effect on these parameters
(p<0.001) and thus included in the model.

This work was co-funded through the European Agricultural Fund for Rural Development (EAFRD) and the
Portuguese Government within the PDR 2020 1.01 Operating Groups initiative, under the project ICas Bísaro
(reference no. PDR2020-101-031029). GP holds a PhD grant from the Animal Science Doctoral Program (AniSci),
operation number NORTE-08-5369-FSE-000040, co-funded by European Social Fund FSE and through National funds
NORTE 2020 and Regional Operational Programme of North 2014/2020.
 Oral Communication 20
Environmental conditions in pigs facilities located in Southern Europe

T. Morgado1, V.F. Cruz2, F. Baptista3, R. Charneca4

1
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Pólo da Mitra,
Departamento de Engenharia Rural, Ap. 94, 7002-554 Évora, Portugal.

2
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Pólo da Mitra,
Departamento de Engenharia Rural, Ap. 94, 7002-554 Évora, Portugal.

3
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Pólo da Mitra,
Departamento de Engenharia Rural, Ap. 94, 7002-554 Évora, Portugal.

41
ICAAM (Instituto de Ciências Agrárias e Ambientais Mediterrânicas), Universidade de Évora, Pólo da Mitra,
Departamento de Medicina Veterinária, Ap. 94, 7002-554 Évora, Portugal.

Email: [email protected]

In a world with increasing demanding for quality food, produced taking in consideration the
animal welfare and by principles of ethics and by social and environmental responsibility,
requires a special care with costs rationalization and increased efficiency. The constant need
for change and growth, resize and reorganize makes pig farming an excellent case study for
challenges in animal welfare. Pig production is an important sector in Portuguese economy,
and almost half of the national pig herd (45%) is produced in Alentejo region. This region is
where the climate variables are most extreme; demanding high monitoring of environmental
and physiological variables to ensure sustainable and safe production. Nowadays the
environmental control systems are limited to analysing temperatures and humidity, and
actuate to opening windows and cooling or heating systems. These control systems forget the
real impact in animal welfare. Therefore, there is a need to incorporate new variables to be
monitored, in real-time, that are related to animal welfare, like physiological indicators of
welfare.
The first activity of this project is the study of which variable/indicator are more indicated to
evaluate the welfare in growing pigs. In a small lab 2 trials were performed, in each one with 7
growing gilts (±45kg starting weight) submitted to different environmental conditions
(summer, winter and comfort). In each situation animal adaptation/stress was evaluated
trough several indicators like physiological (e.g. cortisol levels), behavioural (e.g. video records)
and productive (e.g. growth rate). In addition, environmental conditions were monitored
(exterior and interior) and tested the interconnection between all the collected data in order
to adjust environment according to animal responses in an automatic and real time basis. The
second phase of the project is in a commercial farm, where the animals are submitted to real
conditions. The aim of this phase is to analyze the influence of environmental parameters on
the welfare of growing and fattening pigs and to test the equipment and methodology for
collecting and monitoring information on indicators of animal welfare, provided by the animal
itself, in real time.
 Poster 10
Bioprotective solution against pathogens: selection of protective starter cultures
for meat products

M.J. Fraqueza, A. Chaves, C. Rebelo, S. Gonçalves, A. Martins, M.H. Fernandes, M.J.

Fernandes, A.S. Barreto

CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de

Medicina Veterinária, Universidade de Lisboa, 1300-477 Lisboa

The potential presence of pathogens on meat products needs to be controlled. The

main control measures are the implementation of HACCP method coupled with
technological strategies namely bioprotective solutions by the use of protective starter
cultures. The aim of this work was to select presumptively safe Lactobacillus strains
isolated from traditional dry fermented sausages regarding their potential production of
bacteriocins. Lactobacillus sakei (n= 32) and Lactobacillus plantarum (n=82) were
isolated from meat products, equipment surfaces and ingredients on different meat
industries. All isolates were assessed regarding antibiotic susceptibility. Disc
susceptibility testing was performed according to CLSI for nine antibiotics belonging to
different classes. The PCR detection for tet(L), tet(K) and tet(M) genes was performed.
Genomic typing was done by PCR-fingerprinting using the primers (GTG)5 and M13. L.
plantarum and L. sakei isolates genetic background to express phenotypic
bacteriocinogenic ability was achieved by applying PCR for known sakacin (sakA, P
and Q) and plantaricin genes. Bacteriocinogenic potential was also assessed by Skalka
qualitative test. The majority of isolates L. plantarum and L. sakey were susceptible to
tetracycline (75% and 81%) and erythromycin (71% and 97%). The presence of tetM
and tetL genes linked to mobile elements, were not frequent. However, among L.
plantarum the tetK gene was detected on 44% of the isolates. The potential for
producing Sakacin was detected only in three L. sakey isolates (L1B8, L3B8 and P2B3)
with sakA and sakQ genes while the majority of the L. plantarum presented genes
plnC, N, R, K, L for Plantaricin. L. plantarum P3B8 was selected to further studies as a
safe strain with plantaricin genes and high bacteriocinogenic activity against L.
monocytogenes and S. aureus.

Keyword: Bacteriocins, Protective starter, lactobacilli, Safety, Meat products

This study was supported by Fundação para a Ciência e a Tecnologia (FCT, Lisbon,
Portugal) through CIISA project (UID/CVT/00276/2013) and project PTDC/AGR-
ALI/119075/2010.
 Poster 11
Combined effect of nisin loaded pectin nanoparticles and high hydrostatic pressure
on the extension of alheira’s shelf life
A. Borges1, T.Krivorotova2,J. Sereikaite2, L. T. Gama1, M. H. Fernandes1, M. J. Fernandes1,
A. Barreto1, M. J. Fraqueza1.
1
CIISA. Faculty of Veterinary Medicine. University of Lisbon. Lisbon, Portugal; 2Department
of Chemistry and Bioengineering, Vilnius Gediminas Technical University, Vilnius, Lithuania.

Alheira is a traditional Portuguese smoked and fermented meat sausage produced in Trás-
os-Montes, a region in the north of Portugal. Its industry represents an important economic
resource for the region. Bad practices can occur during alheira’s manufacture, storage and
cooking, exposing consumers to microbial hazards. Considering these facts and with the aim
of promoting its sustainable production, it is important to apply alternative technologies such
as high hydrostatic pressure (HHP) and nisin loaded pectin nanoparticles (NLPN) to assure
alheira’s safety and increase its shelf life while maintaining their organoleptic authenticity.
Three batches of vacuum-packaged alheira were produced according to 4 conditions:
without treatment, NLPN treatment, HHP processing (600MPa for 390 seconds at 10°C) and
a combination of both treatments. The addition of NLPN was done during the mixing step of
the process and the HHP processing was done after packaging. Microbiological analysis
was carried out 1 day after treatments and then every month during a storage time of 5
months under 4ºC. C. perfringens and Listeria spp. were not detected in any of the samples
analysed. The total aerobic microorganisms at 30ºC and lactic acid bacteria counts were
over 7-8 log cfu/g before the application of technologies in study. The application of HHP
significantly (P<0.05) reduced the levels of all microorganisms analysed and this effect was
maintained over the time. The benefit of the NLPN addition was only verified in the
coagulase-negative Staphylococci counts, maintaining this effect during the storage
time. Moreover, the application of HHP on alheira produced with NLPN showed a synergistic
effect, with better results when compared with those obtained when the technologies were
applied alone (P<0.05). Overall, HHP combined with NLPN had a synergistic effect, and was
considered the most suitable process for preserving and extending alheira’s shelf life. This
study emphasizes the usefulness of using the synergism of two technologies to extend the
shelf life of a fermented sausage and assure safety.
Keywords: Emergent Technologies, Alheira, Fermented Meat Product, Shelf life,
Synergism.

This work was conducted with the financial support of the project PRODER medida 4.1-
Cooperação para a inovação no. 56345 “Novos produtos derivados da alheira” and CIISA-
Project UID/CVT/00276/2013.
 Poster 12
Chemical hazards screening in dry fermented sausages from Mediterranean influence
countries
Maria J. Fernandes1, Susana P. Alves1, Cristina M. Alfaia1, Biljana D. Škrbic2, Jelena R.
Živancev2, Rui J.B. Bessa1, Maria J. Fraqueza1
1
CIISA - Centro de Investigação Interdisciplinar em Sanidade Animal, Faculdade de
Medicina Veterinária, Universidade de Lisboa, 1300-477 Lisboa; 2University of Novi Sad,
Faculty of Technology, 21000 Novi Sad, Serbia

Chemical compounds such as polycyclic aromatic hydrocarbons (PAHs), biogenic amines

(BAs) and heavy metals are hazards identified and related with meat products. The
introduction of these different compounds in dry fermented sausages (DFS) could be linked
to raw materials and auxiliary ingredients used, to a specific step of their technological
process, or by environmental contamination. This work aims to evaluate the presence of
chemical hazards (PAHs, BAs and heavy elements) on commercially-available dry
fermented sausages from different origins and provide preventive recommendations to
assure products safety. Samples (n=20) were purchased at retail level in different markets of
Lisbon (Portugal) and Novi Sad (Serbia). The levels of PAHs were determined by gas-
chromatography coupled to mass spectrometry (GC–MS) while BAs and heavy elements
were analyzed using reversed-phase high performance liquid chromatography (RP-HPLC)
and atomic absorption/emission spectrometry, respectively. The most abundant PAHs in
DFS were those with low molecular weight, comprising more than 90% of total PAHs.
Serbian and most of the Portuguese DFS well meet the EU legislation for the PAHs content.
The levels of BAs in DFS from distinct origin (Portugal and Serbia) show a wide variation,
however, the major BAs found were tyramine (32.6–206 mg/kg and 5.0–246 mg/kg,
respectively) and putrescine (3.5–401 mg/kg and 1.4–291 mg/kg, respectively) followed by
cadaverine (0.6–202 mg/kg and 1.6–291 mg/kg, respectively). Histamine never reached
considerable levels in both DFS. Contents of heavy elements in DFS from Portuguese and
Serbian markets indicated that these potential hazards were present but at low levels. The
most prevalent heavy element in DFS from both origins was lead (0.01–0.34 mg/kg). In
summary, the data demonstrated that the health risks derived from the intakes of these
chemical compounds present in both Portuguese and Serbian DFS is not a major concern
for consumers. Nonetheless, additional research still has to be done regarding the selection
of autochthonous starters and smoking step optimization to minimize the contents of
potential toxic PAHs and BAs in dry meat sausages.
Keywords: Dry fermented sausage; polycyclic aromatic hydrocarbons; biogenic amines;
minerals; safety.
This study was supported by the Ministry of Education, Science and Technology Development of the
Republic of Serbia through Project No. 172050, by Fundação para a Ciência e a Tecnologia (FCT,
Lisbon, Portugal) through CIISA project (UID/CVT/00276/2013), Project Agreement FCT – 6818 and
by PTDC/AGR-ALI/119075/2010 and by Ministério da Agricultura, do Mar, do Ambiente e do
Ordenamento do Território through PRODER-PA Project No. 13017. Individual fellowship to SPA
(SFRH/BPD/76836/2011) from FCT is gratefully acknowledged.
 Poster 13

Optimization of the smoking process to control polycyclic aromatic hydrocarbons in a dry-cured meat sausage

S.P. Alves1, M.J. Fernandes1, M. Elias2, M. Laranjo2, R.J.B. Bessa1, A.S. Barreto1, M.J. Fraqueza1

1
CIISA-Centro de Investigação Interdisciplinar em Sanidade Animal. Faculty of Veterinary Medicine. University of
Lisbon, Portugal.

2
ICAAM-Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Pólo da Mitra, Ap. 94,
7006-554 Évora, Portugal

Polycyclic aromatic hydrocarbons (PAH) are potential chemical hazards related with smoked meat products. The
composition and concentration of PAH in meat sausages depends on multiple factors throughout smoking process.
This study aims to optimize smoking conditions for a high calibre dry-cured meat sausage (DCMS) and assess the
presence of PAH. Three batches of DCMS were prepared from commercial crossbred pigs, seasoned, stuffed and
smoked with natural smoke generated by burning wood from Ilex aquifolium, followed by a curing period of 45 days.
Conditions for smoking process until 35% weight loss; were: a) no smoking; b) 20h of smoking; c) 60h of smoking,
d) continuous smoking until cured. PAH from DCMS were extracted according to Alves et al. (2017) and determined
by gas-chromatography mass-spectrometry (GC-MS) using a mixture of deuterated labelled PAH as internal
standard. No heavy PAH, including the benzo[a]pyrene, were detected in sausages without a smoking period.
However, benzo[a]pyrene was detected in meat sausages submitted to continuous smoking or in sausages with 60h of
smoking, although its concentration did not reach the maximum legal limit of 5 g/kg established by the European
Union. The maximum level of 30 mg PAH4 (sum of Benzo[a]pyrene, Benzo[a]anthracene, Benzo[b]fluoranthene,
and Chrysene) per kg of smoked meat and smoked meat products (EC 835/2011) was not reached in any of the
DCMS. However, DCMS with extended smoking times (i.e. 60h of smoking) showed a trend (P=0.061) towards
higher levels of PAH4 compared with sausages submitted to 20h of smoking. Thus, to maintain typical organoleptic
characteristics of DCMS, smoking periods as long as 60h are acceptable to avoid the accumulation of toxic levels of
PAH in high calibre DCMS.

Acknowledgements: Project GO77 PDR2020-1.0.1-FEADER-031359 and CIISA project (UID/CVT/00276/2013).

M. Laranjo acknowledges a Post-Doc research grant from FCT(SFRH/BPD/108802/2015).

Meat Product, Safety, Polycyclic aromatic hydrocarbons.

List of Participants
 Nome Apelido Afiliação Email
Agostinho Serranito Universidade de Évora [email protected]
Ana Amaral Universidade de Lisboa [email protected]
Ana Galveias Universidade de Évora [email protected]
Anabela Filipe Universidade de Évora [email protected]
Ana Maria Burgos Universidad de Castilla-La Mancha [email protected]
Ana Patrícia Poeiras Universidade de Évora [email protected]
Ana Teresa Ribeiro Instituto Politécnico de Santarém [email protected]
André Albuquerque Universidade de Évora [email protected]
Antonio Almeida Universidade de Évora [email protected]
Beatriz Lara Espinar Universidad de Castilla-La Mancha [email protected]
Catarina Esgalhado Universidade de Évora [email protected]
Christiane Ramirez dos Santos Universidade de Évora [email protected]
Clarisse Mourinha Universidade de Évora [email protected]
Elsa Lopes Instituto Politécnico de Portalegre [email protected]
Elyane Borges Dias Universidade de Évora [email protected]
Emanuel Carreira Universidade de Évora [email protected]
Fernanda Firmino Fiúza Universidade de Évora [email protected]
Francesco Valerio Universidade de Évora [email protected]
Gabriela Carneiro Murta Universidade de Évora [email protected]
Gabriele Guerrini Universidade de Évora [email protected]
Gustavo Paixão Universidade de Trás-os-Montes e Alto Douro [email protected]
Hugo Viriato Universidade de Évora [email protected]
Inês de Sousa Rocha Universidade de Coimbra [email protected]
Jonathan Concas Università Politecnica delle Marche [email protected]
José Farias Universidade de Évora [email protected]
José Rico Universidade de Évora [email protected]
Juca Cunha Universidade de Évora [email protected]
Leonel Nunes Universidade de Évora [email protected]
Luís Coelho Universidade de Évora [email protected]
Luís Santa Maria Instituto Politécnico de Beja [email protected]
Luísa Coelho Universidade do Algarve [email protected]
Márcia Araújo Universidade do Porto [email protected]
Maria Inês Rouxinol Universidade de Évora [email protected]
Maria João Fraqueza Universidade de Lisboa [email protected]
Martinho Murteira Universidade de Évora [email protected]
Mauro Raposo Universidade de Évora [email protected]
Miguel Ferro Universidade de Évora [email protected]
Milton Gussule Universidade de Évora [email protected]
Nuno Guiomar Universidade de Évora [email protected]
Paola Hernandez Universidade de Évora [email protected]
Patrícia Carmona Pires Universidade de Évora [email protected]
Pedro Barbosa Universidade de Évora [email protected]
Raydson Pires da Cruz Universidade de Évora [email protected]
Rogério Louro Universidade de Évora [email protected]
Rui Jorge Raimundo Universidade de Évora [email protected]
Sara Ricardo Rodrigues Universidade de Évora [email protected]
Susana Saraiva Dias Universidade de Évora [email protected]
Taiana Homobono Universidade de Évora [email protected]
Taiana Araújo Conceição Universidade de Évora [email protected]
Teresa da Silva Morgado Universidade de Évora [email protected]
Teresa Louro Universidade de Évora [email protected]