icfc 2021 book of abstracts

4th INTERNATIONAL CONFERENCE ON

FOOD CONTAMINANTS:

CHALLENGES ON EARLY-LIFE

EXPOSURE

ICFC 2021 Book of Abstracts

27-28 September

Lisbon, Portugal

27 – 28 SEPTEMBER 2021

PORTUGAL

CONFERENCE THEME

Challenges on early-life exposure

ICFC2021 Book of Abstracts

Edited by

Elsa Vasco, Carla Martins, Inês Serrenho, Ricardo Assunção, Paula Alvito

Published by

Departamento de Alimentação e Nutrição

Instituto Nacional de Saúde Doutor Ricardo Jorge

Avenida Padre Cruz

1649-016 Lisboa

Production services: INSA

Cover Designer: Inês Serrenho

Setember 2021: First edition

ISBN

978-989-8794-82-6 [Suporte: Eletrónico]; [Formato: n.d.]

ICFC 2021 | Table of Contents

1

Acknowledgments………………………………………………………………. 2

Welcome message…………………………………………………………...... 3

Organizing Committee…………………………………………………………. 4

Scientific Committee……………………………………………………………. 5

Invited Speakers………………………………………....…………………….. 6

Programme……………………………………………………………………… 9

Invited Lectures and Oral Communications…………………………………. 11

Abstracts with poster submission……………………………………………... 28

ICFC 2021 | Acknowledgements

2

The Organizing Committee of this event expresses the profoundest gratitude to all those who contributed and supported this

event.

Thanks are due to the invited speakers for accepting to participate actively in the conference and share their knowledge, to

the Sientific Committee members for all support on Abstracts and Best Poster Award selection and to Conference Chairs for

their contribute to make the conference attractive and dynamic. Porto Ideias, the responsible for the website and participants

management, is also acknowledge for their important contribute and collaborative atmosphere.

This event succeeds other events held in Lisbon (ICFCF2015), Braga (ICFC2017) and Aveiro (ICFC 2019) under the seal of

the National Institute of Health Doutor Ricardo Jorge (INSA). Therefore, the Organization expresses its recognition to the role

played by INSA, namely the Chaiman and Vice-Chairman of the Executive Board, Dr Fernando Almeida e Dra Cristina Abreu

Santos, respectively.

Institutional Supporters

Sponsors

Thanks are due to FCT/MCTES for the financial support to earlyMYCO project (PTDC/MEDTOX /28762/2017) and to CESAM

(UID/AMB/50017/2019), through national funds.

ICFC 2021 | Welcome message

3

Dear Participants,

On behalf of the National Institute of Health Doctor Ricardo Jorge (INSA), we are honored and delighted to welcome you to

the 4th International Conference on Food Contaminants (ICFC2021), this time held by virtual transmission.The ICFC2021 is

organised by INSA and will focus on the topic “Challenges on early-life exposure”.

Exposure to hazardous chemicals during first 1000 days of life has been linked to an increased risk of a number of health

disorders.There is growing evidence that early-life exposure to chemical hazards can lead to non-communicable diseases

(NCDs) in adulthood including cancer, diabetes, obesity and auto-immune diseases. Because cells are rapidly dividing and

organ systems are developing during childhood and adolescence, exposure to carcinogens during these early life stages is a

major risk factor for cancer later in life. Exposure during early-life due to food ingestion of toxic chemicals seems to have a

critical role on the rapid epidemiological rise of obesity as well as a trans-generational transmission of risk of later obesity.

Some reports provide evidence of a link between early-life adverse events (ELAE)-induced intestinal barrier disruption,

inflammation, epigenetic modifications, and the occurrence of late metabolic disorders (MD), including obesity and type 2

diabetes (T2D). Changes in neurobehavior, sexual development, the prevalence of asthma and allergy, among others, have

shown to be associated with chemical contaminants exposure at early-life stages.

Early-life is a critical sensitive window of susceptibility. Therefore, this conference will aim to gather research scientists, health

and food professionals, regulatory entities, and students to discuss different perspectives on this particular period of life in

order to debate i) the risks associated with exposing vulnerable populations to chemicals, in particular, food contaminants and

ii) toxic effects of early-life exposure to chemicals.

This conference is promoted within the framework of the earlyMYCO national project, funded by the Foundation for Science

and Technology (FCT), Portugal, aiming to assess the impact of early-life exposure to mycotoxins and their potential impact

on human health.

We hope that ICFC2021 represents an opportunity to network, discuss new ideas and develop future research actions, in an

attempt to contribute to elucidate strategies promoting the evidence based crucial for policy maker decisions in order to protect

human health of citizens, namely the vulnerable population groups as children and pregnant women.

Prevention needs to shift focus from individual responsibility to societal responsibility and an understanding that effective

prevention of NCDs ultimately relies on improved contaminants management to reduce exposure to modifiable risks.

We wish you an excellent Conference!

Fernando de Almeida

Chairman of the Executive Board of the National Institute of Health Doctor Ricardo Jorge, I.P.

Paula Alvito

Scientific Researcher at Food and Nutrition Department

National Institute of Health Doctor Ricardo Jorge, I.P.

ICFC2021 Conference Chair

ICFC 2021 | Organizing Committee

4

Organizing Committee

Carla Martins

(ENSP, Lisbon, Portugal)

Elsa Vasco

(INSA, Lisbon, Portugal)

Inês Serrenho

(INSA, Lisbon, Portugal)

Paula Alvito

(INSA, Lisbon, Portugal and CESAM, UA, Aveiro, Portugal)

Ricardo Assunção

(CESAM, UA, Aveiro, Portugal)

CESAM – Centre for Environmental and Marine Studies; ENSP - NOVA National School of Public Health; INSA – National Institute of Health Doutor Ricardo

Jorge; UA – University of Aveiro;

ICFC 2021 | Scientific Committee

5

Scientific Committee

Armando Venâncio (UM, Braga, Portugal)

Artur Alves (CESAM, UA, Aveiro, Portugal)

Carla Martins (ENSP, Lisbon, Portugal)

Carlos Oliveira (USP, Pirassununga, Brazil)

Elsa Reis Vasco (INSA, Lisbon, Portugal)

Hans Steinkellner (EFSA, Parma, Italy)

Isabel Castanheira (INSA, Lisbon, Portugal)

Maria João Silva (INSA, Lisbon, Portugal)

Marthe de Boevre (UG, Ghent, Belgium)

Paula Alvito (INSA, Lisbon and CESAM, UA, Aveiro Portugal)

Ricardo Assunção (CESAM, UA, Aveiro, Portugal)

Susana Loureiro (UA, Aveiro, Portugal)

CESAM – Centre for Environmental and Marine Studies; EFSA – European Food Safety Authority; ENSP - NOVA National School of Public Health.

INSA – National Institute of Health Doutor Ricardo Jorge; UA – University of Aveiro; UG – University of Ghent; UM – University of Minho; USP - University

of São Paulo;

ICFC 2021 | Invited Speakers

6

Invited Speakers

Hans Steinkellner

EFSA | Italy - holds a PhD in toxicology from the University of

Vienna. He works as a toxicologist in the BIOCONTAM Unit of the

European Food Safety Authority (EFSA) in Parma since 2014. From

2007 to 2014 he worked in EFSA’s Pesticide Unit. Before joining

EFSA he worked for the European Chemicals Bureau of the Joint

Research Centre (JRC) in Ispra.

Christophe ROUSSELLE

ANSES | France - is graduated from the National Veterinary School

of Lyon. He then get a PhD in Toxicology in Paris (2000 and more

recently an International Master in Public Health in Rennes (2016).

He has been working since more than 15 years in regulatory

toxicology, first in the French Agency for Medicines (AFSSAPS) and

then at the French Agency for Environment and Occupational Health

and Safety (AFSSET) and now at ANSES. He was for more than

10 years at the head of the chemical risk assessment unit dealing

with chemicals in consumer products and also with the Reach

regulation and CLP. Now he is working in the International and

European Affairs Department, managing the Coordination team set

up for the preparation of PARC. He is also an expert in the Scientific

Committee for Consumer Safety at the European level from 2009.

German Cano-Sancho

German Cano-Sancho | INRAE | France - is a research scientist at

the French Institute for Agricultural and Environmental Research

(INRAE). He joined the LABERCA Unit, Nantes (France), after a

multidisciplinary training in dietary chemical exposure, in vitro

modelling and chemical risk assessment at University of Lleida

(Spain) or University of California at Davis (United States), among

others. His current activity at LABERCA includes the application of

integrative approaches to support the identification and

interpretation of exposome-health associations in observational

settings, more specifically related to chemical pollutants through

mass-spectrometry platforms. Methodologically, he is particularly

interested on the application of statistical methods to characterize

the effects of chemical mixtures in observational studies, with

special interest on the toxicant-nutrient and toxicant-metabolome

interfaces.


7

Marta Esteban Lopez

ISCIII | Spain - PhD, is a researcher at the National Center for

Environmental Health of the Instituto de Salud Carlos III (ISCIII),

Spain. She has published 49 peer-reviewed papers and has an H

index of 21. Her research focusses on exposure to environmental

and human biomonitoring and product-use related chemicals.

Luis Pereira da Silva

NMS |Portugal - is a senior pediatrician and neonatologist at the

pediatric Hospital Dona Estefânia, Centro Hospitalar Universitário

de Lisboa Central. Cumulatively, he is Associate Professor of

Pediatrics and of Pediatric Nutrition at NOVA Medical School and

Invited Coordinator Professor of dietetics and nutrition at the Lisbon

School of Health Technology. His main area of interest and research

is perinatal and infant nutrition. Among other positions, he is the

Nutrition and Health Promotion group team leader of the

Comprehensive Health Research Center, and coordinates the

Nutrition Lab of Hospital Dona Estefânia, the Committee on Nutrition

of the Portuguese Neonatal Society, and the Pediatric Section of the

Portuguese Association for Enteral and Parenteral Nutrition.

Claudia Marques

NMS/CINTESIS | Portugal - Nutritionist (0268N), PhD in

Biomedicine by the Faculty of Medicine of University of Porto. She

is an Invited Assistant Professor at NOVA Medical School and a

researcher at Center for Health Technology and Services Research

(CINTESIS). Her research is focused on the effects of dietary

intervations on the composition and function of the intestinal

microbiota and their relationship with health outcomes.

Manuela Ferreira

Manuela Ferreira | University of Coimbra and Champalimaud Foundation | Portugal – graduated in Biochemistry and was awarded a PhD in 2010 from Coimbra University upon developing her thesis work in Cancer Research UK. She joined Henrique Veiga-Fernandes lab in Lisbon (iMM and Champalimaud Foundation) between 2010 and 2020 as postdoctoral fellow. Ferreira made ground-breaking contributions to the understanding of epithelial biology and immune-sensing field, which were published in high impact factor peer-reviewed journals such as Nature, Science, Cell, Nature, Genes & Development, and Cancer Research. Ferreira is invited to review papers and as main arguer for Jury of MSc academic degree. Ferreira leads a group at Centre for Neuroscience and Cell Biology (Coimbra University) since 2020, where she is currently investigating how environmental cues shape immune cells and body barriers at early life.


8

Ana Cristina Santos

ISPUP | Portugal - Completed her Graduation in Nutrition Sciences

from the Faculty of Nutrition and Food Sciences in 1998, the Master

in Public Health by the University of Porto Medical School in 2003,

and the Ph.D. in Public Health in 2008, by University of Porto

Medical School. In 2020, she attained the degree of "Agregado" in

Public Health, by the University of Porto Medical School. She is a

Researcher at of EPIUnit - Epidemiology Research Unit of the

Institute of Public Health from the University of Porto and an Invited

Assistant Professor at the University of Porto Medical School. She

has been the leader of the research Group of Perinatal and Pediatric

Epidemiology of EPIUnit - Epidemiology Research Unit since 2018.

Since 2009, she is the executive coordinator of the birth cohort,

Generation XXI. The scientific activity was mostly based on the

study of obesity, its determinants, and the main metabolic

consequences, mainly studying the dynamics of obesity from

childhood to early adolescence. The life course approach has been

central to the more recent work since the analysis of data from

across the life course enables the examination of the dynamics in

which variables of interest change and interact across a person’s life

span, determinants of these changes, and how the pattern of

change relates to later health.

ICFC 2021 | Programme

9

27.09.2021 – Day 1

Session 1 - From chemicals exposure to the risk of vulnerable populations

Chairs: Paula Alvito & Marthe De Boevre

13.30 – 14.00 Opening session

14.00 - 14.30 L1

EFSA risk assessments of contaminants – Consideration of

vulnerable groups

(Hans Steinkellner, EFSA, Italy)

14.30 - 15.00 L2

European Partnership for the Assessment of Risks from

Chemicals – PARC

(Christophe Rousselle, ANSES, France)

15.00 - 15.30 L3

A novel landscape in chemical risk assessment called

Exposome: current applications and opportunities

(German Cano-Sancho, INRAE, France)

15.30 - 15.45 COFFEE BREAK

15.45 – 16.15 L4

Fish and mercury: the choice matters

(Marta Esteban López, ISCIII, Spain)

16.15 - 16.30

OC1

Exposure to mycotoxins in spanish children through the

analysis of their levels in plasma samples

(Beatriz Arce-López, Brest University, France)

16.30 - 16.45 OC2

Mercury exposure in portuguese adolescents

(Sónia Namorado, INSA, Portugal)

16.45 - 17.00 OC3

Biomonitoring of metals and minerals in urine of Portuguese

school-aged children: from levels to multi-element patterns

(Diogo Pestana, NMS-UNL & CINTESIS, Portugal)

17.00 End of session 1

ICFC 2021 | Programme

10

28.09.2021 – Day 2

Session 2 – Health effects of early-life exposure to chemicals

Chairs: Susana Loureiro & Carlos Oliveira

13:30 – 14.15

14.15 - 14.45

FFT

L5

"Early-life exposure to food contaminants: a real public

health concern?"

(Ricardo Mexia, INSA, Portugal)

Adverse effects of mycotoxin exposure in the first 1000

days of life

(Luís Pereira da Silva, NMS, Portugal)

14.45 – 15.15 L6

Interplay between food contaminants and intestinal

microbiota: effects of early-life exposures on health

(Claúdia Marques, NMS/ CINTESIS, Portugal)

15.15 - 15.45 L7

How maternal exposure to aflatoxina B1 impacts the

development of progeny intestinal immune system?

(Manuela Ferreira, UC/ Champalimaud Foundation,

Portugal)

15.45 - 16.00 COFFEE BREAK

16.00 - 16.30 L8

Cohort studies to assess early-life exposure: Generation

XXI cohort study

(Ana Cristina Santos, ISPUP, Portugal)

16.15 - 16.30

OC4

Children are exposed to higher levels of the dietary

mycotoxin deoxynivalenol and are less capable of

mycotoxin detoxification than adults – evidence from urinary

biomarker studies.

(Silvia Gratz, Aberdeen University, United Kingdom)

16.30 - 16.45 OC5

Toxicological interactions on gastric human cells between

mycotoxins co-occurring in infant and children foods

(Soraia Sá, FFUP/LAQV-REQUIMTE, Portugal)

16.45 - 17.00 Closing session and Best Poster Award

ICFC 2021 | Challenges on early-life exposure

11

INVITED LECTURES

AND ORAL

COMMUNICATIONS


12

Session 1

From chemicals exposure to

the risk of vulnerable populations


13

L1 - EFSA’s risk assessments of contaminants – consideration of vulnerable groups Hans Steinkellner European Food Safety Authority (EFSA), Italy [email protected] The European Food Safety Authority (EFSA) is in charge with providing scientific advice in the field of food and feed safety in

the European Union (EU). EFSA’s Panel on Contaminants in the Food Chain (CONTAM) provides risk assessments on

contaminants in the food chain and undesireable substances such as natural toxicants, mycotoxins and residues of

unauthorised substances. The risk of contaminants in food is characterised by comparing health based guidance values

(HBGV) with dietary exposures. These exposures and the consequent risks are estimated separately for the different age

groups of the population based on their distinct consumption patterns. Vulnerable populations are considered in CONTAM

opinions on an ad hoc basis whenever there is data available indicating a higher susceptibility of certain individuals because

of particular medical /physiological conditions and/or consumption patterns. The HBGV for nickel in food and drinking water

was based on effects seen in nickel-sensitised indivduals which are particularly prone to nickel mediated toxicity. The hazard

characterisation for tropane alkaloids was based on cardiac effects only adverse in people suffering from bradycardia. Risks

from perchlorate in food and drinking water was based on effects seen in iodine deficient individuals. Infants, toddlers and

children are generally more vulnerable towards food contaminants because their caloric intake on a body weight basis and

consequently food consumption is much higher as compared to adults. In a specific exposure scenario developed for the fatty

acid esters of 3-monochloropropanediol (3-MCPD) and glycidol it was shown that infants receiving only infant formula are

particularly at risk. For the opinions on ochratoxin, chlorinated paraffins and hexabromocyclododecanes, perfluoroalkyl

substances, compounds which can also be found in mother’s milk, specific exposure scenarios for exclusively breastfed babies

were developed to cover also the risk for this particularly vulnerable group. Recently, EFSA issued a guidance for the risk

assessment of infants below 16 weeks of age in which application of an additional saftey factor for this group is recommended

because of the high susceptibility of infants towards xenobiotics.

Keywords: EFSA, risk assessment, vulnerable populations


14

L2 – European Partnership for the Assessment of Risks from Chemicals – PARC

Christophe Rousselle

French Agency for Food, Environmental and Occupational Health & Safety (ANSES), France

[email protected]

PARC is an EU-wide research and innovation partnership programme to support EU and national chemical risk assessment

and risk management bodies with new data, knowledge, methods, networks and skills to address current, emerging and novel

chemical safety challenges. PARC will facilitate the transition to next generation risk assessment to better protect human health

and the environment, in line with the Green Deal’s zero-pollution ambition for a toxic free environment and will be an enabler

for the future EU “Chemicals Strategy for sustainability”. It builds in part on the work undertaken and experience acquired in

past and on-going research and innovation actions, but goes beyond by its vocation to establish an EU-wide risk assessment

hub of excellence.

To contribute to several expected impacts of destination 2 “Living and working in a health-promoting environment”, PARC will

organise the activities to reach three specific objectives:

- An EU-wide sustainable cross-disciplinary network to identify and agree on research and innovation needs and to support

research uptake into regulatory chemical risk assessment.

- Joint EU research and innovation activities responding to identified priorities in support of current regulatory risk assessment

processes for chemical substances and to emerging challenges.

- Strengthening existing capacities and building new transdisciplinary platforms to support chemical risk assessment.

The Partnership brings together Ministries and national public health and risk assessment agencies, as well as research

organisations and academia from almost all of EU Member States. Representatives of Directorates-General of the EC and EU

agencies involved in the monitoring of chemicals and the assessment of risks (ECHA, EEA, EFSA) are also participating.

PARC will meet the needs of risk assessment agencies to better anticipate emerging risks and respond to the challenges and

priorities of the new European policies.

Keywords: Chemicals, Risk assessment, New Approach Methodologies, Science to policy dialogue


15

L3 - A novel landscape in chemical risk assessment called Exposome: current applications and opportunities

German Cano-Sancho

National Research Institute for Agriculture (INRAE), France

[email protected]

Humans are exposed nowadays to a growing list of environmental pollutants and food contaminants for which comprehensive

risk assessment is available only for a minority. Traditional chemical risk assessment frameworks based on single-chemicals

and high-dose toxicity testing studies has become limited to address major challenges currently posed by low-dose co-

exposures, in many cases with poor structural and toxicological knowledge. The ‘exposome’ concept has quickly evolved

during the last decades as a novel paradigm to overcome longstanding challenges in public health and providing large

opportunities to advance on chemical risk assessment. Some inspiring exposome frameworks and approaches favoring

comprehensive chemical risk assessment with special focus on food safety are herein presented. Novel technological

advances on high-resolution mass spectrometry will be discussed on the light of high-throughput identification of known, but

also emerging potential chemical hazards in food and biological matrices. Epidemiological settings accommodating novel

methods to characterize and integrate large panels of exposures will be featured to illustrate this transformative effort to capture

the complex interactions between human environments, diets and health with major potential impacts in a new risk assessment

era.

Keywords: exposome, biomarkers, chemical risk assessment, food contaminants, mass spectrometry


16

L4 - Fish and mercury: the choice matters

Esteban-López M, Castaño A.

National Centre for Environmental Health. Instituto de Salud Carlos III. Majadahonda, Spain.

[email protected]

The toxicity of mercury has been known for centuries although the associated risk has been modified as far as the exposure

sources have varied over the time. Currently, the main source of mercury exposure in the general population is diet, mainly

mercury-contaminated fish. Consumption of certain species of fish can lead to high levels of this heavy metal in the organism

what can cause adverse health effects. Methylmercury (MeHg) is the most toxic chemical form of mercury and the predominant

one in fish. MeHg can cause severe damage to the central nervous system with even more dramatic effects if the exposure

occurs during the developmental phase. Therefore, prevention of early-life exposure constitutes a priority in public health.

In recent years, much progress has been made in controlling mercury emissions, manufacture, trade, waste and uses, with

stringent EU regulations and the coming into force of the United Nations (UN) Minamata convention on mercury in 2017.

However, these measures are not sufficient, as they will not result in a direct and immediate reduction of mercury levels in fish.

Additionally, other factors such as climate change, can increase mercury exposure. Therefore, taking into account the current

situation and the benefits associated with the consumption of fish, it is essential to develop strategies at national level, to inform

and educate the vulnerable population in order to reduce the exposure and associated risks, making the most of the benefits

that this food offers. This is especially important in countries with a high fish consumption such as the Mediterranean countries.

Despite the complexity of the issue of environmental exposure to mercury, measures as simple as the directed dietary advice

can contribute to reduce significantly the mercury levels keeping the benefits of eating fish.

Keywords: mercury, fish, human biomonitoring


17

OC1 - Exposure to mycotoxins in Spanish children through the analysis of their levels in plasma samples

Beatriz Arce-Lópeza,b, Elena Lizarragab, Reyes López de Mesac, Elena González-Peñasb

a University Laboratory of Biodiversity and Microbial Ecology, ESIAB, Technopôle de Brest Iroise, University of Brest, EA3882,

29280 Plouzané, France bDepartment of Pharmaceutical Technology and Chemistry, Research Group MITOX, School of Pharmacy and Nutrition,

Universidad de Navarra, 31008 Pamplona, Spain cDepartment of Pediatrics, Clínica Universidad de Navarra, Universidad de Navarra, 31008 Pamplona, Spain

[email protected]

In order to assess human exposure to mycotoxins, human biomonitoring studies (HBM) in biological samples have been

developed [1]. To date, these HBM studies are still not well known in children population.

Objective: The aim of this study was to evaluate the exposure to 19 mycotoxins in plasma samples from healthy and sick

children (digestive, autism spectrum (ASD) and attention deficit hyperactivity (ADHD) disorders) (n=79, 2-16 years) from a

region of the northern Spain.

Methodology: Samples were analysed by a validated LC-MS/MS method, following a sample deproteinization and cleanup in

only one step using Captiva EMR®-lipid (3 mL) cartridges and acetonitrile (with 1% formic acid) [2]. To identify possible

metabolites, an additional enzymatic procedure has been performed using β-glucuronidase/arylsulfatase enzyme.

Results: Ochratoxin A (OTA) was the most prevalent mycotoxin in all groups of children, before and after enzyme treatment,

with higher incidence and levels in healthy children. Ochratoxin B appears only in the samples of healthy children with a low

incidence (11.4%), always co-occurring with OTA. Sterigmatocystin (STER) was detected only after enzymatic hydrolysis with

a high incidence in all groups, especially in sick children (98.7% in healthy children and 100% in patients). None of the other

studied mycotoxins (aflatoxins B1, B2, G1, G2, and M1; T-2 and HT-2 toxins; deoxynivalenol, deepoxy-deoxynivalenol, 3-

acetyldeoxynivalenol, 15-acetyldeoxynivalenol; zearalenone; nivalenol; fusarenon-X; neosolaniol; and diacetoxyscirpenol),

were detected either before or after enzymatic treatment in any of the groups of children.

Conclusions: This is the first HBM report conducted to determine multiple mycotoxin exposure in plasma samples from children

[3]. For this vulnerable population, OTA and STER should be highly considered. This justifies the need for further risk

assessment studies to obtain data on their sources of exposure, their toxicokinetics and the relationship between plasma levels

and toxic effects in children.

Acknowledgements: This work was supported by the Spanish “Ministerio de Economía, Industria y Competitividad, Agencia

Estatal de Investigación” (AGL2017-85732-R) MINECO/AEI/FEDER, UE). The authors are grateful to the volunteers and the

“Clínica Universidad de Navarra” for plasma donation.

Keywords: child; digestive problems; human biomonitoring; mycotoxins


18

OC2 – Mercury exposure in portuguese adolescents

Sónia Namorado1,2,3, Diogo Gomes4, Diogo Iria4, Federico Cestelli4, Afonso Mota4, Bernardo Alves4, João Leal4,

Andreia Luz4, João Gomes4, Rafael Barros5,6, M. Ramiro Pastorinho6,7,8, Ana C. Sousa8,9

1 National Institute of Health Dr. Ricardo Jorge (INSA), Lisbon, Portugal 2 NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, Lisbon, Portugal 3 Comprehensive Health Research Center (CHRC), Lisbon, Portugal 4 Colégio Valsassina, Lisbon, Portugal 5 Health Sciences Research Centre (CICS), Faculty of Health Sciences, University of Beira Interior, Covilhã, Portugal 6 NuESA - Health and Environment Study Unit, Faculty of Health Sciences, University of Beira Interior, Covilhã, Portugal 7 Department of Medical and Health Sciences, School of Health and Human Development, University of Évora, Évora, Portugal 8 Comprehensive Health Research Centre (CHRC), University of Évora, Évora, Portugal 9 Department of Biology, School of Science and Technology, University of Évora, Évora, Portugal

[email protected]

The possible health effects related to exposure to hazardous chemicals have let to citizens interest in environmental health.

Mercury, in particular, has been recognized as a chemical of concern, due to its well-known neurotoxicity and endocrine disrupting

potential. In this work a group of teenagers from a school in Lisbon together with their teachers and researchers from the academia

conducted a human biomonitoring survey on mercury levels in the students from their school.

Recruitment was performed in a private school in Lisbon by a group of students from the 11th and 12th grade under the supervision

of biology teachers and after having received training from researchers from the academia. All participants answered a

questionnaire with specific questions on possible mercury exposure sources (e.g. seafood consumption, dental amalgams). Hair

samples were collected and analysed using atomic absorption spectrometry with thermal decomposition and gold amalgamation

alongside with human hair certified reference material. The study protocol was approved by the Pedagogical Board of the school

and authorized by the National Data Protection Commission.

From the 92 students recruited, with ages ranging from 12 to 18 years, 56.5% were females, most lived in urban areas (91.3%)

and the majority (97.8%) did not have dental amalgam fillings. Participants Body Mass index (BMI) ranged from 15.1 to 28.4 kg/m2,

with a mean of 20.6 ± 2.6 kg/m2. Total mercury in hair ranged from 0.01 to 3.32 μg/g, with a mean of 1.09 ± 0.66 μg/g. Around

40% of the students had mercury levels higher the US EPA reference limit of 1 μg/g, while 9.8% exhibit levels higher than the

reference dose set by the WHO (2 μg/g).

Levels of mercury above the US EPA reference limit are significantly associated with higher consumption of fish (more than once

a week), but not with shellfish and seaweed consumption.

Keywords: mercury, students, citizens science, Portugal.


19

OC3 – Biomonitoring of metals and minerals in urine of Portuguese school-aged children: from levels to multi-element

patterns

Rebelo, I.A, Cruz Fernandes, V.B, Keating, E.C,D, Costa Leite, J.C,E, Diana Teixeira, D.A,F, Luz Maia, M.B, Norberto, S.C,

Pinto, E.G, Moreira-Rosário, A.A,C, Sintra, D.C, Moreira, B.C, Costa, A.C, Silva, S.C, Costa, V.C, Martins, M.C, Castro

Mendes, F.C, Queirós, P.C, Guerra, A.C,H, Fontoura, M.C,I, Palmares Carvalho, I.C,J, Matias Lima, R.K, Martins, C.L,

Delerue-Matos, C.B, Almeida, A.G, Azevedo, L.C,M, Calhau, C.A,C, Pestana, D.A,C

ANutrition & Metabolism, NOVA Medical School, Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Portugal; BREQUIMTE/LAQV – ISEP, Politécnico do Porto, Portugal. CCINTESIS - Center for Health Technology and Services Research, Porto, Portugal. DDepartment of Biomedicine - Biochemistry Unit, Faculty of Medicine of the University of Porto, Porto, Portugal. EFaculty of Health Sciences, Fernando Pessoa University, Porto, Portugal. FComprehensive Health Research Centre. Universidade NOVA de Lisboa. Lisboa. Portugal; GREQUIMTE/LAQV- Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, Porto, Portugal. HDivision of Paediatric Nutrition, Department of Paediatrics, Integrated Paediatric Hospital, Centro Hospitalar São João, Porto.

Faculty of Medicine, University of Porto, Portugal. IDivision of Paediatric Endocrinology, Department of Paediatrics, Integrated Paediatric Hospital, Centro Hospitalar São João,

Porto. Faculty of Medicine, University of Porto, Portugal. JDepartment of Clinical Neurosciences and Mental Health, Faculty of Medicine, University of Porto, Portugal. KDirectorate-General of Education. LObesity research Group, Department of Cancer research and Molecular Medicine, Faculty of Medicine, Norwegian University

of Science and Technology (NTNU), Trondheim, Norway. MDept. Community Medicine, Information and Health Decision Sciences (MEDCIDS), Faculty of Medicine, University of Porto,

Portugal

[email protected]

Introduction: The presence of chemical elements in small amounts in living beings have recognized importance in the physiology

and nutrition of humans. According to WHO, we can classify them as essential elements, probably essential elements and

potentially toxic elements. Disturbances in homeostasis, balance and interaction between elements can result in the development

of dysfunction and pathologies. This study was conducted to assess urinary levels of metals and minerals in a population of

Portuguese school-aged children (6–12 years), as well as to explore the determinants of exposure and multi-element patterns.

Methodology: Briefly, it was conducted a cross-sectional study in three regions of northern Portugal, using a multi-stage sampling

method to provide a representative sample of the population (n= 2018). The levels of major toxic metals (As, Cd, Pb, Ni) and other

metals and micronutrients (Al, Sb, Co, Cu, Sn, Mn, Mo, Se, Tl, I) were measured by ICP-MS, using spot first-morning urine

samples.

Results: In this study it was possible to observe that some elements, namely arsenic, copper, nickel, and thallium presented higher

levels when compared with reference values from other countries. In this context, the regions of Greater Porto and Entre Douro e

Vouga had the presented the higher levels. Furthermore, there were observed differences regarding age, as younger children had

higher levels. Multi-element patterns were also determined and explored, which allowed us to understand possible interactions

and the relationship with the characteristics of the children and food consumption. The region and age, namely younger age, were

shown to be important determinants of pattern adhesion.

Conclusions: These preliminary results emphasize the need to further evaluate the health impacts of both metals and minerals

levels in susceptible populations such as children. In this context, we will further assess the determining factors of exposure and

the potential impacts on health.

Relevance: This data could bring great benefits to the health of the population, through knowledge of the levels of these elements

in Portugal and their potential role in the state of health and disease.

Acknowledgements: The project IoGeneration (153NU2) was supported by EEA Grants and Norway Grants and FEDER through

the operation POCI-01-0145-FEDER-007746 funded by the Programa Operacional Competitividade e Internacionalização –

COMPETE2020 and by National Funds through FCT - Fundação para a Ciência e a Tecnologia within CINTESIS, R&D Unit

(reference UID/IC/4255/2013).

Conflict of interests: The authors declare no competing interests.

Keywords: Endocrine disruptors, biomonitoring, children, elements, metals


20

Session 2

Health effects of

early-life exposure to chemicals


21

FFT - Early-life exposure to food contaminants: a real public health concern?

Paula Alvito1,2, Luis Pereira-da-Silva3, Ricardo Mexia4

1Departamento de Alimentação e Nutrição, Instituto Nacional de Saúde Doutor Ricardo Jorge, Lisboa, Portugal 2Centro de Estudos do Ambiente e do Mar, Universidade de Aveiro, Aveiro, Portugal 3Comprehensive Health Research Centre (CHRC), Medicine of Woman, Childhood and Adolescence, NOVA Medical School,

Universidade Nova de Lisboa 4Departamento de Epidemiologia, Instituto Nacional de Saúde Doutor Ricardo Jorge, Lisboa, Portugal

[email protected]

Exposure to hazardous chemicals during first 1000 days of life has been linked to an increased risk of several health disorders.

Early-life exposure to chemicals is an emerging research field that needs urgent clarification as a contribution to the exposome

research. There is a growing awareness of the scientific community towards this topic; however, a doubt remains whether

early-life exposure could be considered a real public health concern.

To answer this question, two approaches were performed. A preliminary bibliographic search was conducted on published

reviews and original articles at PubMed in 2021, including as key words “early-life exposure”, “food contaminants” and “public

health”, for the last decade (2011-2021). In addition, the opinion of Public Health professionals was gathered through their

representative, in order to discuss about the real concern of the professional health community towards early-life exposure to

food contaminants.

This preliminary approach revealed an increased number of papers reporting the role of non-nutritional components as

contributors to the epidemic of metabolic disorders (MD), including obesity and type 2 diabetes (T2D), with a special emphasis

on food contaminants and social stress. Some reports provide evidence of a link between early-life adverse events (ELAE)-

induced intestinal barrier disruption, inflammation, epigenetic modifications, and the occurrence of late MD. There is rising

evidence of the negative effects of contaminants present in foodstuffs (such as endocrine-disrupting chemicals, heavy metals,

and mycotoxins) following exposure during pregnancy and breastfeeding, which should be considered a major public health

issue. Exposure during early-life due to food ingestion of toxic chemicals seems to have a critical role on the rapid

epidemiological rise of obesity as well as a trans-generational transmission of risk of later obesity.

Convergent research in toxicology, epidemiology, and occupational medicine has documented children's unique sensitivities

to chemical, nutritional, and psychosocial hazards during windows of vulnerability in early development and has shown that

early-life exposures can produce disease and disability in childhood and across the life span.

Acknowledgments: FCT/MCTES for funding with national funds related to the earlyMYCO project (PTDC/MEDTOX

/28762/2017) and to CESAM (UIDP/50017/2020+UIDB/50017/2020).

Keywords: early-life exposure, public health, food contaminants


22

L5 - Adverse effects of mycotoxin exposure in the first 1000 days of life

Luís Pereira-da-Silva, MD, PhD

NOVA Medical School | Faculdade de Ciências Médicas, and Hospital Dona Estefânia, Centro Hospitalar Universitário de

Lisboa Central, Lisbon, Portugal.

[email protected]

The first 1000 days of life, the period between the conception and 24 months’ postnatal age, is highly vulnerable to adverse

environmental effects. Mycotoxins present in food and environment may affect health of the fetus and small child with long-

standing effects.

Maternal exposure to fumonisins and aflatoxins may increase the risks of fetal death, neural tube defects, and fetal growth

restriction. On the other hand, prenatal exposure to aflatoxin was reported to be associated with postnatal growth restriction

and neonatal jaundice.

Exposure of small children to fumonisins and aflatoxins has been associated with reduction in body weight and length.

As available studies in humans are observational, it is difficult to establish a causal relationship between mycotoxins exposure

and clinical manifestations of mycotoxicosis in children, which are nonspecific. In addition, in many studies possible

confounders were not controlled. This topic is an excellent opportunity for further research with impact in public health.

Keywords: Aflatoxins, Fumonisins, Neural tube defect, Infant, Prenatal exposure


23

L6 - Interplay between food contaminants and intestinal microbiota: effects of early-life exposures on health

Cláudia Marques1,2

1Nutrition and Metabolism, NOVA Medical School, Faculdade de Ciências, Universidade NOVA de Lisboa, 1169-056 Lisboa,

Portugal 2CINTESIS - Center for Health Technology Services Research, NOVA Medical School, Faculdade de Ciências, Universidade

NOVA de Lisboa, 1169-056 Lisboa, Portugal

[email protected]

The gut microbiota has opened a new horizon in nutrition and biomedical research by being implicated in metabolic diseases

such as obesity and diabetes. These diseases are also influenced by lifestyle factors such as dietary habits which in turn can

modify the composition of the gut microbiota.

Diet is one of the most important factors that can modulate the gut microbiota. Some dietary components such as fiber and

phytochemicals, as well as fermented foods may stimulate the growth or contain microorganisms that confer health benefits to

the host. On the other hand, diet can also be a source of food additives and contaminants, such as persistent organic pollutants,

that have been shown to decrease the diversity of the bacteria inhabiting our gut, triggering, in turn, metabolic heath

consequences.

A special attention has to be given to the in utero exposure to these compounds and to the first three years of life which

represents a crucial window in our long-term development as well as in gut microbiota acquisition and maturation.

Thus, the gut microbiota can be considered an intermediary of the effects of diet and food contaminants on metabolic health

assuming a special relevance in the prevention, treatment, and monitoring of these diseases. Moreover, mother-to-infant

microbiota transmission may be one of the mechanisms linking the intrauterine environment and the susceptibility to disease

in later (and even in early) life.

Also, it is important to consider that the relationship between food contaminants and the gut microbiota is bidirectional and that

the gut microbiota can also have a role in xenobiotic metabolism either inactivating or metabolizing them into more toxic

metabolites.

Keywords: Diet; Early-life exposure; food contaminants; gut microbiota


24

L7 - How maternal exposure to aflatoxina B1 impacts the development of progeny intestinal immune system?

Bastos-Amador P.1,4 Duarte EL.2,3 Alvito P.4,5 Assunção R.4,5 Ferreira M1,6*

1 Champalimaud Centre for the Unknown, Champalimaud Foundation, 1400-038 Lisboa, Portugal 2 School of Science and Technology, University of Évora, Évora, Portugal 3 MED-Mediterranean Institute for Agriculture, Environment and Development, Évora, Portugal 4 Food and Nutrition Department, National Institute of Health Dr. Ricardo Jorge, Lisbon, Portugal 5 CESAM, Centre for Environmental and Marine Studies, Department of Biology, University of Aveiro, Aveiro, Portugal 6 Center for Neuroscience and Cell Biology, University of Coimbra, UC-Biotech, Biocant Park, Cantanhede, Portugal

[email protected]

Exposure to toxic contaminants during early-life is associated with the development of diseases. Individuals are exposed to

mycotoxins since early stages of life[1]. However how maternal exposure to mycotoxins influences the development and

function of the offspring’s immune system remains largely unexplored. Recently, we showed that in utero maternal exposure

to micronutrients is critical for the development of the immune system, which sets long term immunity if the progeny[2]. Here

we show that presence of aflatoxin B1 in the diet of pregnant murine females affects the development and function of the

intestinal immune system. Notably, maternal exposure to AFB1 promoted an increase of overall T cell population, while it also

resulted in a selective reduction of cytokine-producing innate lymphoid cells group 2 (ILC2) population in intestine of the

progeny. These alterations were associated with decreased expression of Reg3b, Reg3g and Fut2 by the intestinal mucosa of

progeny. Thus, these results indicate that maternal exposure to mycotoxins impacts the development of offspring intestinal

immune system. ILC2 are critical in intestinal epithelial repair, whether mice exposed to AFB1 display defective tissue damage

response needs to be investigated. Also FUT2-dependent fucosylation is key in host–commensal symbiosis suggesting

alterations in the intestinal microbiota. Our work reveals that maternal exposure to dietary contaminants such as mycotoxins

alters the normal development of intestinal immune system framework of the progeny and may have impact in their immune

function. This work was funded by FCT/MCTES through national funds, to earlyMYCO (PTDC/MED-TOX/28762/2017), and

CESAM (UIDP/50017/2020+UIDB/50017/2020).

Keywords: Early-life, Mycotoxins, Intestinal immune system


25

L8 - Cohort studies to assess early-life exposure: Generation XXI cohort study

Ana Cristina Santos

1 EPIUnit – Instituto de Saúde Pública da Universidade do Porto 2 Laboratory for Integrative and Translational Research in Population Health (ITR), Porto 3 Department of Public Health and Forensic Sciences, and Medical Education, University of Porto Medical School

Keywords: Birth cohort, Life course, Epidemiology

[email protected]

The life course perspective has been receiving increasing attention by the recognition that life’s experiences, extending from

antenatal life, through childhood, adolescence, and adulthood, comprise a longitudinal and dynamic process within which the

risk of various chronic diseases evolve. Thus, this perspective recognizes that many diseases processes in later life have their

origins in perinatal or even antenatal experiences, that many disease “risks” evolve during the life course and are continually

influenced by the experiences and by the presence of other factors, and that there is an interacting system of influences that

encompasses genetic, social, and behavioural factors.

Additionally, the life course perspective assumes that the influence of these factors are not merely sequential, additive, but

rather, that there will be interactions between several domains of causal influence. It assumes that people’s life is uniquely

shaped by the timing and sequencing of life events and exposure. More recent evidence, formally modeled trajectories of

exposures, attempting to understand if, beyond different exposures, different patterns of exposure were associated with the

risk of health outcomes. Thus, it encompasses complex pathways of multiple variables influencing the occurrence of diseases

operating across an individual life course and even across generations.

Birth cohorts may be one of the best ways to approach these complex and dynamic processes, following children from birth

along with their lives, providing a unique opportunity to collect information on dynamic variables of interest.

In 2005, started the enrolment for the first Portuguese birth cohort – Geração XXI (Generation XXI). It assembled more than

8500 newborns and their mothers, having collected a vast amount of data that will help in the comprehension of the

development of multiple diseases. This cohort has been evaluated four times since its recruitment, at 4, 7, 10, and 13 years of

age of the children. As most of the birth cohorts established, an impressive amount of information on exposures in early life

was collected, and this will allow studying the effect of these multiple exposures in the future development of diseases.

Keywords: Birth cohort, Life course, Epidemiology


26

OC4 - Children are exposed to higher levels of the dietary mycotoxin deoxynivalenol and are less capable of

mycotoxin detoxification than adults – evidence from urinary biomarker studies.

Gratz S. W., Currie V., Duncan G.

Rowett Institute, University of Aberdeen, UK

[email protected]

Introduction: Mycotoxin contamination of cereal foods is a ubiquitous problem for food safety assessments and dietary exposure to

mycotoxins is highly prevalent around the globe. In temperate regions Fusarium fungi pose the main problems in small grain cereals

and dietary exposure to trichothecene mycotoxins such as deoxynivalenol (DON) has been reported. Upon ingestion, plant-bound

DON can reach the large intestine where microbial activity can release DON and microbial detoxification to de-epoxy deoxynivalenol

(DOM-1) has been reported in some individuals, but not others.

Methodology: In this study we compare the frequency and magnitude of DON exposure and the capability of DON detoxification

between children and adults to assess the risk posed by dietary DON in both population groups. Urinary DON and DOM-1 were

measured in 15 adults and 21 children using Immunoaffinity clean-up and LC-MS/MS quantification including 13C15-labelled internal

standards. Urinary DON and DOM-1 were used to determine dietary DON exposure which was compared to tolerable daily intakes1,2.

Results: In adults and children DON was detectable in 100% of urine samples. However, the frequency of detoxification to DOM-1

was significantly higher in adults (40.7%) compared to children (9.5%). At the same time, children were exposed to levels > TDI in

52% of cases while only 7% of adults exceeded the TDI for DON.

Conclusion and relevance: Our findings clearly demonstrate that children are at a far greater risk of high exposure to DON through

their diet and at the same time lack the capability to detoxify DON to DOM-1 in most cases. More information is needed to fully

understand the extent of DON exposure in children and the contribution of the childhood microbiota to DON release and detoxification.

Keywords: Mycotoxin, exposure, urine, microbiota, children


27

OC5 - Toxicological interactions on gastric human cells between mycotoxins co-occurring in infant and children foods

Sá S.A, Faria M.A, Fernandes J.A, Cunha S.A

ALAQV-REQUIMTE, Laboratory of Bromatology and Hydrology, Faculty of Pharmacy, University of Porto, Portugal

Rua Jorge de Viterbo Ferreira 228, 4050-313 Porto, Portugal

[email protected]

Introduction: Exposure to mycotoxins during childhood can influence health later in life. Enniatin B (ENNB), sterigmatocystin

(STG) and aflatoxin B1 (AFB1) are likely to co-occur naturally in cereal-based products, abundantly consumed by infant and

children. Their combined toxicity could lead to additive, synergistic and/or antagonistic effects, thus the evaluation of these

interactions is of paramount importance to toxicity assessment.

Methodology: Human-derived gastric cells (NCI-N87) were grown in RPMI with 10% FBS, 1% penicillin/streptomycin, incubated

at 37ºC with 5% CO2 until 80% confluence. Cytotoxicity assays were performed in 96-well plates for an exposition period of 72

h using the MTT assay. Cytotoxicity of individual compounds and of mixtures (binary and ternary) was determined for different

levels of concentration (individual: 0.625-80 µM; binary: 2.37-76.36 µM; ternary: 10.38-82.96 µM), following a constant ratio

combination design proposed by Chou-Talatay (Chou 2006). All measurements were done in 6 replicates in two independent

experiments and data reported as mean in cell viability graphs. Compusyn software was used to calculate the inhibitory

concentration at 50% (IC50) for combination indexes (CI).

Results: In terms of individual toxicity, ENNB was the mycotoxin with the highest toxic effect on cells viability, having the lowest

IC50 value of 3.3 µM, followed by STG (6.2 µM) and finally AFB1 (32.3 µM). For binary combinations, ENNB-STG (IC50=12.7

µM) had antagonistic effects (CI>1) at all inhibition levels as well as ENNB-AFB1 (30.8 µM); STG-AFB1 (35.3 µM) had

antagonistic effects at most inhibition levels and a low synergism at IC97. The ternary combination also showed antagonistic

effects at all inhibition levels (IC50=46.3 µM).

Conclusions: ENNB was more cytotoxic alone than combined. The combination ENNB-STG was the most toxic, having

antagonistic effects at all inhibition levels. Of the three mycotoxins under study, ENNB and STG are the ones with the lowest

IC50, therefore they show the highest toxicity. Since they are considered emerging mycotoxins (with no legislation for maximum

values), these mycotoxins may present a serious threat to human health, especially to a group as vulnerable as children.

Relevance: The toxicological interactions identified are of biological importance since these mycotoxins can co-occur in

children foods, therefore they should be taken into account for proper risk evaluation.

Acknowledgements: This work was supported by FCT (Fundação para a Ciência e a Tecnologia) and FEDER through the

scholarship SFRH/BD/143928/2019 with financial support from FCT/MEC through national funds and co-financed by FEDER,

under the Partnership Agreement PT2020., FCT and FEDER. Sara C. Cunha acknowledges FCT for IF/01616/2015 contract.

Miguel A. Faria acknowledges FCT the researcher contract. This research was partially supported by national funds through

FCT within the scope of UIDB/04423/2020 and AgriFood XXI R&D&I project, operation No. NORTE-01-0145-FEDER-000041,

co-financed by the European Regional Development Fund (ERDF) through NORTH 2020 (Northern Regional Operational

Program 2014/2020).

Keywords: mycotoxins, children, citotoxicity, NCI-N87 cells


28

ABTRACTS WITH POSTER SUBMISSION


29

P1 – earlyMYCO – a mother & child pilot study to assess early-life exposure to mycotoxins in Portugal

Martins Ca,b,c,d, Assunção Rd,e, Costa Aa, Serrano Da, Namorado Sb,c, Vidigal Cf, Almeida Ef, Alvito Pc,d, Nunes Ca,b

a NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, Avenida Padre Cruz,

1600-560 Lisboa, Portugal b Comprehensive Health Research Center (CHRC), Universidade NOVA de Lisboa, Campo Mártires da Pátria, 1169-056

Lisboa, Portugal c National Institute of Health Dr Ricardo Jorge, Avenida Padre Cruz 1649-016, Lisbon, Portugal d Centre for Environmental and Marine Studies (CESAM), University of Aveiro, Campus Universitário de Santiago, 3810-193

Aveiro, Portugal e IUEM, Instituto Universitário Egas Moniz, Egas Moniz – Cooperativa de Ensino Superior, CRL, Monte de Caparica 2829 –

511, Caparica, Portugal f ACES Lisboa Central, Regional Health Administration of Lisbon and Tagus Valley (ARSLVT), Lisboa, Portugal

[email protected]

Early-life exposure occurs during gestation through transfer of toxic substances present in the maternal diet to the fetus and

later during lactation, through the breast milk. Mycotoxins are well known carcinogenic, nephrotoxic, hepatotoxic, and

immunosuppressive food chemical contaminants. Recently, human biomonitoring data revealed that Portuguese population is

exposed to mycotoxins. These results emphasized the need for assessing the prenatal and lactation exposure to mycotoxins

in a vulnerable period of life. The earlyMYCO project (PTDC/MED-TOX/28762/2017) aimed, for the first time, to develop and

implement a pilot study to assess the early-life exposure to mycotoxins through a mother & child cohort, and to identify the

associated challenges, thus contributing to the development of future studies and to the knowledge of the exposome of

Portuguese population.

Participants were recruited in primary health care units in Lisbon (Portugal) during pregnancy (1st trimester) and followed-up in

three moments of observation: 2nd trimester of pregnancy (mother), and 1st and 6th months of the child’s life (mother & child),

with the collection of biological samples (blood, urine, breast milk) and the application of sociodemographic and food

consumption questionnaires (food frequency and 24h dietary recall).

All the participants of the earlyMYCO pilot study (n = 19) reported more than nine years of education, with 37% reporting 10-

12 years and 63% reporting more than 12 years. Regarding the household monthly income, 42% reported a monthly income

above 1941€, with most of participants (95%) being workers for remuneration or profit. Dairy products, bread, tea, and coffee

were reported as the food items with higher frequency of consumption. The 24h dietary recall questionnaires confirmed these

results. Detailed analysis of the data gathered allowed also to highlight the high consumption of meat, fish and vegetables.

Regarding infants, 22% were fed with infant formula and 78% were exclusively breastfed.

It is expected that results obtained within earlyMYCO pilot study will contribute to a deeper knowledge on exposure of

vulnerable population groups (pregnant women and infants) and to the understanding of the impact of early-life exposure to

mycotoxins.

Acknowledgments: The earlyMYCO was funded by Foundation for Science and Technology (PTDC/DTP-

MEDTOX/28762/2017), by CESAM (UIDP/50017/2020+UIDB/50017/2020).

Keywords: Human Biomonitoring; Health Impact Assessment; Mycotoxins; Portugal


30

P2 - Early-life exposure to mycotoxins: assessing the risk under the earlyMYCO project

Assunção, R 1,2,3,4*, Martins, C 2,3,5, Nunes, B 2,3,5, Namorado, S 2,3,5, Silva, S 2, Pires, SM 6, Alvito, P 1,2

1 CESAM - Centre for Environmental and Marine Studies, University of Aveiro, Aveiro, Portugal 2 National Institute of Health Dr. Ricardo Jorge (INSA), Lisbon, Portugal 3 NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, Lisbon, Portugal 4 IUEM, Instituto Universitário Egas Moniz, Egas Moniz – Cooperativa de Ensino Superior, CRL, Caparica, Portugal 5 Comprehensive Health Research Center (CHRC), Lisbon, Portugal 6 Division for Diet, Disease Prevention and Toxicology, The National Food Institute, Technical University of Denmark, Lyngby,

Denmark

[email protected]

Exposure to hazardous chemicals during the first 1000 days of life has been associated to a number of health disorders, e.g.

non-communicable diseases. A proper risk assessment built on accurate data assumes particular importance to evaluate the

potential impact that early-life exposure could represent in adulthood. Mycotoxins, secondary fungal metabolites, could have

harmful effects in humans and animals. Recent studies reported that Portuguese young children are exposed to multiple

mycotoxins through food consumption which could constitute a health concern [1]. However, earlier exposure to these

compounds remains unexplored.

earlyMYCO – Early-life exposure to MYCOtoxins and its impact on health, a national funded project (PTDC/MED-

TOX/28762/2017), intends to contribute to clarify this issue evaluating the health effects of early-life exposure of Portuguese

mother-and-child pairs to mycotoxins and assessing the associated risk. The associated health risk will be characterized and

the health impact assessed combining data from mother-and-child pairs, their exposure to mycotoxins and newly obtained

toxicological data. The Estimated Daily Intake (EDI) will be compared with reference dose values.

Preliminary results of exposure to mycotoxins through food consumption (cereal-based foods) in young children (≤ 3 years

old) revealed a potential adverse health effect for percentiles of intake of aflatoxins above or equal to P50 (corresponding to

0.041 ng/kg body weight/day or higher). Additionally, results also suggest that high levels of exposure to mycotoxins in the

early life are more related with processed foods than with breastmilk.

Obtained results will contribute to reach an accurate risk assessment framework and to establish and prioritize preventive

measures to reduce exposure to chemicals and risk, especially for vulnerable population groups as pregnant women and

infants.

This work was funded by FCT/MCTES through national funds, to earlyMYCO (PTDC/MED-TOX/28762/2017), and CESAM

(UIDP/50017/2020+UIDB/50017/2020).

[1] R. Assunção, et. al., Food and Chemical Toxicology 118, 399–408 (2018).

Keywords: Mycotoxins; Early-life exposure; Health; Risk assessment; Vulnerable populations.


31

P3 - Concentration and risk assessment of Lead and Cadmium in rice commercialized in Morocco using GF-AAS

Sifou A. 1, Abida S. 1, El Haroui k. 1

1 Department of Chemistry, Faculty of Sciences, Mohamed V Agdal University, Avenue Ibn Batouta, Rabat, Morocco

[email protected]

INTRODUCTION: Rice is one of the most commonly consumed grains. It could be a good source of nutrients in a diet, but its

consumption could also contribute to exposure to toxic elements. The present work focused on the study of chemical

contamination by metallic trace elements of rice marketed in Morocco The aim of our study was to estimate the content of Cd

and Pb in rice products and to evaluate the health risk indicators related to exposure to toxic elements consumed with rice

products in the Moroccan population.

METHODOLOGY: A total of sixty two (n=62) samples of rice commercialized in different markets in Rabat in Morocco were

surveyed for their lead and cadmium content using Atomic Absorption Spectrophotometer (GF-AAS) after total mineralization

of samples.

RESULTS: Results showed that out of 62 total samples, 47 samples (75.8%) were contaminated with Pb concentrations in the

range of 0.01-1.08 μg/g. The remaining samples (24.2%) were under the detection limit (LOD) of Pb. In the case of Cd, 40

samples (64.5%) were contaminated with Cd levels that ranged between 0.009 and 0.125 μg/g. In the present study, 12

samples (19.35%) of rice are above the maximum limit (0.2 μg/g) set by the European Commission Regulation No 1881/2006

for Pb in cereals. However, for the Cd, 10 samples exceeded the maximum limit set for this element (0.1 μg/g). The estimation

of the provisional weekly intakes of Pb and Cd showed that the risks of development toxicology effects are low.

CONCLUSIONS AND RELEVANCE: The levels compare well with those reported worldwide for similar foodstuffs. From the

results obtained, it can be concluded that the accumulation of these elements may present health hazards to the consumer

due to the toxic effects associated with their long-term ingestion. This is the first study on the co-occurrence of the two trace

elements (Pb and Cd) in rice samples commercialized in Morocco.

Keywords: Lead, Cadmium, rice, GF-AAS, Morocco, risk assessment


32

P4 - Dietary supplements based on Epilobium parviflorum as a source of molds and mycotoxins?

Iwona Ałtyn, Magdalena Twarużek

Kazimierz Wielki University, Faculty of Biological Sciences, Department of Physiology and Toxicology, Chodkiewicza 30, 85–

064 Bydgoszcz, Poland

[email protected]

The first mention of medicinal properties of Epilobium parviflorum Schreb. comes from 1880. Despite the recognized healing

properties of this plant, synthetic drugs have long displaced it from medicine. Phytochemical and pharmacognostic studies

carried out in the last twenty years in Poland and in the world have again shown the therapeutic properties. Hoary willowherb

is abundant in biologically active substances, because it contains β-sitosterols and acids: linolenic, arachidonic, peanut, oily,

palmitoyl, palmitic, stearic as well as myricetin, quercetin and kaemferol. Willow water extracts that have antibacterial,

antifungal, anti-cancer, immunostimulatory, anti-inflammatory and detoxifying effects are especially active.

The aim of this study was to evaluate molds and mycotoxins contamination of Hoary willowherb available in Poland. The

material consisted of 25 samples. Molds and yeasts were determined using YGC medium. The results are expressed as the

number of colony forming units (cfu) per gram of a sample. The identification of molds was done to genus. Mycotoxins

determination was performed using HPLC-FLD method (immunoaffinity column AflaTest and OchraPrep) and HPLC-MS/MS

method (Bond Elut®Mycotoxin column from Agilent - DON, NIV, ZEN, T-2 and HT-2).

Mycological analysis of dietary supplements based on Hoary willowherb showed a significant level of mold content in the

samples (an average of 4.1 x 104 cfu/g). The predominant types were Eurotium (61.9%), Aspergillus (13.27%) and Penicillium

(10.98%).

The most commonly detected mycotoxins were: T-2, occurring in 92%, while toxin HT-2 and ZEN were found in 76% of

samples. The highest contents of mycotoxins were revealed by: T-2max = 79.6 µg/kg, HT-2max = 57.9 µg/kg, ZENmax = 1048.7

µg/kg. Two of tested samples were slightly contaminated with OTA, DON and NIV with a maximum values of 2.49 µg/kg,

345.2 µg/kg and 49.4 µg/kg, respectively. No AF toxin was detected in any Hoary willowherb sample.

Studies confirm the occurrence of mycotoxins in Epilobium parviflorum. Manufacturers should consider and monitor the level

of mycotoxin contamination in both raw material and products.

This study was supported by the Polish Minister of Education and Science, under the program "Regional Initiative of

Excellence" in 2019 - 2022 (Grant No. 008/RID/2018/19)

Keywords: supplement dietary, mycotoxin, molds, contamination


33

P5 - The occurrence of zearalenone in the urine of lactating women

Ropejko K., Twarużek M.

Department of Physiology and Toxicology, Faculty of Biological Sciences, Kazimierz Wielki University, Chodkiewicza 30, 85-

064 Bydgoszcz

[email protected]

Zearalenone is a mycotoxin produced by fungi of the genus Fusarium. These species are mainly Fusarium graminearum,

Fusarium culmorum and Fusarium sporotrichioides. This mycotoxin is called mycoestrogen because it can bind to estrogen

receptors and disturb the hormonal balance and functioning of the endocrine system, causing ovarian and uterine dysfunction

in women, fertility disorders and neoplastic changes in the genital system. Moreover, it exhibits immunotoxic, teratogenic,

cytotoxic and hepatotoxic effects. Zearalenone is found in many foods (cereals, dried fruit, spices) as well as grains and

compound feeds.

The aim of the research was to detect the presence and amount of zearalenone in the urine of lactating women.

After obtaining the consent of the Bioethical Committee of the Nicolaus Copernicus University in Toruń at the Collegium

Medicum im. Ludwik Rydygier in Bydgoszcz (consent number KB323/2018), the research material was obtained. The research

group consisted of 60 women. Urine samples were obtained from 19 women donors of the Women's Milk Bank in Toruń and

41 women who were recruited for this study. Extraction of zearalenone in urine was performed using ZearalaTest

immunoaffinity columns (Vicam), followed by chromatographic analysis by LC MS-MS.

The results of the analysis showed the presence of zearalenone in all 60 (100%) urine samples. The mean zearalenone in

urine was 0.112 ng/ml, with the minimum value 0.006 ng/ml and the maximum value 0.482 ng/ml. 20 out of 60 samples

(33.33%) were above the mean value, while 40 out of 60 samples (66.67%) were below the mean value. Moreover, all the

samples showed the presence of zearalenone metabolites, i.e. α-zearalenol (mean 0.074 ng/ml) and β-zearalenol (mean 0.067

ng/ml) and α-zearalanol (mean 0.023 ng/ml) and β-zearalanol (mean 0.018 ng/ml).

On the basis of the obtained results, it can be concluded that the urine of the women who were tested is slightly contaminated

with zearalenone and its metabolites.

Keywords: mycotoxins, zearalenone, urine, contamination


34

P6 - Fungal contamination and azole-resistance in tea and other medicinal plants commercially available in Portugal

Simões A 1; Faria M 1; Dias M 1; Viegas C 1,2,3; Caetano LA 1,4

1 H&TRC—Health & Technology Research Center, ESTeSL—Escola Superior de Tecnologia da Saúde, Instituto Politécnico

de Lisboa, 1990-096 Lisbon, Portugal; 2 NOVA National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, 1600–560 Lisbon,

Portugal; 3 Comprehensive Health Research Center (CHRC), 1169–056 Lisbon, Portugal; 4 Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, University of Lisbon, Lisbon, Portugal.

[email protected]

Introduction. Tea and infusions are common beverages worldwide, prepared by infusing Camellia sinensis or other medicinal

plants (eg. Melissa officinalis) with boiling water. Contamination by specific strains of filamentous fungi, such as Aspergillus

sp., may originate health risks. While azole antifungal drugs are first-line therapy against fungal infections, the exposure of

medicinal plants to azole fungicides in conventional agriculture can promote azole-resistance, leading to therapeutic failure.

Considering the growing risks to public health, the fungal contamination of tea and medicinal samples commercially available

in Portugal was evaluated.

Methodology. Samples (N=40) of different origins of conventional and biological agriculture were collected at supermarkets

and herbalist shops in January 2021. Infusions were prepared according to ISO 3103:2019. Samples were also extracted as

previously described and inoculated in fungal selective culture media (MEA and DG18) and in Sabouraud dextrose agar (SDA)

medium supplemented with 4 mg/L itraconazole (ITR), 2 mg/L voriconazole (VOR), and 0.5 mg/L posaconazole (POS). Plates

were incubated at optimal conditions for fungal growth. Fungal counts were determined as colony forming units (CFU) per

gram and fungal morphology was analysed by microscopy.

Results. The average mycobiota was 359.1 ± 816.2 (MEA) and 559.1 ± 1263.8 (DG18) CFU/g in extracted samples, and 341.7

± 277.2 (MEA) and 183.3 ± 271.6 (DG18) CFU/g in infusions. Regarding azole-resistance, the most prevalent species were

Aspergillus sp. (48% ITR) and C. sitophila (39% ITR, 47% VOR, 92% POS) in extracted samples, and C. sitophila (50% ITR,

37% VOR, 57% POS) in infusion samples. Among Aspergillus sp., the most prevalent sections were Nigri (96% MEA, 74%

DG18, 72% SDA) and Circumdati (100% ITR) in extracted samples, and Fumigati (46% MEA, 100% SDA) in infusion.

Conclusions. Fungal contaminants were present in tea samples and medicinal plants for infusion, including azole-resistant

fungi.

Relevance. The presence of Aspergillus sp., particularly section Fumigati, resistant to azoles in infusions may represent a

health risk for consumers, especially immunocompromised individuals. Mycotoxins detection and correlational statistical

analysis are ongoing to further determine whether there is a relation between sample origin, fungal contamination and

mycotoxigenic profile.

The authors gratefully acknowledge Instituto Politécnico de Lisboa for funding the Project "Resistant mycobiota and

mycotoxigenic profile of tea and medicinal plants” (IPL/2020/TEAResMyc_ESTeSL), and FCT/MCTES (UIDB/05608/2020;

UIDP/05608/2020).

Keywords: Azole resistance, Fungal contamination, Tea, Medicinal plants, Aspergillus section Fumigati


35

P7 - Climatic variations and exposure assessment of mycotoxins in wheat collected in two Moroccan areas

Aicha El Jai 1,*, Catherine Brabet 2,3, Noel Durand 2,3, Jalila Ben Salah-Abbès 4, Amar Riba 5, Samir Abbès 4,6, Didier

Montet 2,3, Samira Etahiri 1, Abdellah Zinedine 1,**

1 Chouaib Doukkali University, BIOMARE Laboratory, El Jadida 24000, Morocco, 2 UMR 95 QualiSud, CIRAD, 34398 Montpellier Cedex 5, France, 3 Qualisud, Université de Montpellier, Avignon Université, CIRAD, Institut Agro, IRD, Université de La Réunion, France, 4 Laboratory of LGBBV, University of Monastir, Tunisia, 5 Laboratoire LBSM, ENS de Kouba, Algeria, 6 ISBB of Béja, University of Jendouba, Tunisia.

[email protected]

Introduction: Morocco, a Mediterranean and North African country, has a climate with annual variations in recorded temperature

and precipitations. Moroccan cereal production amounted to 2.56 million tons of wheat harvested in 2020. Wheat is an

important cereal crop that significantly contributes to the livelihoods of farming communities and the Moroccan economy. In

this study, the effect of climatic variations on the multi-mycotoxin presence in wheat sampled in two different areas (continental

and a littoral) in the country was investigated as well as the risk assessment for the population.

Methodology: A total of sixty (n=60) wheat samples were sampled during 2019 and 2020 seasons and analyzed for humidity

content, water activity, and for the occurrence of 25 mycotoxins by using liquid chromatography tandem mass spectrometry

(LC/MS-MS).

Results: Analytical results showed that out of 60 total samples, 19 were contaminated by at least one mycotoxin. Wheat

samples collected from the littoral area were found to be contaminated by five mycotoxins: Enniatins (ENB developpes, ENA1

and ENB1), Aflatoxin B1 and Ochratoxin A. While in the continental area, only three mycotoxins: Enniatins (ENB1, ENB and

DON developpes) were found. The highest mycotoxin level was found for ENB (247 µg/Kg) in a sample from the humid littoral

area. The multi-presence of mycotoxins was also observed in the investigated samples.

Conclusion and Relevance: This is the first study assessing the impact of climate variations on multi-occurrence of mycotoxins

in wheat samples harvested from two different climatic areas from Morocco. Mycotoxin levels were below the maximum

regulatory levels established by EU and Moroccan regulations and by this the risk exposure assessment of regulated

mycotoxins is maintained moderate to low. Therefore, a polyphasic identification of toxigenic fungi is essential to seek the fungi

origin of the contaminations.

Keywords: Climatic variations, Wheat, Multi-mycotoxin, Contamination, Morocco.


36

P8 - Screening of biogenic amines in Portuguese red wines. Does Dekkera/Brettanomyces wine spoilage raise the

risk of intake by consumers?

Luís Filipe-Ribeiro1, Juliana Milheiro1, Leonor C. Ferreira1, Elisete Correia2, Fernanda Cosme1*, Fernando M. Nunes1

1Chemistry Research Centre-Vila Real (CQ-VR), Food and Wine Chemistry Lab, University of Trás-os-Montes and Alto Douro,

School of Life Sciences and Environment, 5000-801, Vila Real, Portugal. 2Center for Computational and Stochastic Mathematics (CEMAT), Department of Mathematics, University of Trás-os-Montes

and Alto Douro, 5000-801, Vila Real, Portugal.

[email protected]

Food safety and quality are the main concerns of consumers and health agencies. Histamine is one of the targeted toxins by

the Food and Drug Administration and the European Food Safety Authority. Biogenic amines (BAs) constitute a potential public

health concern due to their physiological and toxicological effects. BAs can be present in grapes [1] or formed by

microorganisms during the winemaking process [2-4]. Dekkera/Brettanomyces, a wine spoilage yeast [5], can produce BAs in

grape juice [6]. Diamines can produce carcinogenic nitrosamines by reaction with nitrite [7]. BAs are important causes of wine

intolerance [8], producing intoxication symptoms [9]. The sensitivity to BAs depends on insufficient amino oxidase activity,

genetic predisposition, alcohol, acetaldehyde, gastrointestinal disease, or inhibition by other amines [10]. In addition, it is worth

mentioning that in particular, vulnerable people who are immune-compromised and elderly, may exhibit intolerance to even

low levels of biogenic amines and suffer more severe symptoms, these individuals are increasing in industrialised nations [11].

Therefore, factors that influence BAs concentrations are of utmost importance for consumer safety, mainly for vulnerable

people. The main objective of this study was to quantify BAs in industrially produced red wines available in Portuguese wineries.

The impact of Dekkera/Brettanomyces in the BAs concentration was also assessed.

Wine sampling was performed using a maximum variance/heterogeneous purposive non-probability technique [12].

Ethylphenols were determined by GC-MS [13] and BAs by dispersive solid-phase extraction and HPLC-DAD after derivatisation

with benzoyl chloride [14]

To better understand the real input of Dekkera/Brettanomyces activity in red wine BAs concentration, a set of 79 Portuguese

red wines produced at an industrial scale were analysed showing a total range of amines (19.6–331 mg/L) and ethylphenols

concentrations (4-ethylphenol 4.5–5604 μg/L and 4-ethylguaiacol 2.3–831.2 μg/L) [15].

Simultaneous determination of BAs and volatile phenols in industrial produced red wines permitted to conclude that the wine

spoilage activity of Dekkera/Brettanomyces with the production of volatile phenols do not significantly contribute to BAs

increase and consequently intake by the consumers.

BAs need to be controlled in order to ensure high levels of wine quality and safety to reduce risk to more vulnerable consumers.

Acknowledgements: We acknowledge at all wine producers and at Nuno Grosso, Emanuel Cardoso and Raquel Moreira from

SAI Enology technical team for obtaining and supplying all the wines used in this study. To SAI Enology for providing FTIR

and sulfur dioxide analyses. JM and LCF acknowledge the financial support provided by the grants PD/BD/135331/2017 and

PD/BD/128270/2017, respectively. This research was funded by Fundação para a Ciência Tecnologia (FCT-Portugal) to CQ-

VR Chemistry Research Centre—Vila Real (UIDB/00616/2020 and UIDP/00616/2020) and by the European Regional

Development Fund through NORTE 2020 (Programa Operacional Regional do Norte 2014/2020) to the project AgriFood XXI

(NORTE-01-0145-FEDER-000041).

Keywords: Red wine, Biogenic amines, Dekkera/Brettanomyces, safety.


37

P9 - Effects of mercury on human reproductive health: a cross-sectional study

Henriques MC1,2, Santiago J1, Bento F3, Oliveira M3, Patrício A3, Herdeiro MT1, Loureiro S2, Fardilha M1

1Department of Medical Sciences, Institute of Biomedicine (iBiMED), University of Aveiro, Aveiro, Portugal 2Department of Biology, Centre for Environmental and Marine Studies (CESAM), University of Aveiro, Aveiro, Portugal 3Hospital Infante D. Pedro, Centro Hospitalar do Baixo Vouga, Aveiro, Portugal

[email protected]

Mercury (Hg) is one of the most prevalent contaminants, generated by anthropogenic activities such as industrial, pharmaceutical,

and agricultural activities. Exposure to Hg has been associated with adverse effects on human reproductive health (e.g.,

miscarriages, spontaneous abortions, stillbirths, and low birth weights). Several studies also showed that increased Hg levels were

associated with male infertility and subfertility status, and that infertile subjects with unexplained infertility showed higher total Hg

(THg) levels in hair than fertile ones. The main goals of this study were to i) assess human exposure to Hg using non-invasive

biological matrices; ii) examine the influence of variables that may contribute to Hg exposure; iii) evaluate the effects of Hg on

human reproductive health; and iv) improve knowledge regarding metal accumulation and distribution on humans.

For that, a cross-sectional study was carried out in eligible women hospitalized for delivery and men attended urology

appointments in Centro Hospitalar do Baixo Vouga in Aveiro, Portugal. A detailed questionnaire regarding sociodemographic,

lifestyle and reproductive health data was completed by participants and non-invasive samples were collected. THg was quantified

in hair, placental and cord tissues using the Advanced Mercury Analyzer (AMA-254, LECO).

Our results showed Hg bioaccumulation in biological samples from female and male participants. Further, a significant positive

correlation was found between THg levels in hair and all matrices analysed, reinforcing the feasibility of the use of human hair to

assess Hg exposure. Higher THg levels were found in biological matrices of women diagnosed with infertility-related conditions

(e.g., endometriosis).

Finally, this study demonstrated that lifestyle habits seemed to play a role in Hg exposure, but further investigations are necessary.

Keywords: exposure assessment; human biomonitoring; health impact assessment; mercury; infertility


38

P10 - Association between bisphenol A levels, seminal parameters, and sperm proteome: a preliminary study

Santiago J A*, Silva JV A,B,C, Simkova M D, Santos MAS A, Vitku J D, Fardilha M A

A Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, Aveiro, Portugal B LAQV/REQUIMTE, Department of Chemistry, University of Aveiro, Aveiro, Portugal C Unit for Multidisciplinary Research in Biomedicine, Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto,

Portugal D Department of Steroids and Proteofactors, Institute of Endocrinology, Prague, Czech Republic

[email protected]

Introduction: Every day we are exposed to environmental chemicals that affect our endocrine system - endocrine disruptors –

like bisphenol A (BPA). BPA leaks from polycarbonate plastics used to line food and drink containers and seems to negatively

affect male fertility. High urine and seminal plasma BPA levels have been associated with low sperm quality and production,

and paternal exposure may contribute to diseases in their offspring. We aimed to investigate the correlation between seminal

plasma BPA concentration and seminal quality in Portuguese men living in the Aveiro region. Additionally, we intended to

identify altered sperm proteins in exposed men that might compromise fertilization and embryogenesis.

Methodology: BPA levels in the seminal plasma of 43 men from the Aveiro region with different degrees of fertility were

determined using liquid-chromatography mass spectrometry (LC-MS/MS). All donors answered a questionnaire including

questions regarding age, height, weight, medication, lifestyle habits, past/recent illnesses, and (in)fertility history. Basic semen

analyses were performed according to WHO’s guidelines. Thirty sperm samples were divided into 6 groups based on BPA

levels and basic semen parameters and their proteome was evaluated by quantitative proteomic analysis.

Results: BPA was detected in 81% of seminal plasma samples and the mean BPA concentration was 0.155 ng/ml ± 0.142

ng/ml. Our data showed that seminal plasma BPA levels of non-normozoospermic men were significantly higher than the levels

of normozoospermic patients (p<0.05). Additionally, seminal plasma BPA was negatively correlated with sperm concentration

(r =−0.334; p<0.05) and total sperm count (r =−0.42; p<0.05). No correlation with age, BMI, or other semen parameter was

observed. Proteomic analyses revealed differentially expressed proteins in different groups like RPL22, YBX2/YBX3 and

SEMA3C that were significantly upregulated in non-normozoospermic samples with BPA levels ≤0.132 ng/mL compared with

normozoospermic samples with the same BPA levels.

Conclusions and relevance: Identical to what has been observed in other study cohorts, BPA appears to negatively impact

spermatogenesis, affecting sperm production and thus, resulting in lower concentration and sperm counts. The identification

of the molecular mechanisms underlying BPA-induced male infertility might explain some situations of fertilization failure and

abnormal embryo development associated to men exposed to BPA.

Keywords: Bisphenol A; seminal plasma; spermatozoa; male infertility; proteome


39

P11 - Cytotoxicity Assessment of Novel Cellulose Nanomaterials on Intestinal Cells

Vital N1,2, Pinto F1, Rolo D1, Pedrosa J3, Lourenço AF4, Ferreira PJT3, Kranendonk M5, Silva MJ1,5, Louro H1,5

1National Institute of Health Doutor Ricardo Jorge, I.P (INSA), Department of Human Genetics, Lisbon, Portugal. 2NOVA Medical School, NOVA University Lisbon, Portugal 3University of Coimbra, CIEPQPF, Department of Chemical Engineering, Coimbra, Portugal 4RAIZ – Institute for Research on Forest and Paper, Aveiro, Portugal 5Centre for Toxicogenomics and Human Health (ToxOmics), NOVA Medical School, NOVA University, Lisbon, Portugal

[email protected]

Several nanomaterials (NMs) are currently being applied in food and food packaging industry and recent reports described the

identification of NMs in baby formula and infant food. These findings raised important concerns about the potential health

effects from early life exposure to NMs, since children are particularly vulnerable to chemicals due to their specific exposure

patterns and physiological factors. Patients with chronic intestinal diseases constitute another susceptible group given the

inflammatory potential of some NMs. Cellulose nanomaterials (CNMs) have found applications in multiple food-related

products, as zero-calorie filler/thickener/stabilizer, or food packaging materials. To contribute to characterize CNMs potential

adverse effects in the human gastrointestinal tract, their cytotoxicity was investigated in intestinal cell lines.

Two micro/nanofibrillated celluloses (CNFs), were synthetized from industrial bleached Eucalyptus globulus kraft pulp using

different pretreatments (enzymatic and TEMPO-mediated oxidation) followed by a high-pressure homogenization process.

Potential cytotoxic effects were evaluated by the MTT assay using two human intestinal cell models (Caco-2 and HT29-MTX-

E12). Since the digestion process may modify the physicochemical properties of NMs, potentially influencing biological

outcomes, the CNFs were subjected to a harmonized in vitro digestion method before cytotoxicity testing.

No cytotoxic effect was observed after 24h exposure to the undigested CNFs in the concentration-range tested (3.1 µg/mL-

200 µg/mL), irrespectively of the cell line used. Similar results were obtained for the digested CNFs for concentrations up to

14.3 µg/mL. The observation that the in vitro digestion mixture was cytotoxic by itself for concentrations above 7.6 % (v/v) (i.e.,

equivalent to CNMs exposure > 14.3 µg/mL) impaired cytotoxicity assessment at higher CNFs concentrations.

Complementary cytotoxicity analyses and genotoxicity characterization, after optimization of the in vitro digestion procedure to

reduce its toxicity, are underway. The results will increase the knowledge on the biological effects of CNMs in the intestine,

contributing to the choice of safer CNMs for food-related applications, particularly for vulnerable population groups, thereby

protecting human health.

Funded by FCT/MCTES through national funds (PTDC/SAU-PUB/29481/2017; PTDC/SAUPUB/ 32587/2017) and

(UIDB/00009/2020; UIDP/00009/2020); FCT/MCTES PhD studentship 2020.07168.BD was attributed to NV. The authors

thank P.Alvito, R.Assunção and M.Silva (Food Safety Department, INSA) for support with the in vitro digestion.

Keywords: Cellulose nanomaterials; Nanomaterials; In vitro simulated digestion; Intestinal epithelial cells; Cytotoxicity


40

P12 - FLEXiGUT: Investigating the impact of environmental and dietary exposure on chronic low-grade gut

inflammation

Pero-Gascon, R.A,*, Kellner, N.A, Nawrot, T.S.B,C, De Boevre, M.A, De Saeger, S.A

A Centre of Excellence in Mycotoxicology and Public Health, Faculty of Pharmaceutical Sciences, Ghent University, 9000

Ghent, Belgium B Centre for Environmental Sciences, Hasselt University, 3590 Diepenbeek, Belgium C Department of Public Health and Primary Care, KU Leuven, 3000 Leuven, Belgium

[email protected]

Introduction: Exposure to environmental and dietary contaminants has been associated with gut microbiota dysbiosis.

Interestingly, the Bacteroides2 enterotype has recently been linked to gut inflammation, revealing the existence of a gut

microbiome-inflammatory axis. The FLEXiGUT project aims to characterize human life course environmental and dietary

exposure to assess and validate its impact on chronic low-grade gut inflammation and related biological processes and

diseases.

Methods: Two Flemish prospective cohorts are used to cover the human life course: the “ENVIRONAGE birth cohort”, a mother-

child cohort which provides follow-up from gestation to the age of 10 years, and the “Flemish Gut Flora Project longitudinal

cohort”, a cohort of adults. Targeted and untargeted analysis of legacy and emerging contaminants, markers of air pollution,

mycotoxins and the metabolome, including objective markers of e.g. food intake, ensure a comprehensive assessment of the

exposures. The associated biological responses are investigated by applying -omics techniques, including metagenomics,

DNA adductomics and metabolomics, as well as assessment of telomere length and measurement of inflammatory markers.

Results: The biomonitoring of contaminants and the gathering of -omics data for the biological samples collected at different

time points (n=400 mother-child pairs and n=400 adults), complemented with the information available from questionnaires,

lifestyle and clinical data, will allow capturing the exposome from prenatal life onwards. An integrative multi-omics data

processing approach will be applied to uncover associations between the exposures and diseases, but also provide insights

into the mechanisms by which the exposure might be exerting its effects.

Conclusions: In FLEXiGUT, environmental and dietary contaminants and -omics profiles are investigated, encompassing both

exposure and effect. The sample size should allow to achieve statistically significant models for -omics validation.

Relevance: FLEXiGUT is the first large-scale exposomics study focused on chronic low-grade gut inflammation. Gut

inflammation is hypothesized to be related to biological processes that cause health deterioration and increase the risk of

chronic diseases and/or accelerated biological ageing.

Keywords: Exposome / gut inflammation / environmental and food contaminants / microbiome /-omics techniques


41

P13 - Risk assessment of exposure to acrylamide in Spanish breastfeeding mothers using human biomonitoring

F. Fernández, S.AB*, Pardo, O.ABC, Coscollà, C.A and Yusà, V.ABD,

AFoundation for the Promotion of Health and Biomedical Research in the Valencian Region, FISABIO-Public Health, Av.

Cataluña, 21, 46020, Valencia, Spain BDepartment of Analytical Chemistry, University of Valencia, Doctor Moliner 50, 46100, Burjassot, Spain CPublic Health Directorate of Valencia, Av. Cataluña, 21, 46020 Valencia, Spain DPublic Health Laboratory of Valencia, Av. Cataluña, 21, 46020 Valencia, Spain

[email protected]

Introduction: Acrylamide (AA) is an organic compound classified as "Probably carcinogenic to humans" (Group 2A) that can

be found principally in processed carbohydrate-rich foods and tobacco smoke. In humans, after exposure, AA is rapidly

metabolized and excreted in urine, predominantly as N-acetyl-S-(2-carbamoylethyl)-l-cysteine (AAMA), N-acetyl-S-(2-

carbamoyl-2-hydroxyethyl)-l-cysteine (GAMA3) and N-Acetyl-3-[(3-amino-3-oxopropyl)sulfinyl]-L-alanine (AAMA-Sul), which

can be used as short-term biomarkers of exposure to AA.

Methodology: In this study, the presence of AA metabolites in urine samples of lactating mothers living in Spain (n = 114) was

analyzed by "dilute and shoot" and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). A daily intake

of AA was estimated and potential predictors of exposure were investigated by multiple regression analysis. In addition, urinary

levels of AA metabolites were put in a risk assessment context using two different approaches: reverse and forward dosimetry.

Results: All urinary metabolites were detected in 100% of the analyzed samples, with geometric means of 70, 33 and 15 ng·ml-

1, for AAMA, AAMA-Sul and GAMA3, respectively. The consumption of coffee, bread and precooked food products were found

to be significant predictors of internal exposure to AA. Mean estimated daily intake (EDI) values of AA were between 1.2 and

1.9 μg AA·kg bw-1·day-1. The risk assessment strategies showed an average margin of exposure (MOE) of 349 and a hazard

quotient (HQ) of 5.5.

Conclusions: Further regulation and risk management activities are necessary to minimize AA exposure in Spain, since a

health concern with respect to non-neoplastic toxicity could not be discarded.

Relevance: This research work is the first study reporting data on AA internal exposure in a Spanish population and putting

AA urinary levels in a risk assessment context using different approaches.

Keywords: Acrylamide; human biomonitoring; urine; risk assessment; lactating mothers


42

P14 - Mycotoxins in pistachio (Pistacia vera L.): occurrence and trends in analytical methodologies

Mateus, A.A,B; Barros, S. B; Pena, A. A; Silva, A. B,C

ALAQV, REQUIMTE, Laboratory of Bromatology and Pharmacognosy, Faculty of Pharmacy, University of Coimbra, Polo III,

Azinhaga de Stª Comba, 3000-548 Coimbra, Portugal; AP: [email protected]; ARSM: [email protected] B National Institute of Agrarian and Veterinary Research (INIAV), I.P., Rua dos Lagidos, Lugar da Madalena, 4485-655, Vairão,

Vila do Conde, Portugal; ASS: [email protected]; SB: [email protected] C Center for Study in Animal Science (CECA), ICETA, University of Oporto, Oporto, Portugal

[email protected]

Introduction: Pistachios (Pistacia vera L.) are an excellent nutritional source, and they have important benefits for human

health, consequently, the consumption of this nut has been increasing. In addition, pistachio is an important source of exposure

to mycotoxins, especially aflatoxins, as a result of fungal contamination mostly in the field due to the early split of shell at the

end of maturation. Aflatoxin B1 (AFB1), represents the biggest concern due to the carcinogenic toxic effects, related to

hepatocellular carcinomas (IARC group 1). Ochratoxin A (OTA) is also found in pistachios, and more recently, some studies

show the presence of emerging mycotoxins, namely beauvericin, enniatins and tenuazonic acid.

Methodologies and scope: The present study carries out an extensive review of the state of the art of the determination of

mycotoxins in pistachios and addresses the main trends in the analytical methodologies to evaluate mycotoxins in dry nuts,

especially pistachios.

Results and Conclusions: The use of reliable analytical methodologies to determine mycotoxins is of paramount importance

for an accurate evaluation of exposure and risk assessment, especially of vulnerable groups of population, like children which

have a mean consumption of pistachios similar with adults, but they have higher exposure expressed on a body weight basis.

Because of the low levels of concentration and the complexity of a high lipid content matrix, most published studies agree that

Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) extraction followed by High-Performance Liquid

Chromatography coupled with Mass Spectrometry (HPLC-MS) for detection/quantification is currently the most suitable

analytical method to carry out the multi-mycotoxins determination in pistachio. This methodology allows low detection levels

with higher sensitivity, selectivity and specificity. Regarding the future trends, detection of multiple mycotoxins employing High-

Resolution Mass Spectrometry (HRMS) hybrid detectors as Double Quadrupole-Time-of-flight (QqTOF) or Quadrupole-orbital

ion trap (QOrbitrap) will have a critical impact in the near future, due to their potential for high-throughput analysis and more

accurate mass measurement. However, screening methods based on immunoassays will continue to be useful due to their

simplicity and rapid response in the determination of mycotoxins, e.g. in situ.

Keywords: Liquid Chromatography; Mass Spectrometry; Mycotoxins; Pistachios; QuEChERS.


43

P15 - Comparison of different d-SPE clean-up sorbents for determination of mycotoxins in pistachio

Mateus, A.A,B; Barros, S. B; Pena, A. A; Silva, A. B,C

A LAQV, REQUIMTE, Laboratory of Bromatology and Pharmacognosy, Faculty of Pharmacy, University of Coimbra, Polo III,

Azinhaga de Stª Comba, 3000-548 Coimbra, Portugal; AP: [email protected]; ARSM: [email protected] B National Institute of Agrarian and Veterinary Research (INIAV), I.P., Rua dos Lagidos, Lugar da Madalena, 4485-655, Vairão,

Vila do Conde, Portugal; ASS: [email protected]; SB: [email protected] C Center for Study in Animal Science (CECA), ICETA, University of Oporto, Oporto, Portugal

[email protected]

Introduction: Pistachio is of the nuts more contaminated by mycotoxins, and their determination is of utmost importance for

evaluation of exposure. This is especially relevant in the case of children, which have a mean consumption similar to adults,

resulting in a have higher exposure due to their lower body weight. The extraction is a critical step to have reliable analytical

methods. Multi-mycotoxin methods for pistachios are based on Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS)

methodology including clean-up step to eliminate interferents that could suppress or enhance the analytical signal. Dispersive

solid phase extraction (d-SPE) is the most used method, with classic sorbents like C18 (octadecyl modified silica) and PSA

(primary secondary amine). However, new sorbents, namely EMR-Lipid (enhanced matrix removal-lipid) and Z-Sep (modified

silica gel with zirconium oxide) are being used, especially for high lipid content matrices. In this study, those four sorbents were

compared to evaluate the most effective for the extraction of mycotoxins and clean-up from pistachio.

Methodology: The evaluation was performed spiking blank pistachio sample with 2 g/Kg of Aflatoxin B1 (AFB1); 4 g/Kg of

Aflatoxin B2 (AFB2), Aflatoxin G1 (AFG1), Aflatoxin G2 (AFG2); 3 g/Kg of ochratoxin A (OTA); 200 g/Kg of fumonisins (FB1

and FB2), zearalenone (ZEA), toxin T2 (T2) and toxin HT-2 (HT2) and submitting the extract to QuEChERS extraction. Four

variables were tested: (1) 1g of EMR-Lipid; (2) 50 and 100 mg of C18, PSA and Z-Sep; (3) addition of 50, 100 and 150 mg of

MgSO4 to 50 mg of sorbents and (4) mixtures in different proportions. Determination was performed using Ultra-High

Performance Liquid Chromatography combined with Time-of-Flight Mass Spectrometry (UHPLC–ToF-MS). Conclusions about

clean-up efficiency were based on the average peak area of each mycotoxin.

Results: Regarding the single use of sorbents, using 100 mg PSA or Z-Sep results in greater peak areas than 50 mg; except

the use of 100 mg C18 that only increased peak areas of some AFs. The addition of MgSO4 results in a slight increase of

signal but using 100 mg of sorbents provided the best results for most mycotoxins. PSA and C18 caused a significant loss in

the analytical signal of OTA. Concerning fumonisins, PSA and Z-Sep give no signal for FB1. EMR-Lipid and C18 give better

analytical signals for both fumonisins.

Conclusions: Multi-mycotoxins methods are a challenge because mycotoxins have different chemical proprieties. Although

EMR-lipid provides the highest analytical signals for most of mycotoxins, for AFs, the mycotoxins with most concern in

pistachio, 100 mg of Z-Sep provides the best analytical signals and was selected to perform d-SPE clean-up for determination

of multi-mycotoxins in pistachio.

Keywords: Mycotoxins; d-SPE; Sorbents; Z-Sep; EMR-Lipid.


44

P16 - Could young athletes be exposed to harmful substances when taking dietary supplements?

Marques. J 1, Capela. J. P. 2,3

1 –Universityof Porto, FacultyofSciences, Porto, Portugal. Email address: [email protected] 2 - Faculdade de Ciências da Saúde, Universidade Fernando Pessoa, Porto, Portugal. 3 - UCIBIO – Applied Molecular BiosciencesUnit,, Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade

de Farmácia, Universidade do Porto, Porto, Portugal.

[email protected]

Introduction: When great athletic performance is to be achieved, the use of dietary supplements (DS) becomes a necessity

since common foods no longer quenches all of the athlete’s nutritional needs. In turn this creates a growing consumer

population that is vulnerable from a young age to the ingredients present in these DS. This study aims to find out if there are

any doping substances, or dangerous amounts of any other component, stated on the labels of the analyzed DS.

Methodology: After refining both dietary brands and supplements, 40 dietary supplements with the full formula available, were

selected and divided into pre-workout and post-workout. The minimum and maximum dosages of selected ingredients were

statistically described as mean±SD.

Results: Regarding caffeine, the pre-workout group displayed higher mean caffeine (241±86 mg) than the post-workout group

(183±68 mg). Concerning creatine, the pre-workout group displayed lower mean creatine (3106±1079 mg) than the post-

workout group (4137±4177 mg). As for the salt content, the post-workout group displayed a much higher mean salt

(2155±4486 mg) than the pre-workout group (464±605 mg). Citrus aurantium extract, Yohimbe extract, Garcinia cambogia

extract and Maca root extract were also reported in some of the analyzed DS.

Conclusions: Consumption recommendations on the labels of reviewed DS could lead to excessive consumption of some not

yet fully tested ingredients, and consequently harm the athlete or consumer.

Relevance: DS are becoming every day products not only for athletes, but also for the common individual. In the meanwhile

DS brands and producers launch new products with scarcely tested ingredients that are available for purchase to anyone,

including minors.

Keywords: Dietary Supplements, Toxicity, Nutrition Sciences.


45

P17 – The interaction between fungi and insects and its contribute to maize flour food safety

Paula Alvito 1,2, Sónia Duarte 3,4, Ana Magro 3, Joanna Tomás 3, Carolina Hilário 3, Ricardo Boavida Ferreira 3,4 and

Maria Otília Carvalho 3,4

1Food and Nutrition Department, National Health Institute Dr. Ricardo Jorge (INSA), 1600-609 Lisboa, Portugal; 2Centre for Environmental and Marine Studies (CESAM), University of Aveiro, 3810-193 Aveiro, Portugal 3Instituto Superior de Agronomia, Universidade de Lisboa, Tapada da Ajuda, 1349-017 Lisboa, Portugal; 4LEAF—Linking Landscape, Environment, Agriculture and Food, Tapada da Ajuda, 1349-017 Lisboa, Portugal

[email protected]

Globally, 140 million children under five years are stunted, and wasting still impacts the lives of over 47 million children,

especially during the complementary feeding period. Improved formulations of complementary foods (CFs) with animal-source

foods (ASFs) is an important strategy to improve infant and young child feeding. However, ASFs are expensive in many food-

insecure settings where edible insects abound. Some insects as some species belonging to Tenebrionidae family (order

Coleoptera) are edible and could, in future, be used as CFs. It is therefore important to characterize the effects of other

organisms present in flours that could interact with insects, such as mycotoxigenic fungi, that produce mycotoxins. Insects may

trigger fungi growth and mycotoxin production therefore it will be important to evaluate the possible influence of insects on the

production of mycotoxins (e.g. aflatoxins) by mycotoxin producing fungi in maize flour, as a contribution to a better

understanding of this complex interaction and to the safety of future alternative CFs.

Maize was collected directly from fields was stored at −4 ◦C and then ground and sieved to obtain maize flour. The maize flour

was incubated under appropriate conditions of 30 ◦C ± 2 ◦C and 70 % ± 5 % RH for eight weeks on its own (control), inoculated

with: A. flavus conidia (fungus assay); T. castaneum adults (insect assay); or both organisms (mixed assay). Aflatoxin analysis

were performed in all samples by HPLC-FD.

Results showed that, when both organisms were together, the mycotoxins detected in maize flour were far higher than when

the fungi were on their own, suggesting that the presence of insects may contribute positively to fungi development and

mycotoxin production.

This is the first study on this issue. Further investigation would benefit from clarification on the mechanisms leading to the

nature of the detected interactions and to contribute to the safety of alternative foods, specially those that could be used for

children consumption.

Acknowledgements: FCT/MCTES for funding with national funds related to the SafeGrains project (PTDC/ASP-

PLA/28350/2017) and to CESAM (UIDP/50017/2020+UIDB/50017/2020).

Keywords: insects, cereals, mycotoxins, food safety


46

P18 - Biomonitoring of T-2 toxin in human urine and retrospective analysis of its metabolites through a high-resolution

mass spectrometry methodology

Narváez A.A, Izzo L.A, Castaldo L.A, Rodríguez-Carrasco Y.B, Ritieni A.A

A Department of Pharmacy, Università di Napoli Federico II, Via D. Montesano, 49 - 80131 Napoli, Italy. B Department of Food Chemistry and Toxicology, University of Valencia Av/ Vicent A. Estellés, s/n, 46100, Burjassot, Spain

[email protected]

T-2 toxin (T-2) is a type A trichothecene produced by several Fusarium species that is able to induce genotoxicity, cytotoxicity

and immunotoxicity in humans and animals. In response to the health concern derived from T-2 dietary exposure, authorities

have set a tolerable daily intake (TDI) value for the sum of T-2 and HT-2 at 0.1 µg/kg body weight. Nevertheless, other phase

I and phase II metabolites have been also reported after in vivo studies. Considering that human biomonitoring (HBM) represent

the most accurate approach for exposure assessments, the aim of this study was to validate an ultra-high performance liquid

chromatography coupled to Q-Orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap-HRMS) methodology for the

simultaneous detection of T-2 and its major metabolites in human urine samples using a combined targeted/untargeted

strategy. After that, the validated methodology was applied to 50 human urine samples.A salt-assisted procedure using

acetonitrile as solvent of extraction followed by a clean-up step with C18 were applied. Extracts were subjected to UHPLC-Q-

Orbitrap-HRMS analysis for further processing.

Results showed a good performance in terms of linearity, selectivity, trueness and precision for T-2 and HT-2, with limits of

quantification at 0.2 and 0.4 ng/mL, respectively. T-2 and HT-2 were quantified in 36% and 46% of the samples at a mean

level of 1.31 ng/mL and 1.25 ng/mL, respectively. Retrospective analysis revealed a vast occurrence of phase I metabolites,

mainly 3-hydroxy-T-2 (98%) and T-2 triol (94%), whereas phase II metabolites were not as frequent, being T-2-3-glucuronide

(8%) the most relevant compound.

The presence of phase I metabolites in almost all assayed samples regardless the low occurrence of T-2 pointed towards two

main events: firstly, phase I metabolism appeared to be the preferential biotransformation pathway and, secondly, a potentially

complete biotransformation of T-2. In light of these results, the sum of T-2, HT-2 and main phase I metabolites could stand as

a potential biomarker of exposure. Nevertheless, the correlation between ingestion and urinary excretion is still unknown, so

further toxicological studies are required.

This is the first analytical methodology for detecting the simultaneous presence of T-2 and its metabolites in human urine

samples.

Acknowledgments: This research was funded by the Spanish Ministry of Science and Innovation project (PID2020-115871RB-

I00-ALI) and by the Generalitat Valenciana (Spain) GV/2020/020 (Conselleria d’Innovacio, Universitats, Ciencia i Societat

Digital, Generalitat Valenciana).

Keywords biomarkers; exposure; high resolution mass spectrometry; T-2 toxin; metabolites


47

P19 - Development of a multi-residue method and risk characterization for pharmacologically active substances,

mycotoxins, and pesticides occurring in infant milk formulations

Izzo L. a, Narváez A.a, Castaldo L.a, Gaspari A.a, Rodríguez-Carrasco Y.b, Grosso M.c,d, and Ritieni A.a,e

a Department of Pharmacy, University of Naples “Federico II”, Via Domenico Montesano 49, 80131 Naples, Italy;

[email protected] (A.N.); [email protected] (L.C.); [email protected] (A.G.); [email protected]

(A.R.); b Laboratory of Food Chemistry and Toxicology, Faculty of Pharmacy, University of Valencia, Av. Vicent Andrés Estellés s/n,

46100 Burjassot, Valencia, Spain; c Department of Molecular Medicine and Medical Biotechnology, School of Medicine, University of Naples “Federico II”,

[email protected] (M.G.); d CEINGE-Biotecnologie Avanzate, 80131 Naples, Italy; e Health Education and Sustainable Development, Federico II University, 80131 Naples, Italy.

[email protected]

Infant milk formulations (IMFs) are complementary food that occupies a special place in childhood feeding which becomes an

essential product when breastfeeding is unavailable. Despite that, these formulations could serve as a vehicle for the

transmission of toxic substances.

In this study, a multiclass method based on QuEChERS approach was developed for the simultaneous determination of

contaminants (n=382) occurring in IMFs, including veterinary drug residues (VDs), mycotoxins, and pesticides.

The validated method was applied for the analysis of 18 IMFs. Dexamethasone was detected in 16.6% samples (range 0.905–

1.131 ng/mL), and procaine benzyl penicillin in one sample at a concentration of 0.295 ng/mL. Zearalenone was found in

55.5% samples (range 0.133-0.638 ng/mL) and α-zearalenol in 16.6% samples (range 1.534-10.408 ng/mL). Risk

characterization was also evaluated. Taking into consideration the highest contaminant concentration found, for a child 1-3

years old of 12 kg the calculated tolerable daily intake (TDI) percentages are in a range of 0.93-30.84%; whereas for infants

0-12 months of 5 kg the calculated TDI percentages are in a range of 2.23-74.01%.

In the worst-case scenario, the percentage found does not represent a health hazard but a significant contribution to TDI. Risk

characterization is an indispensable aspect to safeguard public health and to avoid the appearance of adverse effects due to

dietary exposure.

Keywords: infant milk formulation; risk characterization; tolerable daily intake; contaminants; UHPLC-Q-Orbitrap HRMS

mailto:[email protected]


48

P20 - Risk assessment associated to ethanol and residual solvents in liquid herbal dietary supplements for pediatric

population

Vojvodic S1,2, Torovic Lj1,3, Kusonic D1, Srdjenovic Conic B1,4

1University of Novi Sad, Faculty of Medicine, Department of Pharmacy, Hajduk Veljkova 3, 21000 Novi Sad, Serbia 2University Business Academy in Novi Sad, Faculty of Pharmacy, Novi Sad, Trg Mladenaca 5, 21000 Novi Sad, Serbia 3Institute of Public Health of Vojvodina, Futoška 121, 21000 Novi Sad, Serbia 4University of Novi Sad, Faculty of Medicine, Center for Medical and Pharmaceutical Investigations and Quality Control, Hajduk

Veljkova 3, 21000 Novi Sad, Serbia

[email protected]

Availability of herbal dietary supplements and belief in their efficacy and safety contributed to the increased use of these

products, emphasizing a need for evaluation of exposure to ethanol and other organic volatile compounds used as extraction

solvents, especially in early life, since children are particularly vulnerable.

Sixty-five liquid herbal dietary supplements for pediatric population were purchased from pharmacies in Novi Sad (Serbia)

during 2021, and analyzed for the presence of ethanol and residual solvents using HSS-GC-MS method.

Ethanol was not quantified in only one of 18 products for infants or toddlers less than two years old, although ethanol should

not be included in these products. Actual content of ethanol was higher than labeled in 9 of 26 samples with specified quantity.

WHO proposed requirement for ethanol content in products intended for children less than 6 years old (<0.5%) was not met

by 28 samples and in products for children 6-12 years old (<5%) by 5 samples. Calculated blood alcohol concentrations

exceeded the relevant toxicological levels for 24 samples following a single dose, and for 9 samples in case of accidental

poisoning with the entire package. Ethanol was considered as a residual solvent in 5 „alcohol free“ products and 31 product

where ethanol content was not mentioned. In 10 of these 36 samples ethanol content exceeded the permitted daily exposure

(PDE) level up to 33-fold in infants, 42-fold in toddlers, 22-fold in preschool children, 29-fold in children and 19-fold in

adolescents. Regarding the residual solvents, acetone and 1-butanol were not quantified, while 1-propanol was quantified in

5 samples and 2-propanol in one, both in low concentrations. Methanol was quantified in 38 samples and one sample was not

in compliance with Ph.Eur.9 requirement for methanol content (<0.05%) and ingestion of the same sample exceeded the PDE

level.

Obtained results are emphasizing a need for raising awareness among healthcare professionals and users in order to minimize

health concern for pediatric population associated with the use of herbal dietary supplements due to ethanol intake, calling for

the establishment of strict guidelines for ethanol content and labeling.

Keywords: ethanol, residual solvents, dietary supplements


49

P21 - Assessment of risk associated to elemental impurities in liquid herbal dietary supplements for pediatric

population

Torović LjA,B, Vojvodić SA,C, Srdjenović-Čonić BA,D, Lukić DB, Bijelović SA,B

AUniversity of Novi Sad, Faculty of medicine, Novi Sad, Serbia; BInstitute of public health of Vojvodina, Novi Sad, Serbia; C University business academy in Novi Sad, Faculty of pharmacy, Novi Sad, Serbia; DUniversity of Novi Sad, Faculty of medicine, Center for medical and pharmaceutical investigations and quality control, Novi

Sad, Serbia

[email protected]

Increased usage of dietary supplements emphasized a need for assessment of risk associated to concurrent exposure to

harfulm chemicals, including toxic elements, especially for vulnerable populations.

A total of 35 liquid herbal dietary supplements intended for pediatric population, claiming support for respiratory and immune

systems, was taken from the pharmacies in Novi Sad (Serbia) and analysed for 24 elements using ICP-MS after microwave

digestion. Elements' intake was calculated on the basis of producers' usage instructions and risk was evaluated by comparison

of exposure with oral permitted daily exposure (PDE), oral reference dose (RfD), and/or benchmark dose (BMDL) of elements.

Study results showed that a daily dose of any of tested supplements would not cause exposure higher than the corresponding

PDE levels. Daily contribution higher than 5% of PDE was reported for four elements, with a maximum of 47% Pb for infants,

67% Cu and 39% Pb for toddlers, 52% Cu, 20% Pb and 6.8% As for preschool children, 36% Cu and 14% Pb for children, and

31% Cu, 27% Pb and 8.0% Hg for adolescents. Conversely, one sample caused exceedance of RfDs for Cu (101%) and Zn

(113%) if consumed by toddlers. The highest contribution to the RfD reached 66% for W, 16% for Hg, and 15% for Tl, all for

adolescents, whereas exposure to other elements remained below 10% of respective RfDs in all population groups. However,

when all hazard quotients (%RfD values for individual elements) for a sample were summed to obtain a hazard index (HI),

values greater than 100% were obtained for one of the investigated samples, intended for toddlers (229%), preschool children

(177%), children (123%), and adolescents (105%). More realistic exposure scenario, asuming two weeks exposure during a

year (recommended duration of usage was very rarely declared), resulted with a substantial decrease of both HQ and HI

values, indicating no health concern. Exposures to As and Pb were below the respective BMDLs in both exposure scenarios.

Risk management decisions based on relevant risk assement data and directed towards mittigation of early-life exposure to

harmful chemicals are pivotal for protection of childrens' health.

Keywords: pediatric population, dietary supplements, elements, risk assessment


50

P22 - Efficacy of bioactive components of bottle gourd oilseed extracts on the antimicrobial activity and toxigenic

fungi

Ahmed N. Badr1 and Adel G. Abdel-Razek2

1 Food Toxicology and Contaminants Dept., National Research Centre, Dokki 12622, Cairo, Egypt. 2 Fats and Oils Dept., National Research Centre, Dokki 12622, Cairo, Egypt.

[email protected]

Introduction: The bottle gourd seeds are surrounded by innumerable bioactive components of phyto-chemicals. This work aim

to evaluate the effect of bioactive components of bottle gourd seed extracts on the antimicrobial activity and toxigenic fungi

compared to the standard antibiotics.

Materials and Methods: Bottle-gourd seeds were prepared using two extraction types (Polar and non-polar). A simple

extraction technique was applied to gain polar eco-friendly iso-propanol extract utilizing ultra-sonication assisted method. The

extract was evaluated for its antioxidant potency, total phenolic, and total flavonoid content. The non-polar extract of seeds

was gained using petroleum ether (40-60). The fatty acid profile of oil was determined using GC. Minimal inhibition

concentration (MIC) and minimal fungicidal concentration (MFC) were determined. The inhibition impact of extracts on toxigenic

fungi and their mycotoxin secretion were evaluated.

Results: The fatty acid profile of bottle-guard oil showed a variation for its content of fatty acids, particularly (C20:1 and C22:1).

The MIC and MFC showed a variation between the two extract types with distinguishing for polar extract against the pathogens.

Three sterols were detected in this oil, with high content of B-sitosterols. The oil also was recorded by valuable B-carotene

content and tocopherols. Otherwise, the polar extract is represented by high antioxidant activity. Total phenolic contents were

shown at 15.5 and 267 mg of GAE/g for oil and polar extract respectively. The total flavonoid contents were 2.95 mg catechol

/g oil and 14.86 mg catechol /g for isopropyl extract of bottle guard seeds. The highest inhibition % of DPPH was shown by

polar extract of seeds than oil. The antimicrobial activity recorded a promising effect for the polar extract against bacterial

pathogen strains and toxigenic fungi compared to the standard antibiotics. Conclusion: The Bottle gaurd seed extracts as a

non-traditional bioactive source, are considered as a novel promising solution, which could participate in increasing the safety,

shelf-life, and security of the food products.

Relevance: Application of bottle gaurd extracts will provide a novel source of component faced the risk of vulnerable

populations.

Keywords: Bottle gourd oilseeds, Bioactive components, antimicrobial activity, Toxigenic fungi.


51

P23 - Determination of Acrylamide in potato chips commercialized in Morocco by high-performance liquid

chromatography–diode array detector (HPLC-DAD)

Abida S. 1, El hamidi A. 1, Sifou A. 1

1 Department of Chemistry, Faculty of Sciences, Mohamed V Agdal University, Avenue Ibn Batouta, Rabat, Morocco

[email protected]

Introduction: Nowadays acrylamide (AA) is known not only as synthetic material used in industry, but also as food contaminant

present in a wide range of frequently consumed foods, which forms during heat-induced process (due to Maillard reaction)

mostly in potato chips, biscuits, coffee, fried nuts and cereals, while it is absent in raw foodstuffs. Due to its genotoxicity and

carcinogenicity, AA was classified as a Group 2A carcinogen by the International Agency for Research on cancer and a

Category 2 carcinogen and Category 2 mutagen by the European Union, which caused worldwide concern. It was put into the

list of substances of ‘‘very high concern’’ by the European Chemical Agency in 2010. Therefore, the food industry faces a

challenge to modify the processes or to change the product parameters without compromising the quality of their foods.

Methodology: In this work a simple and rapid method was developed for the determination of AA in fifty-six (n=56) potato chips

samples commercialized in different markets in Rabat in Morocco. AA was co-extracted with methanol and then purified and

enriched by an Oasis HLB solid-phase extraction cartridge, simultaneously analyzed by high-performance liquid

chromatography (HPLC) and detected with a diode array detector (DAD), respectively, at their optimal wavelength.

Results: The limit of detection and the limit of quantification for this method were 1.09 and 4 μg/kg, respectively. The mean

recoveries of AA obtained by using spiked samples ranged between 75% and 110%, which shows good extraction efficiency.

AA concentrations in potato chips samples are between 25 and 1468 μg/kg and the arithmetic mean AA content resulted 370

μg/kg. 34 of the samples (i.e. 60%) reported AA level above 500 μg/kg (benchmark level settled for the EU Regulation

2158/2017).

Conclusions and relevance: These results revealed reasonable levels of AA in this foodstuff, which indicated the general risk

of consumer exposure. This is the first study on the occurrence of the AA in potato chips samples commercialized in Morocco.

Keywords: Acrylamide (AA), Maillard reaction, HPLC–DAD, potato chips, Morocco


52

P24 - Monitoring of acrylamide formation in biscuits and its mitigation

Siopa, J.1, Cosme, F.1,2, Nunes, F.M.1,3

1 Centro de Química de Vila Real (CQ-VR) – Food and Wine Chemistry Lab, Universidade de Trás-os-Montes e Alto Douro,

Quinta de Prados, 5001-801, Vila Real, Portugal 2 Departamento de Biologia e do Ambiente, Universidade de Trás-os-Montes e Alto Douro, Quinta de Prados, 5001-801, Vila

Real, Portugal 3 Departamento de Química, Universidade de Trás-os-Montes e Alto Douro, Quinta de Prados, 5001-801, Vila Real, Portugal

[email protected]

Acrylamide (2-propenamide) is a Maillard reaction product (MRP) and it is mainly generated through the reaction of the carbonyl

group of reducing sugars and the amine group of asparagine [1]. In 2002, acrylamide was observed in heat-processed foods

for the first time, and since then, many studies were made to understand the impact of this MRP on consumer’s health.

Acrylamide is considered potentially carcinogenic and neurotoxic and it is present in a large variety of widely consumed

products, such as fries and potato chips, breakfast cereal, biscuits, bread, coffee, and many others [2]. Therefore, it is important

to monitor the acrylamide formation and to study strategies to mitigate acrylamide in those foods as much as possible, without

negatively impact its sensory characteristics.

In this work, a method to quantify acrylamide was developed and optimized for its application in biscuits and cookies by GC-

SIM-MS, through the combination of methods previously described in the literature. Wheat flour and biscuit dough were studied

regarding their sugars and amino acids content. Biscuits were also evaluated regarding the amino acids and sugars content,

in order to study the extension of acrylamide formation. The effectiveness of asparaginase as an acrylamide mitigating agent

was also tested. Samples were previously prepared with L-asparaginase, incorporating the enzyme in the dough before the

heat processing, and acrylamide was quantified. The biscuits colour was also evaluated, using the CIELab method, in order to

understand the effects of the treatment on the sensory properties of the final product.

Asparaginase treatment showed to be effective in reducing the acrylamide content, as the final products were slightly lighter

in colour. Asparagine content of the initial ingredients seemed to be the most influencing reagent of acrylamide formation and,

therefore, the use of ingredients with a low content of this amino acid could be another way to prevent large amounts of

acrylamide in the final product.

Biscuits are widely consumed, specially by the younger population. Since acrylamide are potentially carcinogenic, mutagenic

and neurotoxic, it can be a serious threat to the public health. Therefore, monitoring acrylamide formation by the food industries

is extremely necessary.

Acknowledgements: We would like to thank the financial support provided by MobFood project – «Mobilização de

conhecimento científico e tecnológico em resposta aos desafios do mercado Agroalimentar» (POCI-01-0247-FEDER-024524).

We also want to thank the funds from the Portuguese Foundation for Science and Technology (FCT) to CQ-VR (PEst-

OE/QUI/UI0616/2014). Additional thanks to Cerealis company.

Keywords: Maillard reaction, acrylamide, asparagine, asparaginase, biscuits


53

P25 - Detoxification of Aflatoxin B1 and Ochratoxin A by Enterococcus sp strains

Badji T 1, 2; Riba A 2, 3; Brabet C 4 and Durand N 4

1 Laboratory of valorization and conservation of biological resources (VALCOR), Boumerdes, Algeria 2 Laboratoire de biologie des systèmes microbiens, ENS Kouba, Alger, Algeria 3 Département de biologie, faculté des sciences, université M’Hamed Bouguera, Boumerdes, Algeria 4 Centre de coopération internationale en recherche agronomique pour le développement

(CIRAD), UMR –Qualisud, Montpellier, France.

[email protected]

Introduction: Mycotoxins are the secondary fungal metabolites commonly found in foods that can pose a serious health

concerns to both human and animals. This study aims mainly to determine the detoxification properties of two strains of

Enterococcus sp (Ent1 and Ent2). isolated from two different matrices in Algeria, towards Aflatoxin B1 and Ochratoxin A in

vitro.

Methodology: The experiment was carried out in contaminated CPB solution (0.1 M pH 6 or 5) at 25°C over 24 h incubation

period. Subsequently residual mycotoxin concentrations were determined by HPLC for each strain.

Results: The two strains detoxified mycotoxins, with highest efficiency for AFB1. The strain Ent1 remove approximately 16%

OTA, 25% AFB1 and 49% OTA, 73% AFB1, however Ent2 remove 24% OTA, 52% AFB1 and 34% OTA, 54% AFB1 at pH 6

and pH 5 respectively. pH medium effect the amounts of mycotoxins removed by Ent1 strain, whereas pH doesn’t any effect

on the removal ability of Ent2 strain.

Conclusion and relevance: The tested enterococcus sp. strains used in this study are able to remove AFB1 and OTA in tested

conditions. Thus, they can be used as additives to reduce mycotoxins level in contaminated foods.

Keywords: Enterococcus sp. strains, Ochratoxin A, Aflatoxin B1, Removal ability


54

P26- Moisture and mycotoxin content in maize grains and maize silage

Campos, Cláudia A,B,1; Leite, Marta A,C,D; Barbosa, Jorge C; Ramos, Fernando C,D; Afonso, Isabel B; Freitas, Andreia A,C.

A National Institute for Agricultural and Veterinary Research (INIAV), Rua dos Lágidos, Lugar da Madalena, 4485-665 Vila do

Conde, Portugal B Polytechnic Institute of Viana do Castelo, Escola Superior Agrária, Ponte de Lima, 4990-706, Portugal C REQUIMTE/LAQV, R. D. Manuel II, Apartado 55142, Porto, Portugal D Faculty of Pharmacy, Health Science Campus, University of Coimbra, Azinhaga de Santa Comba, 3000-548 Coimbra,

Portugal

[email protected]

Introduction: Maize (Zea mays L.) is one of the most important crops in the world for its use as animal feed and for human

consumption. This cereal is considered a great source of energy, proteins, starch and glucose, and is also of great economic

interest, especially in developing countries and countries with food shortages. However, they are susceptible to contamination

by mycotoxins that can arise at any stage of production, whether in the field and after harvesting or during storage. These

contaminants are harmful to human health, particularly to such vulnerable populations, and to target age groups – children and

elderly people –, whose direct consumption of animal-derived food (e.g. milk), maize and maize-based products (e.g.

processed cereals) is higher. The known individual hazardous effects go from carcinogenicity, immunological toxicity,

neurotoxicity, hepatotoxicity, to reproductive and developmental toxicity, but the emerging issues concerning multiple

mycotoxin exposure due to bioaccumulation since the first stage of the food chain remains an increasing area of concern.

Methodology: In the present work, the moisture content was determined for maize grain and maize silage, given that this is

one of the main factors for the growth of mycotoxigenic fungi and, consequently, production of mycotoxins in such samples.

These toxic compounds were then subjected to an extraction procedure based on modified QuEChERS protocols. The

identification of a wide range of mycotoxins in maize-based samples was performed using a liquid chromatography coupled

with tandem mass spectrometry (UHPLC-MS/MS) methodology.

Results: Regarding the moisture content, maize silage presented an average value of 67.0% (n=11), which is much higher

moisture content than the average value obtained for grain maize (17%, n=8). Concerning the occurrence data, mycotoxins

were identified in both grain and silage samples, being ZEA and FB1 the major occurring mycotoxins.

Conclusions/Relevance: The evaluation of mycotoxins in maize is, therefore, of great importance since it will allow to define

and develop correct prevention and mitigation strategies towards the reduction of this fungi toxins, and, ultimately, the exposure

to multiple mycotoxins throughout the maize value chain.

Keywords: Mycotoxins, maize grains, maize silage, moisture content.


55

P27 - Bisphenol A and its analogues B, E, F and S in infant formulas commercialized in the Brazil: contamination

levels and estimated dietary intake

Petrarca, M.H.A*; Perez, M.A.F.B; Tfouni, S.A.V.A

A Centro de Ciência e Qualidade de Alimentos, Instituto de Tecnologia de Alimentos - ITAL, Avenida Brasil, 2880, 13070-178,

Campinas, SP, Brasil B Centro de Tecnologia de Embalagem, Instituto de Tecnologia de Alimentos - ITAL, Avenida Brasil, 2880, 13070-178,

Campinas, SP, Brasil

[email protected]

Introduction: The occurrence of contaminants, particularly those associated to the endocrine disrupting activity, in foods

intended for infants and young children is an issue of great interest since this consumer group is presumed to be potentially

more sensitive to some toxicological insults when compared to adults due to the higher daily food intake on the body weight

basis. In this way, the objective of the work was to investigate the occurrence of the bisphenols A, B, E, F, and S in infant

formulas commercialized in the Brazil.

Methodology: A sample preparation based dispersive solid-phase extraction with acetonitrile solvent and C18 sorbent was used

for the extraction and cleanup. For identification and quantification purposes, an ultra performance liquid chromatography

system coupled to a triple quadrupole mass spectrometer, with electrospray source operated in negative mode, was employed.

The estimated daily intake (EDI) was calculated as follows: EDI = C x A / b.w., where C is the analyte concentration in the

sample (µg/g), A is the daily consumption of infant formula (g), and b.w. is body weight (kg).

Results: Adequate method performance characteristics were achieved, including linearity in matrix-based calibration curves;

recoveries between 84 and 109% with relative standard deviation values lower than 18%; and suitable limits of detection (5–

10 µg/kg) and quantification (10–20 µg/kg). A total of sixty-one samples were analyzed and bisphenol A (11.3–18.7 µg/kg),

bisphenol E (11.5–14.8 µg/kg), bisphenol F (46.4–198.9 µg/kg), and bisphenol S (10 µg/kg) were found in 11.5%, 3.3%,

19.7%, and 1.7% of the samples, respectively. The EDI values ranged between 0.07 and 2.6 µg/kg b.w.

Conclusions: To the best of our knowledge, this is the first report on simultaneous analysis of bisphenols A, B, E, F, and S in

infant formulas available in the Brazilian market. Although the EDI values were lower than the temporary tolerable daily intake

(t-TDI) of 4 µg/kg b.w. for the bisphenol A established by the European Food Safety Authority, the occurrence of these

chemicals reinforces the need for constant monitoring of bisphenols in infant foods to ensure the quality and safety of these

foodstuffs.

Keywords: Endocrine disrupting compounds; Infant foods; UPLC-MS/MS


56

P28 - Contribution to the study of aflatoxins and ochratoxin producing capacity of toxigenic fungi isolated from

Algerian wheat

Khouni Hayat1, 2;Riba Amar2, 3; Mokran Salim2 and Meklat Atika2

1Laboratory of Valorization and Conservation of Biological Resources (VALCOR), Boumerdes, Algeria 2Laboratoire de Biologie des Systèmes Microbiens, ENS Kouba, Alger, Algeria 3 Département de Biologie, Faculté des Sciences, Université M’Hamed Bouguera, Boumerdes, Algeria

[email protected]

Introduction :Mycotoxins are secondary metabolites of low molecular weight produced by a variety of fungi (mainly Aspergillus,

Fusarium and Penicillium spp.). They present diverse chemical structures, having different biological effects on animals and

humans,such as teratogenicity, carcinogenicity, mutagenicity, immunotoxicity or neurotoxicity. Studies show that mycotoxins

are ubiquitous contaminants. It is estimated that they are present in approximately 25% of cereals consumed worldwide. In

general, mycotoxin contamination is higher when climatic conditions are favourable. Algeria is a North African country whose

climate is characterised by high temperatures and high relative humidity in some areas that seems to stimulate the toxigenic

moulds growth and toxinogenesis, in which cereals and cereal-based products such us wheat represent a staple food for the

population. In this context, the objective of this work is to analyze the fungal flora contaminating Algerian wheat in one hundred

samples of soft wheat and durum wheat, from several wilayas of Algeria harvested in July 2020.

Methodology: The toxigenic molds were isolated on Dichloran Rose Bengal Chloramphenicol Agar (DRBC) medium then

identified and preserved on PDA medium, the aflatoxin and ochratoxin producing power is tested on selective media and

confirmed by thin layer chromatography.

Results: During this study, one hundred and eighty six strains of the genus aspergillus and Penicilluim were isolated and

identified, forty from wheat harvested from fields and one hundred and fifty six from stored wheat. The screening of aflatoxigenic

isolates is based on the demonstration of blue fluorescence under U.V at 365nm on coconut extract (CAM) medium. The

production capacity of the isolates that showed blue fluorescence and an orange-yellow reverse side of the CAM medium was

confirmed by TLC using an aflatoxin standard extracted from the reference strain A.parasiticus .The study of OTA production

capacity was performed on CYA medium for isolates belonging to the genus Penicillium. Thus the analysis by TLC revealed a

rate of 27.04% of which 11.22% are aflatoxigenic and 15.8% of isolates are ochratoxigenic

Conclusion : The presence of aflatoxin and OTA in wheat and its derivatives constitutes a sanitary and economic problem in

the world. In this work, we were particularly interested in studying the aflatoxin and ochratoxinogenic species in wheat in

Algeria.The contamination by fungi was observed in all the analyzed samples of wheat. From these results, we note that more

extended studies and by other methods are necessary to evaluate the risk of mycotoxins.

Keywords: wheat, Aspergillus, Penicilluim, Algeria


57

P29 - Exposure of gastrointestinal tract to the mycotoxin aflatoxin B1: An overlooked impact?

Inês Serrenho 1,2 , Ricardo Assunção 1,3,4,5 and Paula Alvito 1,3

1Departamento de Alimentação e Nutrição, Instituto Nacional de Saúde Doutor Ricardo Jorge, Lisboa, Portugal 2Faculdade de Ciências da Universidade de Lisboa, Lisboa, Portugal 3Centro de Estudos do Ambiente e do Mar, Universidade de Aveiro, Aveiro, Portugal 4 NOVA, National School of Public Health, Public Health Research Centre, Universidade NOVA de Lisboa, Lisboa, Portugal 5 IUEM, Instituto Universitário Egas Moniz, Egas Moniz – Cooperativa de Ensino Superior, CRL, Monte da Caparica, Portugal

[email protected]

Introduction: Human exposure to aflatoxin B1 (AFB1) through contaminated food ingestion and its impact on the gastrointestinal

tract (GIT) is a field of scientific growing interest. Beyond the recognized carcinogenic potential, AFB1 has the ability to disrupt

the intestinal barrier affecting its balance and dynamics, and could promote the development of chronic intestinal inflammation,

among other pathologies. These conditions may affect the general population and could constitute a matter of increased

concern for vulnerable populations, such as pregnant women and children, especially when AFB1 transfer through the placenta

is considered. Since it is expected to occur an increase of AFB1 exposure through food due to climatic change and this could

consequently contribute to an increase in number of GIT pathologies, this study aims to raise awareness of health professionals

for the potential impact of AFB1 in the GIT and associated health effects.

Methodology: The bibliographic search was carried out in 2021, using the Web of Science and PubMed search engines,

including specific key words, for the period 2000-2020. From a total of 265 references found, 208 were excluded (duplicates

and studies outside the intended context) and 57 were analyzed.

Results and Conclusion: The significant impact of AFB1 in the intestinal barrier was evidenced, compromising the intestinal

immunity and, consequently, affecting the health of the considered organism. As it becomes increasingly evident that early

exposure to chemical contaminants can lead to the onset of diseases in adults such as infections, cancer, diabetes and

autoimmune diseases, it is therefore important to develop studies aiming to assess the importance of the environment

intrauterine and first years of life in the health of the child and the future adult. The raise of awareness of health professionals

of a possible increase in number of GIT pathologies associated to exposure to AFB1 arising from climatic change is therefore

crucial to empower health professionals to act based on a deeper scientific knowledge.

Acknowledgments: FCT/MCTES for funding with national funds related to the earlyMYCO project (PTDC/MEDTOX

/28762/2017) and to CESAM (UIDP/50017/2020+UIDB/50017/2020). R.A. thanks for the FCT Individual CEEC 2018 Assistant

Researcher Grant CEECIND/01570/2018.

Keywords: Aflatoxin B1, vulnerable populations, human health, intestinal barrier


58

P30 - Pesticide residue in Brazilian food: thinking about children

Sarmiento-Santos A, J.; NascimentoA, M. B.; AraujoA, L. S.; VaninA, F. M.

A Food Engineering Department, University of São Paulo, Faculty of Animal Science and Food Engineering (USP/FZEA),

Laboratory of Bread and Dough Process (LAPROPAMA), Av. Duque de Caxias Norte 225, 13635-900, Pirassununga, SP,

Brazil.

[email protected]

The risk of contamination with pesticide residues from the consumption of fruits and vegetables is high. Infants, due to their

body weight and higher food intake, when compared to adults, can be considered more sensitive to contamination, justifying

the growing concern with food for this population in the last decades. This is precisely why they are considered the right risk

groups, furthermore when considering the first foods offered during the food introduction, fruits and vegetables, which are the

basis of their solid diet practically up to 2 years old. In 2006, to protect children from ingesting harmful substances, processed

baby foods were regulated and must not contain individual pesticide residues at levels greater than 0.01 mg/kg of food in the

European Union (EU) in accordance with Commission Directive 2006/125/EC. The regulations of other countries besides the

EU do not differ the limit according to the age of the consumer.

Therefore, considering the reference of pesticide residue for baby food in EU and the monitoring programs in Brazil, this

research compared the results of pesticide residue on natural foods, with the child maximum residue level (MRL) established

by the EU. The data used were selected through consultations in the Program for the Analysis of Pesticide Residues in Food

(PARA) of Brazilian National Health Surveillance Agency (ANVISA) and the National Plan for the Control of Residues and

Contaminants in Products of Vegetal Origin (PNCRC/Vegetal) of Brazilian Department of Agriculture, Livestock and Supply

(MAPA), considering the period between 2015 and 2018. Food that are usually not offered for babies were disregarded from

this research.

A total of 5.528 vegetal samples were analyzed by the monitoring programs and 87.1% of these samples were in accordance

with their respective MRL. However, between the foods analyzed by PARA, only garlic, rice, potatoes and chayote had average

results of pesticide residues equal to or below the EU-child-MRL. Besides, only 44.0% of analyzed food had residues with

results less than 0.01 mg/kg of food.

These results evidenced that most fruits and vegetables have pesticide residue above EU recommendations for baby foods.

Acknowledgements: This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior -

Brasil (CAPES) - Finance Code 001

Keywords: Baby food; MRL; monitoring programs; risk groups


59

P31 - Paraben levels in placenta in relation to weight and head circumference at birth in Belgium: an ENVIRONAGE

birth cohort study

Van Overmeire I1, Vrijens K2, De Cremer K1, Neven KY2, Carollo RM2, Van Loco J1, Nawrot TS 2, 3, Vleminckx C1

1Sciensano, Chemical and physical Health Risks, J.Wytsmanstraat 14, 1050 Brussels, Belgium. email:

[email protected] 2Center for Environmental Sciences, Hasselt University, Agoralaan, 3590 Diepenbeek, Belgium. 3Department of Public Health, Environment & Health Unit, Leuven University (KU Leuven), Kapucijnenvoer 35, 3000 Leuven,

Belgium.

[email protected]

Introduction: Parabens are a group of esters of para-hydroxybenzoic acid utilized as antimicrobial preservatives in food,

medicines and many personal care products. Epidemiological studies regarding the adverse effects of parabens on fetuses

are limited. The aim of this study was to determine the association between placental paraben exposure and birth outcomes.

We assessed paraben concentrations in placental tissue, which potentially gives a better understanding of fetal exposure than

the maternal urinary concentrations which are the current golden standard.

Methodology: Placental tissue was collected immediately after birth from 142 mother-child pairs from the ENVIRONAGE birth

cohort. The placental concentrations of four parabens (methyl (MeP), ethyl (EtP), propyl (PrP), and butyl (BuP)) were

determined by ultra-performance liquid chromatography coupled with tandem mass-spectrometry. Generalized linear

regression models were used to determine the association between paraben exposure levels and birth outcomes.

Results: The geometric means of placental MeP, EtP, PrP, and BuP were 1.84, 2.16, 1.68 and 0.05 ng/g tissue, respectively.

The sum of parabens (Σ parabens, including MeP, EtP and PrP) was negatively associated with birth weight in newborn girls

(− 166 g, 95% CI: − 322, − 8.6, p = 0.04) after adjustment for a priori selected covariates. The sum of parabens was negatively

associated with head circumference (− 0.6 cm, 95% CI: − 1.1, − 0.2, p = 0.008) and borderline associated with birth length (−

0.6 cm, 95% CI:-1.3, 0.1, p = 0.08). In newborn girls the placental concentration of EtP was negatively associated with head

circumference (− 0.6 cm, 95% CI:-1.1, − 0.1, p = 0.01) and borderline significantly associated with birth weight and birth length.

Lastly, placental EtP and Σparabens were negatively associated with placental weight in newborn girls but not in newborn boys

(− 45.3 g, 95% CI:-86.2, − 4.4, p = 0.03).

Conclusion: The negative association between maternal paraben exposure and birth outcomes warrants further research and

follow-up over time to determine long term effects of gestational exposure to parabens.

Keywords: paraben exposure, placental paraben concentrations, birth cohort


60

P32 - Preliminary risk assessment of trace elements exposure by consumption of seaweeds

Pinto, F.R.A; Severo, P.B; Mendes, S. C; Martins, L.L.D; Mourato, M. D Gil, M.M. C

AMARE – Marine and Environmental Sciences Centre, Polytechnic of Leiria, Cetemares, Peniche, Portugal BNOVA School of Science and Technology, Caparica, Portugal CMARE – Marine and Environmental Sciences Centre, ESTM, Polytechnic of Leiria, Cetemares, Peniche, Portugal DLEAF – Linking Landscape, Environment, Agriculture and Food, Instituto Superior de Agronomia, University of Lisbon, Lisbon,

Portugal

[email protected]

Seaweeds have a wide application in several industrial sectors like human food. Seaweeds are a low-calorie, highly nutritious

food that provides nutrients such as dietary fibres, essential elements and amino acids, polysaccharides, polyunsaturated fatty

acids and vitamins. Some of these compounds have antimicrobial and antioxidant bioactivity and are not produced by terrestrial

plants. However, seaweeds are also known for their high capacity to accumulate elements that may or may not be beneficial

to human health.

The aim of this study was to assess the trace elements (Cd, Pb, Cr, Fe, Cu and Zn) exposure through the consumption of wild

and farmed seaweeds of the genus Fucus and potential health risk in Portuguese population using a deterministic method.

Mineral elements were quantified by inductively coupled plasma - mass spectrometry (ICP-MS).

The trace elements exposures were estimated by combining the trace elements concentration data with a seaweeds

consumption survey (case study applied in Portugal for a sample of 925 individuals).

It was found that it is very small for the consumption pattern of both farmed and wild seaweeds of Fucus genus in the studied

population categories, suggesting that there is no risk for consumers due to the trace elements.

Both farmed and wild seaweeds of Fucus genus seems to have low levels of studied trace elements and in accordance with

the Recommendation (EU) 2018/464. Yet, the concentration of all elements under study is higher in farmed seaweeds leading

to higher exposures.

Keywords: exposure, risk assessment, Fucus sp., trace elements, food safetyconsumption survey


61

P33 - Genetically Modified Organisms (GMO)-associated herbicides and reproductive tissues: effects on human

sperm mitochondria and prostate cell-based assays

Mikhail M1, Ferramosca A2, Marcoccia D1, Vremere A1, Zara V2, Lorenzetti S1

1 Dept. of Food safety, nutrition and veterinary public health, Istituto Superiore di Sanità (ISS), Rome, Italy 2 Dept. of Biological and Environmental Sciences and Technologies, University of Salento, Lecce, Italy

[email protected]

Introduction. The widespread use of genetically modified organisms (GMOs) led to an increased application of selective

herbicides to which GMOs are resistant, raising concerns about human health upon exposure to them. Glufosinate ammonium

(GA) and glyphosate (GLY), the active principles of the most used GMO-associated formulations, have been investigated for

their potential link to cancer and reproductive toxicity. However, little is known about their effects on sperm quality and male

accessory glands, e.g. the prostate.

Methodology. The effects of GA and GLY on human sperm mitochondrial respiration were investigated by an ex vivo assay

(Ferramosca et al., 2020. Doi: 10.1016/j.reprotox.2020.11.011), whereas their role on the prostate (Lorenzetti et al., 2010. Doi:

10.1016/j.reprotox.2010.03.008) were analyzed by evaluating the prostate-specific antigen (PSA) secretion in the epithelial

cell line LNCaP, derived by a human prostate adenocarcinoma.

GA and GLY treatments (0.1-1000 nM) have been compared to sex steroids’ treatments with testosterone (T), di-

hydroxytestosterone (DHT), 17 -estradiol (E2) and progesterone (P4), depending on the known physiological regulators of

sperms and prostate homeostasis.

Co-incubation experiments were carried out with the two herbicides in presence of each sex steroid at 10 nM, and with the

hormone-like flavonoid quercetin (QRC), an ubiquitous non nutrient in vegetables and fruits.

Results. In human spermatozoa: i) DHT and QRC (≥ 10 nM) significantly reduced mitochondrial functionality; ii) GLY and GA

negatively affected mitochondrial respiration efficiency; iii) in presence of 10 nM DHT, the negative effect of GLY was increased;

iiii) DHT, QRC and GA targeted mitochondria by using a mechanism different from GLY. In the prostate LNCaP epithelial cells,

similarly to DHT, GA induced the secretion of free and total PSA at concentrations higher than 50 nM.

Conclusions and relevance. Both GA and GLY can affect cells of the male reproductive system. Hence, the human health

epidemiological concern about their widespread use can be supported not only by animal studies but also by ex vivo (human

spermatozoa) and in vitro (prostate cell-based assays) studies aiming to identify specific mechanisms and modes of action.

Keywords: spermatozoa, prostate, glufosinate ammonium, glyphosate, quercetin


62

P34 - Aluminium intake through the consumption of selected baby foods and risk characterization in a population of

Brazilian infants aged 0-12 months old

Paivaa,b E. L., Limac, S. G., Marquesc, R. E. F. A., Morganob, M. A., Arisseto-Bragottoa , A. P.

aFaculty of Food Engineering, University of Campinas (UNICAMP), Rua Monteiro Lobato 80, 13083-862 Campinas - SP, Brazil bInstitute of Food Technology (ITAL), Avenida Brasil 2880, C. P. 139, 13070-178 Campinas - SP, Brazil cUniversidade Paulista (UNIP), Av. Comendador Enzo Ferrari 280, 13045-770 Campinas - SP, Brazil

[email protected]

Increased susceptibility to the toxic effects of aluminium (Al), which include anemia, impaired bone formation, disorders in the

digestive, respiratory, and immune systems, as well as neurotoxic effects, is observed in the early stages of life, fetal and early

postnatal periods. In this work, Al dietary intake from selected baby foods was estimated for Brazilian infants from São Paulo

State in order to allow a risk characterization.

The exposure assessment was carried out by combining levels of Al in 171 samples of baby foods (infant formula,

meat/vegetable purees, fruit purees, petit-suisse and cereals), analytically determined by inductively coupled plasma optical

emission spectrometry method, with individual food consumption data, using a deterministic approach. Data on food

consumption were generated using a duplicated 24 h recall applied to the parents of 63 infants aged from 0 to 12 months,

between January and May 2019. Two parameters were used to evaluate the potential risk of exposure to Al: the Provisional

Tolerable Weekly Intake (PTWI) of 2 mg/kg body weight (bw) established by the JECFA and the Tolerable Weekly Intake (TDI)

of 1 mg/kg bw set by the EFSA.

The mean Al intake, calculated using mean Al concentrations, was estimated to be 0.223 mg/kg bw per week, corresponding

to 11.1% of the PTWI and 22.3% of the TWI. When maximum Al concentrations were used, the obtained mean intake was

0.852 mg/kg bw per week, which corresponds to 42.6% and 85.2% of the PTWI and TWI, respectively. In addition, at this

critical scenario, estimated Al intake at 95th percentile was 1.632 mg/kg bw per week, corresponding to 81.6% of the PTWI

and 163.2% of the TWI. The food with the highest frequency of consumption and daily consumed amount was meat/vegetable

purees with values of 80.7% and 142.5g/day, respectively. The contribution of the selected baby foods to the mean Al exposure

were: meat/vegetable purees (54.7%), infant formulas (36.0%), cereal (4.4%), fruit purees (3.3%) and petit-suisse (1.7%).

In conclusion, this study suggests potential concern regarding consumers of highly Al contaminated products and may be used

as a basis for the establishment of risk management actions.

Keywords: infant foods; exposure assessment; contaminants; food safety


63

P35 - Effect of genistein on the behaviour of zebrafish larvae - preliminary data

Ribeiro O.1*, Coimbra A.M. 1, 2, 3 and Carrola J. S. 1, 2

1 School of Life and Environmental Sciences (ECVA), University of Trás-os-Montes and Alto Douro, (UTAD), Vila Real, Portugal 2 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), UTAD, Vila Real, Portugal 3 Inov4Agro – Institute for Innovation, Capacity Building and Sustainability of Agri-food Production

[email protected]

Endocrine disrupting compounds (EDCs) are xenobiotics or natural compounds that mimic or interfere with hormones, and can

affect development, reproduction, behaviour, and may induce carcinogenesis, teratogenesis and obesity. Among the various

types of EDCs, phytoestrogens are produced by plants and present a similar structure to animal oestrogens. Genistein

belonging to the isoflavone class can be found in various soy-based foods and has been detected in effluents from wastewater

treatment plants at mean concentrations of 12.6 ±4.39 ng/L. The increased concentration of this isoflavone in aquatic

ecosystems can have short- and long-term effects on exposed organisms. Thus, the main objective of this work was to study

the effects caused by genistein on spontaneous behaviour and escape response in zebrafish (Danio rerio) larvae exposed

during embryogenesis.

Zebrafish embryos (3 hours post-fertilization) were divided into 3 replicates, each one composed with 30 embryos, and were

exposed for 96 hours to different concentrations of genistein: 20, 200 and 2000 μg/ml. The behavioural assessment of larvae

was performed at 120 hours and different parameters of spontaneous behaviour and escape response to visual stimulus were

evaluated.

No statistically differences were found in the different parameters of spontaneous behaviour, such as speed, total distance

covered, distance to the centre of the well, percentage of time in the upper/lower area of the well, absolute turn angle during

swimming and percentage of active/inactive time on zebrafish larvae exposed to different concentrations of genistein.

Furthermore, no significant effects of genistein on escape behaviour were observed in the red bouncing ball test (aversive

stimulus).

Most EDCs have a non-linear response and the effects observed on organisms differ between low and high concentrations,

which leads to a great concern with exposure to low and very low concentrations, as they can have several effects on the

endocrine metabolism of exposed organisms. Although the results obtained show that exposure to the selected concentrations

of genistein does not affect the behaviour of zebrafish larvae, these studies are important to study potential effect to individual

and population of wild fish.

Acknowledges: This work was supported by National Funds by FCT (Portuguese Foundation for Science and Technology),

under the project CITAB-UIDB/04033/2020 and the program VerãoComCiência summer course “Curso Prático-Aplicação de

modelos biológicos em Estudos de ecotoxicologia’’.

Keywords: Endocrine disrupting compounds, genistein, behaviour, zebrafish larvae, early life exposure


64

P36 - Risk assessment based on the mercury present in the edible tissues of the long-fin squid Loligo forbesi from

Azores archipelago

Vieira, H.C.1, Von Osten, J. R.2, Soares, A.M.V.M.1, Abreu, S. N.1, Morgado, F.1

1 CESAM—Centre for Environmental and Marine Studies, Department of Biology, University of Aveiro, 3810-193 Aveiro,

Portugal 2 EPOMEX Institute, Autonomous University of Campeche, Campeche, Mexico

[email protected]

Nowadays there is widespread recognition that fish and seafood consumption are beneficial to human health due to its high

nutritional value. Fish and seafood are considered an important source of essential nutrients such as omega-3 polyunsaturated

fatty acids (n-3 PUFAs), proteins, iodine, selenium, vitamin D, among other essential elements. Unfortunately, fish and seafood

are also considered as the major pathway of mercury (Hg) exposure in humans.

Due to its high toxicity, even in small amounts the consumption of certain marine species can pose a potential risk to

consumers' health, especially those belonging to vulnerable groups such as fetuses and newborns, since Hg can cross through

the placenta and crosses the blood-brain barrier and can have harmful effects, especially on the brain of the developing fetus.

The combination of growing commercial interest as a fishery resource, its intermediate position in marine food chains acting

as prey and predator, and the ability to accumulate and concentrate contaminants in tissues, makes cephalopods important

players in contaminants transfer along the marine food web.

That way, our study aims to i) assess the Hg concentration in different tissues (mantle and stomach), as well in the stomach

content of the squid Loligo forbesi caught in the Azores Archipelago, ii) evaluate the efficiency of Hg transfer from prey to

predator and iii) estimate the weekly tolerable Hg intake due to squid consumption.

Overall data indicate that stomach tissue (0.1±0.01 µg g-1) has a significantly higher Hg concentration than mantle (0.04±0.001

µg g-1) and stomach contents (0.01±0.001 µg g-1). Furthermore, the BMF (bioaccumulation factor) was > 1 for all samples,

indicating a biomagnification process from prey to predator.

Finally, considering the Hg concentration found in the mantle and the permitted Hg levels, it is advisable to consume up to

1050-1890g of squid per week, depending on the regulatory agency. Thus, our results indicate that, once these doses are

respected, consumption of squids from the Azorean waters do not pose a risk to humans.

Keywords Mercury; Azores archipelago; cephalopods; bioaccumulation factor; food safety


65

P37 - Impact of the co-occurrence of Penicillium sp. mycotoxins on human gastric cells toxicity risk for vulnerable

populations

Sousa Monteiro, C.a,d, Faria, MA.a, Pinto, E.b,c, Cunha, S.a

aLAQV-REQUIMTE, Laboratory of Bromatology and Hydrology, Faculty of Pharmacy, University of Porto, Porto, Portugal

bLaboratory of Microbiology, Biological Sciences Department, Faculty of Pharmacy, University of Porto, Porto, Portugal cInterdisciplinary Centre of Marine and Environmental Research, University of Porto, Matosinhos, Portugal

[email protected]

Introduction: Mycotoxins occurrence in food products is unpredictable and unavoidable worldwide, and responsible for an

economic burden due to financial losses at the international level (1). Moreover, human health could be at risk, especially in

vulnerable populations, such as the elderly, pregnant women, or nutritional deficit cases (2, 3). According to Rapid Alert System

for Food and Feed, mycotoxins are the main hazard cited in EU border rejection notifications. Additionally, the occurrence of

emergent mycotoxins increased the scientific community's concern regarding the possible toxicological interactions which led

to the European Commission request for monitoring recommendations (4). This study evaluated the toxicological interactions

of citrinin (CIT), cyclopiazonic acid (CPZ) and ochratoxin A (OTA) concerning additive, antagonistic and synergistic toxicity

towards human gastric cells.

Methodology: NCI-N87 cells were used as a gastric cell model. Cytotoxicity, of single, binary and ternary mixtures of CIT, CPZ

and OTA was assessed at the concentrations of 28.1-80.0, 0.625-40.0, 0.313-80.0μM, respectively, by measuring the

mitochondrial activity of viable cells using the MTT test. The theoretical Combination Index method by Chou-Talalay was

employed to evaluate the combined effects of these toxins and establish the type of interaction (5).

Results: Ochratoxin A was the mycotoxin with the highest toxic effect on cell viability, having the lowest IC50 value (10.1μM),

followed by CPZ (17.1μM) and CIT (68.8μM). Antagonistic effects occurred when exposing CIT-CPZ and CIT-OTA at all

concentrations, and CPZ-OTA in almost all inhibition levels. Based on binary mixtures, the mycotoxins combination can be

ranked according to their cytotoxicity: CPZ-OTA>CIT-OTA>CIT-CPZ. Regarding the ternary mixture a slight synergism/nearly

additive effect was detected.

Conclusions: The toxicological effect on human gastric cells can significantly differ between mycotoxins when they are exposed

alone or in combinations which is highly probable due to the unavoidable co-occurrence. Further investigation using diverse

human and animal models is needed to deeper understand the actual toxic effects fulfilling the gap of knowledge, namely in

the emergent mycotoxins. This would provide an important contribution to precise and realistic risk assessment, particularly

when considering vulnerable populations exposure.

Acknowledgements: This work was supported by FCT (Fundação para a Ciência e a Tecnologia) and FEDER through the

scholarships SFRH/BD/147313/2019 with financial support from FCT/MEC through national funds and co-financed by FEDER,

under the Partnership Agreement PT2020., FCT and FEDER. This research was partially supported by national funds through

FCT within the scope of UIDB/04423/2020 and AgriFood XXI R&D&I project, operation No. NORTE-01-0145-FEDER-000041,

co-financed by the European Regional Development Fund (ERDF) through NORTH 2020 (Northern Regional Operational

Program 2014/2020). Sara C. Cunha acknowledges FCT for IF/01616/2015 contract. Miguel A. Faria acknowledges FCT the

researcher contract.

Keywords: Mycotoxins; Combination index; Synergism; Antagonism; NCI-N87 cells


66

P38 - The impact of persistent organic pollutants burden in interventions to improve obesity-related metabolic

dysfunction – results from a dietary intervention study

Castela,I.1,2,3; Rodrigues,C.1,2; Ismael,S.1,3; Marques,C.1,3; Silvestre,M.P.1,3; Faria,A.1,2,3; Monteiro,R.4,5,6; Coutinho,S.R.7;

Martins,C.7,8; Fernandes,V.C.9; Domingues,V.9; Calhau,C.1,3; Pestana,D.1,3; Teixeira,D.1,2,3

1Nutrition and Metabolism, NOVA Medical School|Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Lisboa,

Portugal; 2Comprehensive Health Research Centre, Universidade Nova de Lisboa, Lisboa, Portugal; 3CINTESIS – Center for Health Technology and Services Research, Porto, Portugal; 4Departamento de Biomedicina - Unidade de Bioquímica, Faculdade de Medicina, Universidade do Porto, Porto, Portugal; 5Instituto de Investigação e Inovação em Saúde (I3S), Universidade do Porto, Porto, Portugal; 6Unidade de Saúde Familiar Pedras Rubras, Agrupamento de Centros de Saúde Maia-Valongo, Maia, Portugal; 7Obesity Research Group, Department of Cancer Research and Molecular Medicine, Faculty of Medicine, Norwegian

University of Science and Technology, Trondheim, Norway; 8Centre for Obesity, Department of Surgery, St. Olav Hospital,

Trondheim University Hospital, Trondheim, Norway; 9REQUIMTE/ LAQV – Instituto Superior de Engenharia, Instituto Politécnico do Porto, Porto, Portugal

[email protected]

Introduction: Because the toxicodynamic of persistent organic pollutants (POPs) relate directly to the dynamics of adiposity,

POPs might explain puzzling findings in obesity intervention clinical research. This study aimed to evaluate the clinical and

inflammatory characteristics according to the plasma concentration of POPs in obese adults who underwent a weight loss

intervention.

Methodology: Databases and biological samples from a clinical trial (NCT02169778) in Norwegian obese adults were used [1].

Plasma levels of twelve different POPs were determined by gas chromatography with electron-capture detection and

inflammatory cytokines were measured through flow cytometry. Participants were divided into two groups (low vs. high

exposure) according to the median of ∑POPs concentrations at baseline. The comparisons between groups were evaluated

by the Mann-Whitney test.

Results: Among 28 participants, 25 (89.3%) had at least one POP present in plasma at baseline. Median body weight and

waist circumference were significantly lower in participants with higher baseline plasma concentrations of POPs (104.6 vs.

96.0 kg, P=0.011 and 112.5 vs. 98.0 cm, P=0.002, respectively). They also had a lower median concentration of fasting insulin

and an improved β-cell function, with a tendency to have less insulin resistance. Finally, IL-1β, IFN-γ, and IL-6 at baseline were

higher in participants with higher POPs concentrations. After a diet-induced weight loss, the high exposure group continued to

present an imbalance in inflammatory profile with significantly lower levels of the anti-inflammatory cytokine IL-10 (30.8 vs 9.1

pg/mL, P=0.014) and tended to have lower body weight change, as compared to individuals less exposure to POPs.

Conclusions: Our results confirm the current evidence showing that plasma concentration of POPs is associate with

inflammation in obesity. This might reinforce the need to assess POPs before weight loss interventions and defining a dietary

intervention also based on it. Instead of only focus reducing caloric intake, promoting food quality by privileging foods with

bioactive compounds that can offset the negative effects of POPs, might be a more accurate strategy in managing obesity.

Relevance: Evaluating POPs before weight loss interventions can be a promising opportunity to define more effective dietary

strategies, enhancing the metabolic benefits expected from it.

Acknowledgements: This abstract was supported by ERDF through the operation POCI-01-0145-ERDF-007746 funded by the

Programa Operacional Competitividade e Internacionalização – COMPETE2020 and by National Funds through FCT -

Fundação para a Ciência e a Tecnologia within CINTESIS, R&D Unit (reference UID/IC/4255/2013) and CHRC

(UIDP/04923/2020).

Conflict of interests: The authors declare no competing interests.

Keywords: Inflammation, obesity, persistent organic pollutants, weight loss


67

P39 - Environmental pollution as a concerning source of dietary toxics for pregnant women, infants and children

Ribeiro O.1, Ribeiro C.2,3 Gonçalves. C.1, 4, 5 and Carrola J. S.1,5

1 School of Life and Environmental Sciences (ECVA), University of Trás-os-Montes and Alto Douro, (UTAD), Vila Real, Portugal 2 Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Edifício do Terminal de Cruzeiros do

Porto de Leixões, Matosinhos, Portugal 3 TOXRUN – Toxicology Research Unit, University Institute of Health Sciences, CESPU, CRL, 4585-116 Gandra, Portugal. 4 CIAFEL – The Research Centre in Physical Activity, Health and Leisure, University of Porto, 4200-450, Porto, Portugal 5 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB), Vila Real, Portugal

[email protected]

Despite European legislation, low levels of environmental pollutants are still present and can pose risk to ecological health,

and to human beings via food exposure. Pesticides and synthetic fertilizers, veterinary and human pharmaceuticals, corrective

biosolids and sludge application, reuse of wastewater, industrial compounds, among others can reach air, irrigation water,

agricultural soils and later end up in drinking water and foods (vegetable, fruits, dairy products, meat, and fisheries) contributing

to total food contaminants which can be increased, later, by toxicants produced during food processing, packaging, and

cooking.

Chronic low-dose exposure to persistent and nonpersistent pollutants through food is considered a risk to human health,

especially for the developing fetus, and early postnatal development, due to its rapid growth and incomplete protective

mechanism. Persistent organic pollutants, POPs (organochlorine compounds and polybrominated diphenyl ethers) are

connected to restricted fetal growth, immunotoxicity, neurotoxicity, reduced fertility, and obesogenic effects. Polychlorinated

biphenyls (PCBs) are linked to chloracne, immune and carcinogenic effects. Toxic metals (mercury, cadmium, arsenic and

lead) can disturb child’s cognitive, motor, and behavioral development. Polycyclic aromatic hydrocarbons (PAHs) in foods

appear to be genotoxic and carcinogenic. Furthermore, some toxicants like pharmaceutical and personal care products

(PPCPs), dioxins/furans (PCDD/Fs), and plasticizers can cause endocrine disruption and transgenerational effects rising the

long-term effects and concerns.

Although some contaminants have focal effects, many are transported globally, and international measure should be

implemented to ensure trade of safer food products, including education for safer crop cultivation and livestock rearing.

However, is impossible to eradicate completely food toxics thus during sensitive windows of development is imperative to

reduce contaminated foods ingestion, like marine (some fish and seafoods), terrestrial animal origin foods, non-organic meat,

canned foods, fast foods and foods stored in plastic packaging. Also, the knowledge of toxicokinetic, categories and levels of

contaminants in foods play a pivotal role in toxicity assessment to reduce cumulative disease risk assessment and chronic

health problems diet-related. Besides, balance and healthy diet advices are important to enable more practicable consumers

daily food choices and food consumption rates to reduce the exposure and health risk, mainly for pregnant women and children.

Keywords: environmental pollution, food toxicants, pregnant women, children


68

P40 - Non-environmental dietary toxics exposure of vulnerable groups - pregnant women and children

Liz Martins M.1,2 Ferreira R1, Peixoto N1., Cruz A.R1 and Carrola J.S.1,2

1 Escola das Ciências da Vida e do Ambiente (ECVA), Universidade de Trás-os-Montes e Alto Douro, (UTAD), Vila Real,

Portugal 2 Centro de Investigação e Tecnologias Agroambientais e Biológicas (CITAB), UTAD, Vila Real, Portugal

Human diet contributes to natural toxins and to a considerable part of toxics input, and frequent consumption of certain foodstuff

is a health risk a concern. Pregnant women, infants and children are daily exposed to cocktail of toxics and can be affected by

cumulative exposure sources, including diet toxics, that can give rise to subtle and long-term effects. Particularly, early life

exposure to toxic plays a critical part in the development of chronic-related diseases, not only for offspring but also their next

generations. Food toxics from non-environmental sources are added during industrial food processing, packaging and cooking.

During traditional direct smoking carcinogenic compounds are usually formed, namely polycyclic aromatic hydrocarbons

(PAHs) like the carcinogenic benzo-a-pyrene (BaP).

Moreover, children consume around 2-3 more food packaged in plastic than adults, presenting a higher exposure to substances

leaching from plastic food contact. Bisphenol (BPA) can be release from plastic or canned foods – tinned food linings. BPA is

an endocrine disrupter causing several health problems even at very low levels (impaired brain development, postnatal

changes in DNA and cancer). Additionally, fast food use thick paper packaging contains per- and polyfluoroalkyl substances

(PFAS) linked immune system suppression. Commercial food contact materials (FCMs) and microwavable plastic food

containers (MPFCs) are extensively and subsequently migration of toxic compounds and microplastics (MPs) can occurs.

Cooking process, mainly at high temperatures produce toxic compounds, like foods glycation end products linked with

premature aging and cancer. Other toxic compounds are formed, mainly polycyclic aromatic hydrocarbons (PAHs),

heterocycles amines (HA) and AA. Young people are more exposed through fast food and snacks. The last one is probable

human carcinogen found in bread, potato crisps and French fried, biscuits, and baby foods. Grilled processing at high

temperatures (barbecued very well-done) could lead to production of HCAs and PAHs

It’s imperative to optimize processing, packaging methodologies and cooking process to reduce toxicants in foodstuff and

reduce consumption of specific foods to reduce exposure. Guidelines for consumers, mainly pregnant women and parents,

presenting practical daily life advices could be useful.

Keywords: non-environmental toxics, diet toxicants, pregnant women, children


69

P41 - Aflatoxin B1 and early life gut microbiota: preliminary results under earlyMYCO project

Inês Silva1, Elsa L. Duarte1,2*, Patrícia Bastos-Amador3, Manuela Ferreira3,4, Paula Alvito 5,6, Ricardo

Assunção5,6 , Cátia Salvador7 & Ana Teresa Caldeira 1,7,8

1Escola de Ciências e Tecnologia, Universidade de Évora, Évora, Portugal 2MED-Mediterranean Institute for Agriculture, Environment and Development, Évora, Portugal 3Champalimaud Centre for the Unknown, Lisboa, Portugal 4 Centre for Neuroscience and Cell Biology, University of Coimbra, UC Biotech, Biocant Park, Cantanhede, Portugal 5 Food and Nutrition Department, National Institute of Health Dr. Ricardo Jorge, Lisboa, Portugal 6 CESAM - Centre for Environmental and Marine Studies, Department of Biology, University of Aveiro, Aveiro, Portugal & Food and Nutrition Department, National Institute of Health Dr. Ricardo Jorge, Lisbon, Portugal. 7 Hercules Laboratory, Universidade de Évora, Portugal 8City U Macau Chair in Sustainable Heritage, Universidade de Évora, Portugal [email protected]

Aflatoxin B1 (AFB1) produces acute or chronic deleterious health effects in humans and animals. Still, long-term effects derived from initial exposure in early-life, a critical period for colonization and development of gut microbiota, has not been fully evaluated, . particularly, effects on gut microbiota and immunity system. This study, performed under the earlyMYCO project, investigated the impact of maternal exposure to AFB1 on early-life microbiota in a mouse model. Females were fed jelly pellets containing 400 µg/kg AFB1 diluted in DMSO (treated animals n=6) or DMSO vehicle alone (control group n=6) during pregnancy and lactation. Faeces from the offspring were collected immediately after weaning and faecal DNA was extracted and purified. Bacterial taxa diversity and relative abundance were assessed by High-Throughput Sequencing performed in an Illumina Miseq® sequencer, targeting the V3 and V4 regions of the 16S rRNA gene. Operational taxonomic units (OTUs) were determined by clustering reads to 16S reference databases. A hundred and twenty-four (N=124) bacterial genera were found in both groups, 5 were only present in AFB1 treated group and 27 exclusively in control groups. A hundred and fifty-one (N=151) bacterial species were common to both groups, 15 species exclusively found in AFB1 litters and 34 species were exclusively found in control litters. Although present in both groups, Akkermansia muciniphila and Bacteroides acidifaciens were significantly higher in controls. A. muciniphila colonizes the intestinal tract in childhood and regulates mucus thickness, intestinal barrier integrity and is involved in immune modulation. B. acidifaciens participates in the metabolism of lipids and sugars and activates some cytokines and immune cell receptors. Sulfidogenic bacteria, recently related to inflammatory bowel disease, such as Desulfovibrio piger and Bilophila wadsworthia were exclusivly found in the treated litters. Early-life gut microbiome triggers the gut immune defences, but is far less stable than the adult microbiome. These preliminary results open an extensive field to further investigate the association between mycotoxins and microbiome, as the latest is increasingly recognized as a major player in a wide spectrum of diseases. This work was funded by FCT/MCTES through national funds, to earlyMYCO (PTDC/MED-TOX/28762/2017), and CESAM

(UIDP/50017/2020+UIDB/50017/2020).

Keywords: AFB1, early-life exposure, microbiota

icfc 2021 book of abstracts

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