Infection by Campylobacter is the most notified gastrointestinal disease in New Zealand. Reliable recovery and identification of campylobacters is challenging. Improved and validated methods are needed to increase the power of subtyping and epidemiological studies to trace the sources and transmission routes of Campylobacter. An enrichment-PCR method for the isolation and detection of C. jejuni and C. coli was developed and sensitivity levels determined in 13 environmental matrices, including animal faeces, food and water. Less than ten cells per sample of either C. jejuni or C. coli could be detected, except for rabbit faeces where the minimum number of cells detected per sample was greater than ten cells for C. coli (range 3-32 cells). The sensitivity of the method was comparable to that determined for the conventional methods in the same matrices. Application of the method to retail chicken carcasses (n =204) determined a prevalence of 27.5% C. jejuni and 1% C. coli. River water assays (n = 293) found 55.3% of samples to contain C. jejuni and 4.1% C. coli. Furthermore, the enrichment-PCR assay was shown to identify up to three subtypes in individual water samples. It was proposed that the identification of non-dominant subtypes carried by a chicken carcass may aid the identification of subtypes implicated in human cases of campylobacteriosis. An average of twenty-three C. jejuni isolates from each of ten retail chicken carcass were subtyped by PFGE using the two restriction enzymes SmaI and KpnI. Fifteen subtypes, in total, were identified from the ten carcasses. One subtype was identified on three carcasses. Five carcasses carried a single subtype, three carcasses carried two subtypes each and two carcasses carried three subtypes each. Some of the subtypes carried by an individual carcass were shown to be clonally related raising the question of in vivo recombination events during host passage. Comparison of C. jejuni subtypes from chickens with those isolated from human clinical cases revealed three of the fifteen subtypes correlated with those from human cases. None of the minority subtypes were identified in human case isolate data, suggesting that the lack of identification of non-dominant subtypes from chicken carcasses may not hinder the investigation of campylobacteriosis outbreaks.

Infection by Campylobacter is the most notified gastrointestinal disease in New Zealand. Reliable recovery and identification of campylobacters is challenging. Improved and validated methods are needed to increase the power of subtyping and epidemiological studies to trace the sources and transmission routes of Campylobacter. An enrichment-PCR method for the isolation and detection of C. jejuni and C. coli was developed and sensitivity levels determined in 13 environmental matrices, including animal faeces, food and water. Less than ten cells per sample of either C. jejuni or C. coli could be detected, except for rabbit faeces where the minimum number of cells detected per sample was greater than ten cells for C. coli (range 3-32 cells). The sensitivity of the method was comparable to that determined for the conventional methods in the same matrices. Application of the method to retail chicken carcasses (n =204) determined a prevalence of 27.5% C. jejuni and 1% C. coli. River water assays (n = 293) found 55.3% of samples to contain C. jejuni and 4.1% C. coli. Furthermore, the enrichment-PCR assay was shown to identify up to three subtypes in individual water samples. It was proposed that the identification of non-dominant subtypes carried by a chicken carcass may aid the identification of subtypes implicated in human cases of campylobacteriosis. An average of twenty-three C. jejuni isolates from each of ten retail chicken carcass were subtyped by PFGE using the two restriction enzymes SmaI and KpnI. Fifteen subtypes, in total, were identified from the ten carcasses. One subtype was identified on three carcasses. Five carcasses carried a single subtype, three carcasses carried two subtypes each and two carcasses carried three subtypes each. Some of the subtypes carried by an individual carcass were shown to be clonally related raising the question of in vivo recombination events during host passage. Comparison of C. jejuni subtypes from chickens with those isolated from human clinical cases revealed three of the fifteen subtypes correlated with those from human cases. None of the minority subtypes were identified in human case isolate data, suggesting that the lack of identification of non-dominant subtypes from chicken carcasses may not hinder the investigation of campylobacteriosis outbreaks.

<p>Prisustvo organskih zagađujućih supstanci (farmaceutski aktivnih komponenata i prirodnih toksina‐mikotoksina) u uzorcima životne sredine i namirnicama je u porastu kao posledica novih industrijskih procesa i ostalih antropogenih aktivnosti, kao i klimatskih promena. Takođe veliku pažnju javnosti privlače i neorganske zagađujuće supstance kao &scaron;to su te&scaron;ki elementi. S obzirom da zagađujuće supstance imaju negativan uticaj na životnu sredinu i zdravlje ljudi, u svetu se preduzimaju mere u cilju smanjenja stepena izloženosti toksičnim jedinjenjma i posledicama izlaganja. Trenutno, jedan od najvećih izazova, jeste procena rizika povezana sa velikim brojem zagađujućih supstanci u tragovima ili u tzv. ultratragovima, uključujući &ldquo;novo&rdquo; otkrivena zagađujuća jedinjenja, a jedan od osnovnih trendova je razvoj i primena brzih i efikasnih metoda za njihovu analizu u ispitivanim uzorcima na bazi naprednih hromatografskih i spektrometrijskih tehnika.<br />Tehnike bazirane na tečnoj hromatografiji sa različitim masenim analizatorima za<br />kvantifikaciju organskih zagađujućih supstanci kao i metode zasnovane na atomskoj<br />apsorpcijonoj spektrometriji za određivanje ultratragova neorganskih zagađujućih supstanci postale su referentne na međunarodnom nivou. Ovakve napredne spregnute tehnike postale su važne za identifikaciju, kvantifikaciju i praćenje različitih zagađujućih supstanci u uzorcima životne sredine i namirnicama i proceni njihovog &scaron;tetenog uticaja na zdravlje čoveka. S obzirom da su u literaturi retka istraživanja koja se bave razvojem i primenom metoda zasnovanih na naprednim hromatografskim i spektrometrijskim tehnikama i određivanju organskih i neorganskih zagađujućih supstanci u matriksima životne sredine i namirnicama sa prostora zapadnog Balkana, a uzimajući u obzir njihovu važnost, specifični ciljevi disertacije su:<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐rezidualne&ldquo;<br />metode zasnovane na UHPLC‐QqLIT‐MS/MS za analizu 81‐e farmaceutski aktivne<br />komponente (PhAC) u otpadnoj, povr&scaron;inskoj, podzemnoj i pijaćoj vodi i po prvi put<br />dobijanje sveobuhvatnih rezultata njihovog prisustva u različitim tipovima vode sa<br />područja Srbije;<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;) provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐toksin&ldquo;<br />metode za analizu 8 Fusarium mikotoksina u uzorcima ozime p&scaron;enice različitih sorti<br />zasnovane na HPLC‐QqQ‐MS/MS radi određivanja regionalnih razlika između žitnih<br />regiona kao i otpornosti ispitivanih sorti p&scaron;enice na kontaminaciju Fusarium<br />toksinima;<br />&bull; modifikacija postojeće &bdquo;multi‐toksin&ldquo; metode zasnovane na UHPLC‐QqQ‐MS/MS za<br />analizu 11 osnovnih mikotoksina u uzorcima bra&scaron;na i njena unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti, kao i provera kroz interlaboratorijsko poređenje,<br />radi dobijanja podataka za procenu &scaron;tetnog uticaja ispitivanih mikotoksina na<br />zdravlje populacije;<br />&bull; razvoj &bdquo;multi‐toksin&ldquo; (vi&scaron;ekomponentne) i &bdquo;multi‐matriks&ldquo; (za vi&scaron;e matriksa) metode<br />bazirane na UHPLC‐QqQ‐MS/MS za analizu 10 mikotoksina u različitim vrstama<br />ko&scaron;tuničavog voća čija provera kvaliteta je zasnovana na intralaboratorijskoj proveri<br />tačnosti i preciznosti dobijenih rezultata;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &bull; primena postojeće analitičke procedure zasnovane na naprednoj tehnici pripreme<br />(mikrotalasnoj digestiji) različitih uzoraka biljnog i životinjskog porekla i provera<br />kvaliteta metode identifikacije i kvantifikacije zasnovane na atomskom apsorpcionom spektrometru sa grafitnom kivetom (GFAAS) radi dobijanja sveobuhavatnih rezultata o prisustvu te&scaron;kih elemenata (arsena, olova i kadmijuma) radi procene izloženosti stanovni&scaron;tva Srbije toksičnim neorganskim elementima.<br />Postignuti rezultati predstavljaju jedinstvene rezultate za područje Srbije dobijene<br />primenom naprednih spregnutih tehnika koje imaju značajnu ulogu u praćenju prisustva većeg broja organskih i neorganskih zagađujućih supstanci u izabranim uzorcima životne sredine i namirnica, (regulisanih postojećim zakonodavstvom) radi procene stepena zagađenosti ili u slučaju jedinjenja koja nisu regulisana zakonodavstvom radi sticanja novih saznanja o njihovom prisustvu i proceni mogućeg negativnog uticaja na životnu sredinu i zdravlje populacije.</p> / <p>The presence of organic pollutants in environmental samples and food (pharmaceutically active components and natural toxins‐mycotoxins) is increased as a result of new industrial processes and other anthropogenic activities, as well as climate change. Similarly heavy elements as inorganic pollutants have attracted worldwide attention. Since, these pollutants have negative impact on environment and human health, extremely efforts are undertaken in the world to reduce the level of exposure to these pollutants and consequences of the exposure. Currently, one of the highest challenges is to assess the risk associated with a large number of pollutants in trace or ultra trace levels, including &quot;new&quot; (emerging) discovered pollutants, and one of the main trends is development and implementation of fast and efficient methods for their analysis on the basis of advanced chromatographic and spectrometric techniques. Therefore, coupled techniques have become important for the identification, quantification and monitoring of various pollutants in environmental samples and food and assessment of their hazard impact on human health. Since, there are scarce data about the development and application of advanced methods based on chromatographic and spectrometric techniques for determination of organic and inorganic pollutants in environmental matrices and food from the Western Balkan, and taking into account their importance, specific objectives of<br />the dissertation were: &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐residual&quot; method based on UHPLC‐QqLIT‐MS/MS for analysis of 81 pharmaceutically active components (PhAC) in wastewater, surface, underground and drinking water due to obtained for the first time comprehensive results of their presence in different types of water from Serbia; &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐toxin&quot; method for the analysis of 8 Fusarium mycotoxins in samples of different winter wheat cultivars based on HPLC‐QqQ‐MS/MS to determine the differences among wheat-growing regions as well as the resistance of the analysed wheat cultivars towards Fusarium toxins; &bull; modification of existing &quot;multi‐toxin&quot; method based on UHPLC‐QqQ‐MS/MS for analysis of 11 principal mycotoxins in samples of flour and its internal (&quot;in‐house&quot;) quality control as well as verification through the interlaboratory comparison, in order to obtain data for assessing the hazard effect of these mycotoxins on the health of the population; &bull; the development of &quot;multi‐toxin&quot; and &quot;multi‐matrix&quot; method based on UHPLC‐QqQMS/ MS for the analysis of 10 mycotoxins in various types of nuts based on intralaboratory verification of the accuracy and precision of the obtained results; &bull; application of analytical procedure based on advanced preparation technique (microwave digestion) and atomic absorption spectrometer with a graphite furnace (GFAAS) and its verification in order to obtain comprehensive results on the presence of heavy elements (arsenic, lead and cadmium) in different samples of plant and animal origin to assess the exposure of the Serbian population to toxic inorganic elements. The obtained results are unique for the Serbia. They are obtained by applying advanced coupled techniques that have a significant role in monitoring the presence of a numerous organic and inorganic pollutants in analyzed samples of the environment and food. The presented results contribute to the assessment of pollution degree and in the case of new (emerging) not regulated polutant they might give new information about the possible negative impact on the environment and health of the population.</p>

O aleitamento materno é um instrumento afetuoso que transfere nutrientes, vitaminas e imunidade da mãe para o filho. No entanto, também pode transferir diversos contaminantes ambientais, destacando os metais pesados. Como parte da dieta, as mães ingerem água e alimentos produzidos em suas localidades, podendo através dessas fontes ficarem expostas a metais pesados. Alguns metais pesados são essenciais para o organismo, mas outros de origem antropogênica são deletérios. Os objetivos do trabalho em Conceição das Alagoas, MG, Brasil, foram: avaliar as concentrações dos metais pesados no colostro, leite de transição e leite maduro das lactantes; avaliar as concentrações dos metais pesados na água de abastecimento para essa população; medir e avaliar as concentrações de metais presentes no solo agrícola e hortas do município; verificar se existe uma relação entre as concentrações de metais pesados (Cd, Cr, Cu, Hg, Mn, Ni, Pb, Sn, Zn) encontrados no leite materno, na água para consumo, bem como no solo local. Foram coletadas 29 amostras de leite de doadoras. A coleta do colostro ocorreu até 7 dias depois do parto, a coleta do leite de transição ocorreu até o 14º dia e a coleta do leite maduro ocorreu por volta do 28º dia. Foram coletadas 44 amostras de água de residências das lactantes e em pontos diferentes e aleatórios da cidade. As amostras do solo foram coletadas em 23 pontos, entre áreas agrícolas, hortas e a margem do rio Uberaba. Os metais pesados no leite e água foram analisados por ICP-MS e no solo por FRX. As concentrações de metais no leite e na água foram calculadas em ?g/L e expressos pelas medianas. Para o solo, as concentrações foram calculadas em mg/Kg, e também foram expressos pelas medianas. Os perfis de concentração foram analisados através do teste de hipótese de uma população multinomial, utilizando uma distribuição qui-quadrado (x2) (p a=0,05). Os valores de todos os metais pesados no leite se mantêm constantes entre as 3 coletas, exceto o Zn e o Cu que apresentam suas concentrações menores no leite maduro. As concentrações de metais pesados na água se mostraram menores que os recomendados pela OMS e pelo Ministério da Saúde/Brasil, exceto para o cádmio, que se mostrou acima desses níveis, fato que pode ser explicado pela presença geogênica ou pelo uso de alguns defensivos agrícolas na região. Há um perfil semelhante quando são comparadas as concentrações em porcentagem de metais pesados encontrados no leite humano e na água consumida pelas mães (?2=14,36). As concentrações em porcentagem de metais no leite humano e no solo mostraram diferentes perfis (?2=635,05). As concentrações em porcentagem de metais no solo e na água também apresentaram perfis diferentes (?2=721,78). Baseado nestes dados, a análise estatística evidencia uma semelhança entre o perfil da distribuição de metais pesados no leite materno e na água de abastecimento, assinalando que alterações nas concentrações de metais pesados na água podem interferir nas concentrações no leite humano. Esta relação não foi estabelecida entre a distribuição dos metais do solo e do leite humano. As relações encontradas nas matrizes estudadas apresentam relevância, considerando que podem apresentar parâmetros das interações humanas e ambientais. Estudos mais abrangentes devem ser realizados para confirmação da associação entre o perfil das concentrações de metais em água potável e leite materno, visando a confirmação de um indicador ambiental de contaminação por metais que pode ser utilizado em bancos de leite humano. / Breastfeeding is an affective instrument that transfers nutrients, vitamins and immunity from mother to child. However, it also represents a way to transfer environmental contaminants, especially heavy metals. As part of the diet of mothers, they drink water and eat food grown in local gardens, and may be exposed through these sources to heavy metals. Some heavy metals are essential for the organism, but others, from anthropogenic source, are deleterious. The objectives of this research in Conceição das Alagoas, Minas Gerais, Brazil, were: to assess the concentrations of heavy metals in colostrum, transitional milk and mature milk in lactating women, to evaluate the concentrations of heavy metals in the water supply for this population; measure and evaluate the concentrations of metals present in agricultural land and gardens in the city; verify if there is a relationship between the concentrations of heavy metals (Cd, Cr, Cu, Hg, Mn, Ni, Pb, Sn, Zn) found in breast milk, in the water supply and in local soil. There were collected 29 samples from donors. The collection of colostrum occurred until 7 days after delivery, the collection of the transition milk occurred up to 14 days and mature milk collection occurred around day 28. It was collected 44 water samples from homes of the mothers and from different random points in the city. Soil samples were collected at 23 points, including farmland, gardens and the margin of Uberaba river. Heavy metals in milk and water were analyzed by ICP-MS in soil by XRF. The concentrations of metals in milk and water were calculated in ?g/L and expressed by medians. For soil, the concentrations were calculated in mg/kg, and were also expressed by medians. The concentration profiles were analyzed by testing the hypothesis of a multinomial population, using a Chi-square (?2) (p a = 0.05). The amounts of all heavy metals in milk remains constant among the three samples, except Zn and Cu which have their lower concentrations in mature milk. The concentrations of heavy metals in water were lower than those recommended by WHO and the Ministry of Health/Brazil, except for Cd, which was above those levels, which may be explained by the geogenic presence or the use of some pesticides in the region. There is a similar profile when comparing the concentrations in percentage of heavy metals found in human milk and water consumed by mothers (?2=14.36). The concentrations in percentage of metals in human milk and in soil showed different profiles (?2=635.05). The concentrations in percentage of metals in soil and water profiles were also different (?2=721.78). Based on these data, the statistical analyses showed a clear similarity between the profile of heavy metal distribution in breast milk and the water supplied, showing that changes in the concentrations of heavy metals in water can affect the concentrations in human milk. This relationship was not established between the profile of heavy metals from soil and human milk. The relations found in the matrices studied are relevant, considering parameters that may present environmental and human interactions. More studies should be performed to confirm the association between the profile of metal concentrations in drinking water and breast milk, seeking confirmation of an environmental indicator of metal contamination that can be used in human milk banks.

O aleitamento materno é um instrumento afetuoso que transfere nutrientes, vitaminas e imunidade da mãe para o filho. No entanto, também pode transferir diversos contaminantes ambientais, destacando os metais pesados. Como parte da dieta, as mães ingerem água e alimentos produzidos em suas localidades, podendo através dessas fontes ficarem expostas a metais pesados. Alguns metais pesados são essenciais para o organismo, mas outros de origem antropogênica são deletérios. Os objetivos do trabalho em Conceição das Alagoas, MG, Brasil, foram: avaliar as concentrações dos metais pesados no colostro, leite de transição e leite maduro das lactantes; avaliar as concentrações dos metais pesados na água de abastecimento para essa população; medir e avaliar as concentrações de metais presentes no solo agrícola e hortas do município; verificar se existe uma relação entre as concentrações de metais pesados (Cd, Cr, Cu, Hg, Mn, Ni, Pb, Sn, Zn) encontrados no leite materno, na água para consumo, bem como no solo local. Foram coletadas 29 amostras de leite de doadoras. A coleta do colostro ocorreu até 7 dias depois do parto, a coleta do leite de transição ocorreu até o 14º dia e a coleta do leite maduro ocorreu por volta do 28º dia. Foram coletadas 44 amostras de água de residências das lactantes e em pontos diferentes e aleatórios da cidade. As amostras do solo foram coletadas em 23 pontos, entre áreas agrícolas, hortas e a margem do rio Uberaba. Os metais pesados no leite e água foram analisados por ICP-MS e no solo por FRX. As concentrações de metais no leite e na água foram calculadas em ?g/L e expressos pelas medianas. Para o solo, as concentrações foram calculadas em mg/Kg, e também foram expressos pelas medianas. Os perfis de concentração foram analisados através do teste de hipótese de uma população multinomial, utilizando uma distribuição qui-quadrado (x2) (p a=0,05). Os valores de todos os metais pesados no leite se mantêm constantes entre as 3 coletas, exceto o Zn e o Cu que apresentam suas concentrações menores no leite maduro. As concentrações de metais pesados na água se mostraram menores que os recomendados pela OMS e pelo Ministério da Saúde/Brasil, exceto para o cádmio, que se mostrou acima desses níveis, fato que pode ser explicado pela presença geogênica ou pelo uso de alguns defensivos agrícolas na região. Há um perfil semelhante quando são comparadas as concentrações em porcentagem de metais pesados encontrados no leite humano e na água consumida pelas mães (?2=14,36). As concentrações em porcentagem de metais no leite humano e no solo mostraram diferentes perfis (?2=635,05). As concentrações em porcentagem de metais no solo e na água também apresentaram perfis diferentes (?2=721,78). Baseado nestes dados, a análise estatística evidencia uma semelhança entre o perfil da distribuição de metais pesados no leite materno e na água de abastecimento, assinalando que alterações nas concentrações de metais pesados na água podem interferir nas concentrações no leite humano. Esta relação não foi estabelecida entre a distribuição dos metais do solo e do leite humano. As relações encontradas nas matrizes estudadas apresentam relevância, considerando que podem apresentar parâmetros das interações humanas e ambientais. Estudos mais abrangentes devem ser realizados para confirmação da associação entre o perfil das concentrações de metais em água potável e leite materno, visando a confirmação de um indicador ambiental de contaminação por metais que pode ser utilizado em bancos de leite humano. / Breastfeeding is an affective instrument that transfers nutrients, vitamins and immunity from mother to child. However, it also represents a way to transfer environmental contaminants, especially heavy metals. As part of the diet of mothers, they drink water and eat food grown in local gardens, and may be exposed through these sources to heavy metals. Some heavy metals are essential for the organism, but others, from anthropogenic source, are deleterious. The objectives of this research in Conceição das Alagoas, Minas Gerais, Brazil, were: to assess the concentrations of heavy metals in colostrum, transitional milk and mature milk in lactating women, to evaluate the concentrations of heavy metals in the water supply for this population; measure and evaluate the concentrations of metals present in agricultural land and gardens in the city; verify if there is a relationship between the concentrations of heavy metals (Cd, Cr, Cu, Hg, Mn, Ni, Pb, Sn, Zn) found in breast milk, in the water supply and in local soil. There were collected 29 samples from donors. The collection of colostrum occurred until 7 days after delivery, the collection of the transition milk occurred up to 14 days and mature milk collection occurred around day 28. It was collected 44 water samples from homes of the mothers and from different random points in the city. Soil samples were collected at 23 points, including farmland, gardens and the margin of Uberaba river. Heavy metals in milk and water were analyzed by ICP-MS in soil by XRF. The concentrations of metals in milk and water were calculated in ?g/L and expressed by medians. For soil, the concentrations were calculated in mg/kg, and were also expressed by medians. The concentration profiles were analyzed by testing the hypothesis of a multinomial population, using a Chi-square (?2) (p a = 0.05). The amounts of all heavy metals in milk remains constant among the three samples, except Zn and Cu which have their lower concentrations in mature milk. The concentrations of heavy metals in water were lower than those recommended by WHO and the Ministry of Health/Brazil, except for Cd, which was above those levels, which may be explained by the geogenic presence or the use of some pesticides in the region. There is a similar profile when comparing the concentrations in percentage of heavy metals found in human milk and water consumed by mothers (?2=14.36). The concentrations in percentage of metals in human milk and in soil showed different profiles (?2=635.05). The concentrations in percentage of metals in soil and water profiles were also different (?2=721.78). Based on these data, the statistical analyses showed a clear similarity between the profile of heavy metal distribution in breast milk and the water supplied, showing that changes in the concentrations of heavy metals in water can affect the concentrations in human milk. This relationship was not established between the profile of heavy metals from soil and human milk. The relations found in the matrices studied are relevant, considering parameters that may present environmental and human interactions. More studies should be performed to confirm the association between the profile of metal concentrations in drinking water and breast milk, seeking confirmation of an environmental indicator of metal contamination that can be used in human milk banks.

Depuis plusieurs décennies la production mondiale de plastiques ne cesse d’augmenter, menant à une contamination des écosystèmes aquatiques à l’échelle de la planète qui a été récemment estimée à plus de cinq mille milliards de débris de plastiques flottants à la surface des océans. Les microplastiques (particules de plastique < 5 mm), introduits dans l’environnement aquatique directement en tant que microparticules (granulés plastiques industriels, cosmétiques, fibres textiles) ou lors de la fragmentation de plus gros débris plastiques, représentent une préoccupation scientifique et sociétale grandissante. Ces travaux de thèse se sont focalisés sur la contamination par les microplastiques de la rade de Brest (Bretagne, France), un système côtier macrotidal où l’activité anthropique est importante. Les objectifs de ces travaux sont (1) d’évaluer la contamination des matrices environnementales (eau de surface, sédiment subtidal et biote) par les microplastiques, et (2) d’identifier leur rôle potentiel en tant que vecteurs de contaminants chimiques et de bactéries dans la rade de Brest.Pour cela, des développements analytiques ont été nécessaires afin d’améliorer leur extraction des matrices environnementales ainsi que leur caractérisation morphologique et chimique via la microspectrométrie Raman. Les observations in situ ont montré que l’ensemble de l’écosystème de la rade de Brest est contaminé par les microplastiques, avec des concentrations de 0,24 ± 0,35, et 0,97 ± 2,08 (moyenne ± écart-type). L’eau de surface et le sédiment sont contaminés par le polyéthylène, le polypropylène et le polystyrène. Leur distribution à la surface de l’eau semble être liée à l’urbanisation très présente au nord de la rade, mais également à l’hydrodynamisme de cette région marine. Les premiers résultats concernant les bivalves marins ont montré un niveau relativement faible de contamination par les microplastiques (0,01 ± 0,04 et 0,08 ± 0,34 pour les moules et les coques, respectivement), cependant cela est probablement dû aux choix méthodologiques appliqués ici (exclusion des fibres). Les observations in situ ont montré que certains polluants organiques (HAP, PCB et pesticides) étaient détectés sur les microplastiques flottants à des valeurs (non détecté – 49763 ng.g-1, moyenne ± écart-type) similaires de celles retrouvées dans les sédiments et les bivalves locaux, ce qui suggère un risque faible dans le transfert des contaminants chimiques vers les organismes marins en cas d’ingestion. Enfin, les résultats concernant la colonisation bactérienne des microplastiques flottants ont montré des communautés distinctes de celles retrouvées dans l’eau de mer environnante, et l’identification du genre Vibrio en tant que biomarqueur discriminant de cette matrice. Dans l’ensemble, ces travaux fournissent une première évaluation approfondie de l’état de contamination de la rade de Brest par les microplastiques ainsi que de solides recommandations méthodologiques pour des travaux futurs. / World production of plastics has increased steadily for the past decades leading to a major contamination of the worldwide aquatic ecosystems recently estimated at more than five trillion plastic pieces floating the surface of the oceans. Microplastics (plastic particles < 5 mm) are introduced into aquatic environments directly as industrial raw material (plastic pellets, cosmetics, clothing) or indirectly via the fragmentation of larger plastics. This emerging contaminant represents an increasing ecological concern for science and society. The present study focused on the microplastic contamination of the Bay of Brest (Brittany, France), a macrotidal coastal ecosystem characterized by intense anthropogenic activity. The main objectives were: (1) to evaluate the contamination of environmental matrices (surface water, subtidal sediment and biota) by microplastics, and (2) to identify their potential role as vector of chemicals and bacteria in the bay of Brest.Methodological developments were first conducted to improve microplastic extraction from environmental matrices as well as their rapid morphological and chemical identification by Raman micro-spectrometry. The field investigations showed that the ecosystem of the bay of Brest is contaminated by microplastics with mean concentrations of 0.24 ± 0.35, and 0.97 ± 2.08 (mean ± standard deviation) in surface water and sediment, respectively. Microplastic contamination in surface water and sediment was dominated by polyethylene, polypropylene and polystyrene microparticles.Spatial microplastic distribution appeared to be related to proximity to urbanized areas and to hydrodynamic in the bay. Preliminarily results of microplastic contamination in marine bivalves demonstrated relatively low contamination (0.01 ± 0.04, and 0.08 ± 0.34 for mussels and cockles, respectively) by microplastics (mainly polyethylene and polypropylene fragments), however this could be partly related to the methodological limitation identified here (e.g. exclusion of fibers). Organic pollutant (PAH, PCB and pesticides) were detected on floating microplastics at levels (not detected – 49,763 ng g-1, mean ± SD) similar to those measured in sediment and bivalves suggesting low risks in transferring hazardous chemicals in local marine organisms upon microplastic ingestion. Finally, distinct bacterial community assemblages were demonstrated on microplastics as compared with surrounding surface water; the Vibrio genus was identified as a discriminant biomarker of the plastic matrix. Overall, this work provides a first and thorough assessment of the microplastic contamination in the bay of Brest and solid methodological recommendations for further work.

[pt] A Baía de Guanabara é um ecossistema costeiro cujas alterações na bacia de drenagem ao longo dos anos levaram a sua condição atual de degradação ambiental. As razões diagnósticas e abordagens para a diferenciação de fontes de hidrocarbonetos baseiam-se em dados da literatura e a aplicação destas ao ambiente de estudo é de difícil extrapolação. Este estudo objetiva uma avaliação crítica destas ferramentas através da análise de dois tipos de aportes, o material particulado atmosférico (PM2,5) da Região Metropolitana do Rio de Janeiro e amostras de petróleo e seus produtos de origem brasileira, interpretando os resultados separadamente e relacionando-os aos dados obtidos em amostras de sedimentos desta mesma baía. A concentração de hidrocarbonetos na atmosfera é considerada baixa, onde a precipitação mostrou-se o principal fenômeno atmosférico a influenciar na concentração de material particulado e de hidrocarbonetos. As emissões veiculares, principalmente derivadas de veículos a gasolina, são a principal fonte de hidrocarbonetos para a atmosfera. A concentração de hidrocarbonetos em sedimentos corrobora o forte aporte crônico ao qual a baía está sujeita, sendo a fonte antrópica a principal responsável pelos níveis encontrados. Muitas das razões diagnósticas e seus limites estabelecidos em literatura não correspondem aos valores encontrados nas diferentes matrizes analisadas, existindo também sobreposição de valores entre estas. Os sedimentos da Baía de Guanabara encontram-se em um estado elevado de degradação que interfere na quantificação de fontes e avaliação de aportes. A análise de componentes principais realizada com a proporção relativa dos HPAs em relação aos compostos de mesma massa mostrou-se a metodologia mais adequada para a avaliação de aportes. Os hidrocarbonetos policíclicos aromáticos normalmente não quantificados em estudos ambientais, especialmente os de massa 278 e 276, mostraram maior capacidade em discriminar e separar as amostras geograficamente em relação à origem da contaminação. / [en] Guanabara Bay is a coastal ecosystem where changes in the drainage basin over the years led to its current condition of environmental degradation. The diagnostic reasons and approaches for differentiating hydrocarbon sources are based on literature data and their application in the environment is difficult to extrapolate. This study aims a critical evaluation of these tools by analyzing two types of contributions, the atmospheric particulate matter (PM2,5) in the Metropolitan Region of Rio de Janeiro and samples of petroleum and their products of Brazilian origin, interpreting the results separately and linking them to data obtained in sediment samples from the same bay. The hydrocarbons concentration in the atmosphere is relatively low, where precipitation proved to be the main atmospheric phenomena to influence the concentration of particulate matter and hydrocarbons. The vehicular emissions, mainly derived from gasoline of vehicles, are the main source of hydrocarbons to the atmosphere. The concentration of hydrocarbons in sediments corroborates the intense chronic contribution which the bay is subject, presenting the anthropogenic source as the main cause of levels detected. Many of the diagnostic reasons and limits established in the literature do not correspond to the values found in different matrices analyzed, there is also overlap between these values. The sediments of Guanabara Bay are in a high state of degradation that interferes with the sources quantification and assessment of contributions. A principal component analysis performed with the PAHs relative proportion in relation to the compounds with the same molecular mass proved to be the most appropriate methodology for evaluating contributions. The polycyclic aromatic hydrocarbons not usually quantified in environmental studies, especially the ones with mass 278 and 276, demonstrated greater ability to discriminate and separate samples geographically regarding the contamination origin.