Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
The increased use of pesticides has caused concern over the possible direct association of exposure to combinations of these compounds with bee health problems. There is growing proof that bees are regularly exposed to mixtures of agrochemicals, but most research has been focused on managed bees living in farmland, whereas little is known about exposure of wild bees, both in farmland and urban habitats. To determine exposure of wild bumblebees to pesticides in agricultural and urban environments through the season, specimens of five different species were collected from farms and ornamental urban gardens in three sampling periods. Five neonicotinoid insecticides, thirteen fungicides and a pesticide synergist were analysed in each of the specimens collected. In total, 61% of the 150 individuals tested had detectable levels of at least one of the compounds, with boscalid being the most frequently detected (35%), followed by tebuconazole (27%), spiroxamine (19%), carbendazim (11%), epoxiconazole (8%), imidacloprid (7%), metconazole (7%) and thiamethoxam (6%). Quantifiable concentrations ranged from 0.17 to 54.4 ng/g (bee body weight) for individual pesticides. From all the bees where pesticides were detected, the majority (71%) had more than one compound, with a maximum of seven pesticides detected in one specimen. Concentrations and detection frequencies were higher in bees collected from farmland compared to urban sites, and pesticide concentrations decreased through the season. Overall, our results show that wild bumblebees are exposed to multiple pesticides when foraging in agricultural and urban landscapes. Such mixtures are detected in bee tissues not just during the crop flowering period, but also later in the season. Therefore, contact with these combinations of active compounds might be more prolonged in time and widespread in the environment than previously assumed. These findings may help to direct future research and pesticide regulation strategies to promote the conservation of wild bee populations.
ISSN: 0269-7491

hal-01477203
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477203
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01477203/file/Quantifying%20exposure%20of%20wild%20bumblebees_accepted.pdf
DOI : 10.1016/j.envpol.2017.01.001
PUBMED : 28087090

Éditeur(s) : HAL CCSD Elsevier
Résumé : International audience
Because earthworms sustain soil functioning and fertility, there is a need to advance the knowledge of their adaptation potential to chemicals in anthropogenic landscapes. Our hypothesis is that there is acclimation to organic chemicals (pesticides) in earthworms that durably persist under conventional farming in anthropogenic landscapes. The adaptation capability of two populations of earthworms (Aporectodea caliginosa) having a different chemical exposure history, - one originating from 20 years of organic farming (naïve population) and another from 20 years of conventional farming (pre-exposed population) - to cope with soil organic pollutant (Opus®, epoxiconazole a worldwide used fungicide) were investigated. Several complementary metabolic and energetic endpoints were followed, and cast production was assessed as a behavioural biomarker related to earthworms ecological role for the soil. Basal metabolism reflected by respiration rate was increased in both fungicide-exposed worms compared to controls. Glycogen resources were decreased in the same proportion in the two populations but more rapidly for the naïve (7 days) than for the pre-exposed population (28 days). Soluble protein and most amino-acids contents increased in the pre-exposed population only, suggesting a detoxification mechanism. Metabolomic profiles showed a cut-off between fungicide-exposed and control groups in the pre-exposed earthworms only, with an increase in most of the metabolites. Exposure to a low dose of epoxiconazole increased cast production of pre-exposed earthworms, and this resulted in an increase in pesticide disappearance. As far as we know, this is the first study which evidenced there is an acclimation to an agricultural chemical in earthworms derived from conventional farming that also relates to a change in their burrowing behaviour, and for which larger consequences for the soil ecosystem need to be addressed. This original finding is of major interest in the frame of ecosystem resilience to global changes. Whether this physiological adaptation is a general pattern of response against fungicides or other pesticides would need to be confirmed with other molecules and agricultural contexts.
Soil Biology and Biochemistry

hal-01005821
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01005821
https://hal-univ-rennes1.archives-ouvertes.fr/hal-01005821/document
DOI : 10.1016/j.soilbio.2014.01.032

Éditeur(s) : HAL CCSD Springer Verlag
Résumé : International audience
: Human exposure to xenobiotics is usually estimated by indirect methods. Biological monitoring has emerged during the last decade to improve assessment of exposure. However, biomonitoring is still an analytical challenge, because the amounts of sample available are often very small yet analysis must be as thorough and sensitive as possible. The purpose of this work was to develop an untargeted "exposomics" approach by using ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS), which was applied to the characterization of pesticide metabolites in urine from pregnant women from a French epidemiological cohort. An upgradable list of pesticides commonly used on different crops, with their metabolites (more than 400 substances) was produced. Raw MS data were then processed to extract signals from these substances. Metabolites were identified by tandem mass spectrometry; putative identifications were validated by comparison with standards and metabolites generated by experiments on animals. Finally, signals of identified compounds were statistically analyzed by use of multivariate methods. This enabled discrimination of exposure groups, defined by indirect methods, on the basis of four metabolites from two fungicides (azoxystrobin, fenpropimorph) used in cereal production. This original approach applied to pesticide exposure can be extended to a variety of contaminant families for upstream evaluation of exposure from food and the environment.
ISSN: 1618-2642

hal-00875889
https://hal.archives-ouvertes.fr/hal-00875889
DOI : 10.1007/s00216-013-7136-2
PUBMED : 23892877

Résumé : En raison de la nature symbiotique des micro-organismes endophytes, cette étude visait à étudier l'activité antibactérienne, antifongique et cytotoxique chez les métabolites secondaires des extraits endophytes foliaires de plantes de l'Amazonie brésilienne et du Cerrado. Dans ce travail de thèse 147 micro-organismes cultivables ont été isolés (130 champignons, 3 bactéries e 14 champignons non-identifiés ou inconnus) à partir de 28 plantes (4 espèces collectées au Brésil et 24 en Guyane Française). Tous les micro-organismes furent identifiés par analyse moléculaire des régions spécifiques de l'ADNr en utilisant des techniques de séquençage génomique. Des champignons endophytes de l’ordre des Xylariales furent ceux de plus importante fréquence d’isolation dans cette étude, représentés par 25 isolats. Des extraits bruts à l’AcOEt furent produits à partir de cultures de chaque micro-organisme isolé. Une proportion relative signifiante (23,1%) des extraits démontra une activité sur Candida albicans ATCC 10213, pendant que 4% furent actifs sur Staphylococcus aureus ATCC 29213. Le potentiel cytotoxique des extraits fut évalué pour des lignées cellulaires humaines KB (carcinome cervical utérin), MDA-MB-435 (mélanome), et MRC-5 (fibroblastes de poumon normal), résultant en proportion signifiante avec activité d’inhibition de la prolifération cellulaire (24,4%, 23,1% e 16,3%, respectivement).Dix-huit métabolites secondaires furent isolés et identifiés à partir du fractionnement des extraits bruts de huit endophytes. Dix-sept de ces substances furent déjà décrites précédemment dans la littérature: l’acide piliformique (24) et la griséofulvine (25) isolés à partir de Xylaria cubensis SNB-GCI02; La phomopirone (26), la pyrenocine (27), l’alterpérilenol (28) et la novae-zelandine (29), isolés de Lewia infectoria SNB-GTC2402; 5-métilmelleina (31) et la dihidrosporothrioride (32) isolées à partir de Xylaria sp. SNB-GTC2501; La mycoleptodiscine (34) et la mycoleptodiscine B (35) isolées à partir de Mycoleptodiscus sp. SNB-GTC2304; le mycoepoxidiène (36) et la altiloxina A (37) isolés à partir de Diaporthe pseudomangiferae SNB-GSS10; L’acremonisol (40), le semicochliodinol (41) et le cochliodinol (42) isolés à partir de Chaetomium globosum SNB-GTC2114; la colletofragarone A2 (43) isolée à partir de Colletotrichum sp. SNB-GTC0201; la flavoglaucine (44) isolée à partir de Eurotium rubrum BBS01. Parmi ces substances, la flavoglaucine (44) isolée à partir de E. rubrum BBS01 démontra une activité comparable au contrôle fluconazol en C. albicans (CIM de 4 µg.mL-1). La substance (44) présenta IC50 >10 µM sur cellules normales MRC-5, ce qui en fait un candidat pour des études ultérieures. Dans ce travail, fut identifié pour la première fois l’activité de la colletofragarone A2 (43) isolée à partir de Colletotrichum sp. SNB-GTC0201. La substance inédite nommée pyrrocidine C (30) fut isolée à partir de L. infectoria SNB-GTC2402 et identifiée par des analyses spectroscopiques (Casella et al., 2013). La pyrrocidine C (30) fut active en S. aureus ATCC 10213 (CIM de 2 µg.mL-1) et ne fut pas considérées cytotoxique pour les cellules normales MRC-5 (IC50 >10 µM), démontrant une sélectivité dans l’action antimicrobienne. Ces résultats démontrent la grande diversité des endophytes fongiques chez les plantes de l'Amazonie brésilienne et du Cerrado et la grande chimiodiversité associée aux métabolites secondaires de ces micro-organismes. Des champignons endophytes tropicaux, comme ceux présentés dans cette étude, peuvent apparaître comme une nouvelle source de substances antimicrobiennes et cytotoxiques.
Because of the symbiotic nature of endophytes, this survey aims to investigate the probality of discovering antibacterial, antifungal and cytotoxic activities in secondary metabolites of leaf endophytes isolated from plants of Amazon and Cerrado biomes. In this study, 147 cultivable microorganisms were isolated (130 fungi, 3 bacteria and 14 unidentified or unknown microbes) from 28 plant species (4 species collected in Brazil and 24 in French Guyana). All endophytes were identified by molecular analyses of specific rDNA regions, with genomic sequencing techniques. Fungal endophytes belonging to Xylariales order were the most frequently isolated in this study, represented by 25 isolates. Crude AcOEt extracts were produced from cultures of each isolated endophyte. A significant relative proportion (23,1%) of extracts showed activity in Candida albicans ATCC 10213, while 4% were active in Staphylococcus aureus ATCC 29213. The cytotoxic potencial of the extracts was evaluated for human cell lines KB (uterin cervical carcinome), MDA-MB-435 (melanoma) and MRC-5 (normal lung fibroblasts), and a significant proportion of them showed cellular proliferation inhibition (24,4%, 23,1% e 16,3%, respectively).Eighteen secondary metabolites were isolated by the fractionation of eight endophytic extracts. Seventeen of these substances had already been previously described in the literature: piliformic acid (24) and griseofulvin (25) isolated from Xylaria cubensis SNB-GCI02; phomopiron A (26), pyrenocin A (27), alterperilenol (28) and novae-zelandin A (29) isolated from Lewia infectoria SNB-GTC2402; 5-metilmellein (31) and dihidrosporothriorid (32) isolated from Xylaria sp. SNB-GTC2501; mycoleptodiscin A (34) and mycoleptodiscin B (35) isolated from Mycoleptodiscus sp. SNB-GTC2304; mycoepoxidien (36) and altiloxin A (37) isolated from Diaporthe pseudomangiferae SNB-GSS10; acremonisol A (40), semicochliodinol A (41) and cochliodinol (42) isolated from Chaetomium globosum SNB-GTC2114; colletofragaron A2 (43) isolated from Colletotrichum sp. SNB-GTC0201; flavoglaucin (44) isolated from Eurotium rubrum BBS01. Among these substances, flavoglaucin (44), isolated from E. rubrum BBS01, showed comparable antifungal activity with the positive control fluconazol in C. albicans (MIC of 4 µg.mL-1). Flavoglaucin (44) also showed IC50 >10 µM in normal MRC-5 cells, becoming a good candidate for further studies. In this work, the cytotoxic activity of colletofragaron A2 (43), isolated from Colletotrichum sp. SNB-GTC0201 was described for the first time. The unpublished substance named pyrrocidin C (30) isolated from L. infectoria SNB-GTC2402 was identified by spectroscopic analyses (Casella et al., 2013). The pyrrocidin C (30) was active in S. aureus ATCC 10213 (MIC of 2 µg.mL-1), and was not considered cytotoxic for normal MRC-5 cells (IC50 >10 µM), showing selectivity in antimicrobial activity. These results demonstrate the great endophytic fungal diversity in plants of Amazon and Cerrado biomes, along with the chemodiversity associated to the secondary metabolites of these endophytes. Tropical fungal endophytes, like those seen in this work, may emerge as a new source of antimicrobial and cytotoxic substances.
Devido à natureza simbiótica dos micro-organismos endofíticos, este estudo teve como objetivo investigar a atividade antibacteriana, antifúngica e citotóxica em metabólitos secundários de extratos de fungos endofíticos foliares de plantas do bioma Amazônia e Cerrado. Neste trabalho de tese foram isolados 147 micro-organismos cultiváveis (130 fungos, 3 bactérias e 14 fungos não-identificados ou desconhecidos) a partir de 28 plantas (4 espécies coletadas no Brasil e 24 na Guiana Francesa). Todos os micro-organismos foram identificados por análise molecular de regiões específicas de DNAr, com uso de técnicas de sequenciamento genômico. Fungos endofíticos da ordem Xylariales foram os de maior frequência de isolamento neste estudo, representados por 25 isolados. Extratos brutos em AcOEt foram produzidos a partir de culturas de cada micro-organismo isolado. Uma proporção relativa significante (23,1%) dos extratos demonstrou atividade em Candida albicans ATCC 10213, enquanto 4% foram ativos em Staphylococcus aureus ATCC 29213. O potencial citotóxico dos extratos foi avaliado para as linhagens celulares humanas KB (carcinoma cervical uterino), MDA-MB-435 (melanoma), e MRC-5 (fibroblastos de pulmão normal), resultando em proporção significante com atividade de inibição da proliferação celular (24,4%, 23,1% e 16,3%, respectivamente). Dezoito metabólitos secundários foram isolados a partir do fracionamento de oito extratos brutos endofíticos. Dezessete destas substâncias já tinham sido descritas anteriormente na literatura: ácido pilifórmico (24) e griseofulvina (25) isoladas de Xylaria cubensis SNB-GCI02; phomopirona A (26), pyrenocina A (27), alterperilenol (28) e novae-zelandina A (29) isoladas de Lewia infectoria SNB-GTC2402; 5-metilmelleina (31) e Dihidrosporothriolida (32) isoladas de Xylaria sp. SNB-GTC2501; mycoleptodiscina A (34) e mycoleptodiscina B (35) isoladas de Mycoleptodiscus sp. SNB-GTC2304; mycoepoxidieno (36) e altiloxina A (37) isoladas de Diaporthe pseudomangiferae SNB-GSS10; acremonisol A (40), semicochliodinol A (41) e cochliodinol (42) isoladas de Chaetomium globosum SNB-GTC2114; colletofragarona A2 (43) isolada de Colletotrichum sp. SNB-GTC0201; flavoglaucina (44) isolada de Eurotium rubrum BBS01. Dentre estas substâncias, a flavoglaucina (44) isolada de E. rubrum BBS01, demonstrou atividade comparável ao controle fluconazol em C. albicans (CIM de 4 µg.mL-1). Esta substância (44) apresentou IC50 >10 µM em células normais MRC-5, tornando-se candidata para estudos posteriores. Neste trabalho foi identificado pela primeira vez a atividade citotóxica da colletofragarona A2 (43), isolada de Colletotrichum sp. SNB-GTC0201. A substância inédita nomeada pyrrocidina C (30) foi isolada a partir de L. infectoria SNB-GTC2402 e identificada através de análises espectroscópicas (Casella et al., 2013). A pyrrocidina C (30) foi ativa em S. aureus ATCC 10213 (CIM de 2 µg.mL-1), e não foi considerada citotóxica para as células normais MRC-5 (IC50 >10 µM), demonstrando seletividade na ação antimicrobiana. Estes resultados demonstram a grande diversidade fúngica endofítica em plantas do bioma Amazônia e Cerrado, e a quimiodiversidade associada aos metabólitos secundários destes micro-organismos. Fungos endofíticos tropicais, como os vistos neste trabalho, podem emergir como uma nova fonte de substâncias antimicrobianas e citotóxicas.

http://www.theses.fr/2014AGUY0836/document