Agnès Fournier

Transfer kinetics to egg yolk and modeling residue recovered in yolk of readily metabolized molecules: polycyclic aromatic hydrocarbons orally administered to laying hens.

The kinetic of transfer into egg yolk of Persistent Organic Pollutants with contrasting properties regarding biotransformation and bioaccumulation was investigated. Twenty-four Hy line hens, aged 26weeks, were orally administered, during 1 or 3 consecutive days, 6mg of a mixture of three PAHs (phenanthrene, pyrene, benzo[a]pyrene) in equal amounts (w:w) or 3mg of lindane per kg body weight daily. Each day, contaminants were administered 1h after oviposition in one gelatin capsule. Eggs were collected daily during the 12d following the first dosing. Concentrations of PAHs and their main hydroxylated metabolites in yolk were determined using HPLC coupled with fluorimetric detection, while lindane was analysed by GC-ED. At the end of the experiment, hens were sacrified and preovulatory yolks were collected and weighed. All three PAHs were recovered in yolks, mainly as metabolites, indicating an active biotransformation of these compounds in hens. Within 10d, PAHs and metabolites concentrations in yolks were back to initial values. However, the recovery rate of these compounds to yolk calculated over the 12-d experiment, was very low, reaching 0.089%, 0.034% and 0.006% for Phe, Pyr and B[a]P, respectively. Daily deposition of yolk and of PAHs and metabolites were found to be directly proportional, indicating that no significant body storage of these PAHs occurred and that they were rapidly hydroxylated after ingestion. In contrast, lindane and yolk deposition could not be related. Storage of lindane in body fat explains its slower elimination via eggs.

Effects of condensed organic matter on PCBs bioavailability in juvenile swine, an animal model for young children

The exposure assessment of polychlorinated biphenyls (PCBs) contaminated soils is a critical issue in terms of human health, especially since little reliable information on transfer of PCBs to humans via involuntary soil ingestion is available. Indeed, young children with their hand-to-mouth activity may be exposed to contaminated soils. The current study addresses the impact of soil organic matter (OM) condensation on bioavailability of sequestrated NDL-PCBs. Three artificial soils (ASs) were prepared according to OECD guideline 207. One standard soil (SS), devoid of OM, and two amended versions of this SS with fulvic acid (FA) or activated carbon (AC) were prepared to obtain 1% organic mass. This study involved fourteen juvenile male swine as a digestive physiology model of young children. Animals were randomly distributed into 4 contaminated groups (3 replicates) and a control one (2 replicates). During 10d, the piglets were fed AS or a corn oil spiked with 19200 ng of Aroclor 1254 per g of dry matter (6000 ng g(-1) of NDL-PCBs) to achieve an exposure dose of 1200 ng NDL-PCBskg(-1) of body weight per day. After 10d of oral exposure, NDL-PCBs in adipose tissue, liver and muscles were analyzed by GC-MS, after extraction and purification. Two distinct groups of treatments were found: on the one hand oil, SS and FA, on the other hand C and AC. This study highlights that condensed OM (AC) strongly reduces bioavailability whereas the less condensed one (FA) does not seem to have a significant effect. This result has to be considered as a first major step for further relative bioavailability studies involving mixture of different humic substances.

Polychlorinated Biphenyl (PCB) Decontamination Kinetics in Lactating Goats (Capra hircus) Following a Contaminated Corn Silage Exposure

This study aimed to determine the kinetics of contamination and decontamination of PCBs and PCDD/Fs in milk of lactating goats. Four goats were fed during 39 days with corn silage collected in an area accidentally contaminated and then with uncontaminated silage during 20 days. Concentrations of DL-PCBs + PCDD/Fs in milk exceeded rapidly (<15 days) the European limit value and approached steady state after 5 weeks. The decontamination kinetics in milk included first a rapid elimination phase (<10 days) followed by a slower elimination phase of 33, 51, and 59 days for DL-PCBs, NDL-PCBs, and PCDD/Fs, respectively. Therefore, in lactating goats, PCBs and PCDD/Fs contaminated forage raises concerns in terms of food safety. The study also indicates that a decontamination process of lactating animals remains feasible; 20 days was considered to be sufficient to obtain a DL-PCBs + PCDD/Fs level in milk below the regulatory value.

Bioavailability and bioaccumulation of sediment‐bound polychlorinated biphenyls to carp

The relative bioavailability of sediment-bound polychlorinated biphenyls (PCBs 138, 153, and 180) from a local contaminated site was examined using an in vivo carp model. Surface sediment from the PCB-contaminated site and spiked canola oil containing equivalent masses of PCBs were respectively incorporated in the sediment-dosed diets and oil-dosed diets at 3 dose levels resulting in 6 experimental diets. Juvenile carps (n = 90) were divided in 18 tanks (5 fish × 6 treatments × 3 tanks). Fish were fed the control diet during the adaptation period (15 d). Next, 1 fish was sampled in each tank and muscle tissues were combined in control groups. During the exposure period (15 d), the remaining fish were fed with 1 of the 6 experimental diets. At the end of the experiment, fish were sampled and muscle tissues were combined for each tank. The PCBs were monitored in feed and fish muscle. For both the contaminated sediment and spiked canola oil groups, concentrations of PCBs 138, 153, and 180 in muscle linearly increased with concentrations in food, with similar intercepts and slopes. In the present study, the sediment-bound PCBs were as bioavailable as those spiked into canola oil and fed to carp in a standard diet.

Kinetic study of chlordecone orally given to laying hens (Gallus domesticus).

The former use of chlordecone (CLD) in the French West Indies has resulted in long-term pollution of soils. In this area, CLD may be transferred into eggs of hens reared outdoors, through soil ingestion. In order to assess this risk, a kinetic study involving the contamination of laying hens (22 weeks of age) with a diet containing 500 μg CLD kg(-1) during 42 d, followed by a depuration period of 35 d was carried out. Forty-four hens were sequentially slaughtered all over the experimental period and their liver, egg, abdominal fat and serum were collected. Two additional edible tissues, pectoral and leg muscles, were collected in hens slaughtered at the end of the contamination period. The depuration half-life of CLD in liver, egg, abdominal fat and serum was estimated at 5.0 ± 0.38 (mean ± SE), 5.5 ± 0.29, 5.3 ± 0.37 and 5.1 ± 0.66 d, respectively. CLD concentration at the end of the contamination period reached 1640 ± 274, 460 ± 41, 331 ± 23, and 213 ± 8.5 μg kg(-1) fresh matter (FM), respectively. The corresponding concentrations in pectoral and leg muscles were 119 ± 8.4, 127 ± 11 μg kg(-1) FM, respectively. The steady state carry over rate of CLD in eggs reached 43 ± 7.6%. This experiment demonstrates the preferential accumulation of CLD in liver, its significant transfer to eggs and its quite short half-life. It is concluded that raising hens on even mildly contaminated areas would lead to products exceeding the regulatory maximum residue limit of 20 μg CLD kg(-1).

Impact of soil characteristics on relative bioavailability of NDL-PCBs in piglets

Children may be orally exposed to organic pollutants through involuntary soil ingestion. This study was aimed at determining the impact of the characteristics (organic carbon (OC), black carbon (BC), clay contents and pH) of ten contaminated soils on the bioavailability of non-dioxin-like polychlorobiphenyls (NDL-PCBs). Five juvenile male piglets were exposed to increasing amounts of each of the soils. These soil-fed groups were compared by a relative bioavailability approach (RBA) to a reference group fed with corn oil spiked with increasing doses of Aroclor 1254. After 10days of oral exposure, the animals were sacrificed and NDL-PCB concentrations were determined by GC-MS in the adipose tissue. The relative bioavailability (RBA) factors were calculated for PCB 101, 138, 153 and 180. Despite high variations in the amount of black carbon (0.50gkg(-1)-6.0gkg(-1)d.w.) and organic matter (12gkg(-1)-180gkg(-1)d.w.), only 3 soils exhibited a significantly lower RBA for all NDL-PCBs, compared to the oil-group. High levels of OC (>100gkg(-1)) and BC content (3.0gkg(-1)) were related to a significant reduction in RBA. Overall, RBA was higher than 45% independently of the soil and the PCB congener.

Effects of standard humic materials on relative bioavailability of NDL-PCBs in juvenile swine.

Young children with their hand-to-mouth activity may be exposed to contaminated soils. However few studies assessing exposure of organic compounds sequestrated in soil were realized. The present study explores the impact of different organic matters on retention of NDL-PCBs during digestive processes using commercial humic substances in a close digestive model of children: the piglet. Six artificial soils were used. One standard soil, devoid of organic matter, and five amended versions of this standard soil with either fulvic acid, humic acid, Sphagnum peat, activated carbon or a mix of Sphagnum peat and activated carbon (95∶5) (SPAC) were prepared. In order to compare the different treatments, we use spiked oil and negative control animals. Forty male piglets were randomly distributed in 7 contaminated and one control groups (n  = 5 for each group). During 10 days, the piglets were fed artificial soil or a corn oil spiked with 19 200 ng of Aroclor 1254 per g of dry matter (6 000 ng.g−1 of NDL-PCBs) to achieve an exposure dose of 1 200 ng NDL-PCBs.Kg−1 of body weight per day. NDL-PCBs in adipose tissue were analyzed by GC-MS. Fulvic acid reduced slightly the bioavailability of NDL-PCBs compared to oil. Humic acid and Sphagnum peat reduced it significantly higher whereas activated carbon reduced the most. Piglets exposed to soil containing both activated carbon and Shagnum peat exhibited a lower reduction than soil with only activated carbon. Therefore, treatment groups are ordered by decreasing value of relative bioavailability as following: oil ≥ fulvic acid>Sphagnum peat ≥ Sphagnum peat and activated carbon ≥ Humic acid>>activated carbon. This suggests competition between Sphagnum peat and activated carbon. The present study highlights that quality of organic matter does have a significant effect on bioavailability of sequestrated organic compounds.

Modeling PCB transfer into hen eggs: Influence of physiological characteristics of the animal

Laying hens are likely to be exposed to a wide range of persistent organic pollutants (POPs), including polychlorinated biphenyls (PCBs). To improve the safety of poultry farming systems in terms of POPs, the present research focused on assessing the impact of physiological characteristics of the hen on the transfer of ingested PCBs to eggs. Modeling was used as a research tool to explore the impact of some physiological characteristics on the transfer of PCBs in the laying hen. The mathematical model simulates the dynamics of the size of the lipid compartments in the animal and the frequency of laying, with the PCB concentrations in egg yolk and adipose tissue being model outputs. Simulations were run to assess effects of animal characteristics on the transfer of PCBs to eggs. Laying rate proportionally influenced the PCB level of eggs and adipose tissue at steady state. Body fat diluted absorbed PCBs in the absence of laying and significantly influenced the decontamination rate of tissues during depuration after an exposure period. Application of the present model to actual exposure cases highlights its value in improving the support of risk management in livestock farming.

Development and validation of an HPLC-MS/MS method with QuEChERS extraction using isotopic dilution to simultaneously analyze chlordecone and chlordecol in animal livers

A QuEChERS extraction method followed by HPLC-MS/MS analysis was developed to simultaneously analyze chlordecone and its metabolite chlordecol in animal livers. The overall method was validated with accuracy profiles according to the French Standard NF V03-110 and European Union guidelines. The validation was performed on bovine, ovine and porcine liver samples. Linearity, matrix effect, accuracy, within-laboratory repeatability, specificity, LOQ, Q/q relative ion intensities, and uncertainty were reported. Recoveries were between 70% and 120%. LOQs of 1.36 µg chlordecone kg-1 and 2.50 µg chlordecol kg-1 of fresh liver were found. Twelve contaminated livers of bovine, ovine and porcine origin from the French West Indies or samples from in vivo studies were analyzed. In these liver samples from contaminated animals, chlordecone was quantified at concentrations higher than the maximum residue limit and chlordecol in very low amounts in all the samples. In addition, these results confirm that chlordecone can be metabolized in ruminant species.

Activated carbon, a useful medium to bind chlordecone in soil and limit its transfer to growing goat kids

Chlordecone (Kepone) (CLD) is a highly persistent pesticide which was extensively used in the French West Indies; high levels of CLD can still currently be found in large agricultural areas. As CLD transfers from soil to animals mainly via involuntary ingestion, the consumption of foodstuffs derived from animals raised in contaminated areas may significantly contribute to exposure of humans to CLD. The present study was designed to test the efficacy of two different activated carbons (ACs) sources in limiting CLD transfer from soil to animal. Three soils (ASs) were prepared according to the OECD guideline 207. One standard soil (SS) lacking AC, and two modified preparations of SS supplemented with 2% coconut-based activated carbon (ORBO), SSO or with 2% lignite-based one (DARCO), SSD. All three soils were spiked with 10 μg of kepone per g of dry matter and aged for three weeks. This study involved 15 goat kids randomly assigned to the 3 experimental groups (n = 5/group), which were fed the experimental matrices at an exposure dose of 10 μg CLD per kg of body weight per day. After 21 d of oral exposure, CLD in adipose tissue and liver were analysed by LC-MS-MS. A significant decrease of 63.7% and 74.7% of CLD concentrations in adipose tissue and liver, respectively, were obtained from animals exposed using SS containing DARCO as compared to those receiving only SS. Decreases in CLD levels of 98.2% (adipose tissue) and 98.7% (liver) were obtained for animals exposed using SS containing ORBO. This study leads us to conclude that (i) the presence of AC in CLD-contaminated soil strongly reduces CLD bioavailability, and (ii) the efficacy depends on the nature and characteristics of the AC used.