Cumulative risk assessment of the dietary heavy metal and aluminum exposure of Finnish adults.

While directly comparable toxicological data are unavailable, this pilot study used published toxicological endpoints for neurological damage to weigh the relative importance of cadmium, lead, arsenic, mercury, nickel, and aluminum in the mixture Finnish adults receive from their daily diet. In addition, the effects of a selection of these chemicals on cognition, kidney tubular damage, and fertility were assessed using the toxicological endpoints available in the Chemical Mixture Calculator developed by the Technical University of Denmark. Consumption data from the FinDiet 2012 national survey of 25 to 74-year-olds and occurrence data mainly obtained in national monitoring were used to estimate the cumulative dietary exposure, which was found to be so high that the possibility of neurological damage or kidney effects cannot be ruled out for most of the population, particularly fertile age women. For Finns below the age of 65 years, the main sources of cumulative exposure were bread and other cereals, non-alcoholic drinks, and vegetables. When mean exposure was statistically compared between age groups and genders, women aged 25 to 45 years had a statistically significantly higher exposure than men of the same age (P < 0.05) and women aged 46 to 64 years (P < 0.001).

BIKE: Dietary Exposure Model for Foodborne Microbiological and Chemical Hazards.

BIKE is a Bayesian dietary exposure assessment model for microbiological and chemical hazards. A graphical user interface was developed for running the model and inspecting the results. It is based on connected Bayesian hierarchical models, utilizing OpenBUGS and R in tandem. According to occurrence and consumption data given as inputs, a specific BUGS code is automatically written for running the Bayesian model in the background. The user interface is based on shiny app. Chronic and acute exposures are estimated for chemical and microbiological hazards, respectively. Uncertainty and variability in exposures are visualized, and a few optional model structures can be used. Simulated synthetic data are provided with BIKE for an example, resembling real occurrence and consumption data. BIKE is open source and available from github.

Dietary Heavy Metal Exposure among Finnish Adults in 2007 and in 2012.

For the non-smoking and non-occupationally exposed population in Europe, food is the main source of heavy metal exposure. The aim of the study was to estimate the dietary exposure of the Finnish adult population to cadmium, lead, inorganic arsenic, inorganic mercury and methyl mercury as well as nickel using governmental as well as industry data on heavy metal occurrence in foodstuffs and the data from two national food consumption surveys conducted in 2007 and 2012. The sources of heavy metal exposure were estimated for the working-age population (25 to 64 years) and for the elderly (65 to 74 years). Exposure differences between years and between population groups were compared statistically. The mean exposure of women aged 25 to 45 years to cadmium and lead was statistically significantly (p < 0.001) higher, and the methyl mercury exposure lower (p = 0.001) than that of women aged 46 to 64 years. For nickel and inorganic arsenic the differences were lower but still statistically significant (p < 0.05). Between genders, significant difference (p < 0.05) was only seen for lead and nickel. Mean cadmium exposure was significantly higher in 2012 than in 2007. For at least 95% of the adult population, the risk of health damage from mercury or nickel exposure is negligible, but the margin of exposure for lead and inorganic arsenic is small and shows a possible risk of cancer or neurotoxic effects.

Antimicrobial use, biosecurity, herd characteristics, and antimicrobial resistance in indicator Escherichia coli in ten Finnish pig farms.

We investigated connections between antimicrobial use (AMU), biosecurity, and the numbers of pigs and staff in ten Finnish farrow-to-finish herds. Data on AMU in each herd were collected for 12 months. AMU was quantified as treatment incidences per 1000 days at risk (TI) using the consensus defined daily dose calculation. Biosecurity was scored using the Biocheck.UGent™ system. We also examined antimicrobial resistance patterns of indicator E. coli isolated from faeces of selected pigs. In each herd, two groups of five pigs were formed: 1) antimicrobial treatment group (ANT: at least one pig in the litter was identified as sick and treated with antimicrobials) and 2) non-antimicrobial treatment group (NON: the litter was not medicated). Faecal samples were taken from these pigs at 5 and 22 weeks of age, cultured, and indicator E. coli isolates were tested for antimicrobial susceptibilities. The AMU varied considerably between the herds. Altogether, most of the antimicrobial treatment courses were assigned to weaned piglets. When AMU was quantified as TIs, suckling piglets had the highest TI (mean 46.6), which was significantly higher (P < 0.05) than TIs in fatteners and breeders (9.3 and 7.3, respectively). The difference between TI in suckling and TI in weaned piglets (19.1) was not statistically significant. There was a tendency for a negative correlation between the TI in breeders and the number of sows (r = -0.56, P = 0.09). Larger herds had higher external biosecurity scores than smaller herds (LS-means; 72 vs. 66, P < 0.05). The proportions of E. coli isolates resistant to at least one antimicrobial were higher in pigs at 5 weeks than in pigs at 22 weeks of age (Binomial proportion means; 40.5 % vs. 15.5 %, P < 0.05); as well as proportions of isolates resistant to at least three antimicrobial classes (23.0 % vs. 3.7 %, P < 0.01). These proportions did not differ between the ANT and NON groups at either 5 or 22 weeks of age (P> 0.05). We found few connections: enhanced external biosecurity levels found in the large herds co-occurred with lower use of antimicrobials and herds with low biosecurity scores - especially in the internal subcategories - appeared to have higher proportions of resistant isolates. Conclusively, we suggest that enhancing internal biosecurity might contribute to a reduction in the spreading of antimicrobial resistance in pig herds.

Maternal Nitrate and Nitrite Intakes during Pregnancy and Risk of Islet Autoimmunity and Type 1 Diabetes: The DIPP Cohort Study.

BACKGROUND: High dietary intake of nitrate and nitrite might increase the risk of type 1 diabetes. To our knowledge, no earlier prospective study has explored whether maternal dietary intake of nitrate and nitrite during pregnancy is associated with the risk of type 1 diabetes in the offspring. OBJECTIVE: Our aim was to study association between maternal intake of nitrate and nitrite during pregnancy and the risk of islet autoimmunity and type 1 diabetes in the offspring. DESIGN: Children born between 1997 and 2004 at Oulu and Tampere University Hospitals in Finland and carrying increased human leukocyte antigen (HLA)-conferred risk for type 1 diabetes were followed in the Type 1 Diabetes Prediction and Prevention (DIPP) study from 3 mo of age. Islet autoantibodies were screened at 3- to 12-mo intervals from serum samples. Of 4879 children, 312 developed islet autoimmunity and 178 developed type 1 diabetes during a 15-y follow-up. Maternal intake of nitrate and nitrite during the eighth month of pregnancy was assessed after birth using a validated self-administered FFQ. Cox proportional hazards regression was used for the statistical analyses. RESULTS: Maternal intake of nitrate and nitrite during pregnancy was not associated with the child's risk of islet autoimmunity [nitrate: HR 0.99 (95% CI: 0.88, 1.11); nitrite: HR 1.03 (95% CI: 0.92, 1.15)] or type 1 diabetes [nitrate: HR 1.02 (95% CI: 0.88, 1.17); nitrite: HR 0.97 (95% CI: 0.83, 1.12)] when adjusted for energy (residual method), sex, HLA risk group, and family history of diabetes. Further adjustment for dietary antioxidants (vitamin C, vitamin E, and selenium) did not change the results. CONCLUSION: Maternal dietary intake of nitrate or nitrite during pregnancy is not associated with the risk of islet autoimmunity or type 1 diabetes in the offspring genetically at risk for type 1 diabetes.

Dietary heavy metal exposure of Finnish children of 3 to 6 years.

The dietary exposure of Finnish 3-year-old and 6-year-old children to cadmium, lead, arsenic and mercury was determined using concentration data from Finland and individual food consumption data as well as individual weights of the children. Using middle bound estimates, 88% of the 3-year-olds and 64% of the 6-year-olds exceeded the tolerable weekly intake of cadmium. The benchmark dose for neurological damage caused by lead was exceeded by 14% and 1%, while the lowest benchmark dose of inorganic arsenic was exceeded by 43% and 29% for the 3-year-olds and 6-year-olds, respectively. The exposure of both age groups was below the tolerable weekly intake for inorganic mercury and methyl mercury. Although high, the exposures calculated with predominantly national concentration data were lower than previously estimated by EFSA, due to, for example, lower average concentrations in some much-consumed foods. The heavy metal exposure levels of the girls and the boys were also compared. Exposure to cadmium and lead was significantly higher for the boys than for the girls in both age groups, and exposure to inorganic arsenic was significantly higher for the 6-year-old boys than the girls of same age.

Quantitative risk assessment on the dietary exposure of Finnish children and adults to nitrite.

Nitrite intake from the consumption of cured meat and tap water was estimated for Finnish children of 1, 3 and 6 years as well as Finnish adults of 25-74 years. Nitrite content in the foods was measured by capillary electrophoresis, and was then used together with individual food consumption data from the FINDIET 2007 and DIPP studies in a stochastic exposure assessment by a Monte Carlo Risk Assessment (MCRA) program. Nitrite intake from additive sources and tap water was assessed, and more than every 10th child between the ages 3 and 6 years was estimated to have a nitrite intake exceeding the acceptable daily intake (ADI) of nitrite. The high exposure levels were caused by frequent consumption of large portions of sausages, up to 350 g day(-1) or 750 g in 3 days, among the children. Median nitrite intake from cured meat was 0.016, 0.040, 0.033 and 0.005 mg kg(-1) body weight day(-1) for children of 1, 3 and 6 years and adults, respectively. Bayesian estimation was employed to determine safe consumption levels of sausages and cold cuts for children, and these results gave rise to new national food consumption advice.

Separation of steroids using vegetable oils in microemulsion electrokinetic capillary chromatography.

The steroids, hydrocortisone, androstenedione, 17-α-hydroxyprogesterone, testosterone, 17-α-methyltestosterone, and progesterone were separated with microemulsion electrokinetic chromatography (MEEKC) and detected with UV absorption. The microemulsion phases were prepared from both artificial and vegetable oils, from them the first was made of alkane and alcohol and the latter from colza, olive, linseed, and walnut oils. The electrolyte solutions were made to emulsions using sodium dodecyl sulfate and alkaline tetraborate. The solution mixtures made from ethyl acetate, sodium dodecyl sulfate, 1-butanol, acetonitrile, and sodium tetraborate were used as the reference solutions to evaluate the performance of the vegetable oil emulsions. Our study showed that the lipophilic organic phase in the microemulsion did provide resolution improvements but not selectivity changes. The results also correlate with real interactions of the steroids with the lipophilic organic microemulsion phase. The quality of the oils between the manufacturers did not have importance, which was noticed from the equal behavior of the steroids in the vegetable oil emulsions. Detection limits of the steroids in vegetable oil emulsions were at the level of 0.20-0.43µg/L. Thus, they were 2-10 times higher than the concentrations in the partial filling micellar electrokinetic chromatography (PF-MEKC), which we have obtained earlier. The repeatability (RSD%) of the electrophoretic mobilities of the steroids was between 0.50 and 3.70. The RSD% values between the inter-day separations were below 1%, but when walnut and olive oils were used the values exceeded even 10%.

Time-resolved detection of hot electron-induced electrochemiluminescence of fluorescein in aqueous solution.

Strong electrogenerated chemiluminescence (ECL) of fluorescein is generated during cathodic pulse polarization of oxide-covered aluminum electrodes and the resulting decay of emission is so sluggish that time-resolved detection of fluorescein is feasible. The present ECL in aqueous solution is based on the tunnel emission of hot electrons into the aqueous electrolyte solution, which probably results in the generation of hydrated electrons and hydroxyl radicals acting as redox mediators. The successive one-electron redox steps with the primary radicals result in fluorescein in its lowest excited singlet state. The method allows the detection of fluorescein (or its derivatives containing usable linking groups to biomolecules) over several orders of magnitude of concentration with detection limits well below nanomolar concentration level. The detection limits can still be lowered, e.g., by addition of azide or bromide ions as coreactants. The results suggest that the derivatives of fluorescein, such as fluorescein isothiocyanate (FITC), can be detected by time-resolved measurements and thus be efficiently used as electrochemiluminescent labels in bioaffinity assays.

Determination of iridoid glycosides in larvae and adults of butterfly Melitaea cinxia by partial filling micellar electrokinetic capillary chromatography-electrospray ionisation mass spectrometry.

The iridoid glycosides, methyl catalpol, asperuloside, verbenalin, cinnamoyl catalpol, catalpol and aucubin, were studied from both larvae and adults of butterfly Melitaea cinxia. Special emphasis in the study was put on finding a correlation between the iridoid glycoside content in butterflies and plants. An optimised partial filling micellar electrokinetic capillary chromatographic-electrospray ionisation mass spectrometric (PF-MECC-ESI-MS) method was employed for the separation and identification of the six iridoid glycosides. In this work, the isolation and determination of catalpol and aucubin from extracts of both larvae and adults of Melitaea cinxia butterflies is demonstrated. The PF-MECC-ESI-MS method, using the [M+Na]+, [M+Li]+ and/or [M+NH(4)]+ adducts in ESI-MS, was used for quantification of aucubin and catalpol in the insects. In addition, the identification of all analytes was attempted by direct infusion MS/MS analysis. LOQ values for the iridoid glycosides varied between 10 mg/l (for verbenalin) to 50 mg/l (for catalpol and aucubin) corresponding to 0.1% of the samplés dry mass. A correlation was noticed between the concentrations of iridoid glycosides in plants and the concentrations in larvae feeding on them.

Effect of iridoid glycoside content on oviposition host plant choice and parasitism in a specialist herbivore.

The Glanville fritillary butterfly Melitaea cinxia feeds upon two host plant species in Aland, Finland, Plantago lanceolata and Veronica spicata, both of which produce iridoid glycosides. Iridoids are known to deter feeding or decrease the growth rate of many generalist insect herbivores, but they often act as oviposition cues to specialist butterflies and are feeding stimulants to their larvae. In this study, two iridoid glycosides (aucubin and catalpol) were analyzed by micellar electrokinetic capillary chromatography. We measured the spatial and temporal variation of iridoid glycosides in natural populations of the host plants of M. cinxia. We also analyzed the aucubin and catalpol content in plants in relation to their use by ovipositing females, and in relation to the incidence of parasitism of M. cinxia larvae in natural populations. The mean concentrations of aucubin and catalpol were higher in P. lanceolata than in V. spicata, and catalpol concentrations were higher than aucubin concentrations in both host species. Plantago lanceolata individuals that were used for oviposition by M. cinxia had higher aucubin concentrations than random plants and neighboring plants. Additionally, oviposition and random plants had higher catalpol concentrations than neighboring plants, indicating that ovipositing females select for high iridoid glycoside plants or that oviposition induces iridoid glycoside production in P. lanceolata. Parasitism by the specialist parasitoid wasp Cotesia melitaearum occurred most frequently in larval groups that were feeding on plants with low concentrations of catalpol, irrespective of year, population, and host plant species. Therefore, parasitoids appear to avoid or perform poorly in host larvae with high catalpol content.

Analysis of eleven iridoid glycosides by micellar electrokinetic capillary chromatography (MECC) and screening of plant samples by partial filling (MECC)-electrospray ionisation mass spectrometry.

Of ammonium, lithium and sodium salts of dodecyl sulfate studied as surfactants in the separation of iridoid glycosides by micellar electrokinetic capillary chromatography (MECC), the last one gave the best results. Eleven neutral iridoid glycosides were separated by MECC with sodium dodecyl sulfate as surfactant, and the water-micelle partition coefficients of the compounds were calculated. The separation system was coupled via a coaxial sheath flow electrospray interface to a mass spectrometer, and the partial filling technique was used in the on-line analysis. Seven plant species belonging to five genera (Plantago, Veronica, Melampyrum, Succisa and Valeriana) were screened for the iridoid glycosides by the new method that was developed. The findings confirmed those of an earlier study on five of the iridoid glycosides. Some new iridoid glycosides were found in Plantago lanceolata, Veronica spicata and V. chamaedrys.