Management of human health risk in the context of kitchen gardens polluted by lead and cadmium near a lead recycling company

Purpose

At the global scale, gardening activities are often performed in urban areas with a historical background of pollution. In this study, a participatory program was developed with citizens concerned by gardening activities near a 50-year-old regulated lead recycling company, with the aim of co-constructing the tools for the assessment and management of potential sanitary risks induced by historic pollution with persistent (eco) toxic metals: lead and cadmium.

Materials and methods

Soils and vegetables (lettuce, leek, celery, carrot, chard, pumpkin, and celeriac) samples were collected from four kitchen gardens neighboring a 50-year-old secondary lead smelter. Both total and in vitro human bioaccessible metal concentrations in the cultivated plants were measured in relation to soil characteristics.

Results and discussion

The results showed that the soils of these gardens were slightly contaminated by metals (Pb, 77 to 236 mg kg^?1; and Cd, 0.5 to 1 mg kg^?1) in comparison with the natural geologic background. However, significant pollution of vegetables can occur especially with lead (Pb up to 9.8 mg kg^?1 in lettuce) and certainly as a result of direct foliar transfer. The washing of plants before consumption is therefore recommended in the context of atmospheric fallout of ultrafine particles enriched with metals.

Conclusions

Metal bioaccessibility measure integrates the influence of metal type, plant type, and soil physico-chemical properties. Based on the results, it is proposed that human bioaccessible fraction of metals may also be currently taken into account as well as total metal quantities and bioaccumulation factors in risk assessment studies performed in gardens. Overall, this study has led to reflections and functional recommendations aimed at reducing human exposure and to finally developing sustainable gardening practices.

     

Effects of accelerators on mobility of 14C-2,4-dichlorophenoxy butyric acid in plant cuticles depends on type and concentration of accelerator

Effects of diethyl suberate (DESU), diethyl sebacate (DES), dibutyl suberate (DBSU), dibutyl sebacate (DBS), and tributyl phosphate (TBP) on diffusion of 14C-2,4-dichlorophenoxy butyric acid (2,4-DB) across cuticular membranes (CM) was studied. Astomatous CM were isolated enzymatically from Stephanotis floribunda Brongn. leaves, and diffusion was measured at 20 degrees C. The alkyl-substituted dicarboxylic acids constitute a homologous series with carbon numbers increasing from C12 to C18. Molecular weights increased only moderately from 230.0 (DESU) to 314.5 (DBS), while partition coefficients varied over orders of magnitude from 92 (DESU), to 1213 (DES), to 15,988 (DBSU), to 210,762 (DBS). All the above compounds turned out to be accelerators as they increased 2,4-DB mobility by up to 40-fold with accelerator concentrations in the CM ranging from only 9.2 to 105 g kg(-1). Efficacy (2,4-DB mobility in the presence/mobility in the absence of accelerators) increased with increasing concentrations of accelerators in CM or in reconstituted cuticular waxes. Plotting efficacy vs accelerator concentration in the CM resulted in straight lines, and their slopes increased in the order DBS (0.14), DBSU (0.31), DES (0.51), and DESU (0.85). Hence, DESU was the most powerful accelerator in this series as it increased 2,4-DB mobility in the CM about 6 times more than DBSU. Waxes constitute the major barrier in plant cuticles, and plots of efficacy vs accelerator concentration in Stephanotis wax were also linear, but compared to CM slopes were steeper by factors of 3.20 (DBS), 2.97 (DBSU), 2.70 (DES), and 1.62 (DESU). TBP was similarly effective as DESU, but plots of efficacy vs concentration were not linear, and curves approached a plateau at 60-80 g kg(-1). These data are discussed with regard to suitability of these accelerators for formulating systemic pesticides.     

Greek plant extracts exhibit selective estrogen receptor modulator (SERM)-like properties

To prevent bone loss that occurs with increasing age, nutritional and pharmacological factors are needed. Traditional therapeutic agents (selective estrogen receptor modulators or SERMs, biphosphonates, calcitonin) may have serious side effects or contraindications. In an attempt to find food components potentially acting as SERMs, we submitted four plant aqueous extracts derived from Greek flora (Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum) in a series of in vitro biological assays reflective of SERM profile. We examined their ability (a) to stimulate the differentiation and mineralization of osteoblastic cell culture by histochemical staining for alkaline phosphatase and Alizarin Red-S staining, (b) to induce, like antiestrogens, the insulin growth factor binding protein 3 (IGFBP3) in MCF-7 breast cancer cells, and (c) to proliferate cervical adenocarcinoma (HeLa) cells by use of MTT assay. Our data reveal that all the plant extracts studied at a concentration range 10-100 microg/mL stimulate osteoblastic cell differentiation and exhibit antiestrogenic effect on breast cancer cells without proliferative effects on cervical adenocarcinoma cells. The presence of estradiol inhibited the antiestrogenic effect induced by the extracts on MCF-7 cells, suggesting an estrogen receptor-related mechanism. In conclusion, the aqueous extracts derived from Sideritis euboea, Sideritis clandestina, Marticaria chamomilla, and Pimpinella anisum may form the basis to design "functional foods" for the prevention of osteoporosis.     

Zinc Blotting Assay for Detection of Zinc‐Binding Prolamin in Barley (Hordeum vulgare) Grain

In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol‐soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. ⁶⁵Zn overlay and blotting techniques have been widely used for detecting zinc‐binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (⁶⁵ZnCl₂) blotting technique was optimized to detect zinc‐binding prolamins, followed by development of an easy‐to‐follow nonradioactive colorimetric zinc blotting method with a zinc‐sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS‐PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc‐binding specificity of certain proteins was detected either by autoradiography or color formation. The dithizone staining method gave similar reproducibility to the radioactive blotting. The detected zinc‐binding protein was identified as B‐hordein by Western blotting.     

Validation of 2 commercial Neospora caninum antibody enzyme linked immunosorbent assays

This is a validation study of 2 commercially available enzyme linked immunosorbent assays (ELISA) for the detection of antibodies against Neospora caninum in bovine serum. The results of the reference sera (n = 30) and field sera from an infected beef herd (n = 150) were tested by both ELISAs and the results were compared statistically. When the immunoblotting results of the reference bovine sera were compared to the ELISA results, the same identity score (96.67%) and kappa values (K) (0.93) were obtained for both ELISAs. The sensitivity and specificity values for the IDEXX test were 100% and 93.33% respectively. For the Biovet test 93.33% and 100% were obtained. The corresponding positive (PV+) and negative predictive (PV−) values for the 2 assays were 93.75% and 100% (IDEXX), and 100% and 93.75% (Biovet). In the 2nd study, competitive inhibition ELISA (c-ELISA) results on bovine sera from an infected herd were compared to the 2 sets of ELISA results. The identity scores of the 2 ELISAs were 98% (IDEXX) and 97.33% (Biovet). The K values calculated were 0.96 (IDEXX) and 0.95 (Biovet). For the IDEXX test the sensitivity and specificity were 97.56% and 98.53%, whereas for the Biovet assay 95.12% and 100% were recorded, respectively. The corresponding PV+ and PV− values were 98.77% and 97.1% (IDEXX), and 100% and 94.44% (Biovet). Our validation results showed that the 2 ELISAs worked equally well and there was no statistically significant difference between the performance of the 2 tests. Both tests showed high reproducibility, repeatability and substantial agreement with results from 2 other laboratories. A quality assurance based on the requirement of the ISO/IEC 17025 standards has been adopted throughout this project for test validation procedures.     

Experimental Infection of Lymnaea palustris snails with Fascioloides magna

This report describes experimental infection of Lymnaea (Galba) palustris. (Müller, 1774) snails with Fascioloides magna (Bassi, 1875). Of 725 L. palustris, 400 survived the experimental infection and in 172 (43%) the developmental cycle of the fluke was completed. Free emergence of cercariae was obtained was obtained 68 days after infection. The infectivity of metacercariae thus obtained was demonstrated in guinea-pigs, rabbits and a Cameroon goat. In sheep, the infection produced only changes in the organs, but no flukes could be recovered. Considering that L. palustris are found frequently in natural habitats in Czechoslovakia, they should be considered as possible intermediate hosts for F. magna.     

Virulence, cytotoxic and inflammatory activities of Vibrio anguillarum and Aeromonas salmonicida isolated from cultivated salmonid fish in Sweden

Extracellular products in culture filtrates of Aeromonas salmonicida subsp. achromogenes and Vibrio anguillarum isolated from infected fish have been shown to possess skin inflammatory factor. The extracellular products from Vibrio anguillarum were cytotoxic in HeLa and CHO cells. In addition to the skin lesions, the culture filtrates of V. anguillarum caused necrotic reaction on the rabbit skin. Five of 6 strains of V. anguillarum were lethal to mice after intraperitoneal administration of 3×10⁷ CFU. Only 1 strain of 5 A. salmonicida subsp. achromogenes produced extracellular products which elicited cytotoxic effects in the CHO cells. None of the A. salmonicida subsp. achromogenes strains were lethal to mice. The cytotoxins were inactivated when heated at 65°C for 30 min. The results indicate that the thermolabile exotoxins are non-enterotoxic since they failed to stimulate fluid accumulation in the rabbit ileal loop and did not cause elongation of the CHO cells. The rounding off of CHO cells, as well as of HeLa cells indicate that the exotoxins may play an important role in fish diseases.     

Babesia canis canis in dogs from Hungary: detection by PCR and sequencing

Canine babesiosis in Hungary has always been a severe and frequent disease, attributed to infection with Babesia canis transmitted by Dermacentor reticulatus. Identification of the disease agent has been based merely on size and morphology of the intraerythrocytic parasites and no evidence has been found concerning the subspecies (genotype) of B. canis. Therefore, a molecular survey on natural Babesia infection of dogs in Hungary using PCR and sequence analysis was attempted to clarify the subspecies (genotype) and to obtain information on the occurrence of B. canis. A total of 44 blood samples from dogs showing clinical signs of babesiosis were collected. A piroplasm-specific PCR amplifying the partial 18S rRNA gene yielded an approximately 450 bp PCR product in 39 (88.6%) samples. Thirteen positive samples originated from Budapest and 26 from 21 other locations. Five PCR products were chosen randomly for sequencing. The partial 18S rDNA sequences were submitted to GenBank (accession numbers AY611729; AY611730; AY611731; AY611732 and AY611733). The sequences showed 100% homology to one another or differed by one nucleotide. BLAST search against GenBank revealed the highest similarity (99.8 or 100%) with Babesia canis canis. The implication of these data, for the further study and diagnosis of canine babesiosis is discussed.