Capítulo 3. Hacia una definición de la vulnerabilidad territorial desde la cooperación internacional
3.2 Usos del término Vulnerabilidad en relación con el territorio y la cooperación internacional
3.2.3 Vulnerabilidad territorial frente a amenazas naturales
Colloidal characteristics of NP have the most important influence on their distribution in the (aquatic) environment (Wiesner et al. 2009). These properties determine whether NP build stable dispersions, whether they agglomerate or adsorb to surfaces and other particles, or whether they tend to decompose and release potentially critical substances such as toxic ions (Kahru et al. 2008, Navarro et al. 2008a). The colloidal stability therefore determines exposure routes and interactions at the bio-nano interface (Rivera-Gil et al. 2013).
The colloidal properties of NP highly depend on their reactivity, their surface chemistry and the composition of the dispersion medium (Handy et al. 2008a). Usually NP are supplied with
a surface coating which serves to increase the colloidal stability to maintain their nano- specific characteristics in dispersion (Batley et al. 2013, Nowack & Bucheli 2007). NP can be (colloidally) stabilized by increasing their charge to achieve electrostatic repulsion, or they can be surrounded by a sterically stabilizing shell, which suppresses direct contact between the NP (Segets et al. 2011, Studart et al. 2007).
IONP with different coatings were synthesized by Darius Arndt and tested on their colloidal properties in different media including the Daphnia medium Elendt M7. The four colloidally most stable IONP were chosen for ecotoxicological investigations with D. magna. These IONP were present in the water column at least for several days or even months, guaranteeing high exposure potential to the pelagic filter feeding daphnids. One set of IONP was stabilized via electrostatic repulsion with ascorbate (ASC) and citrate (CIT). Sterically stabilized IONP were functionalized with dextrane (DEX) and polyvinylpyrrolidone (PVP). First, the acute toxicity of the four different IONP was investigated over a period of 96 h. Since (negatively) charged NP were already shown to be more (cyto-) toxic than less or neutrally charged NP due to increased reactivity and the potential to form more reactive oxygen species (ROS) (Lee et al. 2013a, Park et al. 2013, Schaeublin et al. 2011), ASC- and CIT-IONP were expected to be more toxic than DEX- and PVP-IONP. However, the greatest immobilization and toxicity was found for ASC- and DEX-IONP which was attributed to an increased agglomeration in the tests. Although ASC/DEX-IONP were tested to be colloidally stable, they quickly agglomerated in the presence of daphnids. It was hypothesized that the colloidal destabilization was related to turbulences induced by the filtering and swimming movement of the daphnids. The agglomeration and adsorption of IONP disturbed the molting of the neonates. The incomplete ecdysis often immobilized the neonates, but usually did not cause their death. Comparable agglomeration of both IONP in the Elendt M7 stocks was only observed after one year. Most frequent cases of incomplete ecdysis were induced by the colloidally instable ASC- and DEX-IONP. IONP possibly adsorbing to the carapace were obviously able to disturb the molting process. This effect was also less frequently observed for CIT-IONP and did not occur for PVP-IONP.
In CIT-IONP additional release of iron ions was measured. However, toxicity could not exclusively be related to free iron since release rates were too low. Toxicity was hypothesized to also originate from the possible formation of ROS. For ASC- and CIT-IONP Arndt et al. (2012) had shown increased production of ROS.
PVP-IONP did not have any significant harmful effects within 96 h at concentrations of up to 100 mg iron L-1. PVP-IONP had the highest long-term stability without any agglomeration or
prevented the IONP from agglomeration (Studart et al. 2007). These IONP are therefore a very promising tool to investigate long-term NP effects without the influence of effects from released (toxic) ions or decreasing colloidal stability.
Consequently, PVP-IONP were chosen for the investigation of the long-term effects of IONP on D. magna. The chronic test aimed to investigate sub-lethal endpoints such as inhibition of reproduction and growth over 21 days. By chance, one batch of PVP-IONP synthesized for the tests showed decreased colloidal stability with slow agglomeration. Although this effect was not desired, it opened up the opportunity to compare long-term effects of equal IONP with different colloidal properties.
Both IONP induced increased mortality at concentrations ≥ 25 mg iron/L, but with stronger effects of instable IONP. Many or even all daphnids died after 5 to 8 days, which indicates that acute tests might not be sufficient for the testing of NM since effects are often delayed compared to those of diluted chemicals. Also the reproduction as well as the development (growth) was significantly reduced under exposure to both IONP. However, stable IONP only induced significant effects at 100 mg iron/L, instable IONP already affected the life history responses of daphnids significantly at 1 mg iron/L.
When NP are present, they are very effectively concentrated by daphnids in their digestive tracts (see above). NP composed of non-toxic materials (such as IONP) might also significantly decrease nutrient uptake (Mendonca et al. 2011) by physically clogging the digestive tract and slowing down the gut passage (Campos et al. 2013). In consequence, many life history effects and even increased mortality observed under chronic IONP exposure should be related to the starving of daphnids as a consequence of insufficient nutrient uptake. For a complete understanding of effects and to confirm the developed hypotheses the accumulation of (IO)NP in daphnids has to be investigated in more detail. Investigations of the intestines of exposed daphnids with histological methods might help to prove the assumptions of disturbed nutrient exchange in the daphnids’ gut.
The increased effects of colloidally instable IONP were explained by the additional flocculation of algae, which were administered as food. The algae flocs were concentrated between the filtering legs of the daphnids, partly completely inhibiting their locomotion. This greatly increased physiological stress, which may have even been amplified by the reduced food acquisition. Furthermore, respiration might also have been affected sicne the filtering current also serves respiration. It was hypothesized that the filtering movements and the associated water current accelerated the release of PVP from the IONP. Released PVP, colloidally instable PVP-IONP or the mixture of both interacted with the algae forming the visible algae agglomerates. However, since the observed effects might exclusively be related
to PVP, more research on the interaction of algae and i-IONP would allow a better understanding of this phenomenon.