Visible intruders: Tracing (micro-) plastic in organic fertilizers.

Agricultural soils have been identified as potential reservoirs for plastic pollution, with adverse effects on soil properties. Primary sources of plastic input in agricultural landscapes are associated with the application of sewage sludge or compost. Understanding the sources and anticipated plastic content is crucial in mitigating plastic pollution in agricultural fields. This study presents one of the first investigations into the plastic content and other impurities, e.g. glass, of seven organic fertilizers (biowaste compost, digested pig slurry, sewage sludge compost, dry chicken manure, green waste compost, sewage sludge, and a mixed digestate comprising pig slurry, chicken manure, and 74 % renewable raw materials). Potentially visible foreign substances were assessed on the surface of each fertilizer pile. No impurities could be detected in digested pig slurry, chicken manure, and mixed digestate. For the remaining fertilizers, visible potential foreign substances were collected, cleaned, visually described, weighed, photographed, size measured, and chemically characterized using ATR-FTIR. The quantification revealed that plastic particles are the most abundant and are contained in all other fertilizers, in contrast to glass and metal. An increasing trend in plastic particle number per m2: green waste < biowaste < sewage sludge compost < sewage sludge, which is about 4 times greater in sewage sludge than in green waste compost, could be observed. However, sewage sludge compost has the largest plastic mass and surface area per square meter. This illustrates that sewage sludge compost application can be a significant entry pathway for visual plastics into agricultural soils.

Comparison of different salt solutions for density separation of conventional and biodegradable microplastic from solid sample matrices.

Microplastics are the new emerging pollutants ubiquitously detectable in aquatic and terrestrial ecosystems. Fate and behavior, as well as ecotoxicity, are of increasing environmental concern, particularly in sediments and soils as natural sinks. For a global environmental risk assessment, reliable and easy to apply analytical methods are mandatory to obtain comparable data. This is based on the isolation of microplastics out of the solid sample matrices prior to instrumental detection. Thus, this study provides an easy to apply approach for density separation. The technique emerged from a comparative study using different salt solutions to isolate conventional, and for the first time biodegradable, microplastics from different solid sample matrices, i.e., sand, artificial soil, and compost. Four solutions (water, sodium chloride, sodium hexametaphosphate, and sodium bromide) of different densities were applied followed by oxidizing digestion. Finally, the impact of the procedures on size and surface properties of microplastics was tested. Dependent on the sample matrix, the highest recovery rates of 87.3-100.3% for conventional polymers, and 38.2-78.2% for biodegradable polymers, were determined with sodium bromide. It could be shown that the type of solid sample matrix influences the recovery rates and has to be considered when choosing a sample preparation technique.

Recent Reticulate Evolution in the Ecologically Dominant Lineage of Coccolithophores.

The coccolithophore family Noëlaerhabdaceae contains a number of taxa that are very abundant in modern oceans, including the cosmopolitan bloom-forming Emiliania huxleyi. Introgressive hybridization has been suggested to account for incongruences between nuclear, mitochondrial and plastidial phylogenies of morphospecies within this lineage, but the number of species cultured to date remains rather limited. Here, we present the characterization of 5 new Noëlaerhabdaceae culture strains isolated from samples collected in the south-east Pacific Ocean. These were analyzed morphologically using scanning electron microscopy and phylogenetically by sequencing 5 marker genes (nuclear 18S and 28S rDNA, plastidial tufA, and mitochondrial cox1 and cox3 genes). Morphologically, one of these strains corresponded to Gephyrocapsa ericsonii and the four others to Reticulofenestra parvula. Ribosomal gene sequences were near identical between these new strains, but divergent from G. oceanica, G. muellerae, and E. huxleyi. In contrast to the clear distinction in ribosomal phylogenies, sequences from other genomic compartments clustered with those of E. huxleyi strains with which they share an ecological range (i.e., warm temperate to tropical waters). These data provide strong support for the hypothesis of past (and potentially ongoing) introgressive hybridization within this ecologically important lineage and for the transfer of R. parvula to Gephyrocapsa. These results have important implications for understanding the role of hybridization in speciation in vast ocean meta-populations of phytoplankton.

Seaweeds early development: detrimental effects of desiccation and attenuation by algal extracts.

The effects of desiccation on the early development stages of Mazzaella laminarioides, Scytosiphon lomentaria and Lessonia nigrescens, algal species with different patterns of distribution across the intertidal zone, were examined in the laboratory. In addition, the protective effect against desiccation was evaluated using algal extracts, including those from Porphyra columbina, a macroalga tolerant to desiccation that lives in the uppermost part of the intertidal zone. Our results showed that M. laminarioides displayed the highest resistance to daily desiccation, followed by S. lomentaria, whereas L. nigrescens was the most susceptible. Spores from L. nigrescens exposed to desiccation, although being able to germinate, ceased further post-germination development. In addition, our results showed that all species exposed to extracts from desiccated P. columbina successfully completed their development and strongly suggest the occurrence of compounds with protective properties that help in attenuating the stress caused by desiccation. Finally, our results indicate that the magnitude of the effects generated by desiccation on the early algal development is related to the position of the species in the intertidal zone, and that the protective effects of P. columbina extracts reveal an exceptional metabolism of this species under desiccation stress.

Tolerance to oxidative stress induced by desiccation in Porphyra columbina (Bangiales, Rhodophyta).

Unravelling the mechanisms underlying desiccation tolerance is crucial in order to understand the position of algal species in the intertidal zone. The alga Porphyra columbina lives in the uppermost part of the rocky intertidal zones around the world and was selected as a model for this study. Naturally desiccated plants were collected during low tide and studied for morphological changes, oxidative burst induction, biomolecule oxidation, antioxidant responses, and photosynthetic status. Naturally hydrated plants collected during high tides were used for comparative purposes. In addition, changes induced by desiccation were assessed in vitro and the capacity to recover from desiccation was determined by rehydrating the fronds in seawater. The global results show that desiccation induces morphological and cellular alterations accompanied by a loss of ∼96% of the water content. Overproduction of reactive oxygen species (ROS) was induced by desiccation and two peaks of H(2)O(2) were detected at 1 and 3 h of desiccation. However, during in vitro rehydration post-desiccation, the ROS quickly returned to the basal levels. At the biomolecular level, only a low production of oxidized proteins was recorded during desiccation, whereas the activity of diverse antioxidant enzymes increased. However, this activity diminished to near basal levels during rehydration. The photosynthetic efficiency (F(v)/F(m)) during desiccation declined by 94-96% of the values recorded in hydrated plants. This reduction was generated by the low levels of trapped energy flux per cross-section (TRo/CS), electron transport flux per CS (ETo/CS), and density of reaction centres (RC/SCo) as well as the chlorophyll content. The inverse pattern was observed for the levels of phycocyanin and phycoerythrin content. F(v)/F(m) and the photosynthetic indicators were restored to normal levels after only 5 min of rehydration. The results indicate that desiccation in P. columbina causes overproduction of ROS that is efficiently attenuated. The morphological and photosynthetic changes could be operating as tolerance mechanisms due to the fact that these responses principally prevent biomolecular alteration and cellular collapse. Thus, the activation of different physiological mechanisms helps to explain the high tolerance to desiccation of P. columbina and, at least in part, the position of this species at the highest level in the intertidal zone.

LAMINARIOCOLAX SP. (PHAEOPHYCEAE) ASSOCIATED WITH GALL DEVELOPMENTS IN LESSONIA NIGRESCENS (PHAEOPHYCEAE)(1).

As part of a long-term study on the biology and ecology of the intertidal kelp Lessonia nigrescens Bory, we report on the occurrence of gall development on this alga, identified the possible causal agent, and assessed the extent of the phenomenon in two wild stands of the host. Our results showed that galls affecting natural populations of L. nigrescens were associated with the infection by a filamentous brown algal endophyte of the genus Laminariocolax. Assignment to Laminariocolax of the endophytes isolated from cultured gall tissue was based on the (i) high internal transcribed spacer 1 (ITS1) sequence similarity and phylogenetic relationship between the Chilean isolates and several species of the genus Laminariocolax endophytic in other kelps, (ii) reproductive and vegetative features of the endophyte in culture, and (iii) anatomical agreement of fully developed galls of Lessonia with those described for other kelp galls caused by endophytic members of Laminariocolax. Unequivocal identification at the species level of the endophytes infecting Lessonia, however, awaits further studies.