Effects of the sunscreen ultraviolet filter oxybenzone (benzophenone-3) on the seagrass Posidonia oceanica (L.) Delile and its associated N2 fixers.

Oxybenzone/benzophenone-3 (BP-3) is one of the most detrimental organic ultraviolet filters for marine biota, leading to legislative measures banning its presence in commercial sunscreen formulations of several countries. It remains poorly explored how this contaminant is currently threatening the persistence of critical ecosystems for conservation in the Mediterranean, such as Posidonia oceanica meadows, but it is essential for promoting sustainable coastal tourism. Our investigation aimed to determine the effects of BP-3 on P. oceanica under a short-term laboratory setup, recreating summer conditions while testing three environmentally relevant concentrations for Mallorca, Spain (minimum: 53.6 ng L-1, maximum: 557.5 ng L-1 and increased: 1115 ng L-1) and a control (0 ng L-1). Primary productivity was unaffected by the treatments, however, a reduction in leaf chlorophyll content and nitrogen fixation activity associated with rhizomes was evidenced under BP-3 addition. This may be related with oxidative damage, as reactive oxygen species production and catalase activity in P. oceanica leaves were the highest even at minimum BP-3 concentrations. Alkaline phosphatase rates showed inverted trends between old leaves and rhizomes, being enhanced in the former under BP-3 addition and reduced in the latter. These results are of great relevance for the future management of P. oceanica meadows, elucidating that even minimum concentrations of BP-3 reported in coastal waters of Mallorca can induce elevated levels of oxidative stress in the seagrass, that lead to impairments in its photosynthetic pigments production and supply of essential nutrients through belowground tissues.

Sunscreen exposure interferes with physiological processes while inducing oxidative stress in seagrass Posidonia oceanica (L.) Delile.

The effects of a commercial sunscreen mixture on the Mediterranean seagrass Posidonia oceanica were investigated, evaluating its response in physiological processes and biochemical indicators of oxidative stress. Short-term laboratory experiments were conducted recreating summer conditions, and two sunscreen concentrations were tested in whole P. oceanica plants placed inside aquaria. Although primary productivity of leaf segments seemed to benefit from sunscreen addition, probably due to inorganic nutrients released, the rest of the biological parameters reflected possible impairments in the overall functioning of P. oceanica as a result of oxidative damages. Chlorophyll production and nitrogen fixation associated with old leaves were inhibited under high sunscreen concentrations, which concurred with elevated reactive oxygen species production, catalase activity and polyphenols content in the seagrass leaves. These results emphasize the importance of directing future investigations on determining which specific components of sunscreen products are likely threatening the wellbeing of critical species, such as P. oceanica.

Mediterranean seagrass Posidonia oceanica accumulates sunscreen UV filters.

Certain ultra-violet filter (UVF) components of solar creams have negative impacts on coral reefs and have been prohibited in international tourism destinations (i.e., Hawaii, Florida, and Palau) to protect coral reefs. In the Mediterranean coasts which are also hotspots of international tourism and where endemic seagrass Posidonia oceanica forms extensive meadows, the accumulation of UVF components have not been studied. We report for the first time, that the rhizomes of P. oceanica internally accumulated UVFs BP3, BP4, AVO, 4MBC and MeBZT and the paraben preservative MePB. The components BP4 and MePB occurred in higher concentrations reaching up to 129 ng g-1 dw and 512 ng g-1 dw, respectively. This work emphasizes the need for more experimental studies on the effects of UVFs on seagrasses and check if we should follow suit to prohibit certain UVFs to protect this species as what has been done in other regions to protect corals.

Everything Is Everywhere: Physiological Responses of the Mediterranean Sea and Eastern Pacific Ocean Epiphyte Cobetia Sp. to Varying Nutrient Concentration.

Bacteria are essential in the maintenance and sustainment of marine environments (e.g., benthic systems), playing a key role in marine food webs and nutrient cycling. These microorganisms can live associated as epiphytic or endophytic populations with superior organisms with valuable ecological functions, e.g., seagrasses. Here, we isolated, identified, sequenced, and exposed two strains of the same species (i.e., identified as Cobetia sp.) from two different marine environments to different nutrient regimes using batch cultures: (1) Cobetia sp. UIB 001 from the endemic Mediterranean seagrass Posidonia oceanica and (2) Cobetia sp. 4B UA from the endemic Humboldt Current System (HCS) seagrass Heterozostera chilensis. From our physiological studies, both strains behaved as bacteria capable to cope with different nutrient and pH regimes, i.e., N, P, and Fe combined with different pH levels, both in long-term (12 days (d)) and short-term studies (4 d/96 h (h)). We showed that the isolated strains were sensitive to the N source (inorganic and organic) at low and high concentrations and low pH levels. Low availability of phosphorus (P) and Fe had a negative independent effect on growth, especially in the long-term studies. The strain UIB 001 showed a better adaptation to low nutrient concentrations, being a potential N2-fixer, reaching higher growth rates (µ) than the HCS strain. P-acquisition mechanisms were deeply investigated at the enzymatic (i.e., alkaline phosphatase activity, APA) and structural level (e.g., alkaline phosphatase D, PhoD). Finally, these results were complemented with the study of biochemical markers, i.e., reactive oxygen species (ROS). In short, we present how ecological niches (i.e., MS and HCS) might determine, select, and modify the genomic and phenotypic features of the same bacterial species (i.e., Cobetia spp.) found in different marine environments, pointing to a direct correlation between adaptability and oligotrophy of seawater.

Differential Effects of Varying Concentrations of Phosphorus, Iron, and Nitrogen in N2-Fixing Cyanobacteria.

Diazotrophs or N2-fixers are one of the most ecologically significant groups in marine ecosystems (pelagic and benthic). Inorganic phosphorus (PO4 3-) and iron (Fe) can limit the growth and N2-fixing capacities of cyanobacteria. However, studies investigating co-limitation of these factors are lacking. Here, we added different concentrations of PO4 3- and Fe in two cyanobacterial species whose relatives can be found in seagrass habitats: the unicellular Halothece sp. (PCC 7418) and the filamentous Fischerella muscicola (PCC 73103), grown under different nitrate (NO3 -) concentrations and under N2 as sole N source, respectively. Their growth, pigment content, N2-fixation rates, oxidative stress responses, and morphological and cellular changes were investigated. Our results show a serial limitation of NO3 - and PO4 3- (with NO3 - as the primary limiting nutrient) for Halothece sp. Simultaneous co-limitation of PO4 3- and Fe was found for both species tested, and high levels of Fe (especially when added with high PO4 3- levels) inhibited the growth of Halothece sp. Nutrient limitation (PO4 3-, Fe, and/or NO3 -) enhanced oxidative stress responses, morphological changes, and apoptosis. Furthermore, an extensive bio-informatic analysis describing the predicted Pho, Fur, and NtcA regulons (involved in the survival of cells to P, Fe, and N limitation) was made using the complete genome of Halothece sp. as a model, showing the potential of this strain to adapt to different nutrient regimes (P, Fe, or N).

"The Good, the Bad and the Double-Sword" Effects of Microplastics and Their Organic Additives in Marine Bacteria.

Little is known about the direct effects of microplastics (MPs) and their organic additives on marine bacteria, considering their role in the nutrient cycles, e.g., N-cycles through the N2-fixation, or in the microbial food web. To fill this gap of knowledge, we exposed marine bacteria, specifically diazotrophs, to pure MPs which differ in physical properties (e.g., density, hydrophobicity, and/or size), namely, polyethylene, polypropylene, polyvinyl chloride and polystyrene, and to their most abundant associated organic additives (e.g., fluoranthene, 1,2,5,6,9,10-hexabromocyclododecane and dioctyl-phthalate). Growth, protein overproduction, direct physical interactions between MPs and bacteria, phosphorus acquisition mechanisms and/or N2-fixation rates were evaluated. Cyanobacteria were positively affected by environmental and high concentrations of MPs, as opposed to heterotrophic strains, that were only positively affected with high concentrations of ~120 µm-size MPs (detecting the overproduction of proteins related to plastic degradation and C-transport), and negatively affected by 1 µm-size PS beads. Generally, the organic additives had a deleterious effect in both autotrophic and heterotrophic bacteria and the magnitude of the effect is suggested to be dependent on bacterial size. Our results show species-specific responses of the autotrophic and heterotrophic bacteria tested and the responses (beneficial: the "good," deleterious: the "bad" and/or both: the "double-sword") were dependent on the type and concentration of MPs and additives. This suggests the need to determine the threshold levels of MPs and additives concentrations starting from which significant effects can be observed for key microbial populations in marine systems, and these data are necessary for effective environmental quality control management.