The association between oxidative processes and physiological responses has received much attention in ecotoxicity assessment. In the Baltic Sea, bloom-forming cyanobacterium Nodularia spumigena is a significant producer of various bioactive compounds, and both positive and adverse effects on grazers feeding in cyanobacteria blooms are reported. To elucidate the effect mechanisms and species sensitivity to the cyanobacteria-dominating diet, we exposed two Baltic copepods, Acartia bifilosa and Eurytemora affinis, to a diet consisting of toxin-producing cyanobacteria N. spumigena and a high-quality food Rhodomonas salina at 0-300 mu g C L-1; the control food was R. salina provided as a monodiet at the same food levels. The subcellular responses to food type and availability were assayed using a suite of biomarkers - antioxidant enzymes [superoxide dismutases (SOD), catalase (CAT), and glutathione S-transferases (GST)] and acetylcholinesterase (AChE). In parallel, we measured feeding activity using gut content (GC) assayed by real-time PCR analysis that quantified amounts of the prey DNA in copepod stomachs. As growth and reproduction endpoints, individual RNA content (a proxy for protein synthesis capacity), egg production rate (EPR), and egg viability (EV%) were used. In both toxic and nontoxic foods, copepod GC, RNA content, and EPR increased with food availability. Antioxidant enzyme activities increased with food availability regardless of the diet type. Moreover, CAT (both copepods), SOD, and GST (A. bifilosa) were upregulated in the copepods receiving cyanobacteria; the response was detectable when adjusted for the feeding and/or growth responses. By contrast, the diet effects were not significant when food concentration was used as a co-variable. A bimodal response in AChE was observed in A. bifilosa feeding on cyanobacteria, with up to 52% increase at the lower levels (5-25 mu g C L-1) and 32% inhibition at the highest food concentrations. These findings contribute to the refinement of biomarker use for assessing environmental stress and mechanistic understanding of cyanobacteria effects in grazers. They also suggest that antioxidant and AChE responses to feeding activity and diet should be accounted for when using biomarker profiles in field-collected animals in the Baltic Sea and, perhaps other systems, where toxic cyanobacteria are common.

Toxin-producing cyanobacteria can be harmful to aquatic biota, although some grazers utilize them with often beneficial effects on their growth and reproduction. It is commonly assumed that gut microbiota facilitates host adaptation to the diet; however, the evidence for adaptation mechanisms is scarce. Here, we investigated the abundance of mlrA genes in the gut of the Baltic copepods Acartia bifilosa and Eurytemora affinis during cyanobacteria bloom season (August) and outside it (February). The mlrA genes are unique to microcystin and nodularin degraders, thus indicating the capacity to break down these toxins by the microbiota. The mlrA genes were expressed in the copepod gut year-round, being >10-fold higher in the summer than in the winter populations. Moreover, they were significantly more abundant in Eurytemora than Acartia. To understand the ecological implications of this variability, we conducted feeding experiments using summer- and winter-collected copepods to examine if/how the mlrA abundance in the microbiota affect: (1) uptake of toxic Nodularia spumigena, (2) uptake of a non-toxic algal food offered in mixtures with N. spumigena, and (3) concomitant growth potential in the copepods. The findings provide empirical evidence that the occurrence of mlrA genes in the copepod microbiome facilitates nutrient uptake and growth when feeding on phytoplankton mixtures containing nodularin-producing cyanobacteria; thus, providing an adaptation mechanism to the cyanobacteria blooms.

Host-microbiome interactions are essential for the physiological and ecological performance of the host, yet these interactions are challenging to identify. Neurotransmitters are commonly implicated in these interactions, but we know very little about the mechanisms of their involvement, especially in invertebrates. Here, we report a peripheral catecholamine (CA) pathway involving the gut microbiome of the model species Daphnia magna. We demonstrate the following: (i) tyrosine hydroxylase and Dopa (3,4-dihydroxyphenylalanine) decarboxylase enzymes are present in the gut wall; (ii) Dopa decarboxylase gene is expressed in the gut by the host, and its expression follows the molt cycle peaking after ecdysis; (iii) biologically active _L-Dopa, but not dopamine, is present in the gut lumen; (iv) gut bacteria produce _L-Dopa in a concentration-dependent manner when provided l-tyrosine as a substrate. Impinging on gut bacteria involvement in host physiology and ecologically relevant traits, we suggest _L-Dopa as a communication agent in the host-microbiome interactions in daphnids and, possibly, other crustaceans.

IMPORTANCE Neurotransmitters are commonly implicated in host-microbiome communication, yet the molecular mechanisms of this communication remain largely elusive. We present novel evidence linking the gut microbiome to host development and growth via neurotransmitter _L-Dopa in Daphnia, the established model species in ecology and evolution. We found that both Daphnia and its gut microbiome contribute to the synthesis of the _L-Dopa in the gut. We also identified a peripheral pathway in the gut wall, with a molt stage-dependent dopamine synthesis, linking the gut microbiome to the daphnid development and growth. These findings suggest a central role of _L-Dopa in the bidirectional communication between the animal host and its gut bacteria and translating into the ecologically important host traits suitable for subsequent testing of causality by experimental studies.

Fossil-made polymers harbor unique bacterial assemblages, and concerns have been raised that ingested microplastic may affect the consumer gut microbiota and spread pathogens in animal populations. We hypothesized that in an ecotoxicity assay with a mixture of polystyrene (PS) and clay: (1) microbiome of the test animals inoculates the system with bacteria; (2) relative contribution of PS and the total amount of suspended solids (SS) select for specific bacterial communities; and (3) particle aggregation is affected by biofilm community composition, with concomitant effects on the animal survival. Mixtures of PS and clay at different concentrations of SS (10, 100, and 1000 mg/L) with a varying microplastics contribution (%PS; 0–80%) were incubated with Daphnia magna, whose microbiome served as an inoculum for the biofilms during the exposure. After 4-days of exposure, we examined the biofilm communities by 16S rRNA gene sequencing, particle size distribution, and animal survival. The biofilm communities were significantly different from the Daphnia microbiota used to inoculate the system, with an overrepresentation of predatory, rare, and potentially pathogenic taxa in the biofilms. The biofilm diversity was stimulated by %PS and decreased by predatory bacteria. Particle aggregate size and the biofilm composition were the primary drivers of animal survival, with small particles and predatory bacteria associated with a higher death rate. Thus, in effect studies with solid waste materials, ecological interactions in the biofilm can affect particle aggregation and support potentially harmful microorganisms with concomitant effects on the test animals.

It is a common view that an organism's microbiota has a profound influence on host fitness; however, supporting evidence is lacking in many organisms. We manipulated the gut microbiome of Daphnia magna by chronic exposure to different concentrations of the antibiotic Ciprofloxacin (0.01-1 mg L-1), and evaluated whether this affected the animals fitness and antioxidant capacity. In line with our expectations, antibiotic exposure altered the microbiome in a concentration-dependent manner. However, contrary to these expectations, the reduced diversity of gut bacteria was not associated with any fitness detriment. Moreover, the growth-related parameters correlated negatively with microbial diversity; and, in the daphnids exposed to the lowest Ciprofloxacin concentrations, the antioxidant capacity, growth, and fecundity were even higher than in control animals. These findings suggest that Ciprofloxacin exerts direct stimulatory effects on growth and reproduction in the host, while microbiome- mediated effects are of lesser importance. Thus, although microbiome profiling of Daphnia may be a sensitive tool to identify early effects of antibiotic exposure, disentangling direct and microbiome-mediated effects on the host fitness is not straightforward.

Increased harmful cyanobacterial blooms and drought are some negative impacts of global warming. To deal with cyanotoxin release during water treatment, and to manage the massive quantities of end-of-life membrane waste generated by desalination processes, we propose an innovative biological system developed from recycled reverse osmosis (RO) membranes to remove microcystins (MC). Our system, named the Recycled-Membrane Biofilm Reactor (R-MBfR), effectively removes microcystins, while reducing the pollution impact of RO membrane waste by prolonging their life span at the same time. This multidisciplinary work showed that the inherent flaw of RO membranes, i.e., fouling, can be considered an advantageous characteristic for biofilm attachment. Factors such as roughness, hydrophilic surfaces, and the role of calcium in cell-cell and cell-surface interactions, encouraged bacterial growth on discarded membranes. Biofilm development was stimulated by using a laboratory-scale membrane module simulator cell. The R-MBfR proved versatile and was capable of degrading 2 mg center dot L-1 of MC in 24 h. The economic feasibility of the scaling-up of the hypothetical R-MBfR was also validated. Therefore, this membrane recycling could be a future green cost-effective alternative technology for MC removal.

Reducing human impacts on drinking water is one of the main challenges for the water treatment industry. This work provides new results to support the recycling of EoL desalination reverse osmosis (RO) membranes for Membranes Biofilm Reactors (MBfRs). We investigate if the controlled-removal of fouling and polyamide layer may favor the use of these membranes in MBfRs. It also would allow establishing a normalized methodology of membrane recycling, regardless of inherited fouling during its lifespan. For this purpose, we transform by chlorination discarded brackish (BWd) and seawater (SWd) membranes into nanofiltration (BWt-NF and SWt-NF) and ultrafiltration (BWt-UF and SWt-UF) membranes. Our results show that chlorine attacks allow the fouling cleaning while improves the hydrophilicity and maintains roughness only in BWt-NF. Therefore, the bacterial deposition in this membrane is greater than the other tested membranes. Besides, the microcystin (MC) degradation capacity of BWt-NF verifies the compatibility of the chemical modification for the biological activity of MC-degrading bacteria. Finally, our results also provide that polyamide thin-film composite (PA-TFC) membranes, originally manufactured for salt rejection during desalination processes, offer competitive gases diffusion at low pressures. Therefore, we conclude that the membrane recycling may provide alternative low cost and gas permeable membranes for MBfRs, according to circular economy principles.

Examination of the diurnal patterns of basic cellular processes in the marine nonheterocystous diazotrophic cyanobacterium Trichodesmium revealed that the division of cells occurred throughout the diurnal cycle, but that it oscillated and peaked at an early stage in the dark period. Transcription of the early cell division gene ftsZ and the occurrence of the FtsZ protein showed a similar diurnal rhythmicity that preceded the division of cells. DNA replication (dnaA gene transcription) occurred before the transcription of ftsZ and hetR, the latter encoding the key heterocyst differentiation protein. Transcription of ftsZ and hetR in turn preceded the development of the nitrogen-fixing diazocytes and nifH transcription, and were at the minimum when diazotrophy was at the maximum. The nifH gene transcription showed a negative correlation to the circadian clock gene kaiC. Together, the data show a temporal separation between cell division and diazotrophy on a diurnal basis.

Cyanobacterial blooms in the Baltic Sea are a common phenomenon and are formed by the heterocystous, filamentous species Nodularia spumigena. The toxicity of these blooms is attributed to the hepatotoxin nodularin, produced by N. spumigena. Little is known regarding the regulatory mechanisms or environmental signaling that control nodularin production. Here we report the characterization of the transcriptional expression pattern of the nodularin synthetase gene cluster (nda) during phosphate depletion, and nitrogen supplementation. Real-time PCR analysis of these genes revealed that while cells continuously expressed the nda cluster, the expression of all nda genes increased when cells were subjected to phosphate depletion, and decreased in the presence of ammonium. In contrast to the shifts in expression, the intracellular and extracellular nodularin concentrations did not vary significantly during the treatments.