Zooplankton taxa repository of DDTtot and sumPCB14: Seasonal and decadal variations in Lake Maggiore.

Lake Maggiore has been the subject of a monitoring program on persistent organic pollutants (DDTs and PCBs) since 1996 when DDT contamination was first detected. In this context, in 2009 we started to estimate the concentration of DDTtot (Dichlorodiphenyltrichloroethane, sum of p,p'-DDT, o,p'-DDT, p,p'-DDD, o,p-DDD, p,p-DDE, o,p-DDE) and sumPCB14 (polychlorinated biphenyls 18, 28, 31, 44, 52, 101, 118, 149, 138, 153, 170, 180, 194 and 209) in zooplankton pelagic organisms preyed on by zooplanktivorous fish (size fraction ≥450 µm). We evaluated taxa specific repositories of DDTtot and sumPCB14, their seasonal variation, their changes over the period 2009-2021 and the potential contribution of different taxa in transferring toxicants to whitefish, based on the Ivlev's Electivity Index. The repository of both POPs was generally higher in spring. A decrease in the zooplankton Standing Stock Biomass (SSB) drove a decline in the zooplankton DDTtot repository over the last six years (2016-2021 SSBmean = 12.5 mg m-3; 2009-2015; SSBmean = 30 mg m-3), despite the concentration being broadly constant during this period. The sumPCB14 repository was generally characterized by lower values during the last six years, but the difference with the previous period was not so marked. Daphnia and cyclopoids were the major contributors to the repository; however, when the whitefish selectivity index was applied, the role of carnivorous Bythotrephes was more important to the detriment of cyclopoids, particularly in summer and winter. Our results are useful to elaborate predictive models on the transfer of POPs along the food chain and highlight not only the importance of freshwater zooplankton in toto, but also that different taxa can have different roles. The increasing importance of microphagous zooplankton, driven by water warming and extended thermal stratification, underlines the need for future studies on the role of small zooplankton as carriers of POPs in freshwater lakes.

Seasonal and plurennial changes of POPs repository in freshwater zooplankton: A 10-year study in the large deep subalpine Lake Maggiore (Italy).

Determining the concentration of POPs (Persistent Organic Pollutants) such as total dichlorodiphenyltrichloroethane (DDTtot) and polychlorinated biphenyls (PCBs) in the zooplankton is essential for evaluating the level of toxicity in the environment and the risk to ecosystem health. The extent to which POPs persist in the environment and affect human health depends on accumulation in the different components of the zooplankton community such as particle feeders and/or predators and their subsequent consumption by fish, which varies seasonally. We analyzed ten-years (2011-2020) of seasonal data on POPs in the crustacean zooplankton community of Lake Maggiore (Italy). The concentration of total DDTs, as sum of all congeners ranged between 23.5 and 156.9 ng/g dry weight, while sumPCB14 ranged from 24.4 and 133.6 ng/g dry weight. We traced particle feeders' vs predatory taxa within the zooplankton community using carbon and nitrogen stable isotopic values. POPs concentration was significantly related to the nitrogen isotopic values. The zooplankton biomass increase in Spring was important, both through this being a period of increased metabolism and activity of planktivorous fish and also as it was the period when the zooplankton repository of POPs was at a maximum. DDTtot stocked in zooplankton ≥450 µm was up to 275 ng/m2, while sumPCB14 was 285 ng/m2 in 2013. A decrease of DDTtot stocked in the zooplankton repository characterized the last five years examined, when mean annual values (14.8-56.9 ng/m2) were less than half those of the previous period (50.7-128.6 ng/m2).

Ten Rules for the Management of Moderate and Severe Traumatic Brain Injury During Pregnancy: An Expert Viewpoint.

Moderate and severe traumatic brain injury (TBI) are major causes of disability and death. In addition, when TBI occurs during pregnancy, it can lead to miscarriage, premature birth, and maternal/fetal death, engendering clinical and ethical issues. Several recommendations have been proposed for the management of TBI patients; however, none of these have been specifically applied to pregnant women, which often have been excluded from major trials. Therefore, at present, evidence on TBI management in pregnant women is limited and mostly based on clinical experience. The aim of this manuscript is to provide the clinicians with practical suggestions, based on 10 rules, for the management of moderate to severe TBI during pregnancy. In particular, we firstly describe the pathophysiological changes occurring during pregnancy; then we explore the main strategies for the diagnosis of TBI taking in consideration the risks related to mother and fetus, and finally we discuss the most appropriate approaches for the management in this particular condition. Based on the available evidence, we suggest a stepwise approach consisting of different tiers of treatment and we describe the specific risks according to the severity of the neurological and systemic conditions of both fetus and mother in relation to each trimester of pregnancy. The innovative feature of this approach is the fact that it focuses on the vulnerability and specificity of this population, without forgetting the current knowledge on adult non-pregnant patients, which has to be applied to improve the quality of the care process.

Causal networks of phytoplankton diversity and biomass are modulated by environmental context.

Untangling causal links and feedbacks among biodiversity, ecosystem functioning, and environmental factors is challenging due to their complex and context-dependent interactions (e.g., a nutrient-dependent relationship between diversity and biomass). Consequently, studies that only consider separable, unidirectional effects can produce divergent conclusions and equivocal ecological implications. To address this complexity, we use empirical dynamic modeling to assemble causal networks for 19 natural aquatic ecosystems (N24◦~N58◦) and quantified strengths of feedbacks among phytoplankton diversity, phytoplankton biomass, and environmental factors. Through a cross-system comparison, we identify macroecological patterns; in more diverse, oligotrophic ecosystems, biodiversity effects are more important than environmental effects (nutrients and temperature) as drivers of biomass. Furthermore, feedback strengths vary with productivity. In warm, productive systems, strong nitrate-mediated feedbacks usually prevail, whereas there are strong, phosphate-mediated feedbacks in cold, less productive systems. Our findings, based on recovered feedbacks, highlight the importance of a network view in future ecosystem management.

Climate warming restructures an aquatic food web over 28 years.

Climate warming can restructure lake food webs if trophic levels differ in their thermal responses, but evidence for these changes and their underlying mechanisms remain scarce in nature. Here we document how warming lake temperatures by up to 2°C, rather than changes in trophic state or fishing effort, have restructured the pelagic food web of a large European lake (Lake Maggiore, Italy). Our approach exploited abundance and biomass data collected weekly to yearly across five trophic levels from 1981 to 2008. Temperature generally had stronger effects on taxa than changes in fish predation or trophic state mediated through primary productivity. Consequently, we found that, as the lake warmed, the food web shifted in numerical abundance towards predators occupying middle trophic positions. Of these taxa, the spiny water flea (Bythotrephes longimanus) most prospered. Bythotrephes strongly limited abundances of the keystone grazer Daphnia, strengthening top-down structuring of the food web. Warmer temperatures partly restructured the food web by advancing peak Bythotrephes densities by approximately 60 days and extending periods of positive population growth by three times. Nonetheless, our results suggested that advances in the timing and size of peak Bythotrephes densities could not outpace changes in the timing and size of peak densities in their Daphnia prey. Our results provide rare evidence from nature as to how long-term warming can favour higher trophic levels, with the potential to strengthen top-down control of food webs.

Trophic transfer of persistent organic pollutants through a pelagic food web: The case of Lake Como (Northern Italy).

Despite DDT and PCB having been banned for about 40 years, they are still detectable in the environment. In the present research we specifically investigated the trophic transfer of these organochlorine contaminants (OC) through a pelagic food web of a deep lake in Northern Italy (Lake Como) over time. Zooplankton and fish were sampled each season of a year and OC concentrations and the carbon and nitrogen isotopic ratios were measured. By using stable isotopes, the direct trophic relationship between pelagic zooplankton and zooplanktivorous fish was confirmed for Alosa agone only in summer. Based on this result, the biomagnification factor normalized on the trophic level (BMFTL) for organic contaminants was calculated. BMFTL values were within the range 0.9-1.9 for DDT isomers and 1.6-4.9 for some PCB congeners (PCB 95, PCB 101, PCB 149, PCB 153, PCB 138 - present both in zooplankton and in fish and representing >60% of the PCB contamination), confirming the biomagnification of these compounds in one of the two zooplanktivorous fish species of the lake.

Plankton dynamics across the freshwater, transitional and marine research sites of the LTER-Italy Network. Patterns, fluctuations, drivers.

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.

Lifetime response of contemporary versus resurrected Daphnia galeata Sars (Crustacea, Cladocera) to Cu(II) chronic exposure.

Resurrecting legacy lineages of organisms from sediment cores of known geological age allows us to understand how environmental change can cause selection pressures that constrain the variation of populations over time. We quantified the lifetime tolerance and effects of Cu(II) exposure on Daphnia galeata in a polluted subalpine lake by comparing extant individuals with those resurrected from ephippia extracted from ca. 30-years-old sediments. Laboratory experiments were conducted using two Cu(II) concentrations, 40 and 10 µg L(-1), corresponding to the levels recorded in the lake, during chemical recovery, when Daphnia first re-appeared and succeeded. Contemporary Daphnia were unable to survive after the 10th day at either of the Cu(II) concentrations, and were unable to successfully reproduce. Daphnia cohorts from the past performed better in low Cu(II) concentrations than in copper-free, control conditions. The copper-adapted, tolerant Daphnia strains grew faster under non-toxic conditions, but were unable to survive new pollution events.

Mitochondrial capture misleads about ecological speciation in the Daphnia pulex complex.

The North American ecological species Daphniapulicaria and Daphniapulex are thought to have diverged from a common ancestor by adaptation to sympatric but ecologically distinct lake and pond habitats respectively. Based on mtDNA relationships, European D. pulicaria is considered a different species only distantly related to its North American counterpart, but both species share a lactate dehydrogenase (Ldh) allele F supposedly involved in lake adaptation in North America, and the same allele is also carried by the related Holarctic Daphniatenebrosa. The correct inference of the species' ancestral relationships is therefore critical for understanding the origin of their adaptive divergence. Our species tree inferred from unlinked nuclear loci for D. pulicaria and D. pulex resolved the European and North American D. pulicaria as sister clades, and we argue that the discordant mtDNA gene tree is best explained by capture of D. pulex mtDNA by D. pulicaria in North America. The Ldh gene tree shows that F-class alleles in D. pulicaria and D. tenebrosa are due to common descent (as opposed to introgression), with D. tenebrosa alleles paraphyletic with respect to D. pulicaria alleles. That D. tenebrosa still segregates the ancestral and derived amino acids at the two sites distinguishing the pond and lake alleles suggests that D. pulicaria inherited the derived states from the D. tenebrosa ancestry. Our results suggest that some adaptations restricting the gene flow between D. pulicaria and D. pulex might have evolved in response to selection in ancestral environments rather than in the species' current sympatric habitats. The Arctic (D. tenebrosa) populations are likely to provide important clues about these issues.

Seasonal fluctuations of DDTs and PCBs in zooplankton and fish of Lake Maggiore (Northern Italy).

Concentrations of DDTs and PCBs were determined in the zooplankton and in three different fish species (shad, whitefish and roach) collected seasonally during 2009 and 2010 in three sites in Lake Maggiore, a south-alpine lake that has been contaminated by DDT since 1996. As previously observed in 2008, even during 2009 DDTs concentrations were higher in zooplankton than in fish, probably due to the very unstable situation of the lake still influenced by local inputs. The situation changed in 2010, when all DDT compounds increased in fish to levels much higher than those measured in zooplankton. Biomagnification was statistically demonstrated for pp'DDE in all the three fish species, indicating a probable signal of recovery of the lake. Although with respect to total PCBs we observed that the contamination levels varied across time periods and across fish species, biomagnification was evident from zooplankton to fish both in 2009 and in 2010. As concern individual PCBs, biomagnification from zooplankton to all three fish species was significant for PCB 153 and PCB 138.

Reticulate evolution of the Daphnia pulex complex as revealed by nuclear markers.

The study of species complexes is of particular interest to understand how evolutionary young species maintain genomic integrity. The Daphnia pulex complex has been intensively studied as it includes species that dominate freshwater environments in the Northern hemisphere and as it is the sole North American complex that shows transitions to obligate parthenogenesis. Past studies using mitochondrial markers have revealed the presence of 10 distinct lineages in the complex. This study is the first to examine genetic relationships among seven species of the complex at nuclear markers (nine microsatellite loci and one protein-coding gene). Clones belonging to the seven species of the Daphnia pulex complex were characterized at the mitochondrial NADH dehydrogenase (ND5) gene and at the Lactate dehydrogenase (LDH) locus. K-means, principal coordinate analyses and phylogenetic network analyses on the microsatellite data all separated European D. pulicaria, D. tenebrosa, North American D. pulex, D. pulicaria and their hybrids into distinct clusters. The hybrid cluster was composed of diploid and polyploid hybrids with D. pulex mitochondria and some clones with D. pulicaria mitochondria. By contrast, the phylogeny of the D. pulex complex using Rab4 was not well resolved but still showed clusters consisting mostly of D. pulex alleles and others of D. pulicaria alleles. Incomplete lineage sorting and hybridization may obscure genetic relationships at this locus. This study shows that hybridization and introgression have played an important role in the evolution of this complex.

Spatial, environmental and anthropogenic effects on the taxon composition of hybridizing Daphnia.

The competitive ability of hybrids, compared with their parental taxa, can cover a wide fitness range from poor to superior. For example communities of the Daphnia galeata-hyalina-cucullata species complex often show hybrid dominance. We tested whether taxa composition of 43 European lakes inhabited by this species complex can be explained by habitat characteristics (e.g. size descriptors, trophy level) or geography. We found that D. galeata occurs more frequently south of the Alps, whereas D. hyalina and D. cucullata are found more in the north. Lakes with D. galeata dominance had higher temperatures whereas D. hyalina dominance could be attributed to low phosphorus loads. The dominance of F1-hybrids, however, was not explainable with current environmental variables. In a subset of 28 lakes, we studied the impact of eutrophication history on F-hybrid success. Lakes with the highest trophic state in the past tended to be dominated by F1-hybrids. Our data demonstrate that human-mediated habitat disturbance (eutrophication) has facilitated hybrid success and altered the Daphnia taxon composition across lakes. At the same time, specific habitat conditions might provide a refuge from hybridization for native genotypes.

Parasite survey of a Daphnia hybrid complex: host-specificity and environment determine infection.

1. Hybridization between species is a common phenomenon in plants and animals. If parasite prevalence differs for hybrids and parental species (i.e. taxa) there may be considerable consequences for relative hybrid fitness. Some studies have investigated hybrid complexes for infection, and complex-specific differences in parasite prevalence have been detected. 2. Based on the results of a field study on a hybridizing Daphnia population from a single lake, it has been hypothesized that permanently over- or under-infected hybrids do not exist. The observed field-patterns can only be temporal because taxa, in addition to single genotypes, might be the subject of parasite driven host frequency-dependent selection. Thus, parasites will track any common taxon within a hybrid complex. 3. In the present study, hybridizing Daphnia populations from 43 lakes were screened for parasite infections to obtain indirect evidence for coevolutionary cycles. It was hypothesized that, due to time lags between the evolution of resistance in host populations and the evolution of the parasite towards tracking of a common host taxon, the same Daphnia taxon will be over-infected in some lakes, while being under-infected in others. 4. Two of the four parasite species were specialists: their prevalence differed among coexisting Daphnia taxa. The varying infection patterns detected across spatially segregated hybridizing Daphnia populations are consistent with theoretical predictions for coevolutionary cycles. Thus the infection patterns, as observed under natural conditions, are temporal and unstable. 5. Additionally, the spatial distribution of the four parasite species was analysed with respect to habitat differences. The results show that the presence of a particular parasite on a host taxon was determined not only by the host-specificity of the parasite, but also by host-habitat relations.

Invasion genetics of the Eurasian spiny waterflea: evidence for bottlenecks and gene flow using microsatellites.

The Eurasian spiny waterflea (Bythotrephes longimanus) is a predacious zooplankter that has increased its range in Europe and is rapidly invading inland water-bodies throughout North America's Great Lakes region. To examine the genetics of these invasions, we isolated five microsatellite DNA loci with between 5 and 19 alleles per locus. We sampled three populations where B. longimanus has been historically present (Switzerland, Italy, and Finland) as well as an introduced European population (the Netherlands) and three North American populations (Lakes Erie, Superior, Shebandowan). Consistent with a bottleneck during colonization (i.e. founder effect), average heterozygosities of the four European populations ranged from 0.310 to 0.599, and were higher than that of three North American populations (0.151-0.220). Pairwise F(ST) estimates among North American populations (0.002-0.063) were not significantly different from zero and were much lower than among European populations (0.208-0.474). This is consistent with a scenario of high gene flow among North American populations relative to that of European ones. Contrary to an invasion bottleneck, however, Erie and Superior populations contained similar numbers of rare alleles as European populations. Assignment tests identified several migrant genotypes in all introduced populations (the Netherlands, Erie, Superior, Shebandowan), but rarely in native ones (Switzerland, Italy and Finland). A large number of genotypes from North America were assigned to our Italian population suggesting a second, previously unidentified, invasion source somewhere in the region of northern Italy. Together, our results support an invasion bottleneck for North American populations that has been largely offset by gene flow from multiple native sources, as well as gene flow among introduced populations.