Assemblage structure and secondary production of mesozooplankton in shallow water volcanic CO2 vents of the Azores

Dissertação de mestrado, Biologia Marinha, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015Atmospheric CO2 concentrations have increased by at least 30% since pre-industrial times due to human activities. Part of this CO2 has been absorbed by oceans, inducing ocean acidification and, therefore, several impacts in the marine biota. Natural shallow-water CO2 vents have generated a substantial interest in recent years as in situ laboratories for ocean acidification studies. The present study was focused on the effects of ocean acidification on mesozooplankton communities. Two active volcanic areas with submarine CO2 emissions, in the islands of São Miguel and Faial, were chosen and independently studied, through the choice of three different sites: Reference, Intermediate and Vent, characterized for their increase in the CO2 degassing activity, and consequent pH decrease. Differences in the abundance, diversity and structure of mesozooplankton among sites were described, along with the characterization of the community in these areas, since previous information is scarce. Differences were found in the composition of the zooplanktonic assemblages among sites in São Miguel and among dates in Faial. Through the nMDS analyses, Radiozoa, Paracalanus parvus and Evadne spinifera appeared more related to vent conditions, while Cirripedia nauplii were closer to the reference conditions. Conditions in Faial did not allow a clear separation among sites in the nMDS analyses, since the strong currents mix the water, dissipating the effect of the gas emissions, and variation among days becomes more important and statistically significant. The RNA:DNA ratio of selected mesozooplankton populations was used, as proxies for physiological condition. RNA:DNA did not show a clear pattern of variation, copepods in São Miguel and cladocerans in Faial had an higher ratio in the reference sites, but with no gradual decrease to the Vent. Fish eggs seem to have an inverted pattern. Additional experiments conducted under natural conditions were performed to determine the egg production rate (EPR) of the dominant free-spawning copepod species, as a proxy for secondary production. In São Miguel, the EPR showed a decreasing trend along the CO2 gradient, with no differences between the exposure times. In Faial, EPR was higher in the reference, but it did not differ significantly from the other sites. This study demonstrated the suitability of the submarine degassing sites in S. Miguel and Faial Islands for investigating the effect of future dissolved CO2 levels in planktonic and pelagic communities of the NE Atlantic. It also provided the first in-situ evidence of a significant decrease of EPR of copepods under near future CO2 levels.Desde os tempos pre-industriais que se registou um incremento do CO2 atmosférico, de pelo menos 30% devido a atividades humanas, principalmente pelo uso de combustíveis fósseis. Os oceanos têm absorvido parte deste CO2 através da sua capacidade de “sumidouro de carbono”, induzindo mudanças na composição química da água do mar (acidificação do oceano), com potenciais impactos nos organismos marinhos. As possíveis consequências ecológicas deste processo têm motivado um incremento no esforço de investigação nos últimos anos. A maioria dos efeitos nos organismos marinhos tem sido observada em experiências no laboratório, onde o CO2 é manipulado em escalas temporais curtas. Não obstante, a necessidade de usar diferentes abordagens, como experiências de campo, em ambientes marinhos específicos que contenham comunidades, mais do que espécies isoladas, tornou-se evidente. Locais de desgaseificação natural de CO2 em águas pouco profundas têm gerado um interesse substancial como laboratórios in situ para estudos de acidificação do oceano. Alterações significativas na estrutura da comunidade bentónica têm sido associadas com estes locais de desgaseificação, mas os efeitos no plâncton permanecem largamente desconhecidos. O presente estudo analisou os efeitos da acidificação do oceano nas comunidades de mesozooplâncton. Os objetivos especificos foram (i): o estudo observacional sobre as comunidades zooplanctónicas, para determinar se existem diferenças consistentes na abundância, diversidade e composição entre os locais afetados por emissões de CO2 e os locais de controlo, sem emissões; e (ii) a relação entre os padrões observados no zooplâncton em relação aos resultados detetados na química da água do mar e do gás das emissões, (iii) a descrição do estado ecofisiológico de grandes grupos de zooplâncton utilizando índices derivados dos ácidos nucleicos e índices de produção de ovos e a sua relação com o possível stress causado pela acidificação. Para atingir estes objetivos foram seleccionadas duas áreas vulcânicas activas, com emanações de CO2 submarinas, uma na ilha de São Miguel e outra na ilha de Faial, no arquipélago dos Açores. Cada ilha foi independentemente estudada, através da escolha de três lugares diferentes: Reference, Intermediate e Vent, ao longo de um gradiente na emissão de CO2, e consequente diminuição de pH. O pH em São Miguel variou de 8.06 no Reference (pCO2 383.80 μatm) a 7.75 no Vent (pCO2 983.10 μatm), enquanto que no Faial diminuiu de 8.13 no Reference (pCO2 319.37 μatm) a 7.88 no Vent (pCO2 894.08 μatm). O CO2 foi o principal constituinte das emissões de gás nas duas ilhas (acima de 98%), minimizando o efeito de outros gases como H2S. Em São Miguel o mesozooplâncton foi amostrado usando arrastos oblíquos com uma rede WP2 de Ø60cm e malhagem de 200 μm, entre os dias 3 e 7 de Julho de 2014, enquanto que no Faial foi usada uma rede com malhagem de 500μm e a amostragem foi feita entre os dias 10 e 15 de Julho de 2014. Foram descritas as diferenças na abundância, diversidade e estrutura do mesozooplâncton entre locais, em paralelo com a caracterização das comunidades nestas áreas, já que a informação prévia era escassa. Foram identificados organismos de nove filos, num total de 71 taxa diferentes, 45 em S. Miguel e 61 no Faial. Em São Miguel foram encontradas diferenças entre locais na abundância total e nas abundâncias de Arthropoda e Chordata, com um incremento no Vent, relativamente ao Intermediate e o Reference. No Faial foram apenas encontradas diferenças na diversidade dos Arthropoda. A análise PERMANOVA ilustrou as diferenças na composição do zooplâncton entre locais, em São Miguel, e entre datas, no Faial. Os principais contribuintes para estas diferenças, realçados pelas anaáises SIMPER, foram Evadne spinifera, Paracalanus parvus, nauplios de Cirripedia e Radiozoa. Através das análises nMDS, Radiozoa, Paracalanus parvus e Evadne spinifera apareceram mais relacionados às condições do Vent, enquanto que nauplios de Cirripedia estiveram mais perto das condições do Reference. No Faial, os principais contribuintes foram Bassia bassensis, Radiozoa e efiras de Scyphozoa. As condições nesta ilha não permitiram uma separação entre locais nas análises nMDS, dado que as fortes correntes misturaram a coluna de àgua, dissipando o efeito das emissões de gases, e a variação entre dias é aparentemente mais importante e estatisticamente significativa. O racio RNA:DNA de populações de mesozooplâncton seleccionadas foi usado como indicador da condição fisiológica. O RNA:DNA não mostrou um padrão claro de variação: Copepoda em São Miguel e Cladocera no Faial apresentaram valores mais elevados no Reference, mas este não diminuiu de forma gradual ao longo do gradiente de CO2. Os ovos de peixe parecem ter um padrão invertido. Experiências adicionais foram levados a cabo para determinar a taxa de produção de ovos (EPR) das espécies dominantes de copépodes, como indicador da produção secundária. As câmaras de incubação foram colocadas perto do fundo durante períodos de 24 e 72h em São Miguel, e 72h no Faial. Em São Miguel, a EPR mostrou uma tendência de diminuição ao longo do gradiente de CO2, sem diferenças nos tempos de exposição. No Faial, EPR foi maior no Reference, mas não diferiu significativamente dos outros locais. Este estudo demonstrou o potencial das zonas de desgaseificação submarina nas ilhas de S. Miguel e Faial para a investigação dos efeitos dos níveis futuros de CO2 dissolvido nas comunidades planctónicas e pelágicas do Atlântico NE. Este trabalho também permitiu registar a primeira evidência in-situ de uma diminuição significativa da produção secundária em copépodes sob concentrações de CO2 previstas para o final do século

The origin and correlated evolution of symbiosis and coloniality in Scleractinian corals

Symbiosis and coloniality are ecologically important traits for corals of the order Scleractinia. Symbiotic (zooxanthellate) species are highly successful in shallow waters of tropical and subtropical seas and most of them are colonial. On the other hand, azooxanthellate species present wide distribution ranges and expand to the deep-sea at more than 6,000 m depth. These are mostly solitary, with only few species colonial that form extensive deep reefs. Each ecologically distinctive group encompasses half of the biodiversity of the order and they are not grouped into differentiated monophyletic clades. Paleontologists and evolutionary biologists have debated for decades whether modern scleractinian corals have evolved from symbiotic or colonial ancestors and how these traits have evolved and being involved in the diversification process in corals. Previous comparative analyses throw evidence in favor of coevolution of these characters and toward repetitive loss of symbiosis and coloniality. Nevertheless, the discovery of the origin of the group deep in the Paleozoic, with a deep divergent clade composed of only azooxanthellate corals has questioned these findings. With this work, we disentangle the patterns in the evolution of symbiosis and coloniality, testing if they are correlated and if they follow a gradual or episodic mode of evolution. To this end, we first produce the most complete time-calibrated phylogenetic tree for the order Scleractinia, including new sequences of never-before sampled species and genera. These novel sequences contribute to alleviate the current molecular under sampling of azooxanthellate species. Incorporating phylogenetic uncertainty, we obtained strong evidence in favor of a correlated and episodic model of evolution. This model led to the inference of an azooxanthellate and solitary most recent ancestor of scleractinians. Transition rates between the four different combinations of the two traits showed that while coloniality is gained and lost multiple times, symbiosis first appears around 282 Ma and is never lost. Also, coloniality seems to have appeared before symbiosis in azooxanthellate lineages. Thus, azooxanthellate corals, and especially colonial lineages, have been acting as a source of biodiversity for shallow zooxanthellate coral communities, highlighting the uniqueness of shallow and deep species and the need to preserve them

Neuron-derived extracellular vesicles contain synaptic proteins, promote spine formation, activate TrkB-mediated signalling and preserve neuronal complexity

Extracellular vesicles (EVs) play an important role in intercellular communication as carriers of signalling molecules such as bioactive miRNAs, proteins and lipids. EVs are key players in the functioning of the central nervous system (CNS) by influencing synaptic events and modulating recipient neurons. However, the specific role of neuron-to-neuron communication via EVs is still not well understood. Here, we provide evidence that primary neurons uptake neuron-derived EVs in the soma, dendrites, and even in the dendritic spines, and carry synaptic proteins. Neuron-derived EVs increased spine density and promoted the phosphorylation of Akt and ribosomal protein S6 (RPS6), via TrkB-signalling, without impairing the neuronal network activity. Strikingly, EVs exerted a trophic effect on challenged nutrient-deprived neurons. Altogether, our results place EVs in the spotlight for synaptic plasticity modulation as well as a possible therapeutic tool to fight neurodegeneration

Widespread Detection of Yersiniabactin Gene Cluster and Its Encoding Integrative Conjugative Elements (ICEKp) among Nonoutbreak OXA-48-Producing Klebsiella pneumoniae Clinical Isolates from Spain and the Netherlands

In this study, we determined the presence of virulence factors in nonoutbreak, high-risk clones and other isolates belonging to less common sequence types associated with the spread of OXA-48-producing Klebsiella pneumoniae clinical isolates from The Netherlands (n = 61) and Spain (n = 53). Most isolates shared a chromosomally encoded core of virulence factors, including the enterobactin gene cluster, fimbrial fim and mrk gene clusters, and urea metabolism genes (ureAD). We observed a high diversity of K-Locus and K/O loci combinations, KL17 and KL24 (both 16%), and the O1/O2v1 locus (51%) being the most prevalent in our study. The most prevalent accessory virulence factor was the yersiniabactin gene cluster (66.7%). We found seven yersiniabactin lineages-ybt 9, ybt 10, ybt 13, ybt 14, ybt 16, ybt 17, and ybt 27-which were chromosomally embedded in seven integrative conjugative elements (ICEKp): ICEKp3, ICEKp4, ICEKp2, ICEKp5, ICEKp12, ICEKp10, and ICEKp22, respectively. Multidrug-resistant lineages-ST11, ST101, and ST405-were associated with ybt 10/ICEKp4, ybt 9/ICEKp3, and ybt 27/ICEKp22, respectively. The fimbrial adhesin kpi operon (kpiABCDEFG) was predominant among ST14, ST15, and ST405 isolates, as well as the ferric uptake system kfuABC, which was also predominant among ST101 isolates. No convergence of hypervirulence and resistance was observed in this collection of OXA-48-producing K. pneumoniae clinical isolates. Nevertheless, two isolates, ST133 and ST792, were positive for the genotoxin colibactin gene cluster (ICEKp10). In this study, the integrative conjugative element, ICEKp, was the major vehicle for yersiniabactin and colibactin gene clusters spreading. IMPORTANCE; Convergence of multidrug resistance and hypervirulence in Klebsiella pneumoniae isolates has been reported mostly related to sporadic cases or small outbreaks. Nevertheless, little is known about the real prevalence of carbapenem-resistant hypervirulent K. pneumoniae since these two phenomena are often separately studied. In this study, we gathered information on the virulent content of nonoutbreak, high-risk clones (i.e., ST11, ST15, and ST405) and other less common STs associated with the spread of OXA-48-producing K. pneumoniae clinical isolates. The study of virulence content in nonoutbreak isolates can help us to expand information on the genomic landscape of virulence factors in K. pneumoniae population by identifying virulence markers and their mechanisms of spread. Surveillance should focus not only on antimicrobial resistance but also on virulence characteristics to avoid the spread of multidrug and (hyper)virulent K. pneumoniae that may cause untreatable and more severe infections.This study was supported by Plan Nacional de I+D+i 2013-2016 and Instituto de Salud Carlos III, Subdirección General de Redes y Centros de Investigación Cooperativa, Ministerio de Economía, Industria y Competitividad, Spanish Network for Research in Infectious Diseases (REIPI RD16CIII/0004/0002), cofinanced by European Development Regional Fund ERDF “A Way To Achieve Europe,” operative program Intelligent Growth 2014-2020. This study was also supported by a grant from the Instituto de Salud Carlos III (grant MPY 1135/16) and by the Antibiotic Resistance Surveillance Program of the Centro Nacional de Microbiología (Instituto de Salud Carlos III, Ministerio de Economía y Competitividad) of Spain. The Dutch CPE surveillance was funded by Dutch Ministry of Health, Welfare, and Sports.S

Effectiveness and safety of anti-CGRP monoclonal antibodies in patients over 65 years: a real-life multicentre analysis of 162 patients

Background Anti-CGRP monoclonal antibodies have shown notable effectiveness and tolerability in migraine patients; however, data on their use in elderly patients is still lacking, as clinical trials have implicit age restrictions and real-world evidence is scarce. In this study, we aimed to describe the safety and effectiveness of erenumab, galcanezumab and fremanezumab in migraine patients over 65 years old in real-life. Methods In this observational real-life study, a retrospective analysis of prospectively collected data from 18 different headache units in Spain was performed. Migraine patients who started treatment with any anti-CGRP monoclonal antibody after the age of 65 years were included. Primary endpoints were reduction in monthly migraine days after 6 months of treatment and the presence of adverse effects. Secondary endpoints were reductions in headache and medication intake frequencies by months 3 and 6, response rates, changes in patient-reported outcomes and reasons for discontinuation. As a subanalysis, reduction in monthly migraine days and proportion of adverse effects were also compared among the three monoclonal antibodies. Results A total of 162 patients were included, median age 68 years (range 65-87), 74.1% women. 42% had dyslipidaemia, 40.3% hypertension, 8% diabetes, and 6.2% previous cardiovascular ischaemic disease. The reduction in monthly migraine days at month 6 was 10.17.3 days. A total of 25.3% of patients presented adverse effects, all of them mild, with only two cases of blood pressure increase. Headache and medication intake frequencies were significantly reduced, and patient-reported outcomes were improved. The proportions of responders were 68%, 57%, 33% and 9% for reductions in monthly migraine days >= 30%,>= 50%,>= 75% and 100%, respectively. A total of 72.8% of patients continued with the treatment after 6 months. The reduction in migraine days was similar for the different anti-CGRP treatments, but fewer adverse effects were detected with fremanezumab (7.7%). Conclusions Anti-CGRP mAbs are safe and effective treatments in migraine patients over 65 years old in real-life clinical practice

CARB-ES-19 Multicenter Study of Carbapenemase-Producing Klebsiella pneumoniae and Escherichia coli From All Spanish Provinces Reveals Interregional Spread of High-Risk Clones Such as ST307/OXA-48 and ST512/KPC-3

ObjectivesCARB-ES-19 is a comprehensive, multicenter, nationwide study integrating whole-genome sequencing (WGS) in the surveillance of carbapenemase-producing K. pneumoniae (CP-Kpn) and E. coli (CP-Eco) to determine their incidence, geographical distribution, phylogeny, and resistance mechanisms in Spain.MethodsIn total, 71 hospitals, representing all 50 Spanish provinces, collected the first 10 isolates per hospital (February to May 2019); CPE isolates were first identified according to EUCAST (meropenem MIC > 0.12 mg/L with immunochromatography, colorimetric tests, carbapenem inactivation, or carbapenem hydrolysis with MALDI-TOF). Prevalence and incidence were calculated according to population denominators. Antibiotic susceptibility testing was performed using the microdilution method (EUCAST). All 403 isolates collected were sequenced for high-resolution single-nucleotide polymorphism (SNP) typing, core genome multilocus sequence typing (cgMLST), and resistome analysis.ResultsIn total, 377 (93.5%) CP-Kpn and 26 (6.5%) CP-Eco isolates were collected from 62 (87.3%) hospitals in 46 (92%) provinces. CP-Kpn was more prevalent in the blood (5.8%, 50/853) than in the urine (1.4%, 201/14,464). The cumulative incidence for both CP-Kpn and CP-Eco was 0.05 per 100 admitted patients. The main carbapenemase genes identified in CP-Kpn were blaOXA–48 (263/377), blaKPC–3 (62/377), blaVIM–1 (28/377), and blaNDM–1 (12/377). All isolates were susceptible to at least two antibiotics. Interregional dissemination of eight high-risk CP-Kpn clones was detected, mainly ST307/OXA-48 (16.4%), ST11/OXA-48 (16.4%), and ST512-ST258/KPC (13.8%). ST512/KPC and ST15/OXA-48 were the most frequent bacteremia-causative clones. The average number of acquired resistance genes was higher in CP-Kpn (7.9) than in CP-Eco (5.5).ConclusionThis study serves as a first step toward WGS integration in the surveillance of carbapenemase-producing Enterobacterales in Spain. We detected important epidemiological changes, including increased CP-Kpn and CP-Eco prevalence and incidence compared to previous studies, wide interregional dissemination, and increased dissemination of high-risk clones, such as ST307/OXA-48 and ST512/KPC-3

The origin and correlated evolution of symbiosis and coloniality in Scleractinian corals

Symbiosis and coloniality are ecologically important traits for corals of the order Scleractinia. Symbiotic (zooxanthellate) species are highly successful in shallow waters of tropical and subtropical seas and most of them are colonial. On the other hand, azooxanthellate species present wide distribution ranges and expand to the deep-sea at more than 6,000 m depth. These are mostly solitary, with only few species colonial that form extensive deep reefs. Each ecologically distinctive group encompasses half of the biodiversity of the order and they are not grouped into differentiated monophyletic clades. Paleontologists and evolutionary biologists have debated for decades whether modern scleractinian corals have evolved from symbiotic or colonial ancestors and how these traits have evolved and being involved in the diversification process in corals. Previous comparative analyses throw evidence in favor of coevolution of these characters and toward repetitive loss of symbiosis and coloniality. Nevertheless, the discovery of the origin of the group deep in the Paleozoic, with a deep divergent clade composed of only azooxanthellate corals has questioned these findings. With this work, we disentangle the patterns in the evolution of symbiosis and coloniality, testing if they are correlated and if they follow a gradual or episodic mode of evolution. To this end, we first produce the most complete time-calibrated phylogenetic tree for the order Scleractinia, including new sequences of never-before sampled species and genera. These novel sequences contribute to alleviate the current molecular under sampling of azooxanthellate species. Incorporating phylogenetic uncertainty, we obtained strong evidence in favor of a correlated and episodic model of evolution. This model led to the inference of an azooxanthellate and solitary most recent ancestor of scleractinians. Transition rates between the four different combinations of the two traits showed that while coloniality is gained and lost multiple times, symbiosis first appears around 282 Ma and is never lost. Also, coloniality seems to have appeared before symbiosis in azooxanthellate lineages. Thus, azooxanthellate corals, and especially colonial lineages, have been acting as a source of biodiversity for shallow zooxanthellate coral communities, highlighting the uniqueness of shallow and deep species and the need to preserve them.JRC.D.2-Water and Marine Resource

Neuron‐derived extracellular vesicles contain synaptic proteins, promote spine formation, activate TrkB‐mediated signalling and preserve neuronal complexity

Abstract Extracellular vesicles (EVs) play an important role in intercellular communication as carriers of signalling molecules such as bioactive miRNAs, proteins and lipids. EVs are key players in the functioning of the central nervous system (CNS) by influencing synaptic events and modulating recipient neurons. However, the specific role of neuron‐to‐neuron communication via EVs is still not well understood. Here, we provide evidence that primary neurons uptake neuron‐derived EVs in the soma, dendrites, and even in the dendritic spines, and carry synaptic proteins. Neuron‐derived EVs increased spine density and promoted the phosphorylation of Akt and ribosomal protein S6 (RPS6), via TrkB‐signalling, without impairing the neuronal network activity. Strikingly, EVs exerted a trophic effect on challenged nutrient‐deprived neurons. Altogether, our results place EVs in the spotlight for synaptic plasticity modulation as well as a possible therapeutic tool to fight neurodegeneration

Intervención educativa en el primer año de vida

Documento elaborado por los Equipos de Atención Temprana de la Región de Murcia. El trabajo se desarrolla en cuatro capítulos donde se recogen las distintas áreas de estudio: exposición global del trabajo que realizan estos equipos; evolución normal del niño en su primer año de vida; aspectos preventivos en función de factores de riesgo y alto riesgo y orientación familiar; modelos de aplicación, práctica y conclusiones en las que se recogen una serie de propuestas de mejora en la calidad de la atención durante el primer año de vida.MurciaES