Multiplex Amplicon Sequencing For Microbe Identification In Community-based Culture Collections

Microbiome analysis using metagenomic sequencing has revealed a vast microbial diversity associated with plants. Identifying the molecular functions associated with microbiome-plant interaction is a significant challenge concerning the development of microbiome-derived technologies applied to agriculture. An alternative to accelerate the discovery of the microbiome benefits to plants is to construct microbial culture collections concomitant with accessing microbial community structure and abundance. However, traditional methods of isolation, cultivation, and identification of microbes are time-consuming and expensive. Here we describe a method for identification of microbes in culture collections constructed by picking colonies from primary platings that may contain single or multiple microorganisms, which we named community-based culture collections (CBC). A multiplexing 16S rRNA gene amplicon sequencing based on two-step PCR amplifications with tagged primers for plates, rows, and columns allowed the identification of the microbial composition regardless if the well contains single or multiple microorganisms. The multiplexing system enables pooling amplicons into a single tube. The sequencing performed on the PacBio platform led to recovery near-full-length 16S rRNA gene sequences allowing accurate identification of microorganism composition in each plate well. Cross-referencing with plant microbiome structure and abundance allowed the estimation of diversity and abundance representation of microorganism in the CBC.6RepsolRepsol Sinope

Unlocking The Bacterial And Fungal Communities Assemblages Of Sugarcane Microbiome

Plant microbiome and its manipulation herald a new era for plant biotechnology with the potential to benefit sustainable crop production. However, studies evaluating the diversity, structure and impact of the microbiota in economic important crops are still rare. Here we describe a comprehensive inventory of the structure and assemblage of the bacterial and fungal communities associated with sugarcane. Our analysis identified 23,811 bacterial OTUs and an unexpected 11,727 fungal OTUs inhabiting the endophytic and exophytic compartments of roots, shoots, and leaves. These communities originate primarily from native soil around plants and colonize plant organs in distinct patterns. The sample type is the primary driver of fungal community assemblage, and the organ compartment plays a major role in bacterial community assemblage. We identified core bacterial and fungal communities composed of less than 20% of the total microbial richness but accounting for over 90% of the total microbial relative abundance. The roots showed 89 core bacterial families, 19 of which accounted for 44% of the total relative abundance. Stalks are dominated by groups of yeasts that represent over 12% of total relative abundance. The core microbiome described here comprise groups whose biological role underlies important traits in plant growth and fermentative processes.6RepsolRepsol Sinope

Fundamentals of Wireless Communication Link Design for Networked Robotics

This chapter aims to present the fundamentals of the design of wireless communication links for networked robotics applications. First, we provide an overview of networked robotics applications, motivating the importance of the wireless communication link as an enabler of these applications. Next, we review the wireless communication technologies available today, discussing the existent tradeoffs between range, power, and data rate, and introducing the main concepts regarding the design of wireless communication links. Finally, we present a design example of a wireless communication link and the results obtained. We conclude the chapter with a discussion of the results and the challenges faced in the design of wireless communication links for networked robotics

Expansion planning of power distribution systems considering reliability : a comprehensive review

One of the big concerns when planning the expansion of power distribution systems (PDS) is reliability. This is defined as the ability to continuously meet the load demand of consumers in terms of quantity and quality. In a scenario in which consumers increasingly demand high supply quality, including few interruptions and continuity, it becomes essential to consider reliability indices in models used to plan PDS. The inclusion of reliability in optimization models is a challenge, given the need to estimate failure rates for the network and devices. Such failure rates depend on the specific characteristics of a feeder. In this context, this paper discusses the main reliability indices, followed by a comprehensive survey of the methods and models used to solve the optimal expansion planning of PDS considering reliability criteria. Emphasis is also placed on comparing the main features and contributions of each article, aiming to provide a handy resource for researchers. The comparison includes the decision variables and reliability indices considered in each reviewed article, which can be used as a guide to applying the most suitable method according to the requisites of the system. In addition, each paper is classified according to the optimization method, objective type (single or multiobjective), and the number of stages. Finally, we discuss future research trends concerning the inclusion of reliability in PDS expansion planning

Valor do EEG na caracterização e prognóstico de patologias neurológicas em recém-nascidos prematuros Relationship of EEG, neurological diseases and follow-up in preterm newborns

A importância do EEG na investigação de patologias neurológicas no período neonatal tem sido discutido com frequência na atualidade, pois muitas vezes este é o único meio de avaliar a função cerebral em recém-nascidos (RN) com patologias graves ou sob efeito de medicações. O presente estudo foi realizado com 85 RN prematuros que apresentaram patologias neurológicas no período neonatal e foram submetidos a avaliação neurológica e eletrencefalográfica (EEG ou polissonografia -PS). Foram relacionadas alterações do EEG, patologias e prognóstico. Os EEGs foram classificados segundo as alterações da atividade de base, presença de atividade paroxística, organização dos estágios do sono e "maturidade". A patologia mais frequente foi asfixia perinatal (40%), seguida de hemorragia intra-ventricular (HIV, 16%). A queixa mais frequente na indicação do exame foi apnéia (71 %), seguida de convulsões (19%). Foram considerados normais 55% dos exames solicitados com queixa exclusiva de apnéia; dos exames solicitados por convulsões somente 31 % foram normais. A alteração do EEG mais frequentemente encontrada nos RN com asfixia, HIV e desnutrição intra-uterina foi a "imaturidade" da atividade elétrica cerebral. Nos RN com convulsões as alterações mais frequentes do EEG foram atividade paroxística anormal e "imaturidade". Os RN com infecção do sistema nervoso central apresentaram diversas alterações no EEG. Os achados de EEG mais correlacionados a mau prognóstico foram isoeletricidade e atividade paroxística anormal (100% dos casos com presença de ondas agudas positivas).The importance of the EEG for the investigation of neurological diseases in the neonatal period has been largely discussed, since it is often the only way to approach cerebral function in newborns with severe pathologies or under drug effect. The present study was carried out with 85 newborns (NB) who presented perinatal dysfuntions and were submitted to neurological and electroencephalogram (EEG) or polysomnography (PS) evaluation. EEG/PS alterations, pathologies and prognosis were reported. The EEG were classified according to basal activity alterations, presence of paroxysmal activity and sleep stages organization and maturity. The most frequent pathology was perinatal asphyxia (40%) followed by intraventricular hemorrhage (HIV, 16%). The most frequent complaint for exam indication was apnea (71%) followed by convulsion (19%). Fifty-five percent of the exams exclusively required because apnea complaint were considered normal and out of all exams required because seizures only 31 % were normal. The EEG alteration most frequently related to perinatal asphyxia, HIV and intrauterine growth delay was immaturity and in the NB with seizures immature EEG and abnormal paroxysms. Many different alterations were registered in the NB with nervous system infection. The EEG findings more correlated with unfavorable prognosis were isoelectricity and abnormal paroxysmal activity including positive sharp waves (100%)

Genome sequences of a plant beneficial synthetic bacterial community reveal genetic features for successful plant colonization

Despite the availability of data on the functional and phylogenetic diversity of plant-associated microbiota, the molecular mechanisms governing the successful establishment of plant bacterial communities remain mostly elusive. To investigate bacterial traits associated with successful colonization of plants, we sequenced the genome of 26 bacteria of a synthetic microbial community (SynCom), 12 of which displayed robust and 14 displayed non-robust colonization lifestyles when inoculated in maize plants. We examined the colonization profile of individual bacteria in inoculated plants and inspected their genomes for traits correlated to the colonization lifestyle. Comparative genomic analysis between robust and non-robust bacteria revealed that commonly investigated plant growth-promoting features such as auxin production, nitrogen (N) fixation, phosphate acquisition, and ACC deaminase are not deterministic for robust colonization. Functions related to carbon (C) and N acquisition, including transporters of carbohydrates and amino acids, and kinases involved in signaling mechanisms associated with C and N uptake, were enriched in robust colonizers. While enrichment of carbohydrate transporters was linked to a wide range of metabolites, amino acid transporters were primarily related to the uptake of branched-chain amino acids. Our findings identify diversification of nutrient uptake phenotypes in bacteria as determinants for successful bacterial colonization of plants10FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2016/23425-5; 2018/19100-9; 2016/04322-0; 2016/23218-0Repsol; Repsol Sinopec; FAPESPFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2016/23425-5, 2018/19100-9, 2016/04322-0, 2016/23218-0

Síndrome da morte súbita do lactente: aspectos clínicos de uma doença subdiagnosticada

Objetivo: Conhecer as características socioepidemiológicas dos lactentes que foram a óbito por SMSL no nosso meio e estabelecer a prevalência desta síndrome como causa de mortalidade no primeiro ano de vida. Métodos: Estudo observacional, retrospectivo, descritivo, baseado na revisão de laudos de necropsia e na aplicação de questionário a familiares de lactentes que apresentaram óbito domiciliar nos anos de 1997 e 1998 em Porto Alegre. Resultados: Dos 335 óbitos ocorridos em lactentes, 21 foram incluídos nos critérios que definem SMSL, mas nenhum deles teve essa causa registrada em sua declaração de óbito. O perfil dos óbitos por SMSL foi caracterizado por lactentes sem história prévia de apnéias ou qualquer doença aguda que justificasse o óbito. As mortes ocorreram principalmente nos meses frios (outono e inverno) e nenhum destes lactentes dormia em posição supina. As famílias possuíam renda média de 3 salários mínimos, 71% das mães eram tabagistas e 42 % fizeram uso de álcool durante a gestação. A prevalência de SMSL foi estimada em 6,3% do total de óbitos, sendo que o coeficiente de mortalidade específico por essa causa foi de 4,5 óbitos em 10.000 nascidos vivos. Conclusões: O perfil clínico-epidemiológico de risco para SMSL apresentado pelas crianças que foram a óbito por essa causa assemelha-se em diversos aspectos ao descrito na literatura internacional. Observa-se, entretanto, que a SMSL é subdiagnosticada no nosso meio, não constando das estatísticas oficiais de mortalidade infantil. Com isso, não são desenvolvidas campanhas para redução do risco a que as crianças estão expostas