Association of body image perception and (dis)satisfaction with adiposity in adults: The Pró-Saúde study.

INTRODUCTION: Body image distortion and/or dissatisfaction may occur primarily due to body fat accumulation and/or distribution. The aim of this study was to evaluate the frequency of body image perception and (dis)satisfaction categories in adult men and women according to the adiposity classification. METHODS: This is a cross-sectional study (n = 514; 33-79 years; 265 women) that is part of a prospective cohort (Pró-Saúde study). Adiposity measurements were determined by two methods: anthropometry, used to calculate the body mass index (BMI) and dual-energy X-ray absorptiometry (DXA), to estimate body fat percentage. Participants were grouped as "no excess adiposity" and "excess adiposity", considering BMI and body fat percentage (>30% for men, >40% for women). Perception and (dis)satisfaction with body image were evaluated using the Kakeshita scale, composed by 15 silhouettes, developed for the Brazilian population. Degree of distortion (perceived BMI - real BMI) and dissatisfaction (perceived BMI - desired BMI) were calculated. RESULTS: A high proportion of men (58.6%; 74.3%), and especially of women (82.6%; 86.8%), presented body size overestimation and dissatisfaction due to excess weight, respectively. A relevant fraction of the women (32.6%) and men (30.8%) who were dissatisfied due to excess weight did not present excess adiposity, especially if classified by DXA. Variability in degree of distortion was hardly explained by anthropometric and DXA variables in women (<5%) and men (∼22%). Both anthropometric and DXA measurements accounted for ∼30% and ∼50% of the variability in degree of dissatisfaction among women and men, respectively. CONCLUSION: Our results suggest a high frequency of body image distortion in Brazilian adult individuals, as well as dissatisfaction with excess weight, especially among women with excess adiposity. The findings indicate that anthropometric measurements explain much of the variability in degree of body image dissatisfaction in men, with no apparent advantage of the use of more refined DXA measurements.

Long-term occurrence of multiple antimicrobial drug resistant Klebsiella pneumoniae isolates harboring virulent potential in a tertiary hospital from Brazil.

Klebsiella pneumoniae strains are globally associated with a plethora of opportunistic and severe human infections and are known to spread genes conferring antimicrobial resistance. Some strains harbor virulence determinants that enable them to cause serious disease in any patient, both in the hospital and in the community. The aim of this study was to determine the frequency of antimicrobial resistance and virulence traits (by gene detection and string test) among 83 K. pneumoniae isolates obtained from patient cultures of a scholar tertiary hospital in the Midwestern Brazil (Brasília, DF). Antimicrobial susceptibility analysis showed that 94% (78/83) of the isolates presented one of the following resistance profiles: resistant (R, 39), multidrug-resistant (MDR, 29), or extensively drug-resistant (XDR, 10). Several MDR and XDR strains harbored multiple virulence genes and displayed hypermucoviscous phenotype. These characteristics were observed among isolates obtained throughout all the sample collection period (2013 - 2017). The K2 serotype gene, a molecular marker of hypervirulence, was detected in three isolates, one of which classified as XDR. Sequence typing revealed the occurrence of isolates belonged to high-risk (ST13) and multiple resistance-spreading clones (ST105). Thus, our findings showed the occurrence of virulent potential isolates that also presented MDR/XDR phenotypes from 2013 to 2015. This study also indicates the probable convergence of virulence and resistance since at least 2013 in Brazil.

fim3-24/ptxP-3 genotype is associated to whooping cough outbreak in Brazilian Midwest: The selection of Bordetella pertussis strains driven by vaccine immunization.

Whopping cough (or Pertussis) is an acute infectious respiratory disease caused by Bordetella pertussis bacteria. The disease is highly transmissible and can be fatal in children under two years old. Since the introduction of vaccine immunization in 1940, Pertussis incidence decreased worldwide. In Brazil, the immunization was introduced in 1977 using the whole cell (wP) vaccine. Despite the high vaccination coverage, an unexpected increase in the number of observed Pertussis cases was observed in 2012. In this year, 2257 cases were reported exceeding the average incidence rate of <1000 cases per year until 2010. This outbreak reached a peak level in 2014 and ended in 2018 according to the Brazilian National Surveillance System (SINAN). To understand the relationship between the outbreak and the vaccination, bacterial isolates (n = 136) from the Brazilian Midwest region obtained during the outbreak were submitted to genotyping of two vaccine loci: ptxP and fim3. Most of isolates (102) were obtained from nursing children (29 days to 2 years old). Genotyping of 94 isolates revealed that fim3-24/ptxP-3 was the most prevalent genotype (68%) associated with the outbreak peak. Two additional genotypes were also observed: fim3-1/ptxP-3 (15%) and fim3-3/ptxP-3 (17%). Conversely, the fim3-1/ptxP-2 genotype, which is harbored by the strain used in the wP vaccine (Bp137), was not observed. These results showed that B. pertussis circulating strains in the outbreak analyzed were different from the strain used for Pertussis immunization in Brazil. These observations provide insights that could be used to target vaccination programs to prevent future whooping cough outbreaks in Brazil.

Adequacy of basal metabolic rate prediction equations in individuals with severe obesity: A systematic review and meta-analysis.

The determination of energy requirements in clinical practice is based on basal metabolic rate (BMR), frequently predicted by equations that may not be suitable for individuals with severe obesity. This systematic review and meta-analysis examined the accuracy and precision of BMR prediction equations in adults with severe obesity. Four databases were searched in March 2021 and updated in May 2023. Eligible studies compared BMR prediction equations with BMR measured by indirect calorimetry. Forty studies (age: 28-55 years, BMI: 40.0-62.4 kg/m2) were included, most of them with a high risk of bias. Studies reporting bias (difference between estimated and measured BMR) were included in the meta-analysis (n = 20). Six equations were meta-analyzed: Harris & Benedict (1919); WHO (weight) (1985); Owen (1986); Mifflin (1990); Bernstein (1983); and Cunningham (1980). The most accurate and precise equations in the overall analysis were WHO (-12.44 kcal/d; 95%CI: -81.4; 56.5 kcal/d) and Harris & Benedict (-18.9 kcal/d; 95%CI -73.2; 35.2 kcal/d). All the other equations tended to underestimate BMR. Harris & Benedict and WHO were the equations with higher accuracy and precision in predicting BMR in individuals with severe obesity. Additional analyses suggested that equations may perform differently according to obesity BMI ranges, which warrants further investigation.

Scalable and versatile container-based pipelines for de novo genome assembly and bacterial annotation.

Background: Advancements in DNA sequencing technology have transformed the field of bacterial genomics, allowing for faster and more cost effective chromosome level assemblies compared to a decade ago. However, transforming raw reads into a complete genome model is a significant computational challenge due to the varying quality and quantity of data obtained from different sequencing instruments, as well as intrinsic characteristics of the genome and desired analyses. To address this issue, we have developed a set of container-based pipelines using Nextflow, offering both common workflows for inexperienced users and high levels of customization for experienced ones. Their processing strategies are adaptable based on the sequencing data type, and their modularity enables the incorporation of new components to address the community's evolving needs. Methods: These pipelines consist of three parts: quality control, de novo genome assembly, and bacterial genome annotation. In particular, the genome annotation pipeline provides a comprehensive overview of the genome, including standard gene prediction and functional inference, as well as predictions relevant to clinical applications such as virulence and resistance gene annotation, secondary metabolite detection, prophage and plasmid prediction, and more. Results: The annotation results are presented in reports, genome browsers, and a web-based application that enables users to explore and interact with the genome annotation results. Conclusions: Overall, our user-friendly pipelines offer a seamless integration of computational tools to facilitate routine bacterial genomics research. The effectiveness of these is illustrated by examining the sequencing data of a clinical sample of Klebsiella pneumoniae.

Outbreak of extensively drug-resistant Serratia marcescens in an intensive care unit.

We present the investigation and control of an extensively drug-resistant Serratia marcescens outbreak in a 30-bed intensive care unit (ICU). Within 6 weeks, 4 critically ill trauma patients were infected by the same strain. Intensive containment measures limited the spread of this strain while sustaining the capacity of the trauma ICU.

Evaluation of microsatellite allelic patterns to DNA fingerprinting in rubber tree clones

The rubber tree (Hevea brasiliensis) is native to the Amazon region, and it is widely exploited due to natural rubber produced from latex. There are many clonal varieties, without certification tests. In order to determine a genetic certification, 15 clones were genotyped to identify their genetic pattern. Ten microsatellites were used to determine a subset of alleles exclusive for each genetic profile. The genetic estimates obtained were: number of alleles per locus (N), expected (HE) and observed (HO) heterozygosity, Polymorphic Information Content (PIC) and Discriminatory Power (DP). The number of alleles (N) ranged from five to 14, with an average of 9.2. The HE mean (0.80) was higher than HO (0.60), indicating a selection for homozygotes. The locus informativeness was verified with PIC (0.77) and DP (0.90) means showing high polymorphism. The dendrogram represented the formation of three groups related to geographical origin. Clone MDF 180 presented the highest genetic divergence. Two genic pools represented the genetic composition of genotypes. Based on allelic profiles, a set of two microsatellites (A2365 and A2368) was able to distinguish all examined clones. The genetic certification using microsatellite fingerprinting proved to be an alternative to morphological traits.

A Novel Multidrug Resistant, Non-Tn4401 Genetic Element-Bearing, Strain of Klebsiella pneumoniae Isolated From an Urban Lake With Drinking and Recreational Water Reuse.

Antimicrobial resistance (AMR) is an increasing and urgent issue for human health worldwide, as it leads to the reduction of available antibiotics to treat bacterial infections, in turn increasing hospital stays and lethality. Therefore, the study and genomic surveillance of bacterial carriers of resistance in and outside of clinical settings is of utter importance. A colony of multidrug resistant (MDR) bacteria identified as Klebsiella spp., by 16S rDNA amplicon sequencing, has been isolated from an urban lake in Brazil, during a drug-degrading bacterial prospection. Genomic analyses revealed the bacteria as Klebsiella pneumoniae species. Furthermore, the in silico Multilocus Sequence Typing (MLST) identified the genome as a new sequence type, ST5236. The search for antimicrobial resistance genes (ARGs) detected the presence of genes against beta-lactams, fosfomycin, acriflavine and efflux pumps, as well as genes for heavy metal resistance. Of particular note, an extended-spectrum beta-lactamase gene (blaCTX-M-15) has been detected in close proximity to siphoviridae genes, while a carbapenemase gene (KPC-2) has been found in an extrachromosomal contig, within a novel non-Tn4401 genetic element (NTEKPC). An extrachromosomal contig found in the V3 isolate is identical to a contig of a K. pneumoniae isolate from a nearby hospital, which indicates a putative gene flow from the hospital network into Paranoá lake. The discovery of a MDR isolate in this lake is worrisome, as the region has recently undergone periods of water scarcity causing the lake, which receives treated wastewater effluent, and is already used for recreational purposes, to be used as an environmental buffer for drinking water reuse. Altogether, our results indicate an underrepresentation of environmental K. pneumoniae among available genomes, which may hamper the understanding of the population dynamics of the species in the environment and its consequences in the spread of ARGs and virulence genes.

Multidrug-Resistant (MDR) Klebsiella variicola Strains Isolated in a Brazilian Hospital Belong to New Clones.

Klebsiella variicola is mainly associated with opportunistic infections and frequently identified as Klebsiella pneumoniae. This misidentification implies a wrong epidemiology result as well as incorrect attribution to K. pneumoniae as the etiology of some severe infections. Recently, huge efforts have been made to study K. variicola, however, the biological aspects of this species are still unclear. Here we characterized five K. variicola strains initially identified as K. pneumoniae, with a Vitek-2 System and 16S rRNA sequencing. One-step multiplex polymerase chain reaction and Whole Genome Sequencing (WGS) identified them as K. variicola. Additionally, WGS analysis showed that all the strains are closely related with K. variicola genomes, forming a clustered group, apart from K. pneumoniae and K. quasipneumoniae. Multilocus sequence typing analysis showed four different sequence types (STs) among the strains and for two of them (Kv97 and Kv104) the same ST was assigned. All strains were multidrug-resistant (MDR) and three showed virulence phenotypes including invasion capacity to epithelial cells, and survival in human blood and serum. These results showed the emergence of new K. variicola clones with pathogenic potential to colonize and cause infection in different tissues. These characteristics associated with MDR strains raise great concern for human health.

Antimicrobial effects of silver diamine fluoride: An in vivo study.

PURPOSE: To compare the antimicrobial effect of treating dentin caries lesions with silver diamine fluoride (SDF) in different concentrations and chlorhexidine (CHX). METHODS: Children aged 7-10 years presenting one occlusal dentin carious lesion in primary molars were selected, totaling 40 teeth. The sample was randomly divided into four groups: (G1) 38%-SDF + potassium iodide (KI); (G2) 30%-SDF; (G3) 2%-CHX; and (G4) control group. After cleaning the cavity up to firm dentin, a sample of dentin from the pulp wall was collected; the cavity was then treated with the antimicrobial agent tested and, immediately after, another dentin sample was collected. Cavities were restored with high viscosity glass ionomer cement. Microorganisms were counted, and species from the Streptococcus genus were analyzed for susceptibility to antimicrobial agents. Shapiro-Wilk and Levene's tests were used to assess normality and homogeneity, respectively. Student's t-test, two-way ANOVA, and Bonferroni post-test were applied for multiple comparisons. RESULTS: For the overall microorganisms count, it was observed that G1 and G2 presented a statistically lower number of microorganisms following treatment in comparison to G3 and G4 (P< 0.05). When analyzing the Streptococcus spp. and Enterococcus sp. separately, a statistical reduction in the microorganism count before and after the treatment was observed for all groups (P< 0.05), excluding the control group. Among the species tested, S. mutans were the least susceptible to SDF treatments compared to the other species. The treatments with SDF were more effective in reducing microorganisms when compared to CHX. Similarly, the susceptibility of Streptococcus to CHX was lower than that observed for SDF. CLINICAL SIGNIFICANCE: In cases where the dental professional decides to apply an antimicrobial agent prior to the placement of a restoration, silver diamine fluoride proved to be more effective than chlorhexidine, slowing the progression of carious lesions, and possibly preventing future restorative interventions thus improving children's quality of life. It is important to note that clinicians should consider the type of restorative material that will be used due to the possibility that the use of SDF may influence adhesion of the subsequent restoration.

Genetic structure and diversity of native Guadua species (Poaceae: Bambusoideae) in natural populations of the Brazilian Amazon rainforest.

The Southwestern Region of the Brazilian Amazon is formed by forests dominated by bamboos. The genus Guadua is endemic to the Americas, and little is known about the genetic diversity and structure of species of this genus. This study aimed to evaluate the genetic diversity and structure of two native Guadua species in natural populations in the Southwestern region of the Brazilian Amazon. Therefore, the genetic diversity and structure of Guadua aff. chaparensis and Guadua aff. lynnclarkiae were evaluated with the use of microsatellite molecular markers (SSR). It was verified that the average genetic diversity for the populations studied was considered high ( H ^ e =0.5) compared to other species of bamboo. All populations had rare and private alleles, and none of them presented significant values of inbreeding. The populations were divergent ( G ^ ST = 0.46), resulting in a low apparent gene flow. The Bayesian analysis showed that among the 350 individuals analyzed, five groups (K=5) were formed, with little similarity among the groups (Populations), although two of them presented clonal individuals. According to the results obtained, it can be conclude that populations should be treated as having unique characteristics, mainly when accessed for management and for in situ and ex situ conservation studies.

Molecular characterization and core collection evaluation of Manihot esculenta Crantz / Caracterização molecular e avaliação de coleção nuclear de Manihot esculenta Crantz

Cassava is one of the most important subsistence crops in tropical regions. It is necessary to preserve and to know the genetic diversity existent for the adequate use of genetic resources. The evaluation of genetic diversity among genotypes results in information about potential parents in breeding programs, allows duplicates identification, and facilitates germplasm exchange between research institutions. The objective of this study was to characterize the genetic diversity of cassava accessions of North Brazil region. A total of 106 accessions were analyzed using ten microsatellite markers. The genetic parameters estimated were: expected heterozygosity (HE), observed heterozygosity (HO) and polymorphic information content (PIC). Clustering was performed using the UPGMA and Neighbor-Joining (NJ) method. Bayesian analysis, analysis of principal coordinates and identification of a core collection were also used. The ten loci amplified 8,40 alleles on average. The average heterozygosity estimates were: HE = 0.71, HO = 0.58 and PIC = 0.72. Genetic distances ranged from 0.158 to 0.908. Six (5,66%) accesses were redundant. Clustering and dispersion analysis didn't differentiate bitter from sweet cassava, and there wasn't correlation between groups and collect origin. The core collection consisted of 22 individuals that represented 94% of total allelic diversity and 20,75% of the base collection. The results indicate high dissimilarity between the accessions and allowed the detection of redundant genotypes, showing the use of genetic markers as informative tools for the management of collections. (AU)

A mandioca é uma das mais importantes culturas de subsistência em países tropicais. É preciso conservar e conhecer a diversidade genética para o uso adequado dos recursos genéticos. A avaliação da diversidade genética entre os genótipos resulta em informações sobre potenciais genitores em programas de melhoramento, possibilita a identificação de duplicatas, além disso, facilita o intercâmbio de germoplasma entre instituições de pesquisa. O objetivo deste trabalho foi caracterizar a diversidade genética dos acessos de mandioca da Região Norte do Brasil. Foram analisados 106 acessos por meio de dez marcadores microssatélites. Os parâmetros de diversidade genética estimados foram: heterozigosidade esperada (HE), heterozigosidade observada (HO) e conteúdo de informação polimórfica (PIC). Agrupamentos foram realizados pelo método UPGMA e Neighbor-Joining (NJ). Utilizou-se também análises bayesianas, dispersão por coordenadas principais e a identificação de uma coleção nuclear. Os dez locos amplificaram 8,40 alelos em média. A média das estimativas de diversidade foram altas: HE = 0,71, HO = 0,58 e PIC = 0,72. As distâncias genéticas variaram de 0,158 a 0,908. Seis (5,66%) acessos estão redundantes. Os agrupamentos e análises de dispersão não evidenciaram distinção entre variedades bravas e mansas e não foi identificada estrutura genética correspondente a origem dos acessos. A coleção nuclear foi formada por 22 indivíduos, que representaram 94% da diversidade alélica total e 20,75% da coleção base. Os resultados indicam alta dissimilaridade entre os acessos e permitiram a detecção de genótipos redundantes, mostrando o uso de marcadores genéticos como ferramentas informativas para o manejo de coleções. (AU)

High Level of Resistance to Antimicrobials and Heavy Metals in Multidrug-Resistant Pseudomonas sp. Isolated from Water Sources.

Antimicrobial-resistant bacteria (ARB) carrying antimicrobial resistance genes (ARGs) have been increasingly detected in water sources. Pseudomonas sp. are opportunistic pathogens commonly reported in water samples and different antimicrobial resistance mechanisms have been described in Pseudomonas sp., producing multidrug-resistant (MDR) phenotype. Besides, the presence of heavy metal resistance genes (HMRGs) may select ARB, which is worrying. Therefore, this study aimed to characterize the genotypes of Pseudomonas sp. isolated from different water sources. Water samples (i.e., rivers, streams, lakes and sewage treatment plants) were collected from different cities in Brazil. The bacterial identification was performed by sequencing the 16S rDNA and the resistance profile to antimicrobials and heavy metals were determined by minimum inhibitory concentration (MIC). Several ARGs, HMRGs, and plasmids were researched by PCR and the amplicons were sequenced for confirmation. A total of 23 Pseudomonas sp. isolates were obtained and identified as Pseudomonas saponiphila, Pseudomonas hunanensis, Pseudomonas aeruginosa, and Pseudomonas asiatica. These isolates presented high MICs to antimicrobials and heavy metals, being the great majority (n = 21; 91%) classified as MDR. Different clinically important ARGs were detected, such as blaGES, qnrS, qepA, tetB, aac(3')-IIa, and ant(2″)-Ia. The czcA gene was the only HMRG detected and no plasmids were found. To the best of our knowledge, this is the first report of the world of P. saponiphila carrying ARGs (i.e., blaGES, qnrS, aac(3')-IIa, tetB) and QepA-producing P. hunanensis and the first time of P. saponiphila, P. asiatica, and P. hunanensis in Brazil.

Genomic Characterization of a Multidrug-Resistant and Hypermucoviscous/Hypervirulent Klebsiella quasipneumoniae subsp. similipneumoniae ST4417 Isolated from a Sewage Treatment Plant.

Aim: Clinical strains of Klebsiella quasipneumoniae subsp. similipneumoniae have been reported worldwide. Multidrug-resistant (MDR) hypermucoviscous (hm)/hypervirulent (hv) lineages have become a global problem for public health worldwide. Therefore, this study aimed to characterize by whole-genome sequencing a MDR-hm/hv K. quasipneumoniae subsp. similipneumoniae SWT10 strain belonging to the new sequence type ST4417 isolated from a sewage treatment plant. Results: The SWT10 strain was recovered from a sewage treatment plant in Brazil and presented the hm and MDR phenotypes. Resistome analysis showed antimicrobial resistance genes associated with resistance to fluoroquinolones, ß-lactams, tetracyclines, trimethoprim, aminoglycosides, sulfonamides, macrolides, and fosfomycin as well as several heavy metal resistance genes. Virulome analysis showed virulence factors related to hv lineages. Multilocus sequence typing analysis revealed the new ST4417, which was grouped in CC1584 by the minimum-spanning tree. Besides, five plasmid incompatibility groups, two prophage-related sequences, and 66 genomic islands were detected. Conclusion: This study reports for the first time the genome sequence of a MDR-hm/hv K. quasipneumoniae subsp. similipneumoniae recovered from the environment, which contributes to a better understanding about these lineages as well as for surveillance studies worldwide.

Inferring mating system parameters in forage peanut, Arachis pintoi, for Brazilian Amazon conditions

The search for alternatives to increase productivity and sustainability of livestock production in the Amazon region without increasing deforestation is challenging. Mixed pastures of grasses with forage peanut (Arachis pintoi) have shown positive economic impacts. However, gaps in the knowledge of the reproductive biology of A. pintoi have limited the development of new cultivars adapted to the environmental variations in the Brazilian Amazon. Pasture consortiums of Brachiaria humidicola with forage peanuts (cv. Mandobi) resulted in a 42% increase in weight gain productivity. New cultivars better adapted to the Amazon climate should bring even greater gains. We evaluated the mating system in twenty A. pintoi accessions, and approximately 40 offspring per accession genotyped with eight microsatellites (or markers). The parameters of genetic diversity and inbreeding, the outcrossing rate and coancestry were calculated. The observed heterozygosity was significantly higher and the fixation index was significantly lower in adults compared with the offspring. The crossing rate was variable among genotypes (2 to 80%), and the mean outcrossing rate was 36%. These results indicate that pollinator presence in pastures can influence gene flow in A. pintoi more than expected. Arachis pintoi presented a mixed mating system with a predominance of selfing, and families presented inbreeding and different levels of relatedness. New strategies of genotype conservation are needed to avoid pollinator-mediated crossing between accessions. (AU)

Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA / Preservação e maceração de tecido de folíolos de açaí da Amazônia para obtenção de DNA genômico

The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µ L (7 days in transport buffer) to 702.00 ng/µ L (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µ L (30 days in silica gel) to 2,850.00 ng/µ L (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.

O objetivo deste estudo foi testar a eficiência de métodos de preservação e maceração de tecidos de folíolos de Euterpe precatoria para obter DNA genômico para estudos moleculares. Os folíolos de E. precatoria foram coletados no campo experimental da Embrapa Acre, Brasil. O estudo foi conduzido em delineamento inteiramente casualizado com 10 repetições, em esquema fatorial 12 × 2, com 12 tratamentos de armazenamento (fresco; liofilizado 3 dias; geladeira 3, 5, e 7; sílica gel 7, 10, 20 e 30 dias e tampão de transporte 3, 5 e 7 dias) e dois tipos de maceração do tecido foliar (nitrogênio líquido e TissueLyser®). Para a variável concentração de DNA houve diferença estatística entre os tipos maceração e de armazenamento. O macerador TissueLyser® apresentou maiores concentrações de DNA quando comparado ao nitrogênio líquido.Para os tipos de armazenamento verificou-se formação de cinco grupos quando macerados TissueLyser® e dois grupos quando macerados com nitrogênio líquido. As concentrações de DNA variaram de 285,00 ng/µ L (7 dias em tampão de transporte) a 702,00 ng/µ L (30 dias em sílica gel) quando maceradas com nitrogênio líquido. Quando maceradas com macerador TissueLyser® variaram de 572,73 ng/µ L (30 dias em sílica gel) a 2.850,00 ng/µ L (3 dias em liofilizador). A pureza do DNA (A260/A280 nm) variou de 1,30 a 1,70 quando os folíolos foram macerados com nitrogênio líquido e de 1,30 a 1,90 quando macerados em macerador TissueLyser®. Apesar das variações na conservação e concentração dos tecidos foliares, todos os tratamentos foram eficazes para o isolamento do DNA e amplificaram regiões de marcadores microssatélites. Concluiu-se que folíolos de E. precatoria armazenados em liofilizador e macerados com macerador automático resultaram em DNA satisfatório para estudos moleculares.

RR2 dye adsorption to Hymenaea courbaril L. bark activated carbon associated with biofilm.

This study addressed the removal performance of RR2 from aqueous solutions in adsorption columns experiments by comparing the potential of activated carbon alone (ACA) and microbially inoculated (MIAC), prepared from barks of a largely available tree in Brazilian Cerrado biome, Hymenaea courbaril L. or "Jatobá," presenting the kinetics, isotherms, breakthrough curves, and dissolved organic carbon removal. ACA presented strong interaction to RR2 dye, evidenced at the first 20 min when absorbance already attained 66.4%. The removal percentage gradually increased with time and the equilibrium occurred around 91.7% within 120 min. Langmuir model best fitted the isotherm data, indicating a maximum adsorption capacity of 4.068 mg g-1 for the amount of 0.5 g of adsorbent. The Langmuir's model parameters KL, RL, and R2 corresponded to 0.0234 L mg-1, 0.4159, and 0.9663, respectively, indicating a favorable adsorption process (0 < RL < 1). The experiments in adsorption columns revealed maximum adsorption capacities of 14.38 and 11.43 mg g-1 for MIAC and ACA, respectively, where the microbial activity favorably retarded the adsorption breakpoint in approximately 20 min and enhanced the RR2 consumption in 25.8%. Effectiveness of DOC removal attained above 90% for both ACA and MIAC, reducing the content from 86.1 to 7.84 mg L-1 and 4.82 mg L-1, respectively.

A Fatal Bacteremia Caused by Hypermucousviscous KPC-2 Producing Extensively Drug-Resistant K64-ST11 Klebsiella pneumoniae in Brazil.

We report a fatal bacteremia caused by Klebsiella pneumoniae in a 60-70-year-old patient from Brazil. The genomic analysis of three isolates (from blood culture, nasal and anal swabs) showed that the bacteremia was caused by a KPC-2 producing extensively drug-resistant K64-ST11 hypermucousviscous K. pneumoniae (hmKP) harboring several virulence and antimicrobial resistance genes. Although the isolates did not present virulence markers associated with hypervirulent K. pneumoniae (hvKP), they showed invasion and toxicity to epithelial Hep-2 cells; resistance to cell microbicidal mechanisms; and blood and human serum survival, evidencing their pathogenic potential. This study highlights the risk of infection caused by hmKp strains not characterized as hvKP as well as the clinical implications and difficulty of treatment, especially in elderly or immunocompromised patients.

Pollen and seed dispersal of Brazil nut trees in the southwestern Brazilian Amazon / Dispersão de pólen e sementes da castanheira no sudoeste da Amazônia brasileira

Pollen and seed dispersal patterns greatly influence the spatial distribution of plant genetic diversity. Microsatellite-based parentage analysis provides accurate estimates of contemporary gene dispersal. Although most tropical trees have been shown to exhibit widespread pollen dispersal, few studies have estimated contemporary gene dispersal after seedling establishment. Bertholletia excelsa (Lecythidaceae) is pollinated by large-bodied bees, while previous seed-tracking experiments suggest their seeds are mainly dispersed across very short distances by scatter-hoarding rodents, who primarily act as seed predators. Here we used parentage analysis to provide contemporary estimates of pollen and seed dispersal in B. excelsa recruits. We examined six 25-ha plots located in two natural stands in the Acre River valley, in the southwestern Brazilian Amazon. We used 11 microsatellite markers to estimate genetic diversity and fixation index parameters in adults, seedlings and saplings. Genetic diversity was moderate and did not differ across size classes or sampling locations. We assigned pollen and seed parents for < 20% of the recruits, indicating that most events of realized gene flow occurred beyond our 25-ha plots. Only 10 parentage assignments were confirmed with 80% confidence. Pollen distance ranged from 33 to 372 m and seed dispersal from 58 to 655 m. Actual seed-dispersal distances were far greater than the estimates obtained in previous seed-tracking experiments. Thus, studies encompassing larger sampling areas are necessary to determine a more representative spatial scale of B. excelsas pollen and seed dispersal capacity in natural stands.

Os padrões de dispersão de pólen e sementes influenciam a distribuição espacial da diversidade genética. Muitas espécies arbóreas tropicais apresentam ampla dispersão de pólen, mas poucos estudos avaliaram fluxo gênico a partir de plântulas. Bertholletia excelsa (Lecythidaceae) é polinizada por abelhas e as sementes são dispersas por roedores do tipo scatter-hoarders (que estocam recursos em diferentes pontos de sua área de vida), que atuam primariamente como predadores de sementes. Experimentos de remoção de sementes tem mostrado que a dispersão de sementes por esses roedores é espacialmente limitada. Nosso objetivo foi obter estimativas de dispersão de pólen e sementes em B. excelsa a partir da análise de parentesco de regenerantes. Nós estudamos seis parcelas de 25 ha, em duas áreas de floresta nativa no vale do Rio Acre, no sudoeste da Amazônia brasileira. Parâmetros de diversidade genética e índice de fixação foram estimados em adultos, varetas e plântulas com 11 marcadores microssatélites. A diversidade genética foi moderada e não diferiu entre classes de tamanho ou entre localidades. A paternidade foi determinada em menos de 20% dos regenerantes, indicando que a maioria dos eventos de fluxo gênico ocorreu em distâncias maiores que as encontradas nas parcelas de 25 ha. As distâncias de pólen variaram de 33 a 372 m e as de dispersão de sementes variaram de 58 a 655 m. As distâncias de dispersão obtidas neste estudo excedem em muito as estimativas obtidas em experimentos de remoção de sementes. Estudos envolvendo áreas maiores são necessários para que possamos aprofundar nosso conhecimento sobre capacidade de dispersão de pólen e sementes em populações naturais de B. excelsa.

Efficacy of Propolis on the Denture Stomatitis Treatment in Older Adults: A Multicentric Randomized Trial.

Our hypothesis tested the efficacy and safety of a mucoadhesive oral gel formulation of Brazilian propolis extract compared to miconazole oral gel for the treatment of denture stomatitis due to Candida spp. infection in older adults. Forty patients were randomly allocated in a noninferiority clinical trial into two groups. The control group (MIC) received 20 mg/g miconazole oral gel and the study group (PROP) received mucoadhesive formulation containing standardized extract of 2% (20 mg/g) propolis (EPP-AF®) during 14 days. Patients were examined on days 1, 7, and 14. The Newton's score was used to classify the severity of denture stomatitis. The colony forming unity count (CFU/mL) was quantified and identified (CHROMagar Candida®) before and after the treatment. Baseline characteristics did not differ between groups. Both treatments reduced Newton's score (P < 0.0001), indicating a clinical improvement of the symptoms of candidiasis with a clinical cure rate of 70%. The microbiological cure with significant reduction in fungal burden on T14 was 70% in the miconazole group and 25% in the EPP-AF group. The EPP-AF appears to be noninferior to miconazole considering the clinical cure rate and could be recommended as an alternative treatment in older patients.