Genomic analysis on Brazilian strains of Anaplasma marginale / Análise genômica de cepas Brasileiras de Anaplasma marginale

Abstract Anaplasma marginale is a vector-borne pathogen that causes a disease known as anaplasmosis. No sequenced genomes of Brazilian strains are yet available. The aim of this work was to compare whole genomes of Brazilian strains of A. marginale (Palmeira and Jaboticabal) with genomes of strains from other regions (USA and Australia strains). Genome sequencing of Brazilian strains was performed by means of next-generation sequencing. Reads were mapped using the genome of the Florida strain of A. marginale as a reference sequence. Single nucleotide polymorphisms (SNPs) and insertions/deletions (INDELs) were identified. The data showed that two Brazilian strains grouped together in one particular clade, which grouped in a larger American group together with North American strains. Moreover, some important differences in surface proteins between the two Brazilian isolates can be discerned. These results shed light on the evolutionary history of A. marginale and provide the first genome information on South American isolates. Assessing the genome sequences of strains from different regions is essential for increasing knowledge of the pan-genome of this bacteria.

Resumo Anaplasma marginale é um patógeno transmitido por vetores que causam uma doença conhecida como anaplasmose. Até a presente data, não há genomas sequenciados de cepas brasileiras. O objetivo deste estudo foi comparar o genoma completo das cepas brasileiras de A. marginale (Palmeira e Jaboticabal) com os genomas de cepas de outras regiões (cepas dos EUA e Austrália). As sequências dos genomas das cepas brasileiras foram obtidas mediante sequenciamento de nova geração. As "reads" foram mapeadas usando-se como referência o genoma de A. marginale da cepa Florida. Foram identificados polimorfismos de nucleotídeo único (SNPs) e analisadas inserções/deleções (INDELs). As duas linhagens brasileiras se agruparam em um clado particular que, por sua vez, agrupou-se em um grupo maior junto com as linhagens norte-americanas. Além disso, foram identificadas diferenças significativas nas proteínas de superfície entre os dois isolados brasileiros. Esses resultados lançam luz sobre a história evolutiva de A. marginale e fornecem as primeiras informações de genomas de isolados sul-americanos. Avaliar as sequências de genomas de cepas de diferentes regiões é essencial para aumentar o conhecimento do pan-genoma dessa bactéria.

Salivary cortisol and metabolic syndrome component's association / Associação do cortisol salivar com os componentes da síndrome metabólica

ABSTRACT Background: Actually the lifestyle exposes the population to several risk factors related to alimentary habits and less physical activity that contributes to chronic diseases appearance worldwide. Aim: To analyze the association between salivary cortisol and the components of metabolic syndrome. Methods: This is a cross-sectional study. As part of it, 28 individuals aged 30-59 years presenting three or more of the following findings: CA: ≥88 cm for women and ≥102 cm for men; SBP>130 mmHg and DBP>85 mmHg; GL>100 mg/dl; TG>150 mg/dl; HDL<40 mg/dl for men and <50 mg/dl for women. Was performed analysis of salivary cortisol (by radioimmunoassay) from 25 salivary samples collected throughout the day, for evaluating changes in the circadian rhythm of this hormone (8AM, noon and 8PM). Results: 28 evaluated individuals had a mean age of 51.9±7.5 years, mostly women (64.3%) and a mean of BMI 33.6±3.2 kg/m². The cortisol level from the 8AM averaged 18.7±4.8 ng/dlL. Individuals with FPG>110mg/dl, have significantly lower average levels of cortisol than ones with FPG <110 (12.8±5,2 vs. 17.3±4.2). Significant correlations were HOMA vs. WC (r=0,465; p˂0,005) and TG (r=0,473; p˂0,005), WC vs. FG (r=0,446; p˂0,005) and BMI (r=0,730; p˂0.0001); TG vs. HDL (r=0,441 p˂0,005) and FG (r=0,440; p˂0,005). Conclusion: Morning salivary cortisol in subjects with chronically elevated blood glucose can represent a downregulation of the hypothalamic-pituitary adrenal axis. This is an important finding not yet well investigated.

RESUMO Racional: Atualmente o estilo de vida expõe a população a diversos fatores de risco relacionados a hábitos alimentares e à inatividade física, contribuindo para o surgimento de doenças crônicas. Objetivo: Analisar a associação entre o cortisol salivar e os componentes da síndrome metabólica. Métodos: Estudo transversal com 28 indivíduos, idade entre 30 e 59 anos apresentando três ou mais dos seguintes achados: circunferência abdominal ≥88 cm (mulheres) e ≥102 cm (homens); pressão arterial sistólica >130 mmHg e pressão arterial diastólica >85 mmHg; glicemia >100 mg/dl; triglicerídeo >150 mg/dl; lipoproteína de alta densidade <40 mg/dl (homens) e <50 mg/dl (mulheres). Foram realizadas coletas do cortisol salivar nos seguintes horários 8 h, 12 h e 20 h e analisadas por radioimunoensaio. Resultados: A média de idade foi 51,9±7,5 anos, 64,3% eram mulheres e a média do índice de massa corporal foi 33,6±3,2 kg/m². O nível de cortisol salivar às 8 h teve média de 18,7±4,8 ng/dl. Os indivíduos com glicemia de jejum >110 mg/dl, apresentaram níveis médios de cortisol significativamente menores que os com glicemia de jejum <110 mg/dl (12,8±5,2 vs. 17,3±4,2). As correlações significativas foram HOMA vs. circunferência abdominal (r=0,465; p˂0,005) e triglicerídeos (r=0,473; p˂0,005), circunferência abdominal vs. glicemia de jejum (r=0,446; p˂0,005) e índice de massa corporal (r=0,730; p˂0,0001), triglicerídeos vs. lipoproteína de alta densidade (r=0,441 p˂0,005) e glicemia de jejum (r=0,440; p˂0,005). Conclusão: O cortisol salivar pela manhã, em indivíduos com glicemia cronicamente elevada, pode representar uma contraregulação do eixo hipotálamo-hipófise-adrenal, sendo achado importante e pouco investigado.

Carbapenem-resistant Acinetobacter baumannii contamination in an intensive care unit

Abstract INTRODUCTION: Acinetobacter baumannii is a major pathogen causing infections in intensive care units (ICUs). In this study, we aimed to evaluate the presence of A. baumannii in an ICU environment and gloves from ICU workers and to characterize the antimicrobial resistance of the isolates in comparison with those isolated from ICU patients at the same hospital. METHODS: ICU samples were collected from March to November 2010. Isolates biochemically characterized as Acinetobacter calcoaceticus-Acinetobacter baumannii complex were evaluated by PCR targeting the 16S rDNA and bla OXA-51 genes. Antimicrobial susceptibility was determined using the disk diffusion method, and carbapenem-resistant isolates were also evaluated for the minimum inhibitory concentration of imipenem using broth microdilution. The presence of the bla OXA-23 gene was evaluated in isolates with reduced susceptibility to carbapenems. RESULTS: A. baumannii was detected in 9.5% (84) of the 886 samples collected from the ICU environment, including from furniture, medical devices, and gloves, with bed rails being the most contaminated location (23.8%; 20/84). Multidrug-resistant (MDR) A. baumannii was found in 98.8% (83/84) of non-clinical and 97.8% (45/46) of clinical isolates. Reduced susceptibility to carbapenems was detected in 83.3% (70/84) of non-clinical and 80.4% (37/46) of clinical isolates. All isolates resistant to carbapenems harbored bla OXA-23. CONCLUSIONS: We found a strong similarity between the antimicrobial susceptibility profiles of non-clinical and clinical A. baumannii isolates. Such data highlight the ICU environment as a potential origin for the persistence of MDR A. baumannii, and hence the ICU may be a source of hospital-acquired infections caused by this microorganism.

A specific polymerase chain reaction method to identify Stenotrophomonas maltophilia

Stenotrophomonas maltophilia is a multidrug-resistant nosocomial pathogen that is difficult to identify unequivocally using current methods. Accordingly, because the presence of this microorganism in a patient may directly determine the antimicrobial treatment, conventional polymerase chain reaction (PCR) and real-time PCR assays targeting 23S rRNA were developed for the specific identification of S. maltophilia. The PCR protocol showed high specificity when tested against other species of Stenotrophomonas, non-fermentative Gram-negative bacilli and 100 clinical isolates of S. maltophilia previously identified using the Vitek system.

Identification of candida spp. by phenotypic tests and PCR

The correct identification of Candida species is of great importance, as it presents prognostic and therapeutical significance, allowing an early and appropriate antifungical therapy. The purpose of this study was to identify isolates of Candida spp. from oral mucosa of 38 patients with oral candidosis evaluated in 2004 by phenotypic methods and PCR, discriminating C. albicans from the other Candida species. The tests used for phenotypic analysis were germ-tube and chlamydoconidia production, culture in CHROMAgarTM Candida, carbohydrate assimilation test, growth at 45ºC and culture in Tween 80 agar. Genotypic confirmation was performed by PCR. Phenotypic tests showed that 63.2 percent strains formed germ-tubes, 73.7 percent produced chlamydoconidia, and 63.2 percent showed green colonies in chromogenic medium, presumptively indicating C. albicans or C. dubliniensis. The carbohydrate assimilation test confirmed these results. A total of 21 percent strains were identified as C. krusei and 13.2 percent were indicative of C. tropicalis. Of these later strains, three produced chlamydoconidia. The association of other phenotypic tests with culture in Tween 80 agar identified 95.8 percent of strains as C. albicans and 4.2 percent as C. dubliniensis. All 24 strains indicative of C. albicans and C. dubliniensis were confirmed by PCR as C. albicans.