Strain Classification of Mycobacterium tuberculosis Isolates in Brazil Based on Genotypes Obtained by Spoligotyping, Mycobacterial Interspersed Repetitive Unit Typing and the Presence of Large Sequence and Single Nucleotide Polymorphism

International audienceRio de Janeiro is endemic for tuberculosis (TB) and presents the second largest prevalence of the disease in Brazil. Here, we present the bacterial population structure of 218 isolates of Mycobacterium tuberculosis, derived from 186 patients that were diagnosed between January 2008 and December 2009. Genotypes were generated by means of spoligotyping, 24 MIRU-VNTR typing and presence of fbpC103, RDRio and RD174. The results confirmed earlier data that predominant genotypes in Rio de Janeiro are those of the Euro American Lineages (99%). However, we observed differences between the classification by spoligotyping when comparing to that of 24 MIRU-VNTR typing, being respectively 43.6% vs. 62.4% of LAM, 34.9% vs. 9.6% of T and 18.3% vs. 21.5% of Haarlem. Among isolates classified as LAM by MIRU typing, 28.0% did not present the characteristic spoligotype profile with absence of spacers 21 to 24 and 32 to 36 and we designated these conveniently as "LAM-like", 79.3% of these presenting the LAM-specific SNP fbpC103. The frequency of RDRio and RD174 in the LAM strains, as defined both by spoligotyping and 24 MIRU-VNTR loci, were respectively 11% and 15.4%, demonstrating that RD174 is not always a marker for LAM/RDRio strains. We conclude that, although spoligotyping alone is a tool for classification of strains of the Euro-American lineage, when combined with MIRU-VNTRs, SNPs and RD typing, it leads to a much better understanding of the bacterial population structure and phylogenetic relationships among strains of M. tuberculosis in regions with high incidence of TB

SHORT COMMUNICATION - Direct Sensitivity Test of the MB/BacT System

In order to evaluate the direct-method test of sensitivity to drugs used in the principal tuberculosis treatment regimes, in the Organon Teknika MB/BacT system, we tested 50 sputum samples positive to microscopy taken from patients with pulmonary tuberculosis and with clinical indications for an antibiogram, admitted sequentially for examination during the routine of the reference laboratory. The material was treated v/v with 23% trisodium phosphate solution, incubated for 24 h at 35°C, and neutralized v/v with 20% monosodium phosphate solution. The material was then centrifuged and the sediment inoculated into flasks containing Rifampin - 2 μg/ml, Isoniazid - 0.2 μg/ml, Pyrazinamide - 100 μg/ml, Ethambutol - 2.5 μg/ml, Ethionamide - 1.25 μg/ml, and Streptomycin - 2 μg/ml. The tests were evaluated using the indirect method in the BACTEC 460 TB (Becton Dickinson) system as the gold standard. The results showed that the Rifampin test performed best, i.e., 100% sensitivity at 95% Confidence Interval (82.2-100) and 100% specificity at 95% Confidence Interval (84.5-100), followed by Isoniazid and Pyrazinamide. In this experiment, 92% of the materials showed a final reading in 30 days; this period represents the time for primary isolation as well as the results of the sensitivity profile, and is within Centers for Disease Control and Prevention recommendations regarding time for performance of the antibiogram. The inoculated flasks showed no contamination during the experiment. The MB/BacT is shown to be a reliable, rapid, fully automated nonradiometric system for the tuberculosis antibiogram

Direct Sensitivity Test of the MB/BacT System

In order to evaluate the direct-method test of sensitivity to drugs used in the principal tuberculosis treatment regimes, in the Organon Teknika MB/BacT system, we tested 50 sputum samples positive to microscopy taken from patients with pulmonary tuberculosis and with clinical indications for an antibiogram, admitted sequentially for examination during the routine of the reference laboratory. The material was treated v/v with 23% trisodium phosphate solution, incubated for 24 h at 35°C, and neutralized v/v with 20% monosodium phosphate solution. The material was then centrifuged and the sediment inoculated into flasks containing Rifampin - 2 µg/ml, Isoniazid - 0.2 µg/ml, Pyrazinamide - 100 µg/ml, Ethambutol - 2.5 µg/ml, Ethionamide - 1.25 µg/ml, and Streptomycin - 2 µg/ml. The tests were evaluated using the indirect method in the BACTEC 460 TB (Becton Dickinson) system as the gold standard. The results showed that the Rifampin test performed best, i.e., 100% sensitivity at 95% Confidence Interval (82.2-100) and 100% specificity at 95% Confidence Interval (84.5-100), followed by Isoniazid and Pyrazinamide. In this experiment, 92% of the materials showed a final reading in 30 days; this period represents the time for primary isolation as well as the results of the sensitivity profile, and is within Centers for Disease Control and Prevention recommendations regarding time for performance of the antibiogram. The inoculated flasks showed no contamination during the experiment. The MB/BacT is shown to be a reliable, rapid, fully automated nonradiometric system for the tuberculosis antibiogram

Temperaturas basais e exigências térmicas do açaizeiro na Amazônia Oriental

The objective of this work was to determine the base temperatures, thermal time requirements, and length of the main reproductive growth stages of acai palm (Euterpe oleracea) in the northeast of the state of Pará, in eastern Amazon, Brazil. The experiment was carried out from 2017 to 2019 in a 10 ha acai plantation, using the time-series analysis. Plant phenology was monitored weekly, and local weather conditions were monitored daily. The lower and upper base temperatures were of 12.92 and 32.46°C, respectively, for pre-flowering; 13.50 and 32.23°C for flowering; 12.14 and 32.55°C for green fruit stage; 11.64 and 32.78°C for fruit color-changing stage; and 11.23 and 32.94°C for maturation. The thermal time requirement and the average cycle length for the ideal harvest time of acai palm were 3,893.15 degree-days and 283 days, respectively. The thermal time requirement and the duration of the reproductive growth stage for acai palm are influenced by the period of the year and the variability of air temperature, which, when high, reduces the cycle of the crop, and when mild, increases it.O objetivo deste trabalho foi determinar as temperaturas basais, as necessidades térmicas e a duração dos principais estádios fenológicos reprodutivos do açaizeiro (Euterpe oleracea) no Nordeste do estado do Pará, na Amazônia Oriental, Brasil. O experimento foi realizado de 2017 a 2019, em plantio de açaizeiro de 10 ha, tendo-se utilizado análise de série temporal. A fenologia das plantas foi monitorada semanalmente, e as condições meteorológicas dos locais foram monitoradas diariamente. As temperaturas basais inferiores e superiores foram de 12,92 e 32,46°C, respectivamente, na prefloração; 13,50 e 32,23°C na floração; 12,14 e 32,55°C no estádio de frutos verdes; 11,64 e 32,78°C no estádio de mudança de cor dos frutos; e 11,23 e 32,94°C na maturação. A exigência térmica e a duração média para o ponto ideal de colheita do açaí foram de 3.893,15 graus-dias e 283 dias, respectivamente. A exigência térmica e a duração dos estádios da fase reprodutiva do açaizeiro são influenciadas pelo período do ano e pela variabilidade da temperatura do ar, que, quando elevada, reduz o ciclo da cultura, e, quando amena, o estende

First detection of Mycobacterium triplex in Latin America

In this study we describe the first isolation of Mycobacterium triplex in Latin America. This species causes infections in humans, with very few reports from around the world. We isolated two sputum specimens of a patient with a 6-year history of human immunodeficiency and tuberculosis treatment failure. All tests used confirmed M. triplex and the patient responded well to drug therapy for 18 months

Whole genome sequence of Mycobacterium kansasii isolates of the genotype 1 from Brazilian patients with pulmonary disease demonstrates considerable heterogeneity.

Mycobacterium kansasii is an opportunistic pathogen and one of the most commonly encountered species in individuals with lung disease. We here report the complete genome sequence of 12 clinical isolates of M. kansasii from patients with pulmonary disease in Brazil

Testiradan toimintavarmuuden parantaminen

Työntarkoituksena oli parantaa katsastusaseman, sään vaihteluille alttiina olevan, testiradan toimintavarmuutta ja lisätä mittatarkkuutta, sekä toistomahdollisuutta muuttuvista keliolosuhteista huolimatta. Testiradan korjauksen kohteena olevat osat olivat heilahduksenvaimentimien testauslaite ja jarrudynamometri. Työssä käsiteltiin kyseisten mittalaitteiden toimintaperiaatteita, sekä niiden käytöstä aiheutuvaa kulumista. Varsinainen työ oli näiden mittalaitteiden kulumisesta aiheutuneiden mahdollisten mittaustarkkuuteen vaikuttavien tekijöiden poistaminen tai ainakin mittatarkkuuden heikkenemisen minimointi. Lisäksi tavoitteena oli parantaa laitteiden huollettavuutta, sekä pidentää niiden käyttöikää. Parannusten jälkeiseen laitteiston toimivuuteen oltiin tyytyväisiä. Joitain kulumisen mukanaan tuomia mahdollisia puutteita tai uusia parannuskohteita ei korjauksen jälkeen pystytty arvioimaan lyhyestä seurantajaksosta johtuen.The aim of the work was to improve inspection station`s reliability of the test track. It is exposed to variable weather conditions and sometimes in harsh Finnish winter, reliability of measuring and testing brakes and shock absorbers is at stakes. Main goal is to minimize this risk of measuring errors and make testing repeatability higher. Parts of the test track that were repaired were brake dynamometer and shock absorber tester. During this work, basic principles of how these instruments work are explained. Also the most common problems caused by wearing are presented and showed how they can be repaired. Or at least minimize the defect of the wear. Reliability and maintainability should be increased by these repairing’s. After repairs, functionality of the gadgets increased and reliability should come to higher level. Despite lack of time, during following period of surveying the most of the units of measure were just like they should have been after the repairs