Scielo RSS <![CDATA[Memórias do Instituto Oswaldo Cruz]]> vol. 110 num. 5 lang. pt <![CDATA[SciELO Logo]]> <![CDATA[ERRATUM]]> <![CDATA[<italic>Memórias</italic> and the Journal Citation Reports]]> <![CDATA[Impact of the age of <italic>Biomphalaria alexandrina</italic> snails on <italic>Schistosoma mansoni</italic> transmission: modulation of the genetic outcome and the internal defence system of the snail]]> <p>Of the approximately 34 identified <italic>Biomphalaria</italic>species,<italic>Biomphalaria alexandrina</italic>represents the intermediate host of<italic> Schistosoma mansoni</italic>in Egypt. Using parasitological and SOD1 enzyme assay, this study aimed to elucidate the impact of the age of<italic> B. alexandrina</italic>snails on their genetic variability and internal defence against<italic> S. mansoni</italic>infection. Susceptible and resistant snails were reared individually for self-reproduction; four subgroups of their progeny were used in experiment. The young susceptible subgroup showed the highest infection rate, the shortest pre-patent period, the highest total cercarial production, the highest mortality rate and the lowest SOD1 activity. Among the young and adult susceptible subgroups, 8% and 26% were found to be resistant, indicating the inheritance of resistance alleles from parents. The adult resistant subgroup, however, contained only resistant snails and showed the highest enzyme activity. The complex interaction between snail age, genetic background and internal defence resulted in great variability in compatibility patterns, with the highest significant difference between young susceptible and adult resistant snails. The results demonstrate that resistance alleles function to a greater degree in adults, with higher SOD1 activity and provide potential implications for <italic>Biomphalaria</italic>control. The identification of the most susceptible snail age enables determination of the best timing for applying molluscicides. Moreover, adult resistant snails could be beneficial in biological snail control.</p> <![CDATA[T-cell receptor Vβ repertoire of CD8<sup>+</sup> T-lymphocyte subpopulations in cutaneous leishmaniasis patients from the state of Rio de Janeiro, Brazil]]> <p>In human cutaneous leishmaniasis (CL), the immune response is mainly mediated by T-cells. The role of CD8<sup>+</sup> T-lymphocytes, which are related to healing or deleterious functions, in affecting clinical outcome is controversial. The aim of this study was to evaluate T-cell receptor diversity in late-differentiated effector (LDE) and memory CD8<sup>+</sup> T-cell subsets in order to create a profile of specific clones engaged in deleterious or protective CL immune responses. Healthy subjects, patients with active disease (PAD) and clinically cured patients were enrolled in the study. Total CD8<sup>+</sup> T-lymphocytes showed a disturbance in the expression of the Vβ2, Vβ9, Vβ13.2, Vβ18 and Vβ23 families. The analyses of CD8<sup>+</sup>T-lymphocyte subsets showed high frequencies of LDE CD8<sup>+</sup>T-lymphocytes expressing Vβ12 and Vβ22 in PAD, as well as effector-memory CD8<sup>+</sup> T-cells expressing Vβ22. We also observed low frequencies of effector and central-memory CD8<sup>+</sup> T-cells expressing Vβ2 in PAD, which correlated with a greater lesion size. Particular Vβ expansions point to CD8<sup>+</sup> T-cell clones that are selected during CL immune responses, suggesting that CD8<sup>+</sup> T-lymphocytes expressing Vβ12 or Vβ22 are involved in a LDE response and that Vβ2 contractions in memory CD8<sup>+</sup>T-cells are associated with larger lesions.</p> <![CDATA[Detection of influenza B lineages from 2001 to 2013 in a tertiary hospital in the city of São Paulo, Brazil]]> <p>Two antigenically distinct lineages of influenza B viruses, the Victoria-like and Yamagata-like strains, currently circulate among humans. Surveillance from United States of America and Europe over the last 10 years showed that the chance of a correct matching between vaccine and the circulating lineage had been 50%. We investigated influenza B infection in different patient groups (asymptomatic, general community, with comorbidities and hospitalised) attended at a tertiary hospital in the city of São Paulo, Brazil between 2001-2013. All samples were screened for influenza B virus by one-step real-time reverse transcription-polymerase chain reaction. From 2,992 respiratory samples collected, 114 (3.8%) tested positive for influenza B. Teenagers (13-18 years) presented the highest rate of 18.5% (odds ratio 22.87, 95% confidence interval 2.90-180.66, p &lt; 0.001). One hundred nine samples could be characterised: 50 were Yamagata-like and 59 were Victoria-like strains. Mismatching between the vaccine and predominant circulating strain was observed in 2002 and 2013 seasons. Based on data collected during a period of 12 years, we found that influenza B was more frequent in teenagers. Co-circulation of both lineages and mismatch with the vaccine strain can occur. Our data highlighted the importance of quadrivalent vaccines and future analysis of the age groups included in vaccination programs.</p> <![CDATA[Effect of <italic>Leishmania</italic> spp infection on the survival, life expectancy, fecundity and fertility of <italic>Lutzomyia longipalpis</italic><italic>s.l</italic>. and <italic>Lutzomyia pseudolongipalpis</italic>]]> <p>We evaluated the effects of<italic> Leishmania</italic>spp infection on several population parameters of <italic>Lutzomyia longipalpis sensu lato</italic> and<italic>Lutzomyia pseudolongipalpis</italic>, vectors of visceral leishmaniasis in Venezuela, under experimental conditions during the first post-feeding period. Females of both species were allowed to feed and engorge on a suspension of fresh washed human red blood cells in foetal calf serum. These blood cells were either non-infected or infected with one of the four<italic>Leishmania</italic>spp strains and were offered through a chicken skin membrane. The longevity, life expectancy and the fecundity of uninfected flies were similar in both species, but the fertility was significantly lower in uninfected<italic> Lu. longipalpis</italic> females. In all cases, the infection of<italic> Lu. longipalpis</italic> and<italic> Lu. pseudolongipalpis</italic> by the<italic> Leishmania</italic>strains resulted in significant detrimental effects, which exerted a fitness cost expressed by reduced survival and life expectancy, as well as decreased fertility and fecundity compared with the control groups. Nevertheless, differences in these parameters were observed between these vector species depending on whether they were infected with the autochthonous Venezuelan<italic> Leishmania infantum</italic> strain (NESA) or the Brazilian reference strain (PP75). The experimental data obtained agree with field data on the natural infection of these vector species and the significance of this scenario is discussed.</p> <![CDATA[Molecular profiling of drug resistant isolates of<italic>Mycobacterium tuberculosis</italic> in the state of Santa Catarina, southern Brazil]]> <p>Drug resistance is a global threat and one of the main contributing factors to tuberculosis (TB) outbreaks. The goal of this study was to analyse the molecular profile of multidrug-resistant TB (MDR-TB) in the state of Santa Catarina in southern Brazil. Fifty-three MDR <italic>Mycobacterium tuberculosis</italic>clinical isolates were analysed by spoligotyping and a partial region of the<italic>rpoB</italic> gene, which is associated with rifampicin resistance (RMP-R), was sequenced. Some isolates were also distinguished by their mycobacterial interspersed repetitive units (MIRU). S531L was the most prevalent mutation found within <italic>rpoB</italic>in RMP-R isolates (58.5%), followed by S531W (20.8%). Only two MDR isolates showed no mutations within<italic>rpoB</italic>. Isolates of the Latin American Mediterranean (LAM) family were the most prevalent (45.3%) found by spoligotyping, followed by Haarlem (9.4%) and T (7.5%) families. SIT106 was found in 26.4% of isolates and all SIT106 isolates typed by MIRU-12 (5 out of 14) belong to MIT251. There was a high correlation between the S531W mutation and the LAM family mainly because all SIT2263 (LAM9) isolates carry this mutation. Among isolates with the S531W mutation in <italic>rpoB</italic> MIRU demonstrates a cluster formed by four isolates (SIT2263 and MIT163) and very similar profiles were observed between eight of the nine isolates. Better characterisation of TB isolates may lead to new ways in which to control and treat TB in this region of Brazil.</p> <![CDATA[Development of a novel plaque reduction neutralisation test for hantavirus infection]]> <p>In the Americas, hantaviruses cause severe cardiopulmonary syndrome (HCPS) with a high fatality rate. Hantavirus infection is commonly diagnosed using serologic techniques and reverse transcription-polymerase chain reaction. This paper presents a novel plaque reduction neutralisation test (PRNT) for detecting antibodies to Brazilian hantavirus. Using PRNT, plaque detection was enhanced by adding 0.6% of dimethyl sulfoxide into the overlay culture medium of the infected cells. This procedure facilitated clear visualisation of small plaques under the microscope and provided for easy and accurate plaque counting. The sera from 37 HCPS patients from the city of Ribeirão Preto, Brazil was evaluated for the Rio Mamoré virus (RIOMV) using PRNT. Six samples exhibited neutralising antibodies; these antibodies exhibited a low titre. The low level of seropositive samples may be due to fewer cross-reactions between two different hantavirus species; the patients were likely infected by Araraquara virus (a virus that has not been isolated) and RIOMV was used for the test. This assay offers a new approach to evaluating and measuring neutralising antibodies produced during hantavirus infections and it can be adapted to other hantaviruses, including viruses that will be isolated in the future.</p> <![CDATA[Biocontrol evaluation of extracts and a major component, clusianone, from <italic>Clusia fluminensis</italic>Planch. & Triana against<italic>Aedes aegypti</italic>]]> <p>Studies evaluated the effects of hexanic extracts from the fruits and flowers of<italic>Clusia fluminensis</italic> and the main component of the flower extract, a purified benzophenone (clusianone), against <italic>Aedes aegypti</italic>. The treatment of larvae with the crude fruit or flower extracts from <italic>C. fluminensis</italic> did not affect the survival of<italic>Ae. aegypti</italic> (50 mg/L), however, the flower extracts significantly delayed development of Ae. aegypti. In contrast, the clusianone (50 mg/L) isolate from the flower extract, representing 54.85% of this sample composition, showed a highly significant inhibition of survival, killing 93.3% of the larvae and completely blocking development of Ae. aegypti. The results showed, for the first time, high activity of clusianone against Ae. aegypti that both killed and inhibited mosquito development. Therefore, clusianone has potential for development as a biopesticide for controlling insect vectors of tropical diseases. Future work will elucidate the mode of action of clusianone isolated from <italic>C. fluminensis</italic>.</p> <![CDATA[Inosine triphosphatase allele frequency and association with ribavirin-induced anaemia in Brazilian patients receiving antiviral therapy for chronic hepatitis C]]> <p>Inosine triphosphatase (ITPA) single nucleotide polymorphisms (SNPs) are strongly associated with protection against ribavirin (RBV)-induced anaemia in European, American and Asian patients; however, there is a paucity of data for Brazilian patients. The aim of this study was to evaluate the ITPA SNP (rs7270101/rs1127354) frequency in healthy and hepatitis C virus (HCV)-infected patients from Brazil and the association with the development of severe anaemia during antiviral therapy. ITPA SNPs were determined in 200 HCV infected patients and 100 healthy individuals by sequencing. Biochemical parameters and haemoglobin (Hb) levels were analysed in 97 patients who underwent antiviral therapy. A combination of AArs7270101+CCrs1127354 (100% ITPase activity) was observed in 236/300 individuals. Anaemia was observed in 87.5% and 86.2% of treated patients with AA (rs7270101) and CC genotypes (rs1127354), respectively. Men with AA (rs7270101) showed a considerable reduction in Hb at week 12 compared to those with AC/CC (p = 0.1475). In women, there was no influence of genotype (p = 0.5295). For rs1127354, men with the CC genotype also showed a sudden reduction in Hb compared to those with AC. Allelic distribution of rs7270101 and rs1127354 shows high rates of the genotypes AA and CC, respectively, suggesting that the study population had a great propensity for developing RBV-induced anaemia. A progressive Hb reduction during treatment was observed; however, this reduction was greater in men at week 12 than in women.</p> <![CDATA[Urban Chagas disease in children and women in primary care centres in Buenos Aires, Argentina]]> <p>The primary objective of this study was to estimate the prevalence of this disease in women of childbearing age and children treated at health centres in underserviced areas of the city of Buenos Aires. Demographic and Chagas disease status data were collected. Samples for Chagas disease serology were obtained on filter paper and the reactive results were confirmed with conventional samples. A total of 1,786 subjects were screened and 73 positive screening results were obtained: 17 were from children and 56 were from women. The<italic> Trypanosoma cruzi</italic>infection risk was greater in those individuals who had relatives with Chagas disease, who remember seeing kissing bugs, who were of Bolivian nationality or were born in the Argentine province of Santiago del Estero. The overall prevalence of Chagas disease was 4.08%. Due to migration, Chagas disease is currently predominantly urban. The observed prevalence requires health programme activities that are aimed at urban children and their mothers. Most children were infected congenitally, which reinforces the need for Chagas disease screening of all pregnant women and their babies in Argentina. The active search for new cases is important because the appropriate treatment in children has a high cure rate.</p> <![CDATA[Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant <italic>Mycobacterium tuberculosis</italic>isolates]]> <p>The purpose of this study is to evaluate four rapid colourimetric methods, including the resazurin microtitre assay (REMA), malachite green decolourisation assay (MGDA), microplate nitrate reductase assay (MNRA) and crystal violet decolourisation assay (CVDA), for the rapid detection of multidrug-resistant (MDR) tuberculosis. Fifty<italic> Mycobacterium tuberculosis</italic>isolates were used in this study. Eighteen isolates were MDR, two isolates were only resistant to isoniazid (INH) and the remaining isolates were susceptible to both INH and rifampicin (RIF). INH and RIF were tested in 0.25 µg/mL and 0.5 µg/mL, respectively. The agar proportion method was used as a reference method. MNRA and REMA were performed with some modifications. MGDA and CVDA were performed as defined in the literature. The agreements of the MNRA for INH and RIF were 96% and 94%, respectively, while the agreement of the other assays for INH and RIF were 98%. In this study, while the specificities of the REMA, MGDA and CVDA were 100%, the specificity of the MNRA was lower than the others (93.3% for INH and 90.9% for RIF). In addition, while the sensitivity of the MNRA was 100%, the sensitivities of the others were lower than that of the MNRA (from 94.1-95%). The results were reported on the seventh-10th day of the incubation. All methods are reliable, easy to perform, inexpensive and easy to evaluate and do not require special equipment.</p> <![CDATA[Activation and cytokine profile of monocyte derived dendritic cells in leprosy: in vitro stimulation by sonicated <italic>Mycobacterium leprae</italic> induces decreased level of IL-12p70 in lepromatous leprosy]]> <p>Dendritic cells (DCs) play a pivotal role in the connection of innate and adaptive immunity of hosts to mycobacterial infection. Studies on the interaction of monocyte-derived DCs (MO-DCs) using <italic>Mycobacterium leprae</italic> in leprosy patients are rare. The present study demonstrated that the differentiation of MOs to DCs was similar in all forms of leprosy compared to normal healthy individuals. In vitro stimulation of immature MO-DCs with sonicated <italic>M. leprae</italic> induced variable degrees of DC maturation as determined by the increased expression of HLA-DR, CD40, CD80 and CD86, but not CD83, in all studied groups. The production of different cytokines by the MO-DCs appeared similar in all of the studied groups under similar conditions. However, the production of interleukin (IL)-12p70 by MO-DCs from lepromatous (LL) leprosy patients after in vitro stimulation with <italic>M. leprae</italic>was lower than tuberculoid leprosy patients and healthy individuals, even after CD40 ligation with CD40 ligand-transfected cells. The present cumulative findings suggest that the MO-DCs of LL patients are generally a weak producer of IL-12p70 despite the moderate activating properties of<italic>M. leprae</italic>. These results may explain the poor<italic> M. leprae</italic>-specific cell-mediated immunity in the LL type of leprosy.</p> <![CDATA[<italic>Corynebacterium diphtheriae</italic> putative tellurite-resistance protein (CDCE8392_0813) contributes to the intracellular survival in human epithelial cells and lethality of <italic>Caenorhabditis elegans</italic>]]> <p><italic>Corynebacterium diphtheriae</italic>, the aetiologic agent of diphtheria, also represents a global medical challenge because of the existence of invasive strains as causative agents of systemic infections. Although tellurite (TeO3<sup>2-</sup>) is toxic to most microorganisms, TeO3<sup>2-</sup>-resistant bacteria, including <italic>C. diphtheriae</italic>, exist <italic>in nature</italic>. The presence of TeO3<sup>2-</sup>-resistance (Te<sup>R</sup>) determinants in pathogenic bacteria might provide selective advantages in the natural environment. In the present study, we investigated the role of the putative Te<sup>R</sup> determinant (CDCE8392_<italic>813</italic>gene) in the virulence attributes of diphtheria bacilli. The disruption of CDCE8392_0813 gene expression in the LDCIC-L1 mutant increased susceptibility to TeO3<sup>2-</sup> and reactive oxygen species (hydrogen peroxide), but not to other antimicrobial agents. The LDCIC-L1 mutant also showed a decrease in both the lethality of <italic>Caenorhabditis elegans</italic>and the survival inside of human epithelial cells compared to wild-type strain. Conversely, the haemagglutinating activity and adherence to and formation of biofilms on different abiotic surfaces were not regulated through the CDCE8392_0813 gene. In conclusion, the CDCE8392_813 gene contributes to the Te<sup>R</sup> and pathogenic potential of <italic>C. diphtheriae.</italic></p> <![CDATA[Influence of breeding site availability on the oviposition behaviour of <italic>Aedes aegypti</italic>]]> <p>Despite the importance of the mosquito <italic>Aedes aegypti</italic> in the transmission of arboviruses, such as yellow fever, Chikungunya fever and dengue fever, some aspects of their behaviour remain unknown. In the present study, the oviposition behaviour of <italic>Ae. aegypti</italic> females that were exposed to different densities of breeding sites (2, 4, 8 and 16) was evaluated in laboratory and semi-field conditions. The number of breeding sites that were used was proportional to the number available, but tended towards stabilisation. Females used four-six breeding sites on average, with a maximum of 11. A high percentage of eggs was observed in the water, along with the presence of a breeding site termed “favourite”, which received at least 40% of the eggs. The results are discussed in ecological, evolutionary and epidemiological approaches.</p> <![CDATA[Full-length infectious clone of a low passage dengue virus serotype 2 from Brazil]]> <p>Full-length dengue virus (DENV) cDNA clones are an invaluable tool for many studies, including those on the development of attenuated or chimeric vaccines and on host-virus interactions. Furthermore, the importance of low passage DENV infectious clones should be highlighted, as these may harbour critical and unique strain-specific viral components from field-circulating isolates. The successful construction of a functional Brazilian low passage DENV serotype 2 full-length clone through homologous recombination reported here supports the use of a strategy that has been shown to be highly useful by our group for the development of flavivirus infectious clones and replicons.</p> <![CDATA[Effects of penicillin and erythromycin on adherence of invasive and noninvasive isolates of Streptococcus pyogenes to laminin]]> <p>This study investigated the possible relationship between the invasiveness of group A <italic>Streptococcus</italic> (GAS) strains and their abilities to adhere to laminin and assessed the effects of subinhibitory concentrations of penicillin and erythromycin on the ability of GAS to adhere to laminin. The adherence of noninvasive and highly invasive isolates of GAS to laminin was significantly higher than the adherence displayed by isolates of low invasiveness. Antibiotic treatment caused significant reductions in adherence to laminin in all three groups of strains. Penicillin was more successful in reducing the adherence abilities of the tested GAS strains than erythromycin.</p> <![CDATA[p<italic>Tc</italic>GW plasmid vectors 1.1 version: a versatile tool for <italic>Trypanosoma cruzi</italic> gene characterisation]]> <p>The functional characterisation of thousands of <italic>Trypanosoma cruzi</italic> genes remains a challenge. Reverse genetics approaches compatible with high-throughput cloning strategies can provide the tool needed to tackle this challenge. We previously published the p<italic>Tc</italic>GW platform, composed by plasmid vectors carrying different options of N-terminal fusion tags based on Gateway<sup>®</sup> technology. Here, we present an improved 1.1 version of p<italic>Tc</italic>GW vectors, which is characterised by a fully flexible structure allowing an easy customisation of each element of the vectors in a single cloning step. Additionally, both N and C-terminal fusions are available with new tag options for protein complexes purification. Three of the newly created vectors were successfully used to determine the cellular localisation of four <italic>T. cruzi</italic> proteins. The 1.1 version of p<italic>Tc</italic>GW platform can be used in a variety of assays, such as protein overexpression, identification of protein-protein interaction and protein localisation. This powerful and versatile tool allows adding valuable functional information to <italic>T. cruzi</italic>genes and is freely available for scientific community.</p> <![CDATA[Draft genome sequence of a multidrug-resistant <italic>Acinetobacter baumannii</italic> ST15 (CC15) isolated from Brazil]]> <p><italic>Acinetobacter baumannii</italic> is an important pathogen frequently associated with nosocomial outbreaks around the world. In Brazil, <italic>A. baumannii</italic> has become particularly problematic because of its prevalence and the carbapenems resistance. Here, we report the draft genome sequence of a multidrug-resistant <italic>A. baumannii</italic>(ST15/CC15) isolated in 2009 from the state of Espírito Santo (Southeast Brazil). We observed important resistance determinant genes in an estimated genome size of 4,102,788 bp with 3,862 predicted coding regions. A detailed report of the genomic data analysis might help to understand the specific features of highly successful strains belonged to a relevant complex clonal in different Brazilian geographical regions.</p> <![CDATA[Trypanocidal, trichomonacidal and cytotoxic components of cultivated<italic>Artemisia absinthium</italic> Linnaeus (Asteraceae) essential oil]]> <p><italic>Artemisia absinthium</italic> is an aromatic and medicinal plant of ethnopharmacological interest and it has been widely studied. The use of<italic>A. absinthium</italic>based on the collection of wild populations can result in variable compositions of the extracts and essential oils (EOs). The aim of this paper is the identification of the active components of the vapour pressure (VP) EO from a selected and cultivated <italic>A. absinthium</italic>Spanish population (T2-11) against two parasitic protozoa with different metabolic pathways:<italic> Trypanosoma cruzi</italic> and<italic>Trichomonas vaginalis</italic>. VP showed activity on both parasites at the highest concentrations. The chromatographic fractionation of the VP T2-11 resulted in nine fractions (VLC1-9). The chemical composition of the fractions and the antiparasitic effects of fractions and their main compounds suggest that the activity of the VP is related with the presence of<italic>trans</italic>-caryophyllene and dihydrochamazulene (main components of fractions VLC1 and VLC2 respectively). Additionally, the cytotoxicity of VP and fractions has been tested on several tumour and no tumour human cell lines. Fractions VLC1 and VLC2 were not cytotoxic against the nontumoural cell line HS5, suggesting selective antiparasitic activity for these two fractions. The VP and fractions inhibited the growth of human tumour cell lines in a dose-dependent manner.</p>