Genetic diversity of dengue virus serotypes 1 and 2 in the State of Paraná, Brazil, based on a fragment of the capsid/premembrane junction region

INTRODUCTION: The precise identification of the genetic variants of the dengue virus is important to understand its dispersion and virulence patterns and to identify the strains responsible for epidemic outbreaks. This study investigated the genetic variants of the capsid-premembrane junction region fragment in the dengue virus serotypes 1 and 2 (DENV1-2). METHODS: Samples from 11 municipalities in the State of Paraná, Brazil, were provided by the Central Laboratory of Paraná. They were isolated from the cell culture line C6/36 (Aedes albopictus) and were positive for indirect immunofluorescence. Ribonucleic acid (RNA) extracted from these samples was submitted to the reverse transcription polymerase chain reaction (RT-PCR) and nested PCR. RESULTS: RT-PCR revealed that 4 of the samples were co-infected with both serotypes. The isolated DENV-1 sequences were 95-100% similar to the sequences of other serotype 1 strains deposited in GenBank. Similarly, the isolated DENV-2 sequences were 98-100% similar to other serotype 2 sequences in GenBank. According to our neighbor-joining tree, all strains obtained in this study belonged to genotype V of DENV-1. The DENV-2 strains, by contrast, belonged to the American/Asian genotypes. CONCLUSIONS: The monitoring of circulating strains is an important tool to detect the migration of virus subtypes involved in dengue epidemics.

Molecular characterization of Mycobacterium tuberculosis isolates from Tehran, Iran by restriction fragment length polymorphism analysis and spoligotyping

Abstract: INTRODUCTION Characterization of Mycobacterium tuberculosis (MTB) isolates by DNA fingerprinting has contributed to tuberculosis (TB) control. The aim of this study was to determine the genetic diversity of MTB isolates from Tehran province in Iran. METHODS MTB isolates from 60 Iranian and 10 Afghan TB patients were fingerprinted by standard IS6110-restriction fragment length polymorphism (RFLP) analysis and spoligotyping. RESULTS The copy number of IS6110 ranged from 10-24 per isolate. The isolates were classified into 22 clusters showing ≥ 80% similarity by RFLP analysis. Fourteen multidrug-resistant (MDR) isolates were grouped into 4 IS6110-RFLP clusters, with 10 isolates [71% (95% CI: 45-89%)] in 1 cluster, suggesting a possible epidemiological linkage. Eighteen Iranian isolates showed ≥ 80% similarity with Afghan isolates. There were no strains with identical fingerprints. Spoligotyping of 70 isolates produced 23 distinct patterns. Sixty (85.7%) isolates were grouped into 13 clusters, while the remaining 10 isolates (14.2%) were not clustered. Ural (formerly Haarlem4) (n = 22, 31.4%) was the most common family followed by Central Asian strain (CAS) (n = 18, 25.7%) and T (n = 9, 12.8%) families. Only 1strain was characterized as having the Beijing genotype. Among 60 Iranian and 10 Afghan MTB isolates, 25% (95% CI: 16-37) and 70% (95% CI: 39-89) were categorized as Ural lineage, respectively. CONCLUSIONS A higher prevalence of Ural family MTB isolates among Afghan patients than among Iranian patients suggests the possible transmission of this lineage following the immigration of Afghans to Iran.

The occurrence of the Asian Clam Corbicula fluminea in the lowe amazon basin

This paper records for the first time the presence of Corbicula fluminea (Philipi, 1844) in the Brazilian Amazon Basin. This exotic bivalve was found in localities on the Amazonas, Pará and Tocantins rivers. Density and population size structure were measured in some localities. Mean density is between 6.66 and 7.3 individuals m-2. Population size structure and the dates of the first records suggest that the introductions may have occurred between 1997 and 1998. The introductions may have been mediated by ocean-going vessels visiting the ports of Manaus and Belém. The potential impact of the invasion on native freshwater bivalves is discussed along with the need for monitoring and prevention of further introductions of non-indigenous bivalves in Brazil.

Taxonomy, distribution and population structure of invasive Corbiculidae (Mollusca, Bivalvia) in the Suquía River basin, Córdoba, Argentina

Invasive species are one of the most significant causes of biodiversity loss and changes in ecosystem services, which underlines the importance of their detection and their study. The Asian clams (Corbiculidae) are invasive organisms that accidentally entered the La Plata River, Argentina, presumably in the 1960s. The objectives of the present study were to identify the corbiculid species and to determine their distribution at several locations along the Suquía River basin, an extended area in central Argentina. In addition, population structure was evaluated monthly during one year, at a site in the city of Córdoba that is characterized by high human influence. The presence of Corbicula fluminea (Müller, 1774) and Corbicula largillierti (Philippi, 1844) in the Suquía River basin is reported for the first time. The former species was found only in a lentic environment (San Roque reservoir), while the latter was also found along the tributary rivers and brooks of the basin. Corbicula largillierti showed variations in average density between the different sites and also in biomass and size classes throughout the study period at the site at Córdoba city. The relative composition of the sediments, flow fluctuation and human pollution may be responsible for the observed differences.

Are asian monkeys the original mammalian hosts of Trypanosoma conorhini

It is hypothesized that Asian monkeys were the original hosts of Trypanosoma conorhini- because they have been found naturally infected, the vector among rats is a tropicopolitan triatomine bug that belongs to a complex of Asian species, and primates were shown to be more susceptible than rats.

The Evolution of Trypanosomes Infecting Humans and Primates

Based on phylogenetic analysis of 18S rRNA sequences and clade taxon composition, this paper adopts a biogeographical approach to understanding the evolutionary relationships of the human and primate infective trypanosomes, Trypanosoma cruzi, T. brucei, T. rangeli and T. cyclops. Results indicate that these parasites have divergent origins and fundamentally different patterns of evolution. T. cruzi is placed in a clade with T. rangeli and trypanosomes specific to bats and a kangaroo. The predominantly South American and Australian origins of parasites within this clade suggest an ancient southern super-continent origin for ancestral T. cruzi, possibly in marsupials. T. brucei clusters exclusively with mammalian, salivarian trypanosomes of African origin, suggesting an evolutionary history confined to Africa, while T. cyclops, from an Asian primate appears to have evolved separately and is placed in a clade with T. (Megatrypanum) species. Relating clade taxon composition to palaeogeographic evidence, the divergence of T. brucei and T. cruzi can be dated to the mid-Cretaceous, around 100 million years before present, following the separation of Africa, South America and Euramerica. Such an estimate of divergence time is considerably more recent than those of most previous studies based on molecular clock methods. Perhaps significantly, Salivarian trypanosomes appear, from these data, to be evolving several times faster than Schizotrypanum species, a factor which may have contributed to previous anomalous estimates of divergence times.

Mutations in the rpoB gene of rifampicin-resistant Mycobacterium tuberculosis strains isolated in Brazil and France

We evaluated the mutations in a 193bp of the rpoB gene by automated sequencing of rifampicin (RMP)-resistant and susceptible Mycobacterium tuberculosis strains isolated from Brazil (25 strains) and France (37 strains). In RMP-resistant strains, mutations were identified in 100% (16/16) from France and 89% (16/18) from Brazil. No mutation was detected in the 28 RMP-susceptible strains. Among RMP-resistant or RMP-susceptible strains deletion was observed. A double point mutation which had not been reported before was detected in one strain from France. Among French resistant strains mutations were found in codons 531 (31.2%), 526, 513 and 533 (18.7% each). In Brazilian strains the most common mutations were in codons 531 (72.2%), 526 (11.1%) and 513 (5.5%). The heterogeneity found in French strains may be related to the fact that most of those strains were from African or Asian patients.

The utility of rhesus monkey (Macaca mulatta) and other non-human primate models for preclinical testing of Leishmania candidate vaccines

Leishmaniasis causes significant morbidity and mortality, constituting an important global health problem for which there are few effective drugs. Given the urgent need to identify a safe and effective Leishmania vaccine to help prevent the two million new cases of human leishmaniasis worldwide each year, all reasonable efforts to achieve this goal should be made. This includes the use of animal models that are as close to leishmanial infection in humans as is practical and feasible. Old world monkey species (macaques, baboons, mandrills etc.) have the closest evolutionary relatedness to humans among the approachable animal models. The Asian rhesus macaques (Macaca mulatta) are quite susceptible to leishmanial infection, develop a human-like disease, exhibit antibodies to Leishmania and parasite-specific T-cell mediated immune responses both in vivo and in vitro, and can be protected effectively by vaccination. Results from macaque vaccine studies could also prove useful in guiding the design of human vaccine trials. This review summarizes our current knowledge on this topic and proposes potential approaches that may result in the more effective use of the macaque model to maximize its potential to help the development of an effective vaccine for human leishmaniasis.

Paleoparasitological report on Ascaris aDNA from an ancient East Asian sample

In this study, Ascaris DNA was extracted and sequenced from a medieval archaeological sample in Korea. While Ascaris eggs were confirmed to be of human origin by archaeological evidence, it was not possible to pinpoint the exact species due to close genetic relationships among them. Despite this shortcoming, this is the first Ascaris ancient DNA (aDNA) report from a medieval Asian country and thus will expand the scope of Ascaris aDNA research.

Comparison of spoligotyping, mycobacterial interspersed repetitive units typing and IS6110-RFLP in a study of genotypic diversity of Mycobacterium tuberculosis in Delhi, North India

The aim of the present study was to compare polymerase chain reaction (PCR)-based methods - spoligotyping and mycobacterial interspersed repetitive units (MIRU) typing - with the gold-standard IS6110 restriction fragment length polymorphism (RFLP) analysis in 101 isolates of Mycobacterium tuberculosis to determine the genetic diversity of M. tuberculosis clinical isolates from Delhi, North India. Spoligotyping resulted in 49 patterns (14 clusters); the largest cluster was composed of Spoligotype International Types (SITs)26 [Central-Asian (CAS)1-Delhi lineage], followed by SIT11 [East-African-Indian (EAI) 3-Indian lineage]. A large number of isolates (75%) belonged to genotypic lineages, such as CAS, EAI and Manu, with a high specificity for the Indian subcontinent, emphasising the complex diversity of the phylogenetically coherent M. tuberculosis in North India. MIRU typing, using 11 discriminatory loci, was able to distinguish between all but two strains based on individual patterns. IS6110-RFLP analysis (n = 80 strains) resulted in 67 unique isolates and four clusters containing 13 strains. MIRUs discriminated all 13 strains, whereas spoligotyping discriminated 11 strains. Our results validate the use of PCR-based molecular typing of M. tuberculosis using repetitive elements in Indian isolates and demonstrate the usefulness of MIRUs for discriminating low-IS6110-copy isolates, which accounted for more than one-fifth of the strains in the present study.

Interference of genotypes x environments interaction in the genetic control of resistance to Asian rust soybean

The objectives of this work were to identify parents resistant to Asian soybean rust using diallel crosses, obtain information on the genetic control of soybean resistance to the pathogen and verify whether the combining ability estimates interact with the environment (year or time of assessment). The F1 generation was obtained in a greenhouse from crosses between five contrasting parents for the trait resistance to soybean rust, in a complete diallel without reciprocals. Two rust-severity assessments were carried out on individual soybean plants of 25 treatments (parents and F2 and F3 populations) in 2006/2007 and 2007/2008, in an experimental field at Embrapa Soja, Londrina, PR, Brazil. Additive effects predominated in the genetic control of soybean resistance to Asian rust, and the interaction of the segregant populations with the environment, although significant, did not alter the genetic parameter's general combining ability (GCA) and specific combining ability estimates, indicating that estimates obtained in one year and one assessment can be extrapolated to others. BR01-18437 inbred line is resistant to Asian rust and showed high GCA effects. This line should be used as parent if the objective is the resistance to Phakopsora pachyrhizi.

Selection strategies of segregant soybean populations for resistance to Asian rust

The objective of this work was to identify the best selection strategies for the more promising parental combinations to obtain lines with good resistance to soybean Asian rust (Phakopsora pachyrhizi). Two experiments were carried out in the field during the 2006/2007 and 2007/2008 growing seasons, to determine the percentage of infected leaf area of individual plants of five parents and their segregant F2 and F3 populations. The data obtained indicates that additive genetic variance predominates in the control of soybean resistance to Asian rust, and that the year and time of assessment do not significantly influence the estimates of the genetic parameters obtained. The narrow-sense heritability (h²r) ranged from 23.12 to 55.83%, and indicates the possibility of successful selection of resistant individuals in the early generations of the breeding program. All the procedures used to select the most promising populations to generate superior inbred lines for resistance to P. pachyrhizi presented similar results and identified the BR01-18437 x BRS 232 population as the best for inbred line selection.

Biological aspects of Harmonia axyridis in comparison with Cycloneda sanguinea and Hippodamia convergens

The objective of this work was to evaluate the development, survival, reproductive capacity, and longevity of the Asian ladybug Harmonia axyridis in comparison with Cycloneda sanguinea and Hippodamia convergens. Coccinellid larvae and adults were fed daily with Schizaphis graminum. Ten couples of each species were isolated for evaluation of the adult phase. The duration of the larval stage of H. axyridis is the longest (10.2 days) and its adults are the heaviest (29.7 mg) compared with C. sanguinea and H. convergens. The three species showed similar percentages of survival during the developmental stages. An average of 82% of C. sanguinea, H. axyridis, and H. convergens larvae reached adulthood, which indicates that temperature (25°C) and the offered prey are favorable to coccinellid development. Harmonia axyridis produces a higher total number of eggs per female (1,029.2) than the other evaluated species. However, H. axyridis, which lives for an average of 147.2 days, does not show a significantly greater longevity than C. sanguinea (87.2 days) and H. convergens (134.3 days).

The kiwifruit in Chile and in the world

The kiwi is a fruit that, has only been in Europe for about 60 years, which signifies a great potential for the development of its consumption and great difficulty in the educational process of the product. Currently there is a growing increase in world production, which to date is above the growth in demand. This means that the kiwi categories are not growing significantly and, on the contrary, the kiwi is at a crossroad that will be resolved in the coming years. The changing global economic scenario affects high-value products such as kiwis, making it hard to predict their commercial results. The worldwide market has clearly indicated through the demand and the prices that it is interested in the sweeter yellow varieties. Asian markets have been and are expected to continue to grow. The increase in world production has also generated overlap in stocks between the hemispheres, which makes it necessary to closely monitor commercial strategies regarding the start and end of the season. In this way, the market entry of the southern hemisphere kiwifruit is conditioned according to the presence of fruit from the Northern Hemisphere. The unique features and uses of the kiwi make it one of the most interesting fruits present in the world. For this reason it is also a fruit with high purchase value. In spite of the described variables and market trends, there is a growing demand for consuming better quality kiwis and willingness to pay a higher price.

Adjuvant concentrations and uredospore densities on Phakopsora pachyrhizi infection efficiency in soybean

Experiments were carried out in a growth chamber to evaluate the effect of spreader and uredospore concentrations on the efficiency of infection by Phakopsora pachyrhizi, the causal agent of Asian soybean rust. CD 214 RR soybean cultivar was inoculated with the following polyoxyethylene sorbitane monolaurate concentrations: 0, 30, 60, 120, 240, 480 and 960 µL.L-1 water, as well as a fixed uredospore concentration of 2 x 10(4) spores.mL-1. In a second phase, the inoculum concentrations of 0, 5 x 10³, 1 x 10(4), 2 x 10(4), 4 x 10(4), 8 x 10(4) and 16 x 10(4) uredospores.mL-1 were evaluated, and the spreader concentration of 240 µL.L-1, selected in the previous experiment, was fixed. The spreader concentration of 240 µL.L-1 can be used in artificial inoculation studies, as well as up to 4 x 10(4) uredospores.mL-1. In this work, there was a correlation between uredia and lesion density. Thus, the use of lesion density is recommended to assess disease intensity for its accuracy and less time consuming. There was also a positive correlation between uredia and lesion density.