Anti-Sporothrix Antibody Detection in Domestic Cats as an Indicator of a Possible New Occurrence Area for Sporotrichosis in North Brazil

Mycopathologia - Feline sporotrichosis has emerged as an important public health issue in some countries, especially Brazil. Currently, zoonotic transmission of Sporothrix brasiliensis by domestic...     

Low Parasite Load Estimated by qPCR in a Cohort of Children Living in Urban Area Endemic for Visceral Leishmaniasis in Brazil

Background

An important issue associated with the control of visceral leishmaniasis is the need to identify and understand the relevance of asymptomatic infection caused by Leishmania infantum. The aim of this study was to follow the course of asymptomatic L. infantum infection in children in an area of Brazil where it is endemic. The children were assessed twice during a 12-month period.

Methodology

In this population study, 1875 children, ranging from 6 months to 7 years of age, were assessed. Blood samples were collected on filter papers via finger prick and tested by ELISA (L. infantum soluble antigen and rk39). Seropositives samples (n = 317) and a number of seronegatives samples (n = 242) were subjected to qPCR. After 12 months, blood samples were collected from a subgroup of 199 children and tested for Leishmania spp. to follow the course of infection.

Principal Findings

At baseline qPCR testing identified 82 positive samples. The prevalence rate, as estimated for 1875 children based on the qPCR results, was 13.9%. The qPCR testing of whole blood samples collected from a cohort of children after 12 months (n = 199) yielded the following results: of the 44 (22.1%) children with positive qPCR results at baseline, only 10 (5.0%) remained positive, and 34 (17.1%) became negative; and of the 155 (77.9%) children with negative qPCR results, 131 (65.8%) remained negative, and 24 (12.1%) became positive at the follow-up measurement. The samples with positive findings at baseline (n = 82) had a mean of 56.5 parasites/mL of blood; and at follow-up the mean positive result was 7.8 parasites/mL.

Conclusions

The peripheral blood of asymptomatic children had a low and fluctuating quantity of Leishmania DNA and a significant decrease in parasitemia at 1-year follow-up. Quantitative PCR enables adequate monitoring of Leishmania infection.     

Ecto-Nucleotidase Activities of Promastigotes from Leishmania (Viannia) braziliensis Relates to Parasite Infectivity and Disease Clinical Outcome

Background

Leishmania (Viannia) braziliensis has been associated with a broad range of clinical manifestations ranging from a simple cutaneous ulcer to destructive mucosal lesions. Factors leading to this diversity of clinical presentations are not clear, but parasite factors have lately been recognized as important in determining disease progression. Given the fact that the activity of ecto-nucleotidases correlates with parasitism and the development of infection, we evaluated the activity of these enzymes in promastigotes from 23 L. braziliensis isolates as a possible parasite-related factor that could influence the clinical outcome of the disease.

Methodology/Principal Findings

Our results show that the isolates differ in their ability to hydrolyze adenine nucleotides. Furthermore, we observed a positive correlation between the time for peak of lesion development in C57BL/6J mice and enzymatic activity and clinical manifestation of the isolate. In addition, we found that L. (V.) braziliensis isolates obtained from mucosal lesions hydrolyze higher amounts of adenine nucleotides than isolates obtained from skin lesions. One isolate with high (PPS6m) and another with low (SSF) ecto-nucleotidase activity were chosen for further studies. Mice inoculated with PPS6m show delayed lesion development and present larger parasite loads than animals inoculated with the SSF isolate. In addition, PPS6m modulates the host immune response by inhibiting dendritic cell activation and NO production by activated J774 macrophages. Finally, we observed that the amastigote forms from PPS6m and SSF isolates present low enzymatic activity that does not interfere with NO production and parasite survival in macrophages.

Conclusions/Significance

Our data suggest that ecto-nucleotidases present on the promastigote forms of the parasite may interfere with the establishment of the immune response with consequent impaired ability to control parasite dissemination and this may be an important factor in determining the clinical outcome of leishmaniasis.     

In vitro and in vivo activity of meglumine antimoniate produced at Farmanguinhos-Fiocruz, Brazil, against Leishmania (Leishmania) amazonensis, L (L.) chagasi and L (Viannia) braziliensis

The leishmanicidal activity of four batches of meglumine antimoniate, produced in Farmanguinhos-Fiocruz, Brazil (TAMs), was assessed and compared to Glucantime(R)-Aventis Pharma Ltda. Using the amastigote-like in vitro model, the active concentrations of Sb v varied from 10microg/ml to 300 microg/ml for L. (L.) chagasi and from 50microg/ml to 300microg/ml for L. (L.) amazonensis, with no statistically significant differences among the four batches of TAMs and Glucantime(R). The inhibitory concentrations (IC50) determined by the amastigote-infected macrophage model for TAM01/03 and Glucantime(R) were, respectively: 26.3microg/ml and 127.6microg/ml for L. chagasi, 15.4microg /ml and 22.9microg/ml for L. amazonensis, and 12.1microg/ml and 24.2microg/ml for L. (V.) braziliensis. The activities of the four batches of TAMs were confirmed in an in vivo model by assessing, during eight weeks skin lesions caused by L. braziliensis in hamster that were treated with 20mg Sb v/Kg/day for 30 consecutive days. The meglumine antimoniate produced by Farmanguinhos was as effective as the reference drug, Glucantime(R)-Aventis, against three species of Leishmania that are of medical importance in Brazil.     

Habitat attribute similarities reduce impacts of land‐use conversion on seed removal

Changes in land use strongly influence habitat attributes (e.g., herbaceous ground cover and tree richness) and can consequently affect ecological functions. Most studies have focused on the response of these ecological functions to land‐use changes within only a single vegetation type. These studies have often focused solely on agricultural conversion of forests, making it nearly impossible to draw general conclusions across other vegetation types or with other land‐use changes (e.g., afforestation). We examined the consequences of agricultural conversion for seed removal by ants in native grassland, savanna, and savanna‐forest habitats that had been transformed to planted pastures (Brachiaria decumbens) and tree plantations (Eucalyptus spp.) and explored if changes in seed removal were correlated with differences in habitat attributes between habitat types. We found that land‐use changes affected seed removal across the tree cover gradient and that the magnitude of impact was influenced by similarity in habitat attributes between native and converted habitats, being greater where there was afforestation (Eucalyptus spp in grassland and savanna). Herbaceous ground cover, soil hardness, and tree richness were the most important habitat attributes that correlated with differences in seed removal. Our results reveal that the magnitude of impact of land‐use changes on seed removal varies depending on native vegetation type and is associated with the type of habitat attribute change. Our findings have implications for biodiversity in tropical grassy systems: afforestation can have a greater detrimental impact on ecological function than tree loss.     

DOT-dye-immunoassay for the diagnosis of schistosomiasis mansoni.

A new serological assay dot-dye-immunoassay (dot-DIA) was evaluated for the diagnosis of schistosomiasis mansoni. This method consist of four steps: (a) biding of antigens to a nitrocellulose membrane (NC); (b) blocking of free sites of the NC; (c) incubation in specific primary antibody; (d) detection of primary antibody reactivity by color development using second antibody coupled to textile dyes. Sera from 82 individuals, 61 with Schistosoma mansoni eggs in the stool and 21 stool negative were tested by ELISA, dot-ELISA, and dot-DIA. A high level of agreement between the methods tested was observed for all sera tested: ELISA x dot-ELISA: 95.1%, ELISA x dot-DIA: 92.7% and dot-ELISA x dot-DIA: 97.6%. In this study, dot-DIA proved to be a feasible, sensitive, rapid and practical test for the diagnosis of schistosomiasis.     

Geometric morphometrics meets metacommunity ecology: environment and lineage distribution affects spatial variation in shape

Patterns of univariate trait variation across metacommunities are widely explored, as are searches for their underlying causes. Surprisingly, patterns of multivariate shape remain unknown, and the search for drivers of functional traits of communities often neglect the biogeographical distribution of phylogenetic clades. Our aim was to investigate multivariate shape distribution across metacommunities and to determine the main environmental drivers of shape beyond/taking into account the phylogenetic distribution of lineages. We obtained mean skull and mandible shape for 228 species of Neotropical sigmodontine rodents through geometric morphometrics (GM), and then calculated mean shapes for 1° × 1° cells across the Neotropics based on the incidence of sigmodontines. We investigated the effects of lineage distribution on mean trait variation by using phylogenetic fuzzy weighting to calculate principal coordinates of phylogenetic structure (PCPS). Effects of environmental variables on shape variation incorporating phylogenetic composition were realized through redundancy analysis. We found that the different distributions of major lineages throughout the Neotropics were responsible for much of the mean shape variation. The association of landscape features with tribal groupings (Oryzomyini with Amazonia and Phyllotini and Abrotrichini with the Andes) were standouts. Environmental variables and lineage distribution explain the same (i.e. shared) portion of shape variation, suggesting phylogenetic niche conservatism at the metacommunity level. Seasonality in temperature and land cover were the best environmental predictors of mean shape: larger tympanic bullae, incisive foramina, and check teeth are all associated with highly seasonal and less vegetated areas. Our new approach of using GM shape across metacommunities was demonstrably useful in understanding large‐scale biogeographical patterns of shape variation and identifying its underlying causes. The overlap between environmental variables and phylogenetic lineage distribution suggests that a process of niche conservatism is likely: the phenotype–environment correlation is mediated by the differential biogeographical distribution of the main clades.     

Chromosome replication in four species of snakes

Chromosome measurements were performed in four species of snakes related at the level of suborder (Boa constrictor amarali, Xenodon merremii, Philodryas patagoniensis, Bothrops jararaca). The data obtained point out that pairs 1–3 were common to the four snakes and probably inherited from the ancestor of the suborder Serpentes. Pairs 5–8-W were characteristic of each snake; hence, it is possible to assume that they followed evolving after the appearing of the suborder Serpentes. Z-chromosomes were metacentric in B. constrictor amarali, X. merremii and B. jararaca and slightly submetacentric in P. patagoniensis. Area of these chromosomes varied from 8.6–10.6% of the haploid set in the four species studied.-The study of chromosome replication at the end of the S period points out that shared chromosomes have similar patterns of labeling. Therefore, it is proposed that the distribution of late replicating regions and heterochromatin in the genome is phylogenetically transmitted and probably genetically determined.—The analysis of the ending-sequence of chromosome replication shows that sex chromosomes finish earlier than macroautosomes. It is concluded that snakes probably have no mechanism of sex chromosome heterochromatinization in either sex. The absence of late replicating Z-chromosome in the males, favours the hypothesis that no mechanism of sex dosage compensation is acting in the suborder Serpentes.This work was partially supported by Public Health Service Research grant No. GM-14577-03 from the National Institute of General Medical Sciences and by the Fundo de Pesquisas do Instituto Butantan.     

Cytological evidence of constant tetraploidy in the bisexual South American frog Odontophrynus americanus

Odontophrynus americanus has 2n=44 chromosomes in somatic and gonad cells which can be ordered in 11 groups of homologues. In spermatocytes I they form mostly ring quadrivalents. In metaphase II 22 dyads are present. There is no indication of abnormalities in and gonad development.