Annona squamosa, a new host of Ceratocystis fimbriata

Mango branch blight disease, caused by Ceratocystis fimbriata, is endemic to the municipality of São Fidelis in northern Rio de Janeiro State. In addition to mango, C. fimbriata was found associated with sugar apple trees (Annona squamosa) showing symptoms of branch blight in São Fidelis. Sugar apple and mango isolates from the same region had the same morphology and showed similar ITS-rDNA sequences. These sequences were also similar to other Brazilian isolates of C. fimbriata sensu stricto. Cross inoculation of such isolates obtained from diseased sugar apple and mango resulted in diseased symptoms on both plant species. This is the first record of A. squamosa as a host for C. fimbriata.

Quantitative control of Lettuce mosaic virus fitness and host defence inhibition by P1-HCPro

Two Lettuce mosaic virus isolates capable of overcoming the resistance afforded by the resistance gene mo1² in lettuce, LMV-AF199 from Brazil, and LMV-E, an European isolate, were evaluated for the rapidity and severity of symptoms induced on the lettuce variety Salinas 88 (mo1²). The mosaic symptoms on Salinas 88 plants inoculated with LMV-AF199 appeared 7 days post-inoculation (dpi) and 15 dpi for LMV-E. The symptoms induced by LMV-AF199 in this cultivar were also more severe than those induced by LMV-E. In order to identify the region of the viral genome responsible for this phenotype, recombinant viruses were constructed between these isolates and the phenotype of each recombinant was analysed. The region encoding proteins P1 and HcPro from LMV-AF199 was associated with the increased virulence in Salinas 88.

SPECIES RICHNESS AND ABUNDANCE OF LOW-TRUNK HERB EPIPHYTES IN RELATION TO HOST TREE SIZE AND BARK TYPE, EASTERN AMAZONIA

ABSTRACTThe composition and structure of the low-trunk epiphytic herb assembly as well as its vertical distribution were studied. DBH of host tree and bark type influence species richness and abundance in a nonlooded lowland tropical rainforest in Eastern Amazonia (1º57’36"S 51º36’55"W). A total of 37 epiphytic herb species were identified, among which 60% were Araceae. Species richness and abundance of epiphytic herbs showed tendency of positive correlation with host tree size and no relationships with bark type. Low positive correlation may be a by-product of the predominance of trees with smaller diameter in our sample rather than a reflection of neutral relationship. The absence of relationships with bark type may be partially explained by the large number of secondary, generalist, hemi-epiphytes and also may reflect the absence of suitable substrate in trees with smaller diameter.

Phase variation in Flavobacterium psychrophilum : influence on host-pathogen interactions

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Host suitability of weeds and forage species to root-knot nematode meloidogyne graminicola as a funcion of irrigation management

The irrigated rice production can be limited by various phytopathogenic agents, including root-knot nematodes (Meloidogyne spp.). Thus, the aim of this research was to check the host suitability of plant species most often found off-season and during rice cultivation, to root-knot nematode Meloidogyne graminicola, under two irrigation managements. Two experiments were conducted in a completely randomized design. In the first experiment seven plant species that occur in an area of rice cultivation, in fallow, off-season were evaluated. For the second experiment nine weed species infesting the irrigated rice culture were tested in rainfed and flooding conditions. The sixteen species, kept individually in pots with sterilized substrate, were inoculated with 5,000 eggs and second stage juveniles (J2) of nematode. BRS 410 IRGA rice plants inoculated with M.graminicola were used as control. Two months after inoculation, the root system of each plant was evaluated for number of galls and nematode reproduction factor. It was verified that the species of off-season of rice cultivation Sida rhombifolia, Raphanus raphanistrum, Spergula arvensis, Lotus corniculatus and Trifolium repens, and, during the cycle of rice cultivation, Aeschynomene denticulata, Leersia hexandra, are immune to nematode. The plant species off-season, Avena strigosa and Lolium multiflorum and of cultivation, Alternanthera philoxeroides, red rice, Echinochloa crusgalli, Cyperus difformis, Cyperus esculentus, Cyperus iria and Fimbristylis miliacea would behave as hosts of M.graminicola, mostly under rainfed conditions.

Effects of using different host plants on the detected biodiversity of arbuscular mycorrhizal fungi from an agroecosystem

The influence of peanut (Arachis hypogaea L.), sorghum (Sorghum bicolor (L.) Moench) and maize (Zea mays L.) on the development and diversity of arbuscular mycorrhizal fungi (AMF) from an agrosystem was investigated. Soil collected from an agricultural field where maize had been grown was inserted into sowing holes, under the seeds of peanut, sorghum and maize those were subsequently grown in sterilised quartz sand separately in plastic boxes for five months. After this period, collections of roots and rhizospheric soil were made to evaluate the percentages of root colonization (RC), number of spores (NS) and species of AMF. Peanut showed the highest average values for RC and NS: 24.5% and 547.8/100 g of soil, respectively. Maize had an average RC of 19.7% and 415.2 spores/100g soil. Sorghum showed the lowest values: 15.9% for average RC and 349.8 spores/100 g soil. A total of fourteen species of AMF were identified. Seven species were identified from peanut rhizospheres, Entrophospora colombiana being the most abundant and frequent. In sorghum rhizospheres, twelve species were found, Glomus geosporum was the dominant taxon in terms of number of spores and frequency. Ten species were detected in maize with Acaulospora longula being the most abundant and the most frequent. It was observed that peanut was the best plant for promoting the sporulation of AMF, while sorghum favoured the establishment of most AMF species, followed by maize.

Host Parasite Interactions in Damselflies. From Individuals to Populations

The main goal of this thesis is to increase understanding on evolutionary and ecological factors that have contributed to differences in parasite numbers in insects. Furthermore, the thesis addresses the effects of parasites on their hosts. The most important findings were: The Northern damselfly’s (Coenagrion hastulatum) immune response to artificial pathogen increased with increasing parasite numbers (Article I). Marginal, more isolated C. hastulatum populations on the edge of distribution have fewer parasites when compared to distribution’s core populations (Article II). The Banded damselfly Calopteryx splendens individuals with higher homozygosity have more parasites, however, the rate of homozygosity did not differ between populations (Article III). Parasite prevalence was affected by whether the host species occurred in allopatric or sympatric population: sympatric C. splendens populations with sister species the Beautiful damselfly Calopteryx virgo harbored more parasites (Article IV). Parasites were associated with the wing spot size, an ornament under sexual selection, and thus may play an important role in character displacement, i.e. the size of the wing spot (Article V). To conclude with, this thesis brings about new information on the parasite infection patterns in insects, proposing several factors to contribute to these patters, as well as it addresses the effects of parasites on their hosts, from individual to population level.

Effects of host resistance on the isozymatic patterns of Bipolaris sorokiniana (Dematiaceae, Moniliales)

Nine isolates of Bipolaris sorokiniana were inoculated on three cultivars of wheat plants (susceptible, moderately resistant, resistant). Eight days after the inoculation, the isolates were recovered (27 isolates) and the following isozymatic patterns were analyzed: esterase, alkaline phosphatase, acid phosphatase, malate dehydrogenase, and superoxide dismutase. The esterase system was the most polymorphic, and the isolates recovered from the susceptible cultivar showed the highest variability. This is evidence that this cultivar exerts low selection pressure on the pathogen

The use of confocal laser scanning microscopy to analyze the process of parasitic protozoon-host cell interaction

In this communication we review the results obtained with the confocal laser scanning microscope to characterize the interaction of epimastigote and trypomastigote forms of Trypanosoma cruzi and tachyzoites of Toxoplasma gondii with host cells. Early events of the interaction process were studied by the simultaneous localization of sites of protein phosphorylation, revealed by immunocytochemistry, and sites of actin assembly, revealed by the use of labeled phaloidin. The results obtained show that proteins localized in the interaction sites are phosphorylated. The process of formation of the parasitophorous vacuole was monitored by labeling the host cell surface with fluorescent probes for lipids (PKH26), proteins (DTAF) and sialic acid (FITC-thiosemicarbazide) before interaction with the parasites. Evidence was obtained indicating transfer of components of the host cell surface to the parasite surface in the beginning of the interaction process. We also analyzed the distribution of cytoskeletal structures (microtubules and microfilaments visualized with specific antibodies), mitochondria (visualized with rhodamine 123), the Golgi complex (visualized with C6-NBD-ceramide) and the endoplasmic reticulum (visualized with anti-reticulin antibodies and DIOC6) during the evolution of intracellular parasitism. The results obtained show that some, but not all, structures change their position during evolution of the intracellular parasitism.

Apoptosis in parasites and parasite-induced apoptosis in the host immune system: a new approach to parasitic diseases

Apoptosis, a form of programmed cell death (PCD), has been described as essential for normal organogenesis and tissue development, as well as for the proper function of cell-renewal systems in adult organisms. Apoptosis is also pivotal in the pathogenesis of several different diseases. In this paper we discuss, from two different points of view, the role of apoptosis in parasitic diseases. The description of apoptotic death in three different species of heteroxenic trypanosomatids is reviewed, and considerations on the phylogenesis of apoptosis and on the eventual role of PCD on their mechanism of pathogenesis are made. From a different perspective, an increasing body of evidence is making clear that regulation of host cell apoptosis is an important factor on the definition of a host-pathogen interaction. As an example, the molecular mechanisms by which Trypanosoma cruzi is able to induce apoptosis in immunocompetent cells, in a murine model of Chagas' disease, and the consequences of this phenomenon on the outcome of the experimental disease are discussed.

Adhesion of the human pathogen Sporothrix schenckii to several extracellular matrix proteins

The pathogenic fungus Sporothrix schenckii is the causative agent of sporotrichosis. This subcutaneous mycosis may disseminate in immunocompromised individuals and also affect several internal organs and tissues, most commonly the bone, joints and lung. Since adhesion is the first step involved with the dissemination of pathogens in the host, we have studied the interaction between S. schenckii and several extracellular matrix (ECM) proteins. The binding of two morphological phases of S. schenckii, yeast cells and conidia, to immobilized type II collagen, laminin, fibronectin, fibrinogen and thrombospondin was investigated. Poly (2-hydroxyethyl methacrylate) (poly-HEMA) was used as the negative control. Cell adhesion was assessed by ELISA with a rabbit anti-S. schenckii antiserum. The results indicate that both morphological phases of this fungus can bind significantly to type II collagen, fibronectin and laminin in comparison to the binding observed with BSA (used as blocking agent). The adhesion rate observed with the ECM proteins (type II collagen, fibronectin and laminin) was statistically significant (P<0.05) when compared to the adhesion obtained with BSA. No significant binding of conidia was observed to either fibrinogen or thrombospondin, but yeast cells did bind to the fibrinogen. Our results indicate that S. schenckii can bind to fibronectin, laminin and type II collagen and also show differences in binding capacity according to the morphological form of the fungus.

Correlation of mixed lymphocyte culture with chronic graft-versus-host disease following allogeneic stem cell transplantation

The purpose of the present study was to evaluate the mixed lymphocyte culture as a predictive assay of acute and chronic graft-versus-host disease (GVHD). We studied 153 patients who received a first bone marrow transplantation from human leukocyte antigen-identical siblings. Acute GVHD was observed in 26 of 128 (20.3%) patients evaluated and chronic GVHD occurred in 60 of 114 (52.6%). One-way mixed lymphocyte culture (MLC) assays were performed by the standard method. MLC results are reported as the relative response (RR) from donor against patient cells. The responses ranged from -47.0 to 40.7%, with a median of 0.5%. The Kaplan-Meier probability of developing GVHD was determined for patients with positive and negative MLC. There was no significant difference in incidence of acute GVHD between the groups studied. However, the incidence of chronic GVHD was higher in recipients with RR >4.5% than in those with RR <=4.5%. The Cox Proportional Hazards model was used to examine the effect of MLC levels on incidence of chronic GVHD, while adjusting for the potential confounding effect of others suspected or observed risk factors. The relative risk of chronic GVHD was 2.5 for patients with positive MLC (RR >4.5%), 2.9 for those who received peripheral blood progenitor cells as a graft, and 2.2 for patients who developed previous acute GVHD. MLC was not useful for predicting acute GVHD, but MLC with RR >4.5% associated with other risk factors could predict the development of chronic GVHD, being of help for the prevention and/or treatment of this late complication.