Few substitutions affect the bioluminescence spectra of Phrixotrix (Coleoptera: Phengodidae) luciferases: a site-directed mutagenesis survey

Phrixotrix (railroad worm) luciferases produce bioluminescence in the green and red regions of the spectrum, depending on the location of the lanterns, and are the only luciferases naturally producing red bioluminescence. Comparison of the luciferase sequences showed a set of substitutions that could be involved in bioluminescence colour determination: (a) unique substitutions in the red luciferase replacing otherwise invariant residues; (b) conserved basic residues in the green-yellow emitting luciferases; and (c) an additional R353 residue in red-emitting luciferase (Viviani et al., 1999). To investigate whether these sites have a functional role in bioluminescence colour determination, we performed a site-directed mutagenesis. Natural substitutions in the region 220-344 and residues in the putative luciferin-binding site were also investigated. With the exception of the previously identified substitution of R215 and T226 (Viviani et al., 2002), which display dramatic red-shift effects on the spectrum of green-yellow-emitting luciferases, only a few substitutions had a moderate effect on the spectrum of the green-emitting luciferase. In contrast, no single substitution affected the spectrum of the red-emitting luciferase. The results suggest that the identity of the active site residues is not so critical for determining red bioluminescence in PxRE luciferase. Rather, the conformation assumed during the emitting step could be critical to set up proper interactions with excited oxyluciferin. Copyright ©2007 John Wiley & Sons, Ltd.

Oftalmomiíase externa causada por cochliomyia hominivorax

The purpose of this study was to report a case of external ophthalmomyiasis caused by Cochliomyia hominivorax in a 10 years-old child, which presented acute preseptal celulitis and had 17 worms removed from the lacrimal canaliculus and conjunctival cul de sac, bilaterally, with resolution. The authors discuss the diagnosis and treatment of this nosological entity.

Influência do cancro basal em Eucalyptus grandis nas propriedades da madeira e polpação kraft

The objective of this research is to assess the influences of basal canker on wood properties for the kraft pulp production. The material consisted of seeded E. grandis trees classified into 4 levels of basal canker severity (0, 1, 2 and 3) and installed in three soil types classified by texture (AQ1 and AQ2 - 10 to 15% clay, and LEm2 - 26 to 35 clay). The sampling consisted of randomly selecting five trees for each for each canker severity level and soil type, totaling 60 trees (4 canker levels x 3 soil types x 5 trees). These trees were fallen and cut into sections at 0, 25, 50, 75 and 100% of the commercial height for the collection of disk and logs of wood. The results showed that the soil texture influences in the Eucalyptus canker severity and this fact should be considered when assessing the wood properties and their final destination. The texture of the soil and the severe levels of basal canker influence the wood properties, and therefore the kraft pulping.

Estimativas de parâmetros genéticos para caracteres quantitativos em progênies de polinização aberta de Eucalyptus urophylla

Eucalyptus urophylla S. T. Blake is outstanding for valuable wood production potential, adaptation plasticity for different environmental conditions, and for tolerance to canker. However, genetic improvement of this specie in Brazil depends on the existence of genetic variability in the introduced populations. The objective of this study was to investigate the genetic variation and gains in quantitative traits in a population established in 1992 in Selvíria - MS. The progeny trial was established in a partially balanced, 8 x 8 lattice design, with 64 families, collected at Anhembi Experimental Station (IPEF/ESALQ/USP). Each plot was made up of eight trees planted in a 3 × 3 m spacing. Significant among families genetic variations were observed in diameter at breast height (DBH), bark type, stem form, and survival. Estimates of average family heritability were high for all traits, ranging from 0.50 to 0.85. The simulated selection by using multi-effect index showed that under a varying number of plants per family. This method is the most indicated to maximize genetic gains (7.24%) and the effective population size (69.3). These results are a useful support for the transformation of this test into a seedling seed orchard and to become a source of vegetative material to build a clonal seed orchard.

Daños causados por cancro cítrico en la producción de naranja dulce 'Valencia'

The citrus canker, caused by Xanthomonas axonopodis pv. citri, is one of the main citrus diseases and has threatened the world's citriculture. In this study, the damage caused by citrus canker was quantified in a sweet orange culture, cv. Valencia [Citrus sinensis (L.) Osb.], grafted over 15 year-old Rangpur lime (C. limonia Osb.), in the municipality of Guatambu, SC, Brazil, in the crop of 2004/05. Evaluations of the number of fruits per plant (NF), fruit yield per plant (P), citrus canker incidence on the fruit (IFR), and incidence (IL) and severity (SV) of citrus canker on 80 leaves per orange tree were performed in the harvest period, applying diagrammatical scales. Plants had large variation in SV (0.16%-1.09%), IL (12.50%-56.25%) and IFR (12.30%-56.09%). Linear regressions were significant between IL x SV, NF x SV, P x IFR, and NF x IFR. There was no significant difference (p>0.05) between the mean estimated values of IL in different quadrants of the crowns of the orange trees (N, S, L and O). Each 1% increase in IFR reduced 2.16 kg and 21.3 fruits per tree.

Extrato de própolis no controle de helmintoses em bezerros

The use of conventional anthelmintic has been a growing concern among consumers who have become more and stricter toward drug residues in products of animal origin. Thus, this research aimed to evaluate the anthelmintic effect of alcoholic extract of propolis, at 30% of concentration, in naturally infected calves. Twelve calves weighting 60 kg and aging 90 days old were used. The treatments were performed by direct oral administration of 10 mL of EAP. The calves received the extract at every 8 h intervals, for five consecutive days. The results were evaluated comprising the detection of light and heavy eggs counted per gram of feces (EPG), immediately before and after treatment, calculating the percentage of reduction. For the statistical analysis, the results were evaluated by Wilcoxon unilateral test of signed points at 5% probability. Trichostrongylus sp. and Strongyloides sp. were found. After treatment, 83% of animals showed an average decrease of 48.48% per counting of EPG. Therefore, propolis may represent an important adjunctive therapy to control helminthiasis in cattle. It was particularly important in ecological organic and biological-dynamic systems of production, whose use of chemicals is prohibitive and limiting factor for certification.

Rhizoctonia solani GA-3PT es el principal patógeno asociado con el chancro del tallo y la sarna negra de la papa en Colombia

Stem canker and black scurf diseases of potatoes are caused by the basidiomycetous fungus Tanatephorus cucumeris (ana-morphic species complex Rhizoctonia solani). Tese diseases have worldwide distribution wherever potato is grown but their etiology varies depending on the predominance of distinct R. solani anastomosis groups (AGs) in a particular area. Within the species complex, several AGs have been associated with stem canker or black scurf diseases, including AG-1, AG-2-1, AG-2-2, AG-3, AG-4, AG-5 and AG-9. Tis article reports on the most comprehensive population-based study, providing evidence on the distribution of R. solani AGs in Colombian potato fields. A total of 433 isolates were sampled from the main potato cropping areas in Colombia from 2005 to 2009. Isolates were assigned to AGs by conventional PCR assays using specific primers for AG-3, sequencing of the ITS-rDNA and hyphal interactions. Most of the isolates evaluated were assigned to AG-3PT (88.45%), and a few to AG-2-1 (2.54%). Te remaining isolates were binucleate Rhizoctonia (AG-A, E, and I). Pathogenicity tests on the stems and roots of different plant species, including the potato, showed that AG-3PT affects the stems of solanaceous plants. In other plant species, damage was severe in the roots, but not the stems. AG-2-1 caused stem canker of Solanum tuberosum cv. Capiro and in R. raphanistrum and B. campestris subsp. Rapa plantlets and root rot in other plants. Te results of our study indicated that R. solani AG-3PT was the principal pathogen associated with potato stem canker and black scurf diseases of potatoes in Colombia.

Antibacterial activity of alkyl gallates against Xanthomonas citri subsp. citri

The plant-pathogenic bacterium Xanthomonas citri subsp. citri is the causal agent of Asiatic citrus canker, a seriousdisease that affects all the cultivars of citrus in subtropical citrus-producing areas worldwide. There is no curative treatment for citrus canker; thus, the eradication of infected plants constitutes the only effective control of the spread ofX. citri subsp. citri. Since the eradication program in the state of São Paulo, Brazil, is under threat, there is a clear risk of X. citri subsp. citri becoming endemic in the main orange-producing area in the world. Here we evaluated the potential use of alkyl gallates to prevent X. citri subsp. citri growth. These esters displayed a potent anti-X. citri subsp. citri activity similar to that of kanamycin (positive control), as evaluated by the resazurin microtiter assay (REMA). Thetreatment of X. citri subsp. citri cells with these compounds induced altered cell morphology, and investigations of the possible intracellular targets using X. citri subsp. citri strains labeled for the septum and centromere pointed to a commontarget involved in chromosome segregation and cell division. Finally, the artificial inoculation of citrus with X. citri subsp. citri cells pretreated with alkyl gallates showed that the bacterium loses the ability to colonize its host, which indicates the potential of these esters to protect citrus plants against X. citri subsp. citri infection. © 2013, American Society for Microbiology.

Resistance against gastrointestinal nematodes in Crioulo Lageano and crossbred Angus cattle in southern Brazil

Gastrointestinal nematode (GIN) infection is a major cause of production losses in cattle. This study was carried out to evaluate the natural resistance against nematode infection in Crioulo Lageano and crossbred Angus male calves. Crioulo Lageano is a local cattle breed in the state of Santa Catarina, in southern Brazil. Ten weaned calves of each breed were grazed together on pasture and naturally infected with nematodes between July 2009 and December 2010. Once every 28 days, we collected fecal and blood samples for parasitological and immunological tests, as well as recording body weights. After 19 samplings, all animals were slaughtered for quantification and identification of GINs. We found that the animals had been infected with the following nematode species, in decreasing order by the mean number of specimens: Trichostrongylus axei, Cooperia punctata, Ostertagia ostertagi, Haemonchus placei, Oesophagostomum radiatum, and Trichuris spp. There were no significant differences between the Crioulo Lageano and crossbred Angus groups in terms of worm burden or nematode fecal egg count, nor in terms of the mean levels of immunoglobulin (G and A) against C. punctata and H. placei antigens, except in IgA mean level in abomasal mucus against H. placei adult worms that was significantly higher in crossbred Angus cattle (p<0.05). At the end of the study, the crossbred Angus cattle were heavier than were the Crioulo Lageano cattle (mean live weight, 507.35 and 390.3. kg, respectively). Comparative parasitological and immunological evaluation revealed no difference between two breeds in terms of their natural resistance against GINs. © 2012 Elsevier B.V.

Haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai

This study evaluated the haematological response of curimbas Prochilodus lineatus, naturally infected with Neoechinorhynchus curemai (Acanthocephala: Neoechinorhynchidae). Thirty-seven fish were captured in October 2010 from the Mogi Guaçu River, Porto Ferreira, SP, Brazil. Infected fish presented increased mean corpuscular volume of erythrocytes, and lower thrombocyte and higher monocyte counts than uninfected fish. © 2013 The Authors. Journal of Fish Biology © 2013 The Fisheries Society of the British Isles.

Resistência de genótipos de eucalipto a ceratocystis spp

Eucalyptus is the most important plantation forest species in Brazil. Wilt and canker caused by Ceratocystis fimbriata on eucalyptus were first reported in 1998 in plantations of an E. grandis × E. urophylla hybrid in southern Bahia, Brazil. This work aimed at studying the reaction of different eucalyptus genotypes after inoculation with C. fimbriata isolates, in order to find a possible source of resistance. The study included four isolates of Ceratocystis collected from eucalyptus in different regions. One disc of fungal mycelium with 1-cm-diameter (from colonies growing for 10 days on malt extract agar medium-MEA) was inoculated on the stem of thus injured eucalyptus plants (six months old). A cotton wool moistened with sterile distilled water was wrapped with plastic film. Control plants were inoculated with discs of MEA without fungal colonies. The inoculated plants were kept in a greenhouse. Wilt symptoms were observed 90 days after inoculation. The seedlings were cut in the longitudinal direction of the stem in order to observe the colonization of fungus in the plant xylem. We tested twenty eucalyptus genotypes, but only five showed resistance to all isolates of Ceratocystis, belonging to different species of Eucalyptus: E. urophylla (C2 and C9), E. grandis (C3), E. saligna (C6 and C13) Most E. gramdis genotypes were more susceptible to all four fungal isolates. These results support future studies related to eucalyptus resistance to Ceratocystis.

Liposomal-praziquantel: Efficacy against Schistosoma mansoni in a preclinical assay

Currently, schistosomiasis mansoni is treated clinically with praziquantel (PZQ). Nevertheless, cases of tolerance and resistance to this drug have been reported, creating the need to develop new drugs or to improve existing drugs. Considering the small number of new drugs against Schistosoma mansoni, the design of nanotechnology-based drug delivery systems is an important strategy in combating this disease. The aim of this study was to evaluate the activity of PZQ containing liposome (lip.PZQ) on S. mansoni, BH strain. Mice were treated orally with different concentrations of PZQ and lip.PZQ 30 and 45 days following infection. The number of worms, recovered by perfusion of the hepatic portal system, and the number of eggs found in the intestine and liver were analysed. Parasite egg counts were also performed. The most active formulation for all parameters was 300. mg/kg of lip.PZQ, since as it decreased the total number of worms by 68.8%, the number of eggs in the intestine by 79%, and the number of hepatic granulomas by 98.4% compared to untreated controls. In addition, this concentration decreased egg counts by 55.5%. The improved efficacy of the treatment with lip.PZQ, especially when administered 45 days following infection, compared with the positive-control group (untreated) and the groups that received free PZQ, can be explained by greater bioavailability in the host organism; the preferred target of lip.PZQ is the liver, and lip.PZQ is better absorbed by the tegument of S. mansoni, which has an affinity for phospholipids. © 2013 Elsevier B.V.

Localização sub-celular de proteínas marcadas com GFP em Xanthomonas axonopodis pv. citri por microscopia de fluorescência

Pós-graduação em Ciências Biológicas (Microbiologia Aplicada) - IBRC

Proteção de bovinos contra Haemonchus placei e Haemonchus contortus após imunização com antígenos oriundos da membrana intestinal de H. contortus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resposta imunológica de cordeiros às infecções artificiais por Haemonchus contortus e Haemonchus placei

Pós-graduação em Biologia Geral e Aplicada - IBB