Comparison of the genomes of two Xanthomonas pathogens with differing host specificities.

The genus Xanthomonas is a diverse and economically important group of bacterial phytopathogens, belonging to the gamma-subdivision of the Proteobacteria. Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, which affects most commercial citrus cultivars, resulting in significant losses worldwide. Symptoms include canker lesions, leading to abscission of fruit and leaves and general tree decline. Xanthomonas campestris pv. campestris (Xcc) causes black rot, which affects crucifers such as Brassica and Arabidopsis. Symptoms include marginal leaf chlorosis and darkening of vascular tissue, accompanied by extensive wilting and necrosis. Xanthomonas campestris pv. campestris is grown commercially to produce the exopolysaccharide xanthan gum, which is used as a viscosifying and stabilizing agent in many industries. Here we report and compare the complete genome sequences of Xac and Xcc. Their distinct disease phenotypes and host ranges belie a high degree of similarity at the genomic level. More than 80% of genes are shared, and gene order is conserved along most of their respective chromosomes. We identified several groups of strain-specific genes, and on the basis of these groups we propose mechanisms that may explain the differing host specificities and pathogenic processes.

Assessment of the genetic diversity of Xylella fastidiosa isolated from citrus in Brazil by PCR-RFLP of the 16S rDNA and 16S-23S intergenic spacer and rep-PCR fingerprinting.

The genetic diversity among twenty three strains of Xylella fastidiosa, isolated from sweet orange citrus, was assessed by RFLP analysis of the 16S rDNA and 16S-23S intergenic spacer and by rep-PCR fingerprinting together with strains isolated from coffee, grapevine, plum and pear. The PCR products obtained by amplification of the 16S rDNA and 16S-23S spacer region were digested with restriction enzymes and a low level of polymorphism was detected. In rep-PCR fingerprinting, a relationship between the strains and their hosts was observed by using the BOX, ERIC and REP primers. Two major groups were obtained within the citrus cluster and relationships to the geographic origin of the strains revealed. Citrus strains isolated from the States of São Paulo and Sergipe formed one group and strains from the Southern States formed another group. Distinct origins of X. fastidiosa in the Southern and Southeastern States is postulated. The pear isolate was distantly related to all of the other X. fastidiosa strains.

First Report of Xylella fastidiosa on Catharanthus roseus in Brazil.

In 1998, plants of periwinkle (Catharanthus roseus L.) showing small leaves, short internodes, and dieback symptoms were observed in a garden at the Instituto Agronomico do Parana (IAPAR), Londrina, PR, Brazil. Stems of these plants were cut into short sections and the sap extracted from the tissue by squeezing with pliers. The sap was blotted onto a glass slide and examined for the presence of bacteria by light microscopy (×400). Microscopy observations revealed the presence of a large number of slender, rod-shaped bacterial cells. The bacteria present in the stems of periwinkle were isolated on buffered cysteine-yeast extract (BCYE) and periwinkle wilt (PW) agar media. Stems were disinfected in 70% alcohol and cut into short sections, and the sap extracted as described above. The sap was blotted directly onto the media and the plates were incubated at 28°C. Typical colonies of Xylella fastidiosa were observed 10 days after isolation on both media. Indirect immunofluorescence tests with antibody specific to X. fastidiosa and anti-IgG conjugated with tetrametylrhodamine isothiocyanate (TRITC) were carried out with xylem sap of periwinkle stem and the isolated bacteria. In both cases, immunofluorescence tests were positive for X. fastidiosa. These results confirm that periwinkle plants were infected with X. fastidiosa. This is the first report of the association of X. fastidiosa with periwinkle plants in Brazil. However, the symptoms observed for the X. fastidiosa-infected periwinkle plants differed from those described previously in the U.S. (1): those symptoms consisted of marginal chlorosis and occasional vein clearing of leaves and wilting of the plants. Reference: (1) R. E. McCoy et al. Plant Dis. Rep. 62:1022, 1978.

Randomized comparative trial of three antivenoms in the treatment of envenoming by lance-headed vipers (Bothrops jararaca) in São Paulo, Brazil.

In São Paulo City, Brazil, 121 patients with moderately severe envenoming by Bothrops snakes (principally B. jararaca) were randomized for treatment with Brazilian polyspecific Bothrops antivenoms: Instituto Butantan (39 patients), Instituto Vital Brazil (41), Fundação Ezequiel Dias (FUNED) (41). The initial dose was four ampoules (40 ml) in 89 patients with less severe envenoming and eight ampoules (80 ml) in 32 patients with more severe envenoming. A second dose of four ampoules was required in 20 patients. Patients receiving the three antivenoms were comparable in all respects before treatment. There were no deaths. The majority showed rapid clinical improvement, resolution of local envenoming, cessation of bleeding and restoration of blood coagulability. No differences in the efficacy of the three antivenoms were revealed by clinical or laboratory observations, including measures of haematological, haemostatic and biochemical abnormalities. Twelve patients developed abscesses (Butantan 1, Vital Brazil 6, FUNED 5) and seven developed local necrosis (3,1,3). Of 88 patients followed up 20-30 days after the bite 33 (37.5%) still had symptoms or signs of local envenoming, especially swelling. Early (anaphylactic) reactions were unexpectedly frequent after all three antivenoms but were significantly more frequent with Butantan (87%) than with Vital Brazil (37%) or FUNED (56%) antivenoms (p < 0.001). A possible explanation was the higher total protein content and percentage immunoglobulin of Butantan antivenom. The doses of antivenom recommended in Brazil and used in this study may be unnecessarily high, resulting in an unacceptably high incidence of reactions. Results of the study should prompt a critical re-evaluation of antivenom production techniques and dosage recommendations in Brazil.

Comparison of the potency of three Brazilian Bothrops antivenoms using in vivo rodent and in vitro assays. BIASG (Butantan Institute Antivenom Study Group).

Three Brazilian polyspecific Bothrops antivenoms were compared using standard W.H.O. rodent in vivo and in vitro assays of their ability to neutralize the principal venom activities of pooled whole Bothrops jararaca venom. On a volume basis, the antivenoms were equally effective in neutralizing lethal activity in mice, and there were only minor differences in their ability to neutralize venom-induced haemorrhage, necrosis and procoagulant activity. Antivenom efficacy in neutralizing defibrinogenation varied. However, when equal amounts of antivenom IgG were compared, it was found that the FUNED antivenom best neutralized lethality, haemorrhage, necrosis and fibrinogen clotting activity. Vital Brazil and FUNED antivenoms were equally effective in neutralizing plasma coagulant activity but Vital Brazil antivenom was the more effective in neutralizing defibrinogenation.

[Lymnic molluscs from an Alto Araguaia area (Brazil)]. / Moluscos límnicos de uma área do Alto Araguaia (Brasil).

An analysis was carried out to determine the natural population of freshwater molluscs from 5 municipalities within the area of influence of the Couto Magalhães hydroelectric project. These municipalities cover a large area of the State of Goiás and Mato Grosso do Sul (Brazil). In this study 624 molluscs of the following species were examined: Drepanotrema anatinum, D. lucidum, D. depressissimum, Biomphalaria straminea, D. schrammi, Physa marmorata, Lymnaea columella, Pomacea and Eupera. One to their importance in public health, the discovery of Biomphalaria straminea and Lymnaea columella, intermediate hosts, respectively, of Shistosoma mansoni and Fasciola hepatica, deserves special mention.