RESUMO
Fusarium circinatum is thought to have been moved around the world with pine planting stock consisting, most probably, of infected seed. In this effort, we investigate the genetic structure of F. circinatum in Spain and globally. In total, 223 isolates were studied from five regions in northern Spain and eight countries. Eight microsatellite markers revealed 66 multilocus genotypes (MLGs). Minimum spanning network analysis of MLGs by region within Spain as well as globally, discriminant analysis of principal components, and analysis of molecular variance revealed that Spanish populations are significantly differentiated and structured into two distinct groups, each one including one of the dominant genotypes observed. This result suggests that two independent introductions occurred into Spain that subsequently underwent clonal divergence and admixture. This result is further supported by the linkage disequilibrium and clonality observed for F. circinatum populations in northern Spain. The maintenance of differentiation between the clusters could result from the lack of or rare sexual reproduction in Spain. Possible introduction pathways from other countries and subsequent routes of dispersion of F. circinatum in Spain are discussed.
Assuntos
Fusarium/genética , Estruturas Genéticas , Variação Genética , Genética Populacional , Repetições de Microssatélites/genética , Pinus/microbiologia , Chile , DNA Fúngico/genética , Fusarium/isolamento & purificação , Genes Fúngicos Tipo Acasalamento/genética , Genótipo , Geografia , Haplótipos , Japão , Desequilíbrio de Ligação , México , Reação em Cadeia da Polimerase Multiplex , Doenças das Plantas/microbiologia , Portugal , África do Sul , Espanha , Estados Unidos , UruguaiRESUMO
Mesquite (Prosopis pallida (Wildenow) Kunth) is a drought-tolerant tree widely distributed in the northern Pacific Coast of South America. This species prevents soil erosion, provides shade, conserves prairies, supports bee nutrition, and provides fruits for human and animal consumption. Since the spring of 2004, bark lesions and bleeding cankers were observed on trunks and branches of 70% of declining mesquite trees in some parks at Ica in southern Peru. Badly affected trees were killed by the disease. Isolations were made from the edge of necrotic lesions of the inner bark and roots using PARPH medium (2) and incubated at 22°C for 7 days. A Phytophthora species was consistently isolated from lesions of 10 mesquite trees, and six pure cultures (PS-87-PS-92) were obtained by transferring hyphal tips and characterized. Colonies were stellate on V8 juice agar (VJA; 2 g CaCO3, 200 ml of V8 juice, and 15 g of agar in 800 ml of distilled water), uniform to slightly radiate on corn meal agar (Oxoid Ltd., London, England), and knotty on PDA (Biokar Diagnostics, Beauvais, France). On VJA at 22°C, the average radial growth rate for the six isolates was 1.7 mm per day. Colonies grew slowly at 5 and 25°C with 0.4 and 0.7 mm per day growth rate, respectively. There was no growth at 30°C. Catenulate hyphal swellings formed on VJA and liquid media (1.5% sterile soil extract). Sporangia were persistent, ovoid to obpyriform, semipapillate with narrow exit pores (<5.0 µm in diameter), 32.3 to 39.7 × 21.0 to 27.2 µm, with a length/width ratio of 1.4:1 to 1.6:1. Sporangia were produced by cutting 5-mm disks from the advancing margin of a colony on VJA and adding disks to 10 ml of 1.5% sterile soil extract for 4 to 5 days at 22°C under fluorescent light. Isolates were homothallic with spherical oogonia, 32 to 35 µm in width with paragynous antheridia, and aplerotic oospores, 26 to 31 µm. These characteristics fit the descriptions of Phytophthora syringae (Kleb.) Kleb. (1). Sequences of the internal transcribed spacer regions on the isolates and comparison with other sequences in GenBank showed that they were identical to P. syringae (Accession No. AJ854297 from Citrus limon). In 2005, two methods were used to inoculate mesquite with two isolates. One method used two 20-mm-diameter branches of five 5-year-old mesquite trees where a 5-mm wound was made with a cork borer and a 5-mm block of the agar culture was placed under the bark and sealed with Parafilm. Another method used 10 4-month-old potted plants that received a 30-ml drench of a 104 zoospores/ml suspension per plant. Controls received clean agar blocks and a sterile water drench for 10 control pots. Two weeks after inoculation, black areas and resinosis were observed around inoculated wounds. Inoculated branches produced cankers of 4.7 to 6.8 cm2, 4 weeks after inoculations. Twenty days after inoculation of roots, wilting and root rots of seedlings occurred. No symptoms were found on the control plants. P. syringae was reisolated from the diseased branches and root rots and pure cultures were established. This test was repeated for both methods with similar results. To our knowledge, this is the first report of P. syringae in Peru and the first description of this pathogen on mesquite worldwide. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul MN. 1996. (2) S. N. Jeffers and S. B. Martin. Plant Dis. 70:1038, 1986.
RESUMO
In order to evaluate the effectiveness of a new vaccine against human leptospirosis, a prospective cohort study was done with persons in the Cuban province of Holguín who were at risk of becoming ill with leptospirosis. The study included 118,018 persons from 15 to 65 years old who were considered to face either permanent or temporary risk. The vaccinated cohort consisted of 101,137 persons. They received two vaccine doses, 6 weeks apart, of 0.50 mL via deep intramuscular injection into the deltoid muscle of the nondominant arm. The unvaccinated cohort consisted of 16,881 persons. Epidemiological surveillance began 21 days after the application of the second vaccine dose and continued for 1 year. The same criteria for suspected and confirmed cases were maintained throughout the study period. At the end of the surveillance period effectiveness was calculated as being higher than 97%. It is estimated that the vaccination program prevented eight out of ten cases that would have otherwise occurred. Vaccine reactogenicity was also measured in a subsample of 1,500 persons between 15 and 65 years old. The observed symptomatology was low. Slight pain at the injection site was the most frequent symptom (25%). The results of the study indicate the usefulness of the vaccine for disease prevention among people at risk, and its use is thus recommended.
Assuntos
Vacinas Bacterianas/uso terapêutico , Leptospira/imunologia , Leptospirose/prevenção & controle , Adolescente , Adulto , Idoso , Vacinas Bacterianas/efeitos adversos , Vacinas Bacterianas/imunologia , Estudos de Coortes , Cuba , Feminino , Humanos , Esquemas de Imunização , Injeções Intramusculares/métodos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , RiscoRESUMO
A controlled double-blind trial was conducted with the participation of 80 adult volunteers of both sexes, who were randomly divided into groups of 40 individuals each one. The case-base study received the vaccine and the control group was administered placebo to know the safety, the behaviour of reactogenicity, and to star the immunogenicity studies of the first Cuban vaccine against human leptospirosis. The vaccine used in the case-base study was an inactivated and trivalent vaccine containing strains of Leptospira canicola, icterohaemorrhagiae and pomona, since they have the highest circulation in the country. The results obtained showed the inocuity of the vaccine as no adverse severe reactions were detected. The general symptomatology observed was low, where as febricula was the most common general symptom. It appeared during the first 3 days of observation and there were no significant differences between the 2 group. Only a mild pain at the site of the injection was reported as a local symptom, which was more frequent in the vaccinated group than in the control group (7.8 against 1.5%, respectively). The seroconversion obtained was of 29% by microagglutination, and of 34.2% by ELISA. The final results allowed to conclude that this vaccine is safe for human adults at the ages under study, and give the possibility to continue other studies in more advanced stages to complete the requirements for obtaining its license.