Spoligotyping and variable number tandem repeat analysis of Mycobacterium bovis isolates from cattle in Brazil

We performed spoligotyping and 12-mycobacterial interspersed repetitive unit-variable number tandem repeats (MIRU-VNTRs) typing to characterise Mycobacterium bovis isolates collected from tissue samples of bovines with lesions suggestive for tuberculosis during slaughter inspection procedures in abattoirs in Brazil. High-quality genotypes were obtained with both procedures for 61 isolates that were obtained from 185 bovine tissue samples and all of these isolates were identified as M. bovis by conventional identification procedures. On the basis of the spoligotyping, 53 isolates were grouped into nine clusters and the remaining eight isolates were unique types, resulting in 17 spoligotypes. The majority of the Brazilian M. bovis isolates displayed spoligotype patterns that have been previously observed in strains isolated from cattle in other countries. MIRU-VNTR typing produced 16 distinct genotypes, with 53 isolates forming eight of the groups, and individual isolates with unique VNTR profiles forming the remaining eight groups. The allelic diversity of each VNTR locus was calculated and only two of the 12-MIRU-VNTR loci presented scores with either a moderate (0.4, MIRU16) or high (0.6, MIRU26) discriminatory index (h). Both typing methods produced similar discriminatory indexes (spoligotyping h = 0.85; MIRU-VNTR h = 0.86) and the combination of the two methods increased the h value to 0.94, resulting in 29 distinct patterns. These results confirm that spoligotyping and VNTR analysis are valuable tools for studying the molecular epidemiology of M. bovis infections in Brazil.

Simulium (Psilopelmia) virescens, a new black-fly species (Diptera: Simuliidae) from the southwestern region of the state of Bahia, Brazil

The last-instar larva, pupa, male and female of Simulium virescens sp. nov. are described and illustrated. This species has a peculiar larva, which has an elongated head capsule and light-green colour. The first thoracic segment has tubercle on its dorsal region and the third thoracic segment has one pair of tubercles; the first to the fourth abdominal segments have one pair of tubercles on each segment. Until now this new species had only been collected at the type locality, which is on the middle stretch of the Correntina River in the southwestern portion of the state of Bahia, Brazil. Females were voraciously biting humans during the field work. This new species represents the second species of Simulium(Psilopelmia) in Brazil and the first registered outside of the Brazilian Amazon Region.

Antiviral therapy against chronic hepatitis B in Brazil: high rates of lamivudine resistance mutations and correlation with HBV genotypes

The effectiveness of antiviral treatments of chronic hepatitis B has been poorly studied in Brazil. Here, hepatitis B virus (HBV) DNA positivity, drug resistance mutations and their association with HBV genotypes were evaluated in chronically HBV-infected patients under different drug regimens in Brazil. The study involved 129 patients under interferon or nucleos(t)ide analogue therapy for a median treatment time of 12 months. One hundred and five (81%) of these patients were treated with lamivudine (LAM), either in monotherapy or in combination with newer drugs, such as entecavir (ETV) or tenofovir (TDF). High (37.5-100%) rates of HBV DNA positivity were observed with all but one drug regimen (LAM + ETV). However, patients that were treated with ETV alone, TDF alone or with LAM combination therapies had a mean viral load that was 3-4 log lower than patients treated with LAM monotherapy. Of the patients treated with LAM, 47% developed resistance mutations. HBV genotypes A (59.1%), D (30.3%) and F (9.1%) were found. There was no association between the presence of LAM resistance mutations and genotypes, HBeAg status or treatment duration. Nevertheless, the rtM204V mutation was observed more frequently (12/13, 92%) in genotype A than in the others (p = 0.023). Six out of nine isolates that contained the rtM204I mutation belonged to genotype D and half of them displayed a single mutation. Genotype D isolates with the rtM204V variant preferentially displayed a triple mutation, while genotype A preferentially displayed a double mutation (p = 0.04).

Hepatitis B virus genotypes in Southeast Brazil and its relationship with histological features

Data concerning the relationship between hepatitis B virus (HBV) genotypes and liver histology are scarce. The aim of this study was to compare HBV non-B and non-C genotypes according to demographic features, clinical status, HBV-DNA levels and liver histology in Rio de Janeiro. One hundred twenty one consecutive chronic HBV-infected patients were enrolled during two-year period and data were prospectively collected. Sera were tested for HBV genotyping using restriction fragment length polymorphism. Liver biopsy was obtained from patients with either increased alanine aminotransferase (ALT) or HBV-DNA levels. Genotype A was the most common, found in 82 (68%) patients, followed by F in 19 (15%), D in 17 (14%), B in one (1%) and C in two (2%). There was no association between HBV genotypes A, D and F and gender (p = 0.37), age (p = 0.78), race (p = 0.22), mode of infection (p = 0.94), HB "e" antigen status (p = 0.37) and HBV-DNA levels (p = 0.47). The ALT levels were lower in genotype D (75%) compared with A (47%) and F (55%) (p = 0.05). Liver biopsy showed lower inflammation [histological activity index (HAI) = 4] and fibrosis (F) (= 0) scores in genotype D than in genotypes A (HAI = 5, p < 0.001; F = 2, p = 0.008) or F (HAI = 5, p = 0.009; F = 2, p = 0.01). Genotype A was the most prevalent in chronic HBV-infected patients and genotype D patients presented with less intense liver disease.

A new species of Mesocoelium (Digenea: Mesocoeliidae) found in Rhinella marina (Amphibia: Bufonidae) from Brazilian Amazonia

Mesocoelium lanfrediae sp. nov. (Digenea: Mesocoeliidae) inhabits the small intestine of Rhinella marina (Amphibia: Bufonidae) and is described here, with illustrations provided by light, scanning electron microscopy and molecular approachs. M. lanfrediae sp. nov. presents the typical characteristics of the genus, but is morphometrically and morphologically different from the species described previously. The main diagnostic characteristics of M. lanfrediae sp. nov. are (i) seven pairs of regularly-distributed spherical papillae on the oral sucker, (ii) ventral sucker outlined by four pairs of papillae distributed in a uniform pattern and interspersed with numerous spines, which are larger at the posterior margin and (iii) small, rounded tegumentary papillae around the opening of the oral sucker, which are morphologically different from those of the oral sucker itself, some of which are randomly disposed in the ventrolateral tegumentary region of the anterior third of the body. Addionally, based on SSU rDNA, a phylogenetic analysis including Brachycoeliidae and Mesocoeliidae taxa available on GenBank established the close relationship between M. lanfrediae sp. nov. and Mesocoelium sp.

Mecoptera (Insecta) do estado do Maranhão: chave para identificação das espécies e descrição da fêmea de Bittacus latreillei(Collucci & Amorim)

Coletas recentes no estado do Maranhão indicaram que os mecópteros parecem ser abundantes neste estado. Foram estudadas 55 espécimes de quatro espécies: Bittacus diversinervis Souza Lopes & Mangabeira, 1942, B. femoralis Klug, 1836, B. latreillei (Collucci & Amorim, 2000) e Issikiella araguaiensis Penny & Arias, 1983. A maioria dos indivíduos foi coletada entre os meses de outubro e fevereiro, período chuvoso no Maranhão. Uma chave de identificação para essas espécies e a descrição da fêmea de B. latreillei são apresentadas.

Preliminary characterization of Psidium germplasm in different Brazilian ecogeographic regions

The objective of this work was to characterize 119 accessions of guava and 40 accessions of "araçá" sampled in 35 Brazilian ecoregions, according to the International Union for the Protection of New Varieties of Plants (UPOV) descriptors. The majority of "araçá" accessions presented wide spacing of leaf veins, while guava accessions presented medium to close spacing. Most fruits of "araçá" accessions were classified as small, contrasting with medium to large fruits of guava accessions. Most of "araçá" accessions (91%) presented white flesh fruit color, while 58% of guava accessions presented pale pink, pink and dark pink colors. Fruit differences among wild and cultivated Psidium species indicate fruit as the most altered trait under artificial selection.

Root distribution of irrigated grapevine rootstocks in a coarse texture soil of the São Francisco Valley, Brazil

An experiment was carried out to determine the root distribution of four grapevine rootstocks (Salt Creek, Dogridge, Courdec 1613, IAC 572) in a coarse texture soil of a commercial growing area in Petrolina County, São Francisco Valley, Brazil. Rootstocks were grafted to a seedless table grape cv. Festival, and irrigated by microsprinkler. Roots were quantified by the trench wall method aided by digital image analysis. Results indicated that roots reached 1 m depth, but few differences among rootstocks were found. All of them presented at least 90 % of the roots distributed until 0.6 m depth, with a greater root presence in the first 0.4 m. The upper 0.6 m can be taken into account as the effective rooting depth for soil and water management.

Brotação e fertilidade de gemas em uvas sem sementes no Vale do São Francisco

Com o objetivo de avaliar a porcentagem de brotação e índice de fertilidade de gemas de cinco variedades de uvas sem sementes em cinco diferentes datas de poda, realizou-se um experimento no Campo Experimental de Bebedouro, da Embrapa Semi-Árido, em Petrolina-PE. O delineamento experimental foi em parcelas subdivididas, constituídas por cinco variedades de uvas sem sementes como tratamentos principais e cinco diferentes épocas de poda como tratamentos secundários, utilizando-se dez repetições constituídas por uma planta. As variedades utilizadas foram Perlette, Thompson Seedless, Marroo Seedless, Catalunha e Superior Seedless, e as épocas de poda foram: 18 e 19/12/2000; 18 a 21/06/2001; 23 a 29/10/2001; 29/01 a 05/02/2002 e 08 a 12/04/2002. As variáveis estudadas foram a porcentagem de brotação e o índice de fertilidade real das gemas determinado pela razão número de cachos por número de gemas. A fertilidade real de gemas foi analisada entre a 1ª e 10ª gema da vara. Os maiores valores médios para brotação e fertilidade foram observados nas variedades Marroo Seedless e Perlette, sendo que as podas realizadas nos meses de junho de 2001 e abril de 2002 favoreceram o aumento da brotação e da fertilidade das gemas na maioria das variedades.

Caracterização fenológica e requerimentos térmicos de variedades de uvas sem sementes no Vale do São Francisco

O comportamento fenológico e requerimentos térmicos de variedades de uvas sem sementes foram estudados nas condições do Vale do São Francisco durante os anos de 2001-2002, em seis diferentes datas de poda (12/2000, 04/2001, 06/2001, 10/2001, 01-02/2002 e 04/2002). O experimento foi realizado no Campo Experimental de Bebedouro, em Petrolina-PE. As variedades utilizadas foram Superior Seedless (Festival), Thompson Seedless, Catalunha, Perlette e Marroo Seedless, enxertadas sobre porta-enxerto IAC 572 ('Jales'). Avaliou-se a duração em dias dos estádios fenológicos, gemas dormentes (data de poda) a gemas inchadas (1), gemas inchadas a início de brotação (2), início de brotação a 5-6 folhas separadas (3), 5-6 folhas separadas a início de floração (4), início de floração à plena-floração (5), plena-floração a "chumbinho" (6), "chumbinho" à "ervilha" (7), "ervilha" a ½ baga (8), ½ baga a início de maturação (9) e início de maturação à maturação plena (10). Os requerimentos térmicos foram obtidos em termos de graus-dia (GD) necessários para atingir cada fase fenológica a partir da poda. O ciclo fenológico médio variou de 89 dias e 1.315 GD na variedade Superior Seedless a 105 dias e 1.514 GD na variedade Perlette, destacando-se as variedades Superior Seedless e Marroo Seedless como precoces e as demais com ciclo intermediário. Os períodos compreendidos entre o início e final da maturação, ½ baga a início de maturação e 5 a 6 folhas separadas a início de floração apresentaram a maior duração entre todos os subperíodos em todas as variedades.

Guidelines for irrigation scheduling of banana crop in São Francisco Valley, Brazil. I - Root distribution and activity

In order to establish guidelines for irrigation water management of banana cv. Pacovan (AAB group, Prata sub-group) in Petrolina County, northeastern Brazil, the root distribution and activity were measured on an irrigated plantation, in a medium texture soil, with plants spaced in a 3 x 3 m grid. Root distribution was evaluated by the soil profile method aided by digital image analysis, while root activity was indirectly determined by the changing of soil water content and by the direction of soil water flux. Data were collected since planting in January 1999 to the 3rd harvest in September 2001. Effective rooting depth increased from 0.4 m at 91 days after planting (dap), to 0.6 m at 370, 510, and 903 dap, while water absorption by roots was predominantly in the top 0,6 m.

Guidelines for irrigation scheduling of banana crop in São Francisco Valley, Brazil. II - Water consumption, crop coefficient, and physiologycal behavior

The water consumption and the crop coefficient of the banana cv. Pacovan were estimated in Petrolina County, northeastern Brazil, in order to establish guidelines to irrigation water management. Evaluations were carried out since planting in January 1999 to the 3rd harvest in September 2001 on a microsprinkler irrigated orchard, with plants spaced in a 3 x 3 m grid. Average daily water consumption was 3.9, 4.0, and 3.3 mm in the 1st, 2nd and 3rd growing seasons, respectively. Crop coefficient values increased from 0.7 (vegetative growth) to 1.1 (flowering). Even with high soil water availability, transpiration was reduced due to high evaporative demand.

Efeito do ácido giberélico, do bioestimulante crop set e do anelamento na produção e na qualidade da uva 'Thompson Seedless' no Vale do São Francisco

O objetivo do presente trabalho foi avaliar os efeitos do ácido giberélico, do bioestimulante Crop Set e do anelamento sobre o aumento do tamanho de bagas e a produtividade da uva 'Thompson Seedless' no Vale do São Francisco. O experimento foi conduzido durante dois ciclos de produção (2001 e 2002), no Campo Experimental de Bebedouro, pertencente à Embrapa Semi-Árido, em Petrolina-PE. Utilizou-se do delineamento experimental de blocos ao acaso, com 12 tratamentos e 3 repetições. Os tratamentos corresponderam à aplicação de ácido giberélico em cinco fases de desenvolvimento da videira, nas doses de 10 + 15 + 15 +50 + 50 mg.L-1, do bioestimulante Crop Set em duas doses de 0,1 e 0,2% e do anelamento no caule, isolados e combinados entre si. Os tratamentos combinados de anelamento + ácido giberélico e anelamento + ácido giberélico + Crop Set destacaram-se como aqueles que promoveram os maiores peso e tamanho de cachos e de bagas, com diferenças significativas em relação à testemunha. Entretanto, o anelamento não cicatrizou completamente, causando a morte de plantas, recomendando-se cautela na sua realização. Apesar de não se observar efeito significativo dos tratamentos sobre a produtividade, pode-se notar um aumento de 63% para o tratamento anelamento + ácido giberélico em relação ao ciclo de produção de 2001.

Similaridade genética de acessos de mangueira de diferentes origens geográficas avaliadas por marcadores AFLP

As relações genéticas de 105 acessos de diferentes origens geográficas do banco de germoplasma de mangueira da Embrapa foram determinadas com base no marcador AFLP, de forma a orientar trabalhos de melhoramento e manejo de recursos genéticos da espécie para a região Semi-Árida brasileira. Foram ainda incluídos dois acessos de duas espécies do gênero Mangifera, como "outgroup". O DNA dos acessos foi extraído pelo método do CTAB, as reações de AFLP foram realizadas para os iniciadores EcoRI/MseI e as bandas polimórficas foram analisadas para construção de fenograma, baseando-se no coeficiente de similaridade de Jaccard. Foram obtidas 157 e 54 bandas de AFLP polimórficas e monomórficas, respectivamente, em 13 combinações de iniciadores. O valor co-fenético do fenograma foi estimado em 0,81. Foram observados cinco grupos: 1) cultivares como Amrapali, Malika, híbridos Embrapa-Cpac e algumas variedades americanas formando um grupo; 2) grupo formado, predominantemente, por cultivares americanas, com algumas inclusões de híbridos sul-africanos e brasileiros; 3) grande grupo formado por cultivares brasileiras, com algumas inclusões de cultivares australianas, indianas e americanas; 4) grupo formado por algumas variedades tipo Espada, Rosa e acessos de diferentes origens; e 5) grupo formado por M. foetida e M. similis. Os acessos Carabao e Manilla apresentaram a maior similaridade, 97%. Os acessos estudados apresentaram similaridade superior a 51%, evidenciando a alta variabilidade genética da coleção de germoplasma de mangueira estudada.

Concentrações e associações geoquímicas de Pb e Zn em sedimentos do rio São Francisco impactados por rejeitos da produção industrial de zinco

Surface sediments from the River São Francisco were analyzed to investigate the impact, due to the presence of metals (Cd, Cu, Pb, Zn) in wastes from a metallurgical industry in the city of Três Marias/MG, Brazil. The concentrations and geochemical associations of Pb, Zn and trace metals associated with the minerals employed in zinc production were measured. Sediments close to discharge locations were highly contaminated with Pb (332-512 μg g-1) and Zn (7872-10780 μg g-1), with values decreasing rapidly due to dilution and hydraulic sorting. Evaluation of toxicity according to the Consensus-based Sediment Quality Guidelines indicated for Cd, Pb and Zn a high probability of adverse effects on aquatic biota at these sites.