Lower Detection Probability of Avian Plasmodium in Blood Compared to Other Tissues.

We tested whether the probability of detecting avian haemosporidia (Plasmodium and Haemoproteus) using molecular techniques differs among blood, liver, heart, and pectoral muscle tissues. We used a paired design, sampling the 4 tissue types in 55 individuals of a wild South American suboscine antbird, the white-shouldered fire-eye (Pyriglena leucoptera). We also identified parasites to cytochrome b lineage. Detection probability was significantly lower in blood compared to the other 3 tissue types combined. Eight of 22 infections were not detected in blood samples; 4-7 infections were not detected in the other individual tissues. The same parasite lineage was recovered from different tissues.

Vitamins as influenza vaccine adjuvant components

A number of adjuvant formulations were assayed in mice immunized with 3.75 A mu g of A/California/7/2009 (H1N1) pdm09 influenza vaccine with vitamins A, D and/or E in emulsions or B2 and/or B9 combined with Bordetella pertussis MPLA and/or alum as adjuvants. Squalene was used as positive control, as well as MPLA with alum. The immune response was evaluated by a panel of tests, including a hemagglutination inhibition (HAI) test, ELISA for IgG, IgG1, and IgG2a and IFN-gamma, IL-2, IL-6 and IL-10 quantification in splenocyte culture supernatant after stimulus with influenza antigen. Immunological memory was evaluated using a 1/10 dose booster 60 days after the first immunization followed by assessment of the response by HAI, IgG ELISA, and determination of the antibody affinity index. The highest increases in HAI, IgG1 and IgG2a titers were obtained with the adjuvant combinations containing vitamin E, or the hydrophilic combinations containing MPLA and alum or B2 and alum. The IgG1/IgG2a ratio indicates that the response to the combination of B2 with alum would have more Th2 character than the combination of MPLA with alum. In an assay to investigate the memory response, a significant increase in HAI titer was observed with a booster vaccine dose at 60 days after immunization with vaccines containing MPLA with alum or B2 with alum. Overall, of the 27 adjuvant combinations, MPLA with alum and B2 with alum were the most promising adjuvants to be evaluated in humans

Molecular phylogeny of Threskiornithidae (Aves: Pelecaniformes) based on nuclear and mitochondrial DNA.

The family Threskiornithidae includes 13 genera and 32 species, and it is traditionally divided into 2 subfamilies: Plataleinae and Threskiornithinae. We present a phylogenetic reconstruction to test the monophyly of currently accepted subfamilies, including 15 species from both subfamilies and 10 genera of family Threskiornithidae. Phylogenetic trees were inferred on the basis of the mitochondrial 16S rRNA gene and the nuclear intron 7 of ß-fibrinogen. Threskiornithidae was recovered as a monophyletic group. Plataleinae formed a monophyletic group, but nested within Threskiornithinae, which was thus paraphyletic. Two major phylogenetic groups were identified: the 'endemic New World clade', including genera endemic to the American continent, and the 'widespread clade', comprising the remaining species. These phylogenetic groups diverged about 39-42 million years ago, i.e., before the separation of South America and Antarctica. Our results agree with an initial vicariance due to Gondwana break-up and subsequent colonization of species from the Old World to the New World.

Identifying management units in non-endangered species: the example of the sloth Bradypus variegatus Schinz, 1825

In this study we propose the analysis of genetic diversity of the common three-toed sloth, Bradypus variegatus, in an attempt to understand population structure, identify divergent intraspecific units, and contribute to the knowledge of biodiversity in the neotropical forests. We analyzed a 387 bp segment of the mitochondrial DNA control region in 28 individuals distributed in different localities of both Atlantic and Amazon forests. Our results demonstrated that the genetic diversity of B. variegatus is distributed in six management units, MUs. The observed MUs encompass six phylogenetic lineages and represent respectively north and south regions of Atlantic forest, three regions within the Amazon forest, and a transition region between these two biomes. Considering the fact that these MUs are concordant with phylogroups and endemism areas already described for other vertebrate species, we can say that the study of B. variegatus, a widely distributed and not endangered species, can help to identify areas for conservation biology purposes in neotropical rain forests.

Neste estudo nós realizamos a análise da diversidade genética da preguiça comum, Bradypus variegatus, a fim de compreender os padrões de estrutura populacional, identificar unidades intraespecíficas divergentes e contribuir para o conhecimento da biodiversidade nas florestas da região neotropical. Nós analisamos um segmento de 387 pb da região controle do DNA mitocondrial de 28 indivíduos distribuídos em diferentes localidades da Floresta Amazônica e da Mata Atlântica. Os resultados obtidos demonstram que a diversidade genética da espécie pode ser representada em seis diferentes unidades de manejo (UM). Tais UMs englobam seis linhagens filogenéticas e estão localizadas em diferentes regiões geográficas sendo elas, as porções norte e sul da Mata Atlântica, três regiões dentro da área de Floresta Amazônica e uma área de transição entre os dois domínios de mata. As diferentes unidades intraespecíficas de B. variegatus são concordantes com grupos filogeográficos e áreas de endemismo já observadas para outras espécies de vertebrados. Levando em consideração o fato de que estas UMs concordam com filogrupos e áreas de endemismo previamente descritos para outras espécies de vertebrados, o estudo da preguiça comum, uma espécie amplamente distribuída e considerada não ameaçada de extinção, pode auxiliar na identificação de áreas destinadas à conservação biológica ao longo das florestas úmidas da região neotropical.

Identifying management units in non-endangered species: the example of the sloth Bradypus variegatus Schinz, 1825.

In this study we propose the analysis of genetic diversity of the common three-toed sloth, Bradypus variegatus, in an attempt to understand population structure, identify divergent intraspecific units, and contribute to the knowledge of biodiversity in the neotropical forests. We analyzed a 387 bp segment of the mitochondrial DNA control region in 28 individuals distributed in different localities of both Atlantic and Amazon forests. Our results demonstrated that the genetic diversity of B. variegatus is distributed in six management units, MUs. The observed MUs encompass six phylogenetic lineages and represent respectively north and south regions of Atlantic forest, three regions within the Amazon forest, and a transition region between these two biomes. Considering the fact that these MUs are concordant with phylogroups and endemism areas already described for other vertebrate species, we can say that the study of B. variegatus, a widely distributed and not endangered species, can help to identify areas for conservation biology purposes in neotropical rain forests.

Comparative analyses of the complete genome sequences of Pierce's disease and citrus variegated chlorosis strains of Xylella fastidiosa.

Xylella fastidiosa is a xylem-dwelling, insect-transmitted, gamma-proteobacterium that causes diseases in many plants, including grapevine, citrus, periwinkle, almond, oleander, and coffee. X. fastidiosa has an unusually broad host range, has an extensive geographical distribution throughout the American continent, and induces diverse disease phenotypes. Previous molecular analyses indicated three distinct groups of X. fastidiosa isolates that were expected to be genetically divergent. Here we report the genome sequence of X. fastidiosa (Temecula strain), isolated from a naturally infected grapevine with Pierce's disease (PD) in a wine-grape-growing region of California. Comparative analyses with a previously sequenced X. fastidiosa strain responsible for citrus variegated chlorosis (CVC) revealed that 98% of the PD X. fastidiosa Temecula genes are shared with the CVC X. fastidiosa strain 9a5c genes. Furthermore, the average amino acid identity of the open reading frames in the strains is 95.7%. Genomic differences are limited to phage-associated chromosomal rearrangements and deletions that also account for the strain-specific genes present in each genome. Genomic islands, one in each genome, were identified, and their presence in other X. fastidiosa strains was analyzed. We conclude that these two organisms have identical metabolic functions and are likely to use a common set of genes in plant colonization and pathogenesis, permitting convergence of functional genomic strategies.

Genetic diversity in different populations of sloths assessed by DNA fingerprinting

In this study we analyzed a population of Bradypus torquatus with individuals originally distributed in different localities of Bahia, and two populations of B. variegatus with individuals from Bahia and São Paulo States. Using the DNA fingerprinting method, we assessed the genetic variability within and between populations. Analysis of the DNA profiles revealed genetic similarity indices ranging from 0.34 ± 0.07 to 0.87 ± 0.04. Similar low levels of genetic variability were found only in isolated mammalian populations or among related individuals. This study presents the first analyses of genetic diversity in sloth populations.

Neste estudo foram analisadas uma população de Bradypus torquatus composta por indivíduos provenientes de diferentes localidades do Estado da Bahia e duas populações de B. variegatus, uma na Bahia e outra em São Paulo. A estimativa da variabilidade genética dentro e entre as populações foi feita utilizando-se a técnica de DNA fingerprinting. A partir da análise dos padrões de bandas de DNA, foram calculados índices de similaridade genética que variaram de 0,34 ± 0,07 a 0,87 ± 0,04. Valores semelhantes, que indicam baixa variabilidade genética nas populações de preguiças analisadas, foram encontrados somente em populações de mamíferos isolados ou grupos de indivíduos aparentados. Este trabalho apresenta os primeiros resultados no estudo da diversidade genética em populações de preguiças.

Genetic diversity in different populations of sloths assessed by DNA fingerprinting

In this study we analyzed a population of Bradypus torquatus with individuals originally distributed in different localities of Bahia, and two populations of B. variegatus with individuals from Bahia and Sõo Paulo States. Using the DNA fingerprinting method, we assessed the genetic variability within and between populations. Analysis of the DNA profiles revealed genetic similarity indices ranging from 0.34 ± 0.07 to 0.87 ± 0.04. Similar low levels of genetic variability were found only in isolated mammalian populations or among related individuals. This study presents the first analyses of genetic diversity in sloth populations

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.

Genetic diversity in different populations of sloths assessed by DNA fingerprinting.

In this study we analyzed a population of Bradypus torquatus with individuals originally distributed in different localities of Bahia, and two populations of B. variegatus with individuals from Bahia and São Paulo States. Using the DNA fingerprinting method, we assessed the genetic variability within and between populations. Analysis of the DNA profiles revealed genetic similarity indices ranging from 0.34 +/- 0.07 to 0.87 +/- 0.04. Similar low levels of genetic variability were found only in isolated mammalian populations or among related individuals. This study presents the first analyses of genetic diversity in sloth populations.

Safety and immunogenicity of hepatitis B vaccine ButaNG in adults

Recombinant yeast-derived hepatitis B vaccine manufactured by Instituto Butantan was administered in two groups of adult volunteers (I, II) following two different schedules of immunization. In the first trial (10 &mgr;g doses and 0, 1, 3 months vaccination schedule) 106 individuals completed the full immunization program. The results of seroconversion by age group varied from 70 to 100% and the GMT from 46.5 to 124.9 mIU mL-1. In the second trial with 68 individuals (for dosage comparison and 0, 1, 6 months vaccination schedule) indicated that the vaccine formulated in 20 &mgr;g was more effective than in 10 &mgr;g. The adverse reactions observed in the vaccinees were less frequent than the ones previously found since the introduction of similar vaccines.

DNA fingerprinting in the rare black-fronted piping guan Pipile jacutinga (Cracidae, Aves).

Brazilian Cracidae are threatened by heavy environmental degradation and hunting. The Black-fronted piping-guan (Pipile jacutinga) used to inhabit the Atlantic coastal highland forests. Now it occurs in limited forest areas where it is rarely seen. Interative management, including captive breeding, might be an important action for its survival. We present data on DNA fingerprinting using Jeffreys' human minisatellite probes 33.6 and 33.15. Our results show that this technique is useful for estimating the genetic variability of natural populations and may help to maintain the genetic variability of captive bred individuals of this species. A linkage analysis of the fingerprint profiles in a family with 7 chicks was performed (to estimate the number of independently segregating loci detected in this species) and at least 16 highly polymorphic independent loci were identified for each probe.

[Susceptibility of Vero cell line to vaccine strains of the measles virus]. / Suscetibilidade da linhagem de células Vero a cepas vacinais do vírus do sarampo.

Vero cells used by distinct measles vaccine control laboratories had their susceptibility to Moraten, Schwarz and Biken CAM-70 vaccine strains assayed. Of a total of 72 lots of measles vaccine whose potency was titrated by microtechnique in two Vero cell samples (Vero IB and Vero INCQS), 25 had been produced with Moraten strain, 24 with Schwarz and 23 with Biken CAM-70. The statistical analysis of the results demonstrated that both Vero cells assayed presented comparable susceptibility to Moraten and Biken CAM-70 strains. As to the Schwarz strain, Vero IB cells were more susceptible than the other cell sample tested, thus confirming the existence of different sensitivities of Vero cells to some measles vaccine strains, or even to viruses derived from the same strain but with different passage histories. An altered cell susceptibility to virus replication may significantly alter the results in potency testing. Such alteration may be caused not only by the adoption of distinct protocols for the maintenance of cell cultures by different control laboratories but also by the methodology followed in the vaccine titration. In order to minimize the differences existing among the results obtained in the potency testing, it is suggested that all control laboratories should use the same protocols for cell culture maintenance as well as for vaccine potency testing.

[Identification of mycoplasma by the growth inhibition of samples isolated from cell cultures]. / Identificação de microplasmas pela inibição de crescimento de amostras isoladas de culturas celulares.

Cell cultures must be continuously screened for the presence of mycoplasma because, although these microorganisms sometimes pass unnoticed, they may cause chromosomic alterations and interfere with viral replication, antibody and interferon production etc. The International Organization for Mycoplasmology (IOM) recommends the isolation and identification of mycoplasma with a view to the detection of the origin of the infection and the improvement of the quality of the cultures. In this paper, 37 samples belonging to 27 cell lines contaminated with mycoplasma were assayed by the growth inhibition test. It is known that Mycoplasma orale is the most common human mycoplasma contaminant of cell cultures, the major vehicle of contamination being mouth pippeting, while commercial bovine serum in the main source for Mycoplasma arginini and Acholeplasma laidlawii. M. arginini was found in 18 (48.65%) of the cell samples tested, A. laidlawii in 15 (40.55%), and M. orale in two (5.40%). Two other samples could not be identified by the antisera used (antisera against M. arginini, M. orale, Mycoplasma hyorhinis and A. laidlawii) their characteristics being "fried egg" colonies, digitonine sensitivity, Dienes stained, positive glucose catabolism, negative arginini hydrolysis, and negative tetrazolium reduction. No more than one type of mycoplasma was found in each cell culture.(ABSTRACT TRUNCATED AT 250 WORDS)