Integrative taxonomy of Sprentascaris mahnerti and phylogeny of raphidascaridid nematodes (Nematoda: Ascaridoidea: Raphidascarididae) suggest the monophyly of Sprentascaris.

Sprentascaris mahnerti (Nematoda: Raphidascarididae) collected from Loricariichthys labialis (Siluriformes: Loricariidae) in the Pantanal wetlands, State of Mato Grosso do Sul (Brazil), was redescribed using light and scanning electron microscopy (SEM), and genetically characterised along with two other raphidascaridids: Raphidascaroides brasiliensis and Ro. moraveci. Due to the systematic discussion regarding Raphidascaris and Sprentascaris, as well as the poor knowledge about the phylogenetic relationships within Raphidascarididae, phylogenies were reconstructed based on partial sequences of the 18S and 28S nuclear rRNA gene, the nuclear ITS1-5.8S-ITS2 and the cytochrome c oxidase subunit I (cox1) mtDNA. Morphological study of S. mahnerti, confirmed some previously described features, revealed new characteristics and permitted to elucidate some inconsistencies noted in the literature. Morphological and genetic characterisation of S. mahnerti supported its validity. Phylogenetic reconstructions supported the monophyly of Sprentascaris, which has three pairs of interlabial conspicuous cuticular projections as a synapomorphy. The relationships among several lineages of raphidascaridids were unsolved, albeit Goezia and Ichthyascaris formed well-supported monophyletic assemblages, in which the first included species with no relations regarding the habitat of hosts and the geographic origin. The present findings represent one more step towards the understanding of the interrelationships of raphidascaridid nematodes. In this sense, Sprentascaris should be considered valid as an independent lineage from Raphidascaris.

First description of the male of Philometroides acreanensis and phylogenetic assessment of Philometridae (Nematoda: Dracunculoidea) suggest instability of some taxa.

The male of Philometroides acreanensis, parasitic in the anterior intestine external wall of the freshwater catfish Pimelodus blochii, from the Brazilian Amazon, is described for the first time. Additional data on the morphology of females is given. The new morphological data strengthened the validity of the species as well as its first genetic characterization, using three nuclear genetic markers (18S and 28S of the rDNA and ITS1-58S-ITS2), confirmed the high genetic resemblance of male and female specimens. Philometroides acreanensis shows morphological features of the generic diagnosis of Neophilometroides, Alinema, Philometra and Philometroides. Phylogenetic analyses using sequences of the18S rDNA from representatives of Dracunculoidea confirmed the validity of P. acreanensis and its close relatedness with Alinema rather than with other genera. The validity of Philonemidae was confirmed, as was the monophyly of Philometridae and Clavinema. However, Dentiphilometra, Philometra and Philometroides appear not to be monophyletic. Host taxa, habitat and geographic occurrence seem to have some relationship with the evolutionary traits of certain phylogenetic assemblages of philometrids, which were highly supported in the phylogentic reconstructions. Even though interesting aspects of the phylogeny and taxonomy of Philometridae came to light, further integrative approaches should be used that include additional genetic markers, due to the loose boundaries between some genera as observed here.

Insight into role of microbiota-gut-brain peptides as a target for biotechnology innovations.

It has long been understood that some microorganisms may modify their hosts behavior in various systems. Nevertheless, it has only been in recent years that gut microbiota have opened new perspectives to appreciate their potential for affect complex neurological function in mammals. Efforts have demonstrated the ability of these gut-microbiota to impact neurological outcomes, suggested a prominent role for the gut microbiota in the gut-brain interactions, indicating that alterations in bidirectional microbiota-brain-gut may be involved in a number of brain disorders. Further, the identification of bioactive microbial signals, including their immune mediators, gut hormones and/or peptides, during health and disease situations, can serve as a tool for discovering novel activities that influence behavior and neurological function in hosts. Current review aims to provide an overview and shed some light on fundamental characteristics of the gut microbiota in modulating neurological disorders and consequently to draw up alternative strategies for using the gut microbiota or their active molecules as a therapeutic target for future diagnoses.

Avaliação genética das vacinas contra a brucelose bovina comercializadas no Brasil / Genetic evaluation of vaccine against bovine brucellosis marketed in Brazil

A prevenção contra infecções causadas por Brucella abortus em bovinos é realizada por meio da administração das amostras vacinais B19 e RB51. Existem relatos de que estas vacinas podem causar aborto em fêmeas vacinadas. Portanto, toda a ocorrência de aborto em animais vacinados merece um estudo aprofundado sobre a causa. No Brasil, não há registro sobre a origem das amostras B19 e RB51 utilizadas na produção das vacinas comerciais. Assim, um estudo para verificar possíveis mutações em relação às amostras referência USDA B19 e USDA RB51 de B. abortus se faz necessário, devido às amostras vacinais poderem reverter a sua virulência. Objetivou-se com este estudo caracterizar genotipicamente as amostras vacinais B19 e RB51 comercializadas no Brasil. A metodologia utilizada foi a genotipagem de genes marcadores destas amostras vacinais, por meio da amplificação pela reação em cadeia da polimerase. Os resultados obtidos permitiram a identificação do genótipo das vacinas comerciais B19 e RB51 disponíveis e utilizadas em bovinos no Brasil. A ausência de mutações nas vacinas testadas corrobora com a qualidade genética das mesmas, quanto à estabilidade dos genes analisados.

Vaccine strains B19 and RB51 are administered to cattle for prevention against infection by Brucella abortus. However, there are reports that these vaccines can cause miscarriages. Thus, every miscarriage among vaccinated animals should be thoroughly studied to determine the cause. In Brazil, there are no records on the origin of B19 and RB51 samples used in the preparation of commercial vaccines. Therefore, a study is needed to determine possible mutations in relation to the USDA reference samples of B. abortus due to the fact that vaccine samples could revert to the virulence of the disease. The aim of the present study was to perform a genotype analysis of vaccine strains B19 and RB51 used in Brazil. The methodology was based on the genotyping of marker genes of these vaccine strains by amplification using polymerase chain reaction. The results allowed the identification of the genotype of the B19 and RB51 commercial vaccine available for use on cattle in Brazil. The absence of mutations in the samples tested confirmed the genetic quality of the vaccines and stability of genes analyzed.

Polymorphisms of intron 1 and the promoter region at the PRNP gene in BSE-free Caracu cattle.

The infectious prion protein PrP(Sc) is encoded by the PRNP gene. In cattle, insertion/deletion (indel) polymorphisms are among the changes that occur in this gene, the most studied of which are within intron 1 (12 bp) and the promoter region (23 bp). Sequence variants in this gene may affect the formation of PrP(Sc). In the present study, nucleotide variability in specific regions of the PRNP gene in Caracu cattle free of bovine spongiform encephalopathy was investigated to determine the genotypic profile of each animal within the group. Caracu cattle exhibited high allele frequency for the two polymorphic regions studied, 12ins (70 %) and 23ins (72.5 %), genotype frequencies of 50 % for 12ins/ins and 50 % for 23ins/del, and a high frequency of the 12ins-23ins haplotype (57.5 %). Of the 40 animals sampled, 15 had the 12ins-23ins/12ins-23ins diplotype.

Caracterização da imunogenicidade das proteínas recombinantes Virb9, Virb10 e fator termo instável de Elongação de Peptídeos de Anaplasma marginale em camundongos / Characterization of the immunogenicity of recombinant proteins VirB9, VirB10 and Elongation Factor Tu of Anaplasma marginale in mice

Considerando as limitações dos atuais métodos de controle contra a anaplasmose bovina, o desenvolvimento de uma vacina efetiva se faz necessário. A partir do advento da análise genômica e proteômica, novas proteínas de membrana de Anaplasma marginale foram identificadas como possíveis candidatas a componentes de uma vacina, tais como, VirB9, VirB10 e Fator Termo Instavél de Elongação de Peptídeos (EF-Tu). Embora estas proteínas ainda não estejam bem caracterizadas na membrana de A. marginale, a produção destas na forma recombinante (rVirB 9, rVirB10 e rEF-Tu) tem sido realizada, mas as mesmas ainda não foram exploradas em formulações vacinais. Neste trabalho, avaliou-se o uso de rVirB9, rVirB10 e rEF-Tu emulsionadas em adjuvante Montanide em camundongos. Nas condições testadas, verificou-se a indução de forte resposta imune humoral com a produção de IgG1 e IgG2a, sendo que as proporções dos níveis de produção destas subclasses indicam predomínio de IgG1. Entretanto, esplenócitos de animais, que foram injetados com rVirB9 ou rVirB10, produziram interferon-gama acima do limite de detecção do ensaio após estimulação in vitro, sinalizando assim resposta celular específica. Assim, novas avaliações serão realizadas com a finalidade de modular o perfil de resposta imune obtido em bovinos e avaliar a proteção contra A. marginale.

Considering the limitations of current methods of anaplasmosis control, the development of a more effective vaccine is required. Previous studies, using proteomic and genomic approaches, have identified new membrane proteins in Anaplasma marginale that may be vaccine candidates. These include VirB9, VirB10 and elongation factor-Tu (EFTu). Although the role of these proteins in the membrane of A. marginale has not been properly characterized, production of the recombinant proteins rVirB9, rVirB10, and rEF-Tu has been achieved. However, these recombinant proteins have not yet been exploited in vaccine formulations. The present study describes the use of rVirB9, rVirB10 and rEF-Tu, emulsificated in adjuvant Montanide, in mice. A strong humoral immune response was induced under these conditions, with both IgG1 and IgG2a production. The IgG2a/IgG1 ratios revealed a predominance of IgG1. However, splenocytes of the animals that received rVirB9 or rVirB10 produced high levels of gamma interferon after in vitro stimulation, indicating a specific cellular immune response to these proteins. Therefore, further studies are required to adjust the profile of the immune response in order to perform tests of protection against A. marginale in cattle.

Evidência molecular de brucella sp. em ozotoceros bezoarticus (veado campeiro) do Pantanal sul-mato-grossense / Molecular evidence of brucella sp. in deer (ozotoceros bezoarticus) of the southern Pantanal

A presença de Brucella spp. entre animais silvestres pode influenciar a taxa de reprodução destes hospedeiros, além de atuarem como fonte de infecção natural para os animais domésticos e humanos. O objetivo deste estudo foi identificar a presença de Brucella spp. em 44 amostras de sangue de veado campeiro (Ozotoceros bezoarticus) do Pantanal do Sul-Mato-Grossense, utilizando a técnica de PCR. Observou-se que 20,4 por cento (9/44) das amostras foram positivas. A sequência consenso de nucleotídeo obtida no sequenciamento do isolado de veado campeiro apresentou 514 pb e 95 por cento de identidade com virB5 de B. abortus (best hits acesso nr AF226278, e-value 0.0), já na análise filogenética a amostra de Brucella isolada de veado campeiro apresentou-se muito próximo de B. suis. A alta porcentagem de amostras positivas sugere que a brucelose pode ser um problema entre os veados campeiros na área estudada e que estes animais podem representar riscos para outros animais domésticos e silvestres.

The presence of Brucella spp. in wild animals can influence their reproduction rate and may be a source of infection for domestic animals and humans. The objective of this study was to identify the presence of Brucella spp. in 44 blood samples from the deer Ozotoceros bezoarticus in the southern Pantanal of Sul-Mato-Grossense, using the PCR technique. It was seen that 20.4 percent (9/44) of the samples were positive. The consensus sequence was obtained by sequencing these samples, which then showed 514 pb and 95 percent of identity with gene virB5 of B. abortus (best hits accession nr AF226278, e-value 0.0). The phylogenetic analysis of the sample isolated from deer revealed the Brucella to be very close to B. suis. The high percentage of positive samples suggests that brucellosis may be a concern in deer within the studied area, and that these animals may poses a risk for other domestic and wild ones.

Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis.

Babesia bovis is a tick-borne pathogen that remains an important constraint for the development of cattle industries in tropical and subtropical regions of the world. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these phenotypes have a number of drawbacks, which justifies the search for new, more efficient immunogens based mainly on recombinant protein technology. In the present paper, ribosomal phosphoprotein P0 from a Brazilian isolate of B. bovis was produced and evaluated with regard to conservation and antigenicity. The protein sequence displayed high conservation between different Brazilian isolates of B. bovis and several Apicomplexa parasites such as Theileria, Neospora and Toxoplasma. IgG from cattle experimentally and naturally infected with B. bovisas well as IgG1 and IgG2 from naturally infected cattle reacted with the recombinant protein. IgG from cattle experimentally infected with Babesia bigemina cross-reacted with B. bovis recombinant P0. These characteristics suggest that P0 is a potential antigen for recombinant vaccine preparations against bovine babesiosis.

Molecular and antigenic characterisation of ribosomal phosphoprotein P0 from Babesia bovis

Babesia bovis is a tick-borne pathogen that remains an important constraint for the development of cattle industries in tropical and subtropical regions of the world. Effective control can be achieved by vaccination with live attenuated phenotypes of the parasite. However, these phenotypes have a number of drawbacks, which justifies the search for new, more efficient immunogens based mainly on recombinant protein technology. In the present paper, ribosomal phosphoprotein P0 from a Brazilian isolate of B. bovis was produced and evaluated with regard to conservation and antigenicity. The protein sequence displayed high conservation between different Brazilian isolates of B. bovis and several Apicomplexa parasites such as Theileria, Neospora and Toxoplasma. IgG from cattle experimentally and naturally infected with B. bovisas well as IgG1 and IgG2 from naturally infected cattle reacted with the recombinant protein. IgG from cattle experimentally infected with Babesia bigemina cross-reacted with B. bovis recombinant P0. These characteristics suggest that P0 is a potential antigen for recombinant vaccine preparations against bovine babesiosis.

Desenvolvimento e avaliação de uma cepa knockout de Brucella abortus obtida pela deleção do gene virB10 / Development and evaluation of a strain of Brucella abortus gotten by the knockout of the virB10 gene

Brucella spp. são bactérias gram-negativas, intracelulares facultativas que são patogênicas para muitas espécies de mamíferos causando a brucelose, uma zoonose difundida mundialmente. Por isso a busca de alternativas de controle mais eficientes se faz necessário como o desenvolvimento de novas cepas que possam ser testadas como potenciais imunógenos. Neste estudo realizou-se a deleção do gene virB10 da cepa S2308 de Brucella abortus gerando uma cepa knockout provavelmente incapaz de produzir a proteína nativa correspondente. O gene virB10 faz parte de um operon que codifica para um sistema de secreção do tipo IV, essencial para a sobrevivência intracelular e multiplicação da bactéria em células hospedeiras. A deleção foi realizada pela construção do plasmídeo suicida pBlue:virB10:kan e eletroporação deste em células eletrocompetentes de B. abortus S2308, ocorrendo a troca do gene selvagem pelo gene interrompido, com o gene de resistência a canamicina, por recombinação homóloga dupla. Camundongos BALB/c foram inoculados com as cepas S19, RB-51, ΔvirB10 de B. abortus e B. abortus S2308 selvagem; os resultados demonstraram que camundongos BALB/c inoculados com S19 e camundongos BALB/c inoculados com S2308 apresentaram queda mais rápida de linha de tendência, quando comparadas aos demais grupos, para recuperação bacteriana (RB) e peso esplênico (PE) respectivamente. Os grupos que receberam ΔvirB10 S2308 de B. abortus e RB-51 demonstraram comportamento semelhante para ambas as características. Na sexta semana após a inoculação, os resultados para RB (log de UFC ± desvio padrão) e PE (peso esplênico ± desvio padrão), respectivamente, mostraram: grupos inoculados com as cepas S2308 (4,44±1,97 e 0,44±0,11), S19 (1,83±2,54 e 0,31±0,04), RB-51 (0,00±0,00 e 0,20±0,01) e ΔvirB10 S2308 (1,43±1,25 e 0,19±0,03). Considerado o clearance bacteriano, todos os grupos diferiram...

Brucella spp. are intracellular facultative gram-negative bacteria which are pathogenic for many species of mammals, causing brucellosis, a worldwide spread zoonosis. Therefore the search for more efficient alternatives of control, as the development of new potential immunogens is necessary. In this study, we knockouted virB10 from Brucella abortus S2308 strain, generating a mutant strain probably incapable to produce the corresponding native protein. The gene virB10 is part of an operon that codifies for type IV secretion system, which is essential for the intracellular survival and multiplication of the bacteria in host cells. The knockout was carried through by the construction of the suicidal plasmid pBlue: virB10: kan and eletroporation in eletrocompetent cells of B. abortus S2308, leading to the exchange of the wild gene for the interrupted gene, containing the gene of resistance to kanamycin, for double homologous recombination. BALB/c mice were inoculated with S19, RB-51, ΔvirB10 strains of B. abortus and S2308 wild strain; the results demonstrated that the BALB/c mice inoculated with S19 and BALB/c mice inoculated with S2308 presented faster fall of trend line, when compared with the too much groups, for bacterial recovery (BR) and esplenic weight (EW) respectively. The groups that received ΔvirB10 S2308 B. abortus and RB-51 demonstrated similar behavior for both the characteristics. In the sixth week postinoculation, the results for BR (log UFC ± standart deviations) and EW (esplenic weight ± standart deviations), respectively, showed: groups inoculated with strains S2308 (4,44±1,97 and 0,44±0,11), S19 (1,83±2,54 and 0,31±0,04), RB-51 (0,00±0,00 and 0,20±0,01) and ΔvirB10 S2308 (1,43±1,25 and 0,19±0,03). Considered the bacterial clearance, all the groups differed statistical from the group that received S2308 (p<0,0001), the group inoculated...

[Detection of antibodies against Anaplasma sp. in small ruminants from the semi-arid region against Pernambuco State, Brazil]. / Detecção de anticorpos para Anaplasma sp. em pequenos ruminantes no semi-árido do Estado de Pernambuco, Brasil.

This paper reports the detection of antibodies against Anaplasma sp. in goats and sheep from the semi-arid region from Pernambuco State, Brazil, using an enzyme-linked immunosorbent assay with recombinant MSP5 of Anaplasma marginale. Sera from 243 goats and 68 sheep from Ibimirim municipality were analyzed and frequencies of antibodies of 11.93% (29/243) and 16.17% (11/68) were found for goats and sheep, respectively. The epidemiological relevance of the findings was discussed.

Comparação genotípica de isolados de Corynebacterium pseudotuberculosis de caprinos e ovinos do sertão de Pernambuco / Genotypic comparison between Corynebacterium pseudotuberculosis samples obtained from sheep and goats with caseous lymphadenitis, raised in the semi-arid region of Pernambuco

Objetivou-se com este estudo comparar genotipicamente 35 isolados de Corynebacterium pseudotuberculosis recuperados de conteúdo de abscessos de caprinos e ovinos com linfadenite caseosa, procedentes de cinco municípios localizados no Sertão de Pernambuco, Brasil. Utilizou-se a técnica de fingerprint RFLP-PCR com as enzimas de restrição Hpy-Ch4 e Msp1 aplicada ao gene rpoB e as enzimas Pst I e Msp I para o gene pld. Não houve diferença nos padrões de fragmentos de bandas entre os isolados, independente da espécie hospedeira ou da área geográfica estudada, definindo-se um padrão genotípico homogêneo de C. pseudotuberculosis responsável por abscessos superficiais na região.(AU)

The objective was to genotypically compare 35 samples of Corynebacterium pseudotuberculosis obtained from abscesses of sheep and goats diagnosed with caseous lymphadenitis originated from 5 different municipalities in the semi-arid region of Pernambuco, Brazil. The RFLP-PCR technique with Hpy-Ch4 and Msp I and Pst I Msp I restriction enzimes was used to fingerprint the genes rpoB and pld, respectively. The results demonstrate that there was no difference on the fragments banding pattern among samples, independently of the host species or geographic area studied, defining a homogeneous profile of C. pseudotuberculosis responsible for superficial abscesses for the region.(AU)

Detecção de anticorpos para Anaplasma sp. em pequenos ruminantes no semi-árido do estado de Pernambuco, Brasil / Detection of antibodies against Anaplasma sp. in small ruminants from the semi-arid region against Pernambuco State, Brazil

Neste trabalho é descrita a detecção de anticorpos para Anaplasma sp. em caprinos e ovinos da região do semi-árido do Estado de Pernambuco, Brasil, utilizando-se um ensaio de imunoadsorção enzimática baseado em MSP5 recombinante de Anaplasma marginale. Foram analisados soros de 243 caprinos e 68 ovinos provenientes do município de Ibimirim, e observadas freqüências de anticorpos de 11,93 por cento (29/243) e 16,17 por cento (11/68) para caprinos e ovinos, respectivamente. A importância epidemiológica dos achados foi discutida.

This paper reports the detection of antibodies against Anaplasma sp. in goats and sheep from the semi-arid region from Pernambuco State, Brazil, using an enzyme-linked immunosorbent assay with recombinant MSP5 of Anaplasma marginale. Sera from 243 goats and 68 sheep from Ibimirim municipality were analyzed and frequencies of antibodies of 11.93 percent (29/243) and 16.17 percent (11/68) were found for goats and sheep, respectively. The epidemiological relevance of the findings was discussed.

IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu.

Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and subtropical regions of the world. Immunization with purified outer membranes of this organism induces protection against acute anaplasmosis. Previous studies, with proteomic and genomic approach identified 21 proteins within the outer membrane immunogen in addition to previously characterized major surface protein1a-5 (MSP1a-5). Among the newly described proteins were VirB9, VirB10, and elongation factor-Tu (EF-Tu). VirB9, VirB10 are considered part of the type IV secretion system (TFSS), which mediates secretion or cell-to-cell transfer of macromolecules, proteins, or DNA-protein complexes in Gram-negative bacteria. EF-Tu can be located in the bacterial surface, mediating bacterial attachment to host cells, or in the bacterial cytoplasm for protein synthesis. However, the roles of VirB9, VirB10, and TFSS in A. marginale have not been defined. VirB9, VirB10, and EF-Tu have not been explored as vaccine antigens. In this study, we demonstrate that sera of cattle infected with A. marginale, with homologous or heterologous isolates recognize recombinant VirB9, VirB10, and EF-Tu. IgG2 from naturally infected cattle also reacts with these proteins. Recognition of epitopes by total IgG and by IgG2 from infected cattle with A. marginale support the inclusion of these proteins in recombinant vaccines against this rickettsia.

IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu

Anaplasma marginale is an important vector-borne rickettsia of ruminants in tropical and subtropical regions of the world. Immunization with purified outer membranes of this organism induces protection against acute anaplasmosis. Previous studies, with proteomic and genomic approach identified 21 proteins within the outer membrane immunogen in addition to previously characterized major surface protein1a-5 (MSP1a-5). Among the newly described proteins were VirB9, VirB10, and elongation factor-Tu (EF-Tu). VirB9, VirB10 are considered part of the type IV secretion system (TFSS), which mediates secretion or cell-to-cell transfer of macromolecules, proteins, or DNA-protein complexes in Gram-negative bacteria. EF-Tu can be located in the bacterial surface, mediating bacterial attachment to host cells, or in the bacterial cytoplasm for protein synthesis. However, the roles of VirB9, VirB10, and TFSS in A. marginale have not been defined. VirB9, VirB10, and EF-Tu have not been explored as vaccine antigens. In this study, we demonstrate that sera of cattle infected with A. marginale, with homologous or heterologous isolates recognize recombinant VirB9, VirB10, and EF-Tu. IgG2 from naturally infected cattle also reacts with these proteins. Recognition of epitopes by total IgG and by IgG2 from infected cattle with A. marginale support the inclusion of these proteins in recombinant vaccines against this rickettsia.

Morphology and morphometry of three Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae) isolates.

In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropédica (22 degrees 48' S; 43 degrees 41' W), in the state of Rio de Janeiro; Cruzeiro (22 degrees 33' S; 44 degrees 57' W), in the state of São Paulo; and Santa Bárbara do Tugúrio (21 degrees 15' S; 43 degrees 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a=md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed.

[Transcription of genes of membrane proteins of Brazilians isolates of Anaplasma marginale]. / Transcrição de genes de proteínas de membrana de isolados brasileiros de Anaplasma marginale.

This work shows the transcription profile of membrane protein genes in three Brazilian isolates of Anaplasma marginale (Rio Grande do Norte, Pernambuco-Zona da Mata, and Pernambuco-Sertão). RNA was purified from cattle blood experimentally-infected with the three isolates of A. marginale. After reverse transcription, genes omp1, 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, and 14; opag1-3; virB3, 9, and 10; am097, 197, 254, 854, and 956 were amplified by PCR, with specific primers. Transcripts were detected for all genes, except omp2, 3 e opag3 in all isolates and for omp7 in one out of the three isolates analyzed. Absence of transcription for opag3 and omp7 diverge from the North American isolates of A. marginale. Reasons for such differences were discussed.

Morphology and morphometry of three Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae) isolates / Morfologia e morfometria de três isolados de Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae)

In this work, three isolates of Plasmodium juxtanucleare have been analyzed based on morphological, morphometric and parasitic parameters. Each isolate was sampled from naturally infected adult chicken (Gallus gallus) from rural areas of three Brazilian municipalities: Seropédica (22º 48' S; 43º 41'W), in the state of Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57'W), in the state of São Paulo; and Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), in the state of Minas Gerais. The blood samples taken from each infected chicken were inoculated in three groups of ten young chicken (21 days old). Blood smears of the experimentally infected chicken were sampled every two days until the 69th day in order to evaluate the parasitemia. For the morphological-descriptive and morphometric analyses, we measured 30 individuals from each of the intraerythocytic states, measures of the major (MD) and minor diameters (md), the estimation of morphometric index (Mi=md/MD) and size (T=pab, a= md/2; b=MD/2). The results indicated low and homogeneous parasitemia rates in the three strains, which showed differences among shape and size of the parasitic stadia displayed.

Neste trabalho, três isolados de Plasmodium juxtanucleare foram analisados com base na morfologia, morfometria e parâmetros parasitológicos. Cada isolado foi coletado de aves (Gallus gallus) adultas infectadas naturalmente de áreas rurais de três municípios brasileiros: Seropédica (22º 48' S; 43º 41' W), no estado do Rio de Janeiro; Cruzeiro (22º 33' S; 44º 57' W), no estado de São Paulo; e Santa Bárbara do Tugúrio (21º 15' S; 43º 27' W), no estado de Minas Gerais. As amostras de sangue coletadas de cada ave infectada foram inoculadas em três grupos de dez aves (21 dias de idade). Esfregaços sangüíneos das aves infectadas experimentalmente foram realizados de dois em dois dias durante um período de 69 dias para avaliar a parasitemia. Para análises morfofisiológica e morfométrica, foram mensurados 30 indivíduos de cada estágio intraeritrocítico. Foram tomadas as medidas do diâmetro maior (DM) e diâmetro menor (dm), com os quais foi estimado o índice morfométrico (Mi=md/MD) e o tamanho (T=pab, a= md/ 2; b=MD/2). Os resultados indicaram uma parasitemia homogênea entre os três isolados, havendo diferenças apenas nas formas e tamanhos dos estádios parasitários.

Transcrição de genes de proteínas de membrana de isolados brasileiros de Anaplasma marginale / Transcription of genes of membrane proteins of Brazilians isolates of Anaplasma marginale

Este trabalho demonstra o padrão de transcrição de genes de proteínas de membrana em três isolados brasileiros de A. marginale (Rio Grande do Norte, Pernambuco-Zona da Mata e Pernambuco-Sertão). O RNA foi purificado a partir de sangue de bovinos infectados experimentalmente com os três isolados de A. marginale. Após transcrição reversa, os genes omp1, 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13 e 14; opag1-3; virB3, 9, 10; am097, 197, 254, 854 e 956 foram amplificados por PCR, com oligonucleotídeos iniciadores específicos. Detectaram-se transcritos para todos os genes analisados, exceto omp2, 3 e opag3 em todos os isolados e do gene omp7 em um dos isolados estudados. A ausência de transcrito para os genes opag3 e omp7 diverge do observado em isolados americanos da riquétsia. Possíveis razões para essas diferenças são discutidas.

This work shows the transcription profile of membrane protein genes in three Brazilian isolates of Anaplasma marginale (Rio Grande do Norte, Pernambuco-Zona da Mata, and Pernambuco-Sertão). RNA was purified from cattle blood experimentally-infected with the three isolates of A. marginale. After reverse transcription, genes omp1, 2, 3, 4, 5, 7, 8, 9, 10, 11, 12, 13, and 14; opag1-3; virB3, 9, and 10; am097, 197, 254, 854, and 956 were amplified by PCR, with specific primers. Transcripts were detected for all genes, except omp2, 3 e opag3 in all isolates and for omp7 in one out of the three isolates analyzed. Absence of transcription for opag3 and omp7 diverge from the North American isolates of A. marginale. Reasons for such differences were discussed.

Ocorrência de Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) em Canis familiaris (Linnaeus) de zona rural do município de Piraí, Rio de Janeiro, Brasil / Occurrence of Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) on Canis familiaris (Linnaeus) from rural areas of the county of PiraÝ, State of Rio de Janeiro, Brazil

A ocorrência de Rhopalopsyllus lutzi lutzi (Baker) (Siphonaptera, Rhopalopsyllidae) foi assinalada em Canis familiaris (Linnaeus) de áreas rurais do município de Piraí, estado do Rio de Janeiro, Brasil. No período de junho 2001/novembro 2003, 51 sifonápteros foram capturados em oito cães procedentes de duas propriedades rurais do município de Piraí. Os exemplares coletados foram acondicionados em álcool etílico 70 por cento, levados ao Laboratório de Parasitologia Animal da Universidade Federal Rural do Rio de Janeiro para contagem e sexagem. Os exemplares foram identificados como Rhopalopsyllus lutzi lutzi Baker, 1904 (machos=18 e fêmeas=33). R. lutzi lutzi é, pela primeira vez, assinalada em cães domésticos naturalmente infestados em áreas rurais do estado do Rio de Janeiro.