Carina Elisei

Caracterización morfológica y morfométrica de Plasmodium juxtanucleare (Apicomplexa: Plasmodiidae)

Se realizo el analisis morfometrico y morfologico de un aislado de (MSS-47) de Plasmodium juxtanucleare. Fueron evaluados, la parasitemia, las medidas del diametro menor (dm), el diametro mayor (DM) y el indice morfometrico (IM = dm/DM) de formas intraeritrociticas, aves inoculadas experimentalmente. En 106 trofozoitos se observaron formas puntiformes, ovoides, anilladas, elipticas, piriformes y redondeadas, con predominio de las formas ovoides y elipticas. La media de los trofozoitos fue de 1.802 ± 0,313 x 1.230 ± 0,2 mm, con un IM = 0,623 ± 0,110 mm. En 98 esquizontes, se observaron formas ovoides, elipticas piriformes y ameboides, con predominio de formas ovoides y piriformes. La media de los esquzontes fue de 2.809 ± 0,187 x 1.820 ± 0,386 mm, con un IM = 0,643 ± 0,120. En 18 macrogametocitos, se observaron formas ovoides, redondeadas, elipticas y piriformes, con predominio de las formas ovoides y elipticas. La media de los macrogametocitos fue de 3.559 ± 0,883 x 2.363 ± 0,207 mm, con un IM = 0,766 ± 0,078 mm. En 34 microgametocitos, se observaron formas ovoides, redondeadas, elipticas y piriformes, con predominio de las formas ovoides y elipticas. La media de los microgametocitos fue de 3.891 ± 0,946 x 2.386 ± 0,783 mm, con un IM = 0,505 ± 0,103 mm. Por lo tanto, el aislado MSS-47 de P. juxtanucleare se puede caracterizar como muy pleomorfico

ELISA com MSP5 recombinante truncada para detecção de anticorpos contra Anaplasma marginale em bovinos

The objective of this study was the production and solubilization of recombinant truncated MSP5 of Anaplasma marginale and the evaluation of its performance in an enzyme-linked immunosorbent assay (ELISA), to detect antibodies against the rickettsia in cattle. The fragment of msp5 gene, except the hydrophobic N-terminal region, was amplified by PCR, cloned in pTrcHis-TOPO plasmid and expressed in Escherichia coli. Solubilization of the recombinant protein was evaluated in different pHs and concentrations of urea. The sensibility and specificity of the assay were evaluated with 66 sera from cattle experimentally-infected and 96 sera from cattle free of A. marginale defined by polymerase chain reaction for msp5 gene. Serum samples from 1,666 cattle from Brazil - states of Rio Grande do Sul (73), Mato Grosso do Sul (91), Pernambuco (86), Bahia (314) and Minas Gerais (267), Uruguay (32) and Costa Rica (803), were tested by ELISAs with recombinant truncated MSP5 and with recombinant MSP1a, and the agreement between both ELISAs was calculated. ELISA with recombinant truncated MSP5 protein detected infected animals with sensibility of 96.97% and specificity of 100%. In cattle experimentally-infected, the ELISA detected antibodies from the 12th day post-infection (DPI) to the end of the experiment, at the 37th DPI. The agreement between the ELISAs with truncated MSP5 and MSP1a antigens was 95.67%, with a kappa index of 0.81. Disagreement results showed significative difference (p <0.001). Antibodies for A. marginale were detected in animals of the all the region analyzed. The ELISA with recombinant truncated MSP5 showed a good performance in ELISA for detention of antibodies against A. marginale, with high sensitivity and specificity, representing an important tool for the diagnosis of anaplasmose bovine in epidemiological studies.

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-Sertao). 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

From June/2001 to November/2003, 51 fleas were collected on eight dogs from two rural properties of the county of Pirai, State of Rio de Janeiro. After preservation in 70% ethanol, the fleas were counted and sexed in the Laboratorio de Parasitologia Animal da Universidade Federal Rural do Rio de Janeiro. The samples were identificated as Rhopalopsyllus lutzi lutzi (Baker, 1904) (male=18 and female=33). For the first time R. lutzi lutzi was found paraziting dogs in rural areas of the State of Rio de Janeiro.

Criopreservación de Plasmodium (Novyella) juxtanucleare con glicerol

RESUMEN Se evaluo la criopreservacion de Plasmodium juxtanucleare utilizando glicerol bajo cuatro concentraciones (20%, 16%, 12% y 8%). Posteriormente, congelamiento a -196°C, por seis meses, la sangre fue descongelada e inoculada en pollos, tres pollos para cada concentracion de glicerol (CG). Se realizo frotis sanguineos diarios, de las aves, durante 35 dias. El analisis de variables biologicas revelo que la parasitemia fue directamente proporcional a CG, el periodo pre-patente fue de 12,4 ± 2,7 dias, el pico maximo de parasitemia (PMP) fue de 23,5 ± 3,5 dias y la duracion media de la parasitemia fue de 33,6 ± 1,0 dias. La tasa de hemolisis vario de 12,45% a 40% y, fue inversamente proporcional a CG. Hubo diferencia significativa (P < 0,05) en cuanto a PMP; donde el inoculo con glicerol a 12% difirio de los demas, presentando el mayor PMP (30,5 ± 0,5). La parasitemia media, de los grupos experimentales tuvo el inicio entre el septimo y octavo dia post-inoculacion (DPI), durando, en media, hasta el 35o DPI. El grupo que recibio inoculo con glicerol a 20% presento la mayor y mas alta parasitemia. El glicerol a 20% fue la concentracion mas eficiente para la criopreservacion de P. juxtanucleare.