RESUMEN
The bovine tick Rhipicephalus (Boophilus) microplus is found in several tropical and subtropical regions of the world. This parasite transmits pathogens that cause disease, such as babesiosis (Babesia bovis and B. bigemina) and anaplasmosis (Anaplasma marginale). Tick infestations cause enormous livestock losses, and controlling tick infestations and the transmission of tick-borne diseases remains a challenge for the livestock industry. Because the currently available commercial vaccines offer only partial protection against R. (B.) microplus, there is a need for more efficient vaccines. Several recombinant antigens have been evaluated using different immunization strategies, and they show great promise. This work describes the construction and immunological characterization of a multi-antigen chimera composed of two R. (B.) microplus antigens (RmLTI and BmCG) and one Escherichia coli antigen (B subunit, LTB). The immunogenic regions of each antigen were selected and combined to encode a single polypeptide. The gene was cloned and expressed in E. coli. For all of the experiments, two groups (treated and control) of four Angus heifers (3-6 months old) were used. The inoculation was performed via intramuscular injection with 200 µg of purified recombinant chimeric protein and adjuvated. The chimeric protein was recognized by specific antibodies against each subunit and by sera from cattle inoculated with the chimera. Immunization of RmLTI-BmCG-LTB cattle reduced the number of adult female ticks by 6.29% and vaccination of cattle with the chimeric antigen provided 55.6% efficacy against R. (B.) microplus infestation. The results of this study indicate that the novel chimeric protein is a potential candidate for the future development of a more effective vaccine against R. (B.) microplus.
Asunto(s)
Antígenos Bacterianos/inmunología , Rhipicephalus/patogenicidad , Enfermedades por Picaduras de Garrapatas/prevención & control , Vacunas Sintéticas/inmunología , Animales , Reactores Biológicos , Bovinos , Clonación Molecular , Escherichia coli/genética , Femenino , Inyecciones Intramusculares , Enfermedades por Picaduras de Garrapatas/inmunología , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genéticaRESUMEN
From 50 farmed Rhea americana slaughtered for human consumption, adult forms and eggs of Fasciola hepatica were found in 4. The other three livers were free of flukes but did show lesions caused by larval fluke migration. Histological lesions were similar to those caused by flukes in cattle and sheep. The rheas were from an endemic area of ruminant fascioliasis in Southern Brazil. F. hepatica eggs were also found in faecal samples of wild rheas from another endemic area in Southern Brazil. It is likely that the rheas play a role in the transmission of the disease to ruminants and could be jeopardizing the control of this parasitosis in endemic areas. From the best of our knowledge this is the first report of fascioliasis in R. americana.
Asunto(s)
Enfermedades de las Aves/parasitología , Fascioliasis/veterinaria , Reiformes/parasitología , Crianza de Animales Domésticos , Animales , Enfermedades de las Aves/epidemiología , Brasil/epidemiología , Enfermedad Crónica , Fasciola hepatica/aislamiento & purificación , Fascioliasis/diagnóstico , Fascioliasis/epidemiología , Femenino , Hígado/parasitología , Hígado/patología , MasculinoRESUMEN
We report the cloning, sequence characterization and expression analysis of a calreticulin (CRT) coding cDNA of Boophilus microplus. CRT is a calcium-binding protein involved in multiple cell functions and possibly implicated in parasites host immune system evasion. The CRT cDNA sequence and its molecular characterization are described. Sequence similarity and phylogenetic analyses indicate a close relationship to other arthropod CRT sequences. The CRT cDNA was also expressed in a procariotic system and the recombinant protein (rBmCRT) was used to raise antibodies in a rabbit. Expression analyses of the corresponding gene in different developmental stages and tissues were performed by RT-PCR and Western-blot, which indicated a ubiquitous expression of the B. microplus calreticulin gene and demonstrated its presence in saliva. Sera of tick-infested bovines suggested that this protein may not be able to induce an IgG-based humoral response in its natural host.