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      Effect of polymorphisms in the Slc11a1 coding region on resistance to brucellosis by macrophages in vitro and after challenge in two Bos breeds (Blanco Orejinegro and Zebu)

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          Abstract

          The resistance/susceptibility of selected cattle breeds to brucellosis was evaluated in an F1 population generated by crossing animals classified as resistant (R) and susceptible (S) (R x R, R x S, S x R, S x S) based on challenges in vitro and in vivo. The association between single nucleotide polymorphisms identified in the coding region of the Slc11a1 gene and resistance/susceptibility was estimated. The trait resistance or susceptibility to brucellosis, evaluated by a challenge in vitro, showed a high heritable component in terms of additive genetic variance (h 2 = 0.54 ± 0.11). In addition, there was a significant association (p < 0.05) between the control of bacterial survival and two polymorphisms (a 3'UTR and SNP4 located in exon 10). The antibody response of animals classified as resistant to infection by Brucella abortus differed significantly (p < 0.05) from that of susceptible animals. However, there was no significant association between single nucleotide polymorphisms located in the Slc11a1 gene and the antibody response stimulated by a challenge in vivo.

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          Fast and sensitive silver staining of DNA in polyacrylamide gels

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            Fast and sensitive silver staining of DNA in polyacrylamide gels.

            The photochemically derived silver stain of nucleic acids in polyacrylamide gels originally described by Merril et al. (1981, Science 211, 1437-1438) was modified to reduce unspecific background staining and increase sensitivity (down to 1 pg/mm2 band cross-section). Detection limits for double-stranded DNA fragments from HaeIII endonuclease digests of phage phi X174 were maintained despite eliminating oxidation pretreatment of fixed gels and reducing silver nitrate concentration. Preexposure to formaldehyde during silver impregnation enhanced sensitivity and the inclusion of the silver-complexing agent sodium thiosulphate in the image developer decreased background staining. Higher formaldehyde concentration during image development resulted in darker bands with good contrast. The procedure almost halves the number of steps, solutions and experimental time required and can be used for the staining of DNA fragments in polyacrylamide gels bound to a polyester backing film by controlling temperature during image development. We have applied this improved staining procedure for the routine analysis of complex DNA profiles generated by DNA amplification fingerprinting (DAF).
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              Rough vaccines in animal brucellosis: structural and genetic basis and present status.

              Brucellosis control and eradication requires serological tests and vaccines. Effective classical vaccines (S19 in cattle and Rev 1 in small ruminants), however, induce antibodies to the O-polysaccharide of the lipopolysaccharide which may be difficult to distinguish from those resulting from infection and may thus complicate diagnosis. Rough attenuated mutants lack the O-polysaccharide and would solve this problem if eliciting protective immunity; the empirically obtained rough mutants 45/20 and RB51 have been used as vaccines. Strain 45/20 is reportedly unstable and it is not presently used. RB51 is increasingly used instead of S19 in some countries but it is rifampicin resistant and its effectiveness is controversial. Some controlled experiments have found good or absolute protection in adult cattle vaccinated orally (full dose) or subcutaneously (reduced dose) and in one field experiment, RB51 was reported to afford absolute protection to calves and to perform better than S19. Controlled experiments in calves, however, have shown reduced doses of RB51 to be ineffective, full doses only partially effective, and RB51 less effective than S19 against severe challenges. Moreover, other observations suggest that RB51 is ineffective when prevalence is high. RB51 is not useful in sheep and evidence in goats is preliminary and contradictory. Rough mutants obtained by molecular biology methods on the knowledge of the genetics and structure of Brucella lipopolysaccharide may offer alternatives. The B. abortus manBcore (rfbK) mutant seems promising in cattle, and analyses in mice suggest that mutations affecting only the O-polysaccharide result in better vaccines than those affecting both core and O-polysaccharide. Possible uses of rough vaccines also include boosting immunity by revaccination but solid evidence on its effectiveness, safety and practicality is not available.
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                Author and article information

                Journal
                Genet Mol Biol
                GMB
                Genetics and Molecular Biology
                Sociedade Brasileira de Genética (Ribeirão Preto, SP, Brazil )
                1415-4757
                1678-4685
                Jul-Sep 2010
                1 September 2010
                : 33
                : 3
                : 463-470
                Affiliations
                [1 ]simpleGrupo de Recursos Genéticos y Biotecnología Animal, Centro de Biotecnología y Bioindustria, Corporación Colombiana de Investigación Agropecuaria, C.I., Bogotá DC Colombia
                [2 ]simpleDepartamento de Producción Animal, Facultad de Veterinaria, Universidad Complutense de Madrid, Madrid Spain
                [3 ]simpleCentro de Investigación CEISA, Bogotá DC Colombia
                [4 ]simpleGrupo de Recursos Genéticos y Biotecnología Animal, C.I. El Nus, San Roque, Antioquia Colombia
                Author notes
                Send correspondence to Rodrigo Martínez. Animal Genetic Resource Group, Research Center Tibaitatá, Corporación Colombiana de Investigación Agropecuaria km 14, Road to Mosquera Cundinamarca, Bogotá DC, Colombia. E-mail: ramartinez@ 123456corpoica.org.co .
                Article
                10.1590/S1415-47572010000300014
                3036107
                21637417
                559974aa-edf9-4cb8-a82f-18c458f5e8ff
                Copyright © 2010, Sociedade Brasileira de Genética.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 9 February 2009
                : 22 March 2010
                Categories
                Animal Genetics
                Research Article

                Molecular biology
                zebu,brahman breed,blanco orejinegro creole breed,brucella abortus,genetic resistance

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