RESUMO
Brazil is a major producer and exporter of beef, with a herd of approximately 210 million animals. For the meat industry, a reliable animal traceback from its origin to the consumer market is paramount. Of all available identification systems, DNA is the only one that survives the slaughterhouse and reaches the dish of the consumer. DNA polymorphisms are already used for cattle traceback, but primarily for the subspecies Bos taurus taurus. However, in Brazil, another subspecies, B. taurus indicus predominates. We describe here the development of a DNA traceback method designed primarily for B. taurus indicus (Zebu), without leaving B. taurus taurus aside. We used insertion/deletion (indel) polymorphisms, which have the advantage of being simple and easily automatable, since in most cases, the variable loci are biallelic. We studied 94 indels, with a difference of two or more base pairs, in DNA pools of 60 Zebu and 60 taurine animals. A set of 22 indels with heterozygosity greater than 0.3 were selected and used to construct two multiplex PCRs. On the basis of the allelic frequency of these indels, the probability of random match was calculated to be 1.12 x 10(-8) for B. taurus indicus and 1.60 x 10(-6) for B. taurus taurus. Moreover, we estimated that an analysis would cost less than US$15.00 per animal. Thus, this system (MULTINDELS-BOV) is perfectly suited for building large genetic databases and offering viable prospects of a national system for cattle traceback DNA in Brazil.
Assuntos
Bovinos/genética , DNA/genética , Mutação INDEL/genética , Polimorfismo Genético , Alelos , Animais , Eletroforese em Gel de Poliacrilamida , Frequência do Gene/genética , Loci Gênicos , Coloração pela PrataRESUMO
The International Society of Animal Genetics (ISAG) has chosen nine microsatellites (international marker set) as a standard that should be included in all cattle parentage studies. They are BM1824, BM2113, INRA023, SPS115, TGLA122, TGLA126, TGLA227, ETH10, and ETH225. We decided to ascertain whether this microsatellite set could be used to determine ancestral proportions in individual animals of synthetic breeds produced by crossing zebu and taurine cattle. Since the genotypes of these markers are routinely available, this would constitute a practical and cost-free method to estimate the ancestry of synthetic breed animals. Genotypes of 100 Gir and 100 Holstein animals were examined for this ISAG marker set. As expected, there were very significant allele frequency differences between the two breeds at most loci. We also typed 20 Girolando animals for which there was complete genealogical information. "Structure" software easily distinguished Holstein and Gir animals based on their microsatellite genotypes; it also attributed the genomic proportion of zebu and taurine of each of the 20 Girolando animals. The proportion of Holstein ancestry was then regressed on the genealogical data; there was a highly significant correlation (r = 0.84, P < 0.0001). The nine microsatellites that compose the ISAG international marker set were capable of estimating the ancestral Gir and Holstein genomic proportions in individual Girolando animals within narrow confidence limits. This microsatellite set might also be useful for estimating the proportions of taurine and zebu origins in commercial meat products.
Assuntos
Cruzamento , Bovinos/genética , Frequência do Gene/genética , Repetições de Microssatélites/genética , Característica Quantitativa Herdável , Algoritmos , Animais , Teorema de Bayes , DNA/análise , Marcadores Genéticos , Genótipo , Reação em Cadeia da Polimerase/veterinária , Reprodutibilidade dos TestesRESUMO
The International Society of Animal Genetics (ISAG) has chosen nine microsatellites (international marker set) as a standard that should be included in all cattle parentage studies. They are BM1824, BM2113, INRA023, SPS115, TGLA122, TGLA126, TGLA227, ETH10, and ETH225. We decided to ascertain whether this microsatellite set could be used to determine ancestral proportions in individual animals of synthetic breeds produced by crossing zebu and taurine cattle. Since the genotypes of these markers are routinely available, this would constitute a practical and cost-free method to estimate the ancestry of synthetic breed animals. Genotypes of 100 Gir and 100 Holstein animals were examined for this ISAG marker set. As expected, there were very significant allele frequency differences between the two breeds at most loci. We also typed 20 Girolando animals for which there was complete genealogical information. Structure software easily distinguished Holstein and Gir animals based on their microsatellite genotypes; it also attributed the genomic proportion of zebu and taurine of each of the 20 Girolando animals. The proportion of Holstein ancestry was then regressed on the genealogical data; there was a highly significant correlation (r = 0.84, P < 0.0001). The nine microsatellites that compose the ISAG international marker set were capable of estimating the ancestral Gir and Holstein genomic proportions in individual Girolando animals within narrow confidence limits. This microsatellite set might also be useful for estimating the proportions of taurine and zebu origins in commercial meat products.
Assuntos
Animais , Cruzamento , Bovinos/genética , Frequência do Gene/genética , Repetições de Microssatélites/genética , Característica Quantitativa Herdável , DNA , Algoritmos , Teorema de Bayes , Marcadores Genéticos , Genótipo , Reação em Cadeia da Polimerase/veterinária , Reprodutibilidade dos TestesRESUMO
The molecular weight and the electrofocusing profile of human amniotic membrane interferon (IFN-AM) were determined. When submitted to gel filtration, IFN-AM showed a single 26-28 kDa component; in polyacrylamide gel electrophoresis one component of 19,500. In electrofocusing, IFN-AM displayed a terogeneity was reduced by previous treatment of IFN-AM with neuraminidase. IFN-AMis a siaglycoprotein similarto human beta IFN in terms of antigenicity but different from it in electrofocusing profile
Assuntos
Humanos , Membranas Extraembrionárias/metabolismo , Interferons/química , Placenta/análise , Cromatografia em Gel , Eletroforese em Gel de Poliacrilamida , Focalização Isoelétrica , Peso MolecularRESUMO
1. The molecular weight and the electrofocusing profile of human amniotic membrane interferon (IFN-AM) were determined. 2. When submitted to gel filtration, IFN-AM showed a single 26-28 kDa component; in polyacrylamide gel electrophoresis one component of 19,500. 3. In electrofocusing, IFN-AM displayed a heterogeneous profile with 5 to 7 peaks, but different from human alpha or beta IFNs. This heterogeneity was reduced by previous treatment of IFN-AM with neuraminidase. 4. IFN-AM is a sialoglycoprotein similar to human beta IFN in terms of antigenicity but different from it in electrofocusing profile.
Assuntos
Membranas Extraembrionárias/metabolismo , Interferons/química , Placenta/química , Cromatografia em Gel , Eletroforese em Gel de Poliacrilamida , Humanos , Focalização Isoelétrica , Peso MolecularRESUMO
By centrifuging heparinised human blood through a discontinuous Ficoll-Hypaque density gradient, it is possible to separate simultaneously mononuclear cells, neutrophils, and eosinophils.
