ABSTRACT
El control de calidad genético de los animales de experimentación utilizados en los bioterios debe ser prioritario, para asegurar que los estudios realizados tengan reproducibilidad y además validez científica. La presente investigación tuvo como objetivo evaluar la pureza genética de las ratas Wistar Kyoto (WKY) del bioterio de la Facultad de Ciencias Químicas y Farmacia de la Universidad de San Carlos de Guatemala, "Dra. Amarillis Saravia Gómez", las cuales nunca se han caracterizado genéticamente. Para realizar este análisis se seleccionaron los microsatélites D1Mgh6 y D17Mit3, los cuales muestran un alto grado de variabilidad y gran número de polimorfismos, por lo que permiten tener una visión general de las características genéticas de la cepa. Los análisis se realizaron utilizando la técnica de reacción en cadena de la polimerasa (PCR). Los resultados obtenidos del análisis genético demostraron que el fragmento analizado para el microsatélite D17Mit3 de las WKY del bioterio fue el esperado con un peso molecular de 201 pb, que corresponde al tamaño reportado; en la base de datos genómicos de la rata, por los laboratorios Charles River y los Institutos Nacionales de Salud de los Estados Unidos (NIH). Por el contrario, el tamaño del microsatélite D1Mgh6 no corresponde al fragmento de 147 pb esperado. La colonia de ratas WKY se ha manejado con un método de reproducción consanguíneo lo que se confirmó con los resultados, ya que los microsatélites D17Mit3 y D1Mg6 mostraron una condición homogicota y homoalélica, por lo cual estos animales de experimentación pueden ser usados en ensayos como un modelo consanguíneo de características genéticas propias, puesto que no tienen las características genéticas esperadas para la raza WKY.
Genetic quality control of experimental animals used in laboratory animal centers must be priority, to ensure that studies have reproducibility and also scientific validity. This research aimed to evaluate the genetic purity of the Wistar Kyoto rats at bioterio of the Faculty of Chemistry Sciences and Pharmacy at the University of San Carlos of Guatemala, "Dr. Amarillis Saravia Gómez", which never had been genetically characterized. The D1Mgh6 and D17Mit3 microsatellites were analyzed because they have a high grade of polymorphism and allows to have a general vision of genetic characteristicsof the strain, the analysis was performed by the polymerase chain reaction technique (PCR). The results of the genetic analysis showed that the colony of Wistar Kyoto rats corresponds to the estimated size for the microsatellite D17Mit3 (201 bp) as the genomic database rat and reported by Charles River Laboratories and the NIH. However, It doesn't correspond to the estimated size of the microsatellite D1Mgh6 (147 bp). The WKY strain has been manage with an inbreeding method, that was confirmed by the results because D17Mit3 and D1Mg6 showed an homogenic and homoallelic condition, so these experimental animals can be used in trials as a inbreed model, because do not have the genetic characteristics expected for the WKY breed.
ABSTRACT
Laboratory red crucian carp is a native fish in China. It has been an established inbred fish strain,the laboratory red crucian carp C1HD strain. Based on the standardization research and verification test of laboratory red crucian carp,the local standard of Hunan Province,the called"Laboratory Fish Genetic Quality Control of Laboratory Red Crucian Carp C1HD Strain", has been established. This standard includes preface, scope, normative reference documents,terms and definitions, major biological characteristics, mating and breeding method, breeding environment, genetic quality testing and appendix A,B,C and so on.
ABSTRACT
Objective To screen the markers of chromosome 12, which is vacant in the Beijing local standard DB11/T828.3-2011 for genetic quality control of miniature pigs in Beijing, and to study the applicability of the local standard by comparison of two different methods for evaluation of the genetic quality of the same population.Methods According to the literature, we selected four pairs of microsatellite markers of chromosome 12 and studied the polymorphism through monitoring the genetic quality of two populations of China Agricultural University miniature pigs.We screened the highly polymorphic markers of chromosome 12, combined them with the microsatellite primers of the standard 18 pairs of chromosomes to establish the whole chromosome method.We compared and analyzed the applicability of the local standard DB11/T828.3-2011 through monitoring the genetic quality of the same population of miniature pigs with different method.The data were processed and analyzed using software Popgen32.Results All the screened four pairs of microsatellite markers of chromosome 12 were highly polymorphic (PIC>0.5).The local standard showed a chromosome coverage of 94.7%, stability of amplification of 96.0%, and certified that the China Agricultural University miniature pig III was qualified, while the China Agricultural University miniature pig I was not qualified.When the markers of chromosome 12 were added, the whole chromosome method showed result of 100% chromosome coverage and 96.6% amplification stability, both of the two populations of pigs were certified as qualified.Conclusions The four screened markers of chromosome 12 are all highly polymorphic, and provide a support for supplement of the local standard DB11/T828.3-2011.
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Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.
ABSTRACT
Objective To test and analyze the genetic background of highly immunodeficient mice from different sources.Methods Four highly immunodeficient mouse strains from different sources of NOD background were collected. 30 microsatellite DNA sites were detected, and the genotype can be displayed by gel electrophoresis and STR scanning. Results 17 microsatellite sites exhibit polymorphism in 20 mice of the four groups.There were 30 homozygous loci in the mice of groups A and B, and heterozygous in the other two groups.The genetic distance is minimum between groups A and B, showing a higher genetic similarity.Conclusions The genetic backgrounds are different in highly immunodeficient mice from different sources.
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Objective To screen out specific microsatellite markers for use in Tupaia belangeri chinensis genetic testing. Methods Firstly to screen about 700 microsatellite loci from whole genome.Secondly to choose about 100 better loci without defect factors.Lastly 46 primers were designed by 33 tree shrew’ s microsatellite loci obtained from whole genome and other references.Agarose gel electrophoresis and polyacrylamide gel electrophoresis were used for PCR products, and better loci based on electrophoresis results were chosen.Then STR scan was used to select the microsatellite loci combination for genetic testing.Results Twenty-two microsatellite loci were selected with a significant Stutter peak on STR scanning.Comparing the alternative loci and ultimately selected loci, there were two loci available in the five alternative loci of T.glis.The coincidence rate between T.glis and T.b.chinensis was 40%.There were two loci available in the five alternative loci of T.minor, and the coincidence rate between T.minor and T.b.chinensis was 40%.There were two loci available in the three alternative loci of T.belangeri, and the coincidence rate between T.belangeri and T.b. chinensis was about 70%.Conclusions The 22 microsatellite loci screened in this study are well applied for genetic testing of Tupaia belangeri chinensis, therefore, provide a scientific basis for the genetic quality monitoring of tree shrews.
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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.