Progress 09/01/08 to 08/31/11
Outputs
OUTPUTS: Project TSTAR-122 included the objectives of analyzing the segregation pattern for CAPN and CAST polymorphisms in purebred Senepol cattle (bulls and cows) and its crossbreds within the Caribbean Basin, Southern USA, Central and South America, and or investigating the potential association between these polymorphisms with economically important beef traits. Segregation analysis of CAPN haplotypes was performed in order to determine the number of bulls present in each class of group of animals from Puerto Rico (n = 94). In that population, the more representative haplotypes (CAPN1-316-4751, n = 9) were RY, GY, and GT. Surprisingly, such genetic profiles were associated with weights at birth, 205 d, 240 d, and at slaughter; sensory tenderness (0 and 14 d of aging), bone percentage; and with individual muscles weights (Biceps femoris and Longissimus dorsi). The segregation pattern of CAST SNP, previously reported by Barendse (2002), was determined within our cattle population. For that SNP, it is now well documented that the presence of the T allele is associated with a reduction of 0.15 kg in shear force at 14 d postmortem (Van Enenneman et al., 2007). The same was observed in the Senepol x Charolais crossbreds harvested by our group (n = 42). However, our results demonstrates that such reduction in shear force was greater (0.35 kg) after an aging period of 24 h, all of which compares favorably with a report by Casas et al. (2006) at 14 d postmortem. Meanwhile, we have identified a novel polymorphism in CAST (AA, AB, BB genotypes). A total of 125 bulls (from 50% to purebred Senepol or Charolais) were genotyped to determine their segregation and potential associations with growth and meat quality traits. Overall, the frequency of the allele labeled as A (cytosine deletion) was higher (0.85) than the B (cytosine insertion) allele (0.15). In bulls of Senepol descent, allele frequencies were 0.91A/0.09B, whereas in those with Charolais blood, frequencies were 0.70A/0.30B. Genotypic frequencies were 0.83AA, 0.17AB, 0.0BB and 0.48AA, 0.44AB, 0.08BB for Senepol and Charolais bulls, respectively. Because the BB genotype was observed in only two Charolais bulls, it was excluded for corresponding statistical analyses (n = 55 for growth; n = 24 for meat quality traits). At birth, AB genotype bulls were heavier than AA bulls (P<0.05). However, the AA bulls presented higher body weight and weight gain at 205 and 240 d (P<0.05). After a 24 h aging period, lower Warner Bratzler shear force (WBS) was found in AA bulls (4.48 kg) than in AB bulls (6.05 kg) in the Longissimus dorsi muscle for a difference between genotypes of 1.57 kg (P<0.01). The difference in WBS after 14 d of aging was 0.60 kg (AA = 3.13 kg; AB = 3.73 kg, P>0.05). These results indicate that the Senepol breed has a higher AA genotype frequency than the Charolais, and may suggest a tendency for Senepol animals to gain weight efficiently and produce tender beef. Therefore, this study suggests that this calpastatin polymorphism might be useful in animal selection because of its significant relationship with important beef traits. PARTICIPANTS: PD: Dr. Melvin Pagan: grant and project management, supervision graduate and undergraduate students. In charge of writing reports required by the USDA, data mining and abstracts submission. Genotype/phenotype association study. CO-PD: Dr. Danilo Cianzio: management of the University of Puerto Rico beef cattle herd located at Montana Farm and correspondent collection of growth data. CO-PD: Dr. Guillermo Ortiz Colon: Establishment and validation of techniques used to determine intramuscular fat, myofibrillar fragmentation, and expression (immunoblot) of CAST, CAPN, Desmin, Vinculin, and Troponin T in control and aged beef. Evaluation of the effect of marker genotype/haplotype, and supplementation treatment on carcass and muscle fat. Undergraduate research associates training and supervision. Prof. Americo Casas: scientist in charge of a concurrent strategic supplementation experiment, harvesting of bulls, assortment and organization of carcass data, aging of beef. Prof. Aixa Rivera: coordination of sensory evaluation panels and Warner-Bratzler shear force analysis. Dr. Catherine Ernst: collaborator, Department of Animal Science, Michigan State University. CAST genomic sequence organization for submission to the National Center for Biotechnology Information (NCBI) of the National Institute of Health (NIH). Deborah Velez, Ileana Rivera, Delvin Marrero, Lorraine Lopez/Graduate Research Assistants Natalia Vega, Paollette Rivera, Laura del Valle, Michelle M. Martinez, Beatriz I. Velez, Laura Emmanuelli, Alexandra E. Rodriguez, Deiam Ortiz, Bianca Diaz/Undergraduate Research Assistants. Senepol Cattle Breeders Association (SCBA), ASOSENEPOL Colombia, ASOSENEPOL Venezuela, ASOSENEPOL Panama. TARGET AUDIENCES: (1)Target audiences:The principal target audience of this project was the directory of all six regions of the Senepol Cattle Breeders Association. However,emphasis was provided to region one which encompasses the following countries:US VirginIslands,Panama,Venezuela,Dominican Republic,Colombia,Canada,Brazil,Costa Rica,Guatemala,South Africa,and Puerto Rico. Preliminary results from this project were presented in meetings celebrated in St.Croix,Panama, Venezuela,and Colombia. Through the XXI Meeting of the Latin American Association of Animal Production,results of this project were shown to people from Central and South America,and the Caribbean Basin. Additional data was presented at the national meeting of the American Society of Animal Science in Denver,Colorado. This project was in a 12 months non cost extension. That period of time was required to coordinate all the proposed genotyping and overall analyses,because the proposed objectives included a multinational collaboration with coordination efforts that were completed in June 2011 at the SCBA convention held in Anasco,Puerto Rico.(2)Efforts:Board members of SCBA and international Senepol breeders experienced at first hand,and for the first time,the nature of the integration and use of molecular genetics data (other than parentage testing) in their current animal breeding and selection strategies. Everything complemented the knowledge of the expected progeny differences for economically important growth traits. Efforts were directed to obtain samples for DNA isolation from Colombia,Venezuela,Panama,Dominican Republic,Costa Rica,Bolivia,St.Croix,Florida and other southern states of USA. Individual Senepol breeders were contacted and assistance was requested to the International Senepol Cattle Breeders Association (SCBA). Information regarding cattle inventory,animal's pedigree and phenotypic traits (production records) for each herd was provided by SCBA (EPDs for birth weight,weaning weight,milk, milk/growth,and yearling weight are routinely calculated for registering animals). Very importantly,novel associations with the candidate genes under study were found. Although most of the research was performer using cattle located in Puerto Rico (INPE-UPRM),genotypic data (CAPN1,CAST,and Leptin) was also provided by AsoSenepol Colombia for a total of 226 animals (bulls and cows) registered at SCBA. This information provided a more solid base of evaluation and comparison of such marker segregation to ultimately determine its usefulness in Marker Assisted Selection and Marker Assisted Management Program in the respective herds/countries. Also, Molecular Value Predictions,or in other words, Molecular Breeding Values (MVP,Pfizer Animal Genetics) for feed efficiency,tenderness and quality grade were obtained by using a panel of 56 markers and traditional GeneStar alleles (tenderness: T1-T3 and quality grade: T1-T4) analysis were coordinated for a total of 186 SCBA registered bulls (most of which appears in the Senepol Sire Summary and are use in artificial insemination and embryo transfer) from countries like Brazil,Colombia,St. Croix,Puerto Rico, and the continental US as a project conclusion. PROJECT MODIFICATIONS: No major modifications were performed.
Impacts
This project of knowledge enhancement corresponds to a validation of molecular markers identified in CAPN1 (316, 4751) using both a genotype/haplotype approach and in CAST (published and unpublished SNPs), taking advantage of growth, carcass, meat quality, and biochemical data available for purebred Senepol cattle and their crossbreds. Single nucleotide polymorphism (SNP) reported at exon 9 of bovine CAPN1 gene [cytosine (C) to guanine (G)} predicts an amino acid (AA) change [alanine(Ala) to glycine(Gly)] in the primary structure of micro-calpain, specifically at residue 316. Such SNP/AA substitution has been associated with differences in Warner-Bratzler shear force among animals. Today the consensus is that cattle with the cytosine/alanine molecular profile posses a predisposition to produce tender beef. The current hypothesis is that a potential conformational change in micro-calpain could affect its proteolytic capacity. In the present project we examined, by means of protein modeling, potential structural changes in the proteolytic domain (labeled II) of bovine micro-calpain by comparing the C/Ala and G/Gly molecular scenario. Analysis of domain II tertiary structure (3-D) indicates that the presence of Ala (CC genotype) could result in a higher degree of steric impediment. Moreover, the difference in methyl group (CH3) in Ala and a proton in Gly could result in a higher movement of water and more desirable domain flexibility. Therefore, such conformational change might be responsible for changes in enzyme efficiency during the postmortem aging of beef. Furthermore, additional evidence suggests that a significant interaction (P = 0.05) was observed between CAPN1-316 and beef aging (0 and 14 d) for Warner-Bratzler shear force. Therefore, an extensive analysis of the usefulness of molecular markers identified at the CAPN1 and CAST loci was performed. Ultimately, this project's results indicate that in order for marker assisted selection and management to be implemented; more attention should be given to CAPN1-316 and the novel CAST polymorphisms because of their associations with multiple traits of economic importance. Then, favorable alleles can be incorporated by using Senepol germplasm while paying attention to individual SNPs, haplotypes (CAPN1 and CAST, individually), or the complete combination [CAPN1 (n = 2) plus CAST (n=2)].
Publications
- J. Bosques, M. Pagan, A. Casas, & D. Cianzio. 2011 Effects of polymorphisms at micro-Calpain and Calpastatin over economically important traits of beef cattle from Puerto Rico. J. Agri. Univ. P.R. Submitted.
- J. Bosques, M. Pagan, A. Casas, & D. Cianzio. 2011. Segregation of polymorphisms at micro-Calpain and Calpastatin in beef cattle from Puerto Rico. J. Agri. Univ. P.R. Submitted.
- M. Pagan. Feed Efficiency vs. Intramuscular Fat Deposition: A Molecular Case Study in the Senepol Breed. 34th Senepol Cattle Breeders Association (SCBA) Annual Convention. Rincon Beach Resort. Anasco, PR. June 22-25, 2011.
- G. Ortiz. Comparison of the myofibrillar fragmentation index (MFI) in Senepol x Charolais crossbreds segregating a insertion/deletion polymorphism in Calpastatin. 34th Senepol Cattle Breeders Association (SCBA) Annual Convention. Rincon Beach Resort. Anasco, PR. June 22-25, 2011.
- P. Rivera, J. Bosques, A. Casas, D. Cianzio & M. Pagan. 2010. Breed distribution of genotypes corresponding to candidate genes located at bovine chromosome 1, 5, 7, and 29. SOPCA 2010.
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Progress 01/01/10 to 12/31/10
Outputs
OUTPUTS: Project TSTAR-122 includes the objectives of analyzing the segregation pattern for CAPN and CAST polymorphisms in purebred Senepol cattle (bulls and cows) and their crossbreds within the Caribbean Basin, Southern USA, Central and South America; it also investigates the potential association between these polymorphisms with economically important beef traits. Segregation analysis of CAPN haplotypes was performed to determine the number of bulls present in each class, using a group of animals from Puerto Rico (n = 94). In that population, the more representative haplotypes (CAPN1-316-4751, n = 9) were RY, GY, and GT. Surprisingly, such genetic profiles were associated with weights at birth, 205 d, 240 d, at slaughter; sensory tenderness (0 and 14 d of age); bone percentage; and with individual muscle weights (Biceps femoris and Longissimus dorsi). The segregation pattern of CAST SNP, previously reported by Barendse (2002), was determined within our cattle population. For that SNP, is now well documented that the presence of the T allele is associated with a reduction of 0.15 kg in shear force at 14 d postmortem (Van Enenneman et al., 2007). The same was observed in the Senepol x Charolais crossbreds harvested by our group (n = 42). However, our results demonstrate that such reduction in shear force was greater (0.35 kg) after an aging period of 24 h, all of which compares favorably with a report by Casas et al. (2006) at 14 d postmortem. Meanwhile, we have identified a novel polymorphism in CAST (AA, AB, BB genotypes). A total of 125 bulls (from 50% to purebred Senepol or Charolais) were genotyped to determine segregation and potential association with growth and meat quality traits. Overall, the frequency of the allele labeled as A (cytosine deletion) was higher (0.85) than the B (cytosine insertion) allele (0.15). In bulls of Senepol descent, allele frequencies were 0.91A/0.09B, whereas in those with Charolais blood, frequencies were 0.70A/0.30B. Genotypic frequencies were 0.83AA, 0.17AB, 0.0BB, and 0.48AA, 0.44AB, 0.08BB for Senepol and Charolais bulls, respectively. Because the BB genotype was observed only in two Charolais bulls, it was excluded for corresponding statistical analyses (n = 55 for growth; n = 24 for meat quality traits). At birth, AB genotype bulls were heavier than AA bulls (P<0.05). However, the AA bulls presented higher body weights and weight gain at 205 and 240 d (P<0.05). After a 24 hr aging period, lower Warner Bratzler shear force (WBS) was found in AA bulls (4.48 kg) than in AB bulls (6.05 kg) in the Longissimus dorsi muscle for a difference between genotypes of 1.57 kg (P<0.01). The difference in WBS after 14 d of aging was 0.60 kg (AA = 3.13 kg; AB = 3.73 kg, P>0.05). These results indicate that the Senepol breed has a higher AA genotype frequency than the Charolais, and these findings may suggest a tendency for Senepol animals to gain weight efficiently and to produce tender beef. Therefore, this study suggests that this calpastatin polymorphism might be useful in animal selection because of its significant relationship with important beef traits. PARTICIPANTS: PD: Dr. Melvin Pagan: grant and project management, supervision of graduate and undergraduate students. In charge of writing reports required by the USDA, data mining and abstracts submission. Genotype/phenotype association study. CO-PD: Dr. Danilo Cianzio: management of the University of Puerto Rico beef cattle herd located at Montana Farm and correspondent collection of growth data. CO-PD: Dr. Guillermo Ortiz Colon: Establishment and validation of techniques used to determine intramuscular fat, myofibrillar fragmentation, and expression (immunoblot) of CAST, CAPN, Desmin, Vinculin, and Troponin T in control and aged beef. Evaluation of the effect of marker genotype/haplotype, and supplementation treatment on carcass and muscle fat. Undergraduate research associates training and supervision. Prof. Americo Casas: scientist in charge of a concurrent strategic supplementation experiment, harvesting of bulls, assortment and organization of carcass data, aging of beef. Prof. Aixa Rivera: coordination of sensory evaluation panels and Warner-Bratzler shear force analysis. Dr. Catherine Ernst: collaborator, Department of Animal Science, Michigan State University. CAST genomic sequence organization for submission to the National Center for Biotechnology Information (NCBI) of the National Institute of Health (NIH). Deborah Velez, Ileana Rivera, Delvin Marrero, Lorraine Lopez/Graduate Research Assistants Natalia Vega, Paollette Rivera, Laura del Valle, Michelle M. Martinez, Beatriz I. Velez, Laura Emmanuelli, Alexandra E. Rodriguez, Deiam Ortiz/Undergraduate Research Assistants. Senepol Cattle Breeders Association (SCBA), ASOSENEPOL Colombia, ASOSENEPOL Venezuela, ASOSENEPOL Panama. TARGET AUDIENCES: The principal target of this project is the directory of all six regions of the Senepol Cattle Breeders Association. However, emphasis will be provided to region one, which encompasses the following countries: US Virgin Islands, Panama, Venezuela, Dominican Republic, Colombia, Canada, Brazil, Costa Rica, Guatemala, South Africa, and Puerto Rico. Preliminary results from this project were presented in meetings celebrated in St. Croix, Panama, Venezuela, and Colombia. Through the XXI Meeting of the Latin American Association of Animal Production, results of this project were presented to people from Central and South America, and the Caribbean basin. Additional data were presented at the national meeting of the American Society of Animal Science to be held in Denver, Colorado. This meeting was held in conjunction with the American Dairy Science Association (ADSA), Poultry Science Association (PSA), and the Mexican Association of Animal Production (AMPA). However, we are currently in a 12-month non cost extension. That period of time is required to complete all the proposed genotyping and analyses since this project includes a multinational collaboration with coordination efforts to be completed by the time of the June 2011 SCBA convention to be held in Rincon/Aguadilla, Puerto Rico) Efforts Experiential learning was provided to a total of three students enrolled in ANIN 4029: Special Projects. Additionally, two students are pursuing their graduate degrees as part of the current project. A total of six undergraduate research assistantships were conferred. PROJECT MODIFICATIONS: No major modifications are expected. Ongoing efforts are directed toward obtaining samples from Colombia, Venezuela, Panama, Dominican Republic, St. Croix, Florida and other southern states of the USA. Individual Senepol breeders had been contacted, and assistance was requested of the International Senepol Cattle Breeders Association (SCBA). Information regarding cattle inventory, animal's pedigree, and phenotypic traits (production records) for each herd was provided. Initial efforts were coordinated during the 2010 SCBA convention held in Baton Rouge, LA (June 16-18, 2010), with the purpose of completing this objective. Also, personnel from the USDA-Animal and Plant Health Inspection Service oriented us in terms of the policies and protocols associated with imports of biological material into the United States and its territories. Very important and novel associations with the candidate genes under study have been found. Most of the research was perform using cattle located in Puerto Rico (INPE-UPRM), however, validation is pending once samples and genotypes are collected from cattle located outside Puerto Rico. This information will provide a more solid base of evaluation of such markers, which ultimately could determine its usefulness in Marker Assisted Selection and Marker Assisted Management Programs in the respective herds/countries. Also, we are considering obtaining Molecular Value Predictions (Pfizer Animal Genetics) for comparison with the ultimate project statistical analysis and segregation of alleles.
Impacts
This project's change of knowledge corresponds to a validation of molecular markers identified in CAPN1 (316, 4751) using both a genotype/haplotype approach and in CAST (published and unpublished SNPs), taking advantage of growth, carcass, meat quality, and biochemical data available for purebred Senepol cattle and their crossbreds. Single nucleotide polymorphism (SNP) reported at exon 9 of the bovine CAPN1 gene [cytosine (C) to guanine (G)} predicts an amino acid (AA) change [alanine(Ala) to glycine(Gly)] in the primary structure of micro-calpain, specifically at residue 316. Such SNP/AA substitution has been associated with differences in Warner-Bratzler shear force between animals. Today the consensus is that cattle with the cytosine/alanine molecular profile possess a predisposition to produce tender beef. The current hypothesis is that a potential conformational change in micro-calpain could affect its proteolytic capacity. In the present project we examined, by means of protein modeling, potential structural changes in the proteolytic domain (labeled II) of bovine micro-calpain by comparing the C/Ala and G/Gly molecular scenarios. Analysis of domain II tertiary structure (3-D) indicates that the presence of Ala (CC genotype) could result in a higher degree of steric impediment. Moreover, the difference in methyl group (CH3) in Ala and a proton in Gly could result in a higher movement of water and more desirable domain flexibility. Therefore, such conformational change might be responsible for changes in enzyme efficiency during the postmortem aging of beef. Furthermore, additional evidence suggests that a significant interaction (P = 0.05) was observed between CAPN1-316 and beef aging (0 and 14 d) for Warner-Bratzler shear force. Therefore, we performed an extensive analysis of the usefulness of molecular markers identified at the CAPN1 and CAST loci. However, this investigation is an ongoing project. So far, it seems that in order to marker assisted selection and management to be implemented, more attention should be given to CAPN1-316 and the novel CAST polymorphisms because of their association with multiple traits of economic importance. Then favorable alleles can be incorporated by using Senepol germplasm paying attention to individual SNPs, haplotypes (CAPN1 and CAST, individually), or the complete combination [CAPN1 (n = 2) plus CAST (n=2)].
Publications
- Melvin Pagan, Natalia Vega, Deborah Velez, Americo Casas, Aixa Rivera, y Danilo Cianzio. Molecular markers within the Calpain/Calpastatin axis and its association with meat quality in Bos taurus cattle. Annual Meeting of the Caribbean Food Crops Society, July 11-17, 2010. Boca Chica, Dominican Republic.
- L. del Valle-Mercado, Natalia Vega, Deborah Velez, A. Casas, D. Cianzio, M. Pagan, G. Ortiz-Colon. A Calpastatin insertion/deletion polymorphism is associated with the myofibril fragmentation index in Bos taurus bulls. Annual Meeting of the Caribbean Food Crops Society, July 11-17, 2010. Boca Chica, Dominican Republic.
- N. Vega, D. Velez, A. Casas, D. Cianzio, C.W Ernst & M. Pagan An insertion/deletion polymorphism at the bovine calpastatin locus is associated with economically important traits. National Meeting of the American Society of Animal Science, July 11-15, 2010. Denver, CO.
- P. Rivera, J. Bosques, A. Casas, D. Cianzio & M. Pagan. A genotype combination approach using micro-Calpain as a candidate gene for growth, carcass, and meat quality in bulls of Senepol and Charolais inheritance. National Meeting of the American Society of Animal Science, July 11-15, 2010. Denver, CO.
- D. Velez, C.W. Ernst, and M. Pagan. Evaluation of insertion/deletion and single nucleotide polymorphisms identified at the bovine Insulin-Like Growth Factor Binding Protein-2 locus. National Meeting of the American Society of Animal Science, July 11-15, 2010. Denver, CO.
- L. del Valle, A. Casas, D. Cianzio, M. Pagan, G. Ortiz-Colon. Myofibrilar fragmentation index of longissimus muscle of Senepol and Charolais crossbred bulls. National Meeting of the American Society of Animal Science, July 11-15, 2010. Denver, CO.
- M. Pagan. Characterization and Validation of Senepol Cattle Germplasm using Molecular Genetics Tools. 1st International Encounter of Senepol Cattle Breeders. Los Potros Ranch, Llano Grande, Antioquia, Colombia, November 19-23, 2009.
- D. Cianzio. The Senepol in Puerto Rico: Past and Future. 1st International Encounter of Senepol Cattle Breeders. Caracas, Venezuela, November 19-23, 2009.
- M. Pagan, E. Jimenez, & B. Pastrana. Molecular modeling of bovine micro-calpain domain II from a nucleotide/aminoacid (CAPN1-316). Proceedings XXI Meeting of the Latin-American Association of Animal Production (ALPA), October 18-23, 2009. San Juan, Puerto Rico.
- P. Rivera, J. Bosques, A. Casas, D. Cianzio & M. Pagan. Assestment of candidates genes located at chromosomes 1, 5, 7, and 29 for marker assited selection in purebred senepol bulls. Proceedings XXI Meeting of the Latin-American Association of Animal Production (ALPA), October 18-23, 2009. San Juan, Puerto Rico.
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Progress 01/01/09 to 12/31/09
Outputs
OUTPUTS: Project TSTAR-122 includes the objectives of analyzing the segregation pattern for CAPN and CAST polymorphisms in purebred Senepol cattle (bulls and cows) and their crossbreds within the Caribbean Basin, Southern USA, Central and South America, and to investigate the potential association between these polymorphisms with economically important beef traits. Segregation analysis of CAPN haplotypes were performed to determine the number of bulls present in each class using a group of bulls from Puerto Rico (n = 94). In those animals the more representative haplotypes (CAPN1-316-4751, n = 9) were RY, GY, and GT. Surprisingly, such genetic profiles were associated with weights at birth, 205 d, 240 d, and at slaughter; sensory tenderness at 0 and 14 d; bone percentage; and with individual muscle weights (Biceps femoris and Longissimus dorsi). The segregation pattern of CAST SNP, previously reported by Barendse (2002), was determined within our cattle population. For that SNP, it is now well documented that the presence of the T allele is associated with a reduction of 0.15 kg in shear force at 14 d postmortem (Van Enenneman et al., 2007). The same was observed in the Senepol x Charolais crossbreds harvested by our group (n = 42). However, our results demonstrate that such reduction in shear force was greater (0.35 kg) after an aging period of 24 h, all of which compares favorably with a report by Casas et al. (2006) at 14-d postmortem. Meanwhile, we have identified a novel polymorphism in CAST (AA, AB, BB genotypes) with corresponding allele frequencies of 0.85 A/0.15 B (n = 66 Senepol, Charolais, and Senepol x Charolais crossbreds). Our data indicate that the A allele (AA vs. AB genotype) is associated with lesser shear force (0.69 kg) at 14-d postmortem (n = 25 Senepol x Charolais crossbreds). Dramatically, past an aging period of only 24 h, the reduction in shear force corresponded to 1.57 kg. The interesting finding of such novel CAST polymorphism, which is a Cytosine insertion/deletion, is that it seems to be associated (data set of 56 bulls) with growth traits (birth weight, body weight and gains at 205 and 240 d, and weaning age). These observations are similar to the corresponding evidence for CAPN1 haplotypes. Currently, an additional group of animals (n = 88) will be genotyped in order to have more comprehensive statistics. Blood will be drawn from around 400 bovines for the same purposes. This work is being done by four undergraduate students. The data generated so far were presented in meetings celebrated in St. Croix, Panama, Venezuela, and Colombia. Multiple collaborations were established with the International Senepol Cattle Breeders Association (SCBA), and its corresponding Central and South American branches, in order to include in this project herds located in countries where the Senepol breed is considered important and in which a considerable amount of animals with production records are available. PARTICIPANTS: PD: Dr. Melvin Pagan: grant and project management, supervision graduate and undergraduate students. In charge of writing reports required by the USDA, data mining and abstracts submission. Genotype/phenotype association study. CO-PD: Dr. Danilo Cianzio: management of the University of Puerto Rico beef cattle herd located at Montana Farm and correspondent collection of growth data. CO-PD: Dr. Guillermo Ortiz Colon: Establishment and validation of techniques used to determine intramuscular fat, myofibrillar fragmentation, and expression (immunoblot) of CAST, CAPN, Desmin, Vinculin, and Troponin T in control and aged beef. Evaluation of the effect of marker genotype/haplotype, and supplementation treatment on carcass and muscle fat. Undergraduate research associates training and supervision. Prof. Americo Casas: scientist in charge of a concurrent strategic supplementation experiment, harvesting of bulls, assortment and organization of carcass data, aging of beef. Prof. Aixa Rivera: coordination of sensory evaluation panels and Warner-Bratzler shear force analysis. Dr. Catherine Ernst: collaborator, Department of Animal Science, Michigan State University. CAST genomic sequence organization for submission to the National Center for Biotechnology Information (NCBI) of the National Institute of Health (NIH). Deborah Velez, Delvin Marrero, Natalia Vega, Paollette Rivera, Laura del Valle. Graduate and undergraduate research associates. Senepol Cattle Breeders Association (SCBA), ASOSENEPOL Colombia, ASOSENEPOL Venezuela, ASOSENEPOL Panama TARGET AUDIENCES: The principal target of this project is the directory of all six regions of the Senepol Cattle Breeders Association. However, emphasis will be provided to region one which encompasses the US Virgin Islands, Panama, Venezuela, Dominican Republic, Colombia, Canada, Brazil, Costa Rica, Guatemala, South Africa, and Puerto Rico. Preliminary results from this project were presented in meetings celebrated in St. Croix, Panama, Venezuela, and Colombia. Through the XXI Meeting of the Latin American Association of Animal Production, results of this project were presented to people from Central and South America, and from the Caribbean basin. Additional data will be presented at the national meeting of the American Society of Animal Science to be held in Denver, Colorado. This meeting will be held in conjunction with the American Dairy Science Association (ADSA), Poultry Science Association (PSA), and the Mexican Association of Animal Production (AMPA). Two student's will pursue summer internships at the Animal Molecular Genetics Laboratory of Dr. Eduardo Casas located at the US Meat Animal Research Center in Clay Center, NE. Experiential learning was provided to a total of three students enrolled in ANIN 4029: Special Projects. Additionally, two students are pursuing their graduate degrees as part of the current project. In the fall of 2010, and for the first time in the history of UPRM, an Animal Biotechnology course will be available for enrollment. PROJECT MODIFICATIONS: Not relevant to this project.
Impacts
This projects change of knowledge corresponds to validation of molecular markers identified in CAPN1 (316, 4751) using both a genotype/haplotype approach and in CAST (published and unpublished SNPs), taking advantage of growth, carcass, meat quality, and biochemical data available for purebred Senepol cattle and their crossbreds. Single nucleotide polymorphism (SNP) reported at exon 9 of bovine CAPN1 gene [cytosine (C) to guanine (G) predicts an amino acid (AA) change [alanine (Ala) to glycine (Gly)] in the primary structure of micro-calpain, specifically at residue 316. Such SNP/AA substitution has been associated with differences in Warner-Bratzler shear force among animals. Today the consensus is that cattle with the cytosine/alanine molecular profile possess a predisposition to produce tender beef. The current hypothesis is that a potential conformational change in micro-calpain could affect its proteolytic capacity. In the present project we examined, by means of protein modeling, potential structural changes in the proteolytic domain (labeled II) of bovine micro-calpain by comparing the C/Ala and G/Gly molecular scenario. Analysis of domain II tertiary structure (3-D) indicates that the presence of Ala (CC genotype) could result in a higher degree of steric impediment. Moreover, the difference in methyl group (CH3) in Ala and a proton in Gly could result in a greater movement of water and more desirable domain flexibility. Therefore, such conformational change might be responsible of changes in enzyme efficiency during the postmortem aging of beef. Furthermore, additional evidence suggests that a significant interaction (P = 0.05) was observed between CAPN1-316 and beef aging (0 and 14 d) for Warner-Bratzler shear force. Therefore, an extensive analysis of the usefulness of molecular markers identified at the CAPN1 and CAST loci was performed. However, this procedure is an ongoing project. So far, it seems that in order for marker-assisted selection and management to be implemented, more attention should be given to CAPN1-316 and the novel CAST polymorphisms because of its association with multiple traits of economic importance. Then favorable alleles can be incorporated by using Senepol germplasm and paying attention to individual SNPs, haplotypes (CAPN1 and CAST, individually), or to the complete combination [CAPN1 (n = 2) plus CAST (n=2)].
Publications
- M. Pagan. Characterization and Validation of Senepol Cattle Germplasm using Molecular Genetics Tools. 1st International Encounter of Senepol Cattle Breeders. Los Potros Ranch, Llano Grande. Antioquia, Colombia. November 19-23, 2009.
- M. Pagan. DNA Research for Senepol. 32nd Senepol Cattle Breeders Association (SCBA) Annual Convention. Carambola Spa and Resort, St. Croix, USVI. June 24-26, 2009.
- D. Cianzio. The Senepol in Puerto Rico: Past and Future. 1st International Encounter of Senepol Cattle Breeders. Caracas, Venezuela. November 19-23, 2009.
- M. Pagan, E. Jimenez, & B. Pastrana. 2009. MOLECULAR MODELLING OF BOVINE MICRO-CALPAIN DOMAIN II FROM A NUCLEOTIDE/AMINOACID SUBSTITUTION (CAPN1-316). Proceedings XXI Meeting of the Latin-American Association of Animal Production (ALPA). October 18-23, San Juan, Puerto Rico.
- P. Rivera, J. Bosques, A. Casas, D. Cianzio & M. Pagan. 2009. ASSESTMENT OF CANDIDATES GENES LOCATED AT CHROMOSOMES 1, 5, 7, AND 29 FOR MARKER ASSISTED SELECTION IN PUREBRED SENEPOL BULLS. PROCEEDINGS XXI Meeting of the Latin-American Association of Animal Production (ALPA). October 18-23, San Juan, Puerto Rico.
- M. Pagan, G. Ortiz Colon, A. Rivera, A. Casas, & D. Cianzio. Association of micro-calpain genotype and breed over the myofibillar fragmentation of the longissimus dorsi muscle of young bulls. Oral Presentation at the 2008 Scientific Reunion and Annual Meeting of the Puerto Rican Society of Agricultural Sciences (SOPCA). November 20-21, 2008. El Faro Hotel, Aguadilla, Puerto Rico.
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