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Revista Colombiana de Ciencias Pecuarias

Print version ISSN 0120-0690On-line version ISSN 2256-2958

Rev Colom Cienc Pecua vol.25 no.2 Medellín Apr./June 2012

 

ARTÍCULO ORIGINAL

 

Allelic frequency of the Kappa–Casein gene in Colombian and creole cattle breeds¤

 

Frecuencia alélica del gen Kappa–Caseína en bovinos criollos y colombianos

 

Freqüência do alelo de gene de Kappa–Caseína em bovinos crioulos e colombianos

 

 

Jaime A Rosero1, MS student; Luz A Álvarez 2*, MS; Jaime E Muñoz2, AgrEng, PhD; Carlos V Durán2, MS; Ángela G Rodas3, MS.

 

* Corresponding author: Luz Ángela Álvarez Universidad Nacional de Colombia, Sede Palmira. email: laalvarezf@palmira.unal.edu.co.

1 Palmira Animal Genetics Resource Group, Dept. of Animal Science, School of Agricultural and Livestock Sciences, National University of Colombia, Palmira Campus .

2 Professor, School of Agricultural and Livestock Sciences, National University of Colombia AA 237, Palmira, Colombia.

3 Research Group on Conservation, Improvement, and Use of ''Hartón del Valle'' Creole Cattle and Other Animal Genetic Resources in Southwest Colombia, School of Agricultural and Livestock Sciences, National University of Colombia, Palmira, Colombia.

 

(Recibido: 20 marzo, 2010; aceptado: 23 agosto, 2011)

 

 


Summary

Colombia has one of the most genetically diverse creole cattle populations, with eight creole breeds and two improved creole (Colombian) breeds. A high demand for meat and milk has led to the inevitable selection of highly productive cattle and the introduction of foreign breeds. Unfortunately, these breeds are often ill–suited for tropical conditions. These factors threaten the size of the creole livestock population, which is considered part of Colombia's national heritage. Objective: to estimate the allelic frequencies of the Kappa–Casein gene (CNS3) in Colombian creole cattle breeds (GCC). Methods: a total of 354 blood samples were taken from 30 animals of each of the following breeds: Blanco Orejinegro (BON), Caqueteño (CQT), Casanareño (CAS), Horned Costeño (Costeño con Cuernos, CCC), Chino Santandereano (ChS), Hartón del Valle (HV), Romosinuano (ROM), and Sanmartinero (SM), each representing the 8 established ''criollo'' (creole) breeds; the Lucerna (LUC) and Velasquez (VEL) representing the two Colombian improved breeds; and Brahman and Holstein as control breeds. DNA was extracted by a salting–out procedure and a 453 bp fragment on chromosome 6 was amplified by PCR. CSN3 alleles were identified using single strand conformation polymorphism (SSCP) and their sequence compared with those of the Genebank for Bos taurus and Bos indicus. Results: higher frequencies for allele variants of CSN3 A (0.39) and B (0.41) were found relative to the frequencies of I (0.038), G (0.095), A1 (0.025), E (0.006), and N (0.006). The allele of interest (CSN3 B) had a high frequency in the CCC (0.81), ROMO (0.66), CQT (0.55), ChS (0.48), and VEL (0.43) breeds. Conclusions: these findings suggest that Colombian creole breeds harbor a high genetic diversity which enriches its gene pool and warrants future conservation efforts to protect its integrity.

Key words: genetic variants, milk proteins, molecular markers, PCR–SSCP


Resumen

Colombia es uno de los países más diversos en recursos genéticos criollos. Posee ochos razas de ganado criollo (GCC) y dos razas de criollo mejorado o razas colombianas. La creciente demanda de alimentos ha generado una forzosa selección de individuos altamente productivos e introducción de razas foráneas (Holstein y Brahman) poco adaptadas a condiciones tropicales, lo que ha puesto en riesgo el tamaño efectivo del ganado criollo, considerado patrimonio nacional. Objetivo: estimar la frecuencia alélica del gen (CNS3) de la Kappa–Caseína en el (GCC). Métodos: se usaron 354 muestras de sangre de ocho razas bovinas criollas (30 individuos por raza): Blanco Orejinegro (BON), Caqueteño (CQT), Casanareño (CAS), Costeño con Cuernos (CCC), Chino Santandereano (ChS), Hartón del Valle (HV), Romosinuano (ROMO) y Sanmartinero (SM), dos colombianas Lucerna (LUC) y Velásquez (VEL) y dos controles Brahman y Holstein. Con el fin de estimar la frecuencia de los alelos k–caseína (k–CN) se amplificó un fragmento de 453 pb para k–CN (cromosoma 6). Los alelos se identificaron mediante la técnica PCR–SSCP. Resultados: se encontró mayor frecuencia para las variantes de k–CN A (0.39) y B (0.41), en comparación a I (0.038), G (0.095), A1 (0.025), E (0.006) y N (0.006). El alelo de interés k–CN B presentó alta frecuencia en las razas CCC (0.81), ROMO (0.66), CQT (0.55), ChS (0.48), y VEL (0.43). Conclusiones: la alta frecuencia del alelo de interés del gen k–CN ratifica al GCC como alternativa viable en esquemas sostenibles de producción de leche de mejor calidad y corrobora la necesidad de evaluación y caracterización de recursos zoogenético, como primer paso para su conservación.

Palabras clave: marcadores moleculares, PCR–SSCP, proteínas leche, variantes genéticas.


Resumo

A Colômbia é um dos países mais diversificado em recursos genéticos crioulos, tem oito bovinos da raça nativa e duas raças melhoradas ou raças crioulo colombiano (GCC). A alta demanda por alimentos tem levado a uma seleção forçada das pessoas altamente produtivas e/ou introdução de raças estrangeiras mal adaptados às condições tropicais, que têm prejudicado o tamanho efetivo de animais considerados património crioulo. Objetivo: estimar a frequência do alelo do gene Kappa–Caseína (CNS3) na (GCC). Métodos: foram utilizados 354 amostras de sangue de oito raças nativas (30 indivíduos por raça): Blanco Orejinegro (BON), Caqueteño (CQT), Casanareño (CAS), Costeño con Cuernos (CCC), Chino Santandereano (ChS), Hartón del Valle (HV), Romosinuano (ROM) e Sanmartinero (SM), dois Colombianas Lucerna (LUC) e Velásquez (VEL) e dois controles (Brahman and Holstein). Para estimar a freqüência de alelos de κ–caseína (CSN3), amplificaram um fragmento de 453 pb para CSN3. Os alelos foram identificados por PCR–SSCP. Resultados: encontramos uma maior freqüência de variantes do CSN3 A (0.39) e B (0.41), comparado com I (0.038), G (0.095), A1 (0.025), E (0.006) e N (0.006). O alelo de interesse CSN3 B apresentou alta freqüência em raças CCC (0.81), ROMO (0.66), CQT (0.55), CHS (0.48) e VEL (0.43). Conclusões: estes resultados sugerem que o CCG é um recurso genético, que abriga uma grande diversidade genética e apoia a necessidade de avaliação e caracterização dos recursos genéticos animais como um primeiro passo para a conservação.

Palavras chave: marcadores moleculares, PCR–SSCP, proteínas do leite, variantes genéticas.


 

 

Introduction

Caseins represent 80% of bovine milk proteins (Fox and Brodkorb, 2008). The CSN3 gene in cattle has been extensively studied and plays a role in stabilizing caseins, interacting with 2–LG, and determining the physical stability of some milk products during heat treatment (Braunschweig et al., 2000; Fox and Brodkorb, 2008). Multiple variants of the CSN3 gene have been found, although the A and B alleles are the most common. Ten other allele variants have been described and include the following: A1 C, E, F1, F2, G1, G2, H, I, and J (Caroli et al., 2009). Of these, the B allele is associated with milk that has a shorter curdling time, a higher protein percentage, a higher stability during freezing, a greater cheese yield, and a more consistent curd formation (Lunden et al., 1997; Wedholm et al., 2006; Heck et al., 2009).

Eight Creole cattle breeds have been recognized in Colombia. These include the Romosinuano (ROMO) and the Horned Costeño (CCC) breeds, which are found in the floodplains and the dry plains in northern Colombia (Atlantic Coast); the Blanco Orejinegro (BON) and the Chino Santandereano (Chino) breeds, which are found in the mild temperate region of the Colombian Andes; the Hartón del Valle (Hartón) breed, which is found in the Cauca River Valley; the Casanareño (CAS) and the Sanmartinero (SM) breeds, which are found in the floodplains and high plains of the Colombian Orinoquía region, respectively; and the Caqueteño (CQT) breed, which is found in the Amazon region of Colombia (Martínez, 2010). In the 1930s and the 1950s, the Lucerna and the Velásquez hybrid bovine breeds were developled. The Lucerna breed originates from the Cauca Valley and is the product of hybridizing the Hartón del Valle, the Holstein, and the Shorthorn breeds. The Velásquez breed originates from the Magdalena Valley and is the product of hybridizing the Romosinuano, the Red Poll, and the Brahman breeds (Pinzón, 1991). Although considered a national resource, these breeds are threatened. Currently their population of 18,231 represents only 0.08% of the country's total bovine population (Martínez, 2010).

Assessments of CSN3 in all but the HV breed have not been previously reported (Naranjo et al 2007; Díaz et al., 2006). The objective of this studywas to estimate the allelic frequencies of the Kappa– Casein gene in eight Creole and two Colombian cattle breeds.

 

Materials and methods

Type of study

A uniform sample size of 30 animals from each of the BON, CQT, CAS, CCC, ChS, HV, ROMO, SM, LUC, and VEL breeds were sampled in different regions of Colombia. These sampling locations are shown in table 1 and figure 1. Special care was taken to select non–blood related animals. For the control, Brahman and Holstein DNA samples from the National University of Colombia's DNA bank were used.

The BON breed is known for its meat and milk production abilities under unfavorable tropical conditions. Its natural habitat is found in the central and western mountain foothills between 800 and 1,800 m above sea level, with temperatures between 18 and 24 ºC (López et al., 2001). In contrast, the CQT breed lives in the Caquetá region with an average annual temperature of 26 °C, and 88% relative humidity (Martínez, 1999). The CAS breed inhabits the floodplains of the Arauca and Casanare provinces. This breed is primarily raised for meat, using traditional extensive production methods (Asocriollo, 2007). In contrast, the CCC breed inhabits the coastal plains with temperatures of approximately 27.5 °C and 1.233 mm average annual precipitation (Wilkins et al., 1993). The CCC breed is considered to be one of the best breeds for milk production in the low–tropics (CORPOICA, 2006). The ChS breed inhabits the Lebrija Valley (Santander province) (MADR–Asocriollo, 2003), and the HV breed inhabits the Cauca River Valley between the river's source in the Colombian Massif and La Virginia region (Risaralda province) and between the hydrological divides of the Central and Western Mountain Ranges (Casas and Valderrama, 1998). Conversely, the ROMO breed is adapted to the Sinú Valley conditions, which has a dry tropical forest (DTF) climate and a mean temperature of 27.5 ºC with 83% relative humidity (Martínez, 1998). The SF breed inhabits the wet tropical and very wet tropical forest sub–regions of the Orinoco, which has aluminum rich soils, a mean temperature of 26 ºC, 87% relative humidity during the rainy season, and 55% relative humidity during the dry season (Martínez and González, 2000). The LUC breed originates from the Lucerna Ranch in Bugalagrande (Cauca Valley province) (Asocriollo, 2007), and the VEL breed originates from the Magdalena River Valley in the Caldas province (Corpoica, 2006).

 

Methods

DNA extraction and quantification. The DNA from 5 mL of blood drawn from the coccygealvein was extracted by using the Salting Outextraction protocol (Miller et al., 1988). The DNAquality was evaluated by using 0.8% agarose gelsin a 0.5 X TBE buffer (0.045 M tris–borate, 0.001M EDTA, pH 8.0) and stained with ethidiumbromide. The DNA was quantified by comparingwith known Lambda phage DNA concentrations.Electrophoresis was performed at 80 volts (V) for45 min, and gels were photographed under UV lightusing a Kodak EDAS 290 digital camera.

CSN3 gene amplification. The 453 bp DNA fragment on chromosome 6 was amplified (Barroso et al., 1998). Between 20 and 50 ng of DNA was mixed with a PCR buffer solution containing 20 pmol/μl of primer (sense, 5' –TGT GCT GAG TAG GTA TCC TAG TTA TGG–3'; antisense, 5'– GCG TTG TCT TCT TTG ATG TCT CCT TAG– 3), 25 mM MgCl , 1.25 µM of dNTP Mix (MBI fermentas–USA), and 2 U of Taq Polymerase (MBI fermentas–USA) with a total volume of 25 µL. The samples were subjected to denaturation for 5 min, followed by 35 cycles for 1 min at 94 °C, 1 min at 65 °C, and 2 min at 72 °C. At the end, a final 5 min extension step at 72 °C in a PTC–100TM thermal cycler was conducted (MJ Research, Inc–USA). 2

Identification of alleles. Alleles were identified using SSCP (Single Strand Conformation Polymorphism) as described by Barroso et al. (1998). Two micro–liters of the PCR product were combined with 8 μl of the denaturing buffer (0.05% xylene cyanol, 0.05% bromophenol blue, 5.5 mM EDTA pH 8.0), denatured at 95 °C for 5 min, and then cooled on ice for 2 min. The samples were loaded into 12 x 28 cm (Camera Biometra, ® Göttingen, Germany) 12% polyacrylamide gels (acrylamide: N, N'–methylene–bis–acrylamide ratio, 100:1) with 5% glycerol and a 0.5 X TBE buffer (0.045 M tris–borate, 0.001 M EDTA, pH 8.0). The gels were run at 180 V for 12 hours (h) at 12 °C. The resulting bands were stained using silver nitrate. The alleles were verified using PCR–RFLP (Polymerase Chain Reaction–Restriction Fragment Length Polymorphism) and the HindIII, HinfI HaeIII, HhaI, MaeII, and MspI restriction enzymes. In addition, the alleles were sequenced and compared with the 22 available GenBank sequences and the X14908 (Alexander et al., 1988) GenBank sequence for Bos taurus, and with the two available sequences for Bos indicus.

Statistical analysis. Allelic frequencies were determined by the direct counting method. The Hardy–Weinberg Equilibrium (HWE) within populations was estimated using the FIS coefficient test (Weir and Cockerham, 1984) and the exact test (Guo and Thompson, 1992). The genetic variability for each breed was calculated from the allele number (AN), the expected heterozygosity (HO), and the observed heterozygosity (HE). The coefficient of differentiation (FST Weir and Cockerham, 1984) and AMOVA method were used as a measure of genetic subdivision and breed differentiation. All data analysis was performed using the Arlequin Version 3.1 software (Excoffier et al., 2006). ST;

 

Results

Seven mobility patterns –corresponding to the A, B, A1, G, I, and E alleles as well as one allele that 1 could not be identified (N)– were detected using PCR–SSCP. With the exception of the N allele, allele identification was verified through PCR– RFLP and sequencing. Figure 2 shows the detected mobility patterns for the CSN3 gene belonging to the A, B, A 1 , G, I, and E alleles.

Table 2 shows CSN3 allelic frequencies by breed. The predominant alleles in the GCC population were the B (0.418 ± 0.20) and the A (0.39 ± 0.019) alleles. Other alleles (G, I, A1, E, and 1 N) had low frequencies and represented only 18% of the total frequency. In contrast, the control breeds had low allelic frequencies for B, with only 0.20 and 0.065 in the Holstein and the Brahman breeds, respectively.

The allele of interest (B) showed greater frequency in the CCC, ROMO, CQT, ChS, and VEL breeds than in the BON, SM, HV, and CAS breeds. The lowest B allele frequency was in the LUC breed. Allele A, which was dominant in the majority of commercial breeds, had a lower frequency in the GCC breed (with the exception of LUC [0.916 ± 0.036] and BON [0.616 ± 0.063]).

The I allele variant was found in six of the eight Colombian breeds and had an average value of 0.038 ± 0.078. In particular, this allele emerged in the SM breed (0.13 ± 0.044). The G allele was detected in eight breeds (0.095 ± 0.01) and had the greatest frequency in the CAS (0.36 ± 0.06), the HV (0.233 ± 0.055), and the VEL (0.116 ± 0.041) breeds. The A allele variant was identified in six breeds (0.025 ± 0.06), but was most present in the CAS breed (0.15 ± 0.046). The E allele variant (0.006 ± 0.003) was found with the same frequency (0.016 ± 0.016) in the CQT, the CAS, the SM, and the VEL breeds. The unidentifiable variant (N) was only present in the ChS and the VEL breeds. 1

In the HOLS control breed, a greater proportionof the A allele variant was found than the B allelevariant. The HOLS control breed also had a low Gallele frequency. The A, B, A , and G allele variants 1 were detected in the BRAH control breed, of whichthe G allele was predominant (0.52 ± 0.07).

Table 3 shows the number of alleles (NA), observed heterozygosity (Ho), expected heterozygosity (He), Fis, and the Hardy Weinberg Equilibrium (HWE). The average number of alleles for the GCC breed was 4.60 ± 1.57. A maximum of six alleles were found in the CQT, CAS, ChS, SM, and VEL breeds, and a minimum of two alleles were found in the ROMO breed.

For the GCC breed, the He oscillated between 0.16 and 0.74 and had an average He of 0.65 (p<0.05). The CAS, ChS, HV, VEL, and SM breeds had the highest genetic diversity (He), and the LUC and CCC breeds had the lowest. The commercial breeds all had a low He. The HWE was not found in the BON, ChS, HV, LUC, and VEL breeds.

The GCC breed showed a low global value for intrapopulational endogamy (FIS) that was highly significant (FIS=0.098, p<0.01), and was substantially different than zero in the ChS and LUC breeds.

The AMOVA revealed that 80% of genetic variance was due to individual variation, while 20% was due to variation among breeds. The global value for the coefficient of genetic differentiation (F ) was 0.203 (p<0.01). This value indicated that significant genetic differences were distinguishable ST among the studied cattle breeds.

 

Discussion

According to the Food and Agriculture Organization of the United Nations (FAO, 2007), the inevitable intensification of livestock systems increasingly depends upon the selection of highly productive species. This selection places less– productive but genetically valuable species at risk. As a result, the Colombian Creole SM, ChS, and CQT breeds are in a critical state, the BON, ROMO, HV, CCC, and VEL breeds are in a vulnerable state, and the LUC and CAS breeds are in a threatened state (Martínez, 1999; MADR, 2003). This situation highlights the need to evaluate the diversity of economically important genes in order to enhance their value and assure their conservation.

The alleles detected in this study demonstrated a run pattern similar to that described by Barroso et al. (1998), with the exception of the unidentified (N) allele. The majority of studies using PCR–RFLP only identified the A and B alleles. However, Díaz et al. (2006) found six alleles in the HV breed and Ceriotti et al. (2004) found A, A B, and H alleles in the Bos indicus breed and A, B, and E alleles in the Bos taurus, 1 breed. Likewise, in Nariño province (Colombia), Solarte et al. (2009) only detected A and B alleles with PCR–SSCP (1,087 Holstein, Jersey, Normande, Brown Swiss, Simmental, Red Pol, Swedish Red, and Montbeliard cattle and their crosses).

According to Prinzenberg et al. (1999), the PCR–SSCP method may introduce genotyping errors because the C allele can be confused with the B allele. Additionally, the E, F, G, H, and A 1 alleles can be confused with the A allele. The latter situation would result in an underestimation of the A allele, an inappropriate rejection of animals, and increased undesired alleles in animals used in industrial milk production. The degree of polymorphism detected in the CSN3 gene confirmed that the PCR–SSCP technique was an efficient method for identifying allele variants, and that the sample size used (30 animals) was adequate for detecting the existing variability. The presence of the seven alleles in the CSN3 gene demonstrates the extensive polymorphism found within the creole breeds. These alleles are associated with a range of habitats and management and production systems.

A high B allele frequency is desirable in industrialized milk production systems. The B allele is related to milk with high protein content, greater freezing–stability, higher cheese production (5–10%), shorter coagulation time, and more consistent curds (Van Eenennaam and Medrano, 1990). It is important to highlight that the B allele is predominant in the GCC breed (0.418 ± 0.20) and is above 55% in the CCC, the ROMO, and the CQT breeds. The low frequency of the B allele in the LUC breed is due to its genetic origins (40% HOLS, 30% Shorthorn, 30% HV) and its selection for achieving high milk yield. These traits are related to the CSN3 A allele. However, the highest frequencies of the B allele were found in some meat producing breeds rather than milk or dual– purpose production breeds. This finding suggests a possible selection for cows based on maternal abilities and is expressed by the shorter standing time for calves raised on milk containing higher protein. For example, in the HV breed milk contained 3.37% protein and was superior to that of the Holstein breed milk. The composition was similar to the Ayrshire and Brown Swiss breeds (Hurtado, 1998).

Creole breeds on the American continent share a common origin with cattle introduced during the Spanish conquest era. The average value of the B allele in the Creole breeds was consistent with the values reported in America, the Caribbean, Cuba (Uffo et al., 2006), Brazil (Kemenes et al., 1999; Lara et al., 2002), Perú (Veli et al., 2004), Argentina (Poli et al., 2002; Lirón et al., 2002; Martínez et al., 2003), Bolivia (Lirón et al., 2002), and Uruguay (Postiglioni et al., 2002). The frequency of the B allele in the Creole breeds was significantly higher (p<0.05) than that of the Holstein breed in Colombian–like conditions (López et al., 1999; Solarte–Portilla et al., 2009; Vivas et al., 2008; Díaz et al., 2006) and the specialized breeds (such as the Holstein, Guernsey, Milking Shorthorn, and Swedish Red) from other latitudes (Van–Eenennaam and Medrano, 1991; Beja–Pereira et al., 2002; Barreras et al., 2001; Tsiaras et al., 2005; Caroli et al., 2004; Van–Eenennaam and Medrano, 1991; Hallén et al., 2009). Commercial breeds that are selected for higher production have a lower proportion of the B allele. This selection has drastically reduced the frequency of the alleles that are associated with milk quality. One method to overcome this problem is to redirect selection schemes for specialized dairy breeds (Heck et al., 2009). This study demonstrates that the GCC breed may be a sustainable alternative for improving milk quality in tropical climates.

The frequency of the A allele (0.39 ± 0.019) coincided with the reported range in the Bolivian Creole breeds (Lirón et al., 2002). However, the frequency was below that of the breeds from Argentina (Poli et al., 2002; Lirón et al., 2002; Martínez et al., 2003), Perú (Veli et al., 2004), Cuba (Uffo et al., 2006), Brazil (Kemenes et al., 1999), Africa, and Italy (Ceriotti et al., 2004).

The less common G, I, A1, E, and N alleles represented 18% of the total frequency, which 1 demonstrates that the Colombian Creole breeds maintain a high diversity in spite of their small population size. In the HV breed, Díaz et al., (2006) found that the G, I, A , and E alleles 1 represented 22% of the total frequency. These findings highlighted the importance of using the SSCP technique to correctly identify CSN3 alleles. Although the A and B alleles were dominant in the GCC breed, the I allele was important in the SM breed (0.13 ± 0.044), and the G allele (listed as rare in B. taurus [Caroli et al., 2009]) was abundant in CAS (0.36), HV (0.23), and VEL (0.11) breeds. While there is no clear evidence regarding the impact of the G allele on milk, Prinzenberg et al. (1999) suggests that it has a negative effect on milk coagulation (similar to that of the A allele).

The A1 allele was identified in seven of the twelve studied breeds and was most prominent in the CAS and BRAH breeds. Prinzenberg and Erhardt (1999) verified its specificity to zebuine cattle breeds. Nevertheless, Barrera et al. (2006) found no genetic proximity to the BRAH breed using the CAS breed microsatellite markers. The VEL breed (25% Red Brahman) showed very low A1 allelic frequency. In addition, the E allele frequency was low in the CQT, CAS, SM, and VEL breeds (0.016). The E allele has been associated with poor coagulation (Ikonen et al., 1997; Hallén et al., 2007) and a higher casein proportion than that of the A allele (Heck et al., 2009). The variant (N), which could not be identified using restriction enzymes or sequencing was only found in the ChS and VEL breeds. Therefore, this band pattern corresponds to an uncharacterized mutation.

The NA (4.60 ± 1.57) was noticeably higher than that reported for the majority of breeds from the Americas (Lirón et al., 2002; Postiglioni et al., 2002; Martínez et al., 2003; Uffo et al., 2006, Lara et al., 2002; Veli et al., 2004; Kemenes et al., 1999). This trend probably occurred because these studies only identified two alleles. The expected heterozygosity (He =0.65) was in accordance with the ranges found in the creole breeds from the Americas (Lirón et al., 2002; Martínez et al., 2003) and the native Portuguese breeds (Beja–Pereira et al., 2002), but was considerably higher than the estimates made for the Colombian Holstein breeds (Solarte–Portilla et al., 2009).

The high degree of polymorphism found in GCC reduces the effects of genetic drift and inbreeding (Harrison and Hastings, 1996), a process that tends to be common in small populations. This process brings about reduced biological performance, which is represented by lower survivorship, reduced reproductive performance, low growth rates, and difficulty in adapting to environmental changes (Saccheri et al., 1998). High variability in the Creole breeds from the Americas has been attributed to a series of causes, including low selection pressure, traditional management practices, introgressive hybridization with commercial breeds (Giovambattista et al., 2001; Lirón et al., 2006), demographic factors caused by European and African genes, and the breadth of geographical origin (Lirón et al., 2006).

The FST value and exact test used to analyze the degree of genetic differentiation among the studied Creole breeds showed significant differences between the GCC breeds. However, intrapopulational endogamy estimated by Fis was not significant.

In conclusion, a high degree of diversity for the CSN3 locus was found in the Colombian Creole Breeds. In addition, a high CSN3 B allele frequency (related to milk quality) was found. A significant percentage was also found for other alleles, which emphasizes the importance of conserving the Creole breeds. These breeds contain a diverse allele reservoir. Accordingly, they represent an opportunity for developing production alternatives that avoid genetic erosion.

 

Acknowledgements

Financing was made possible by the National University of Colombia–Palmira Campus Office of Research (DIPAL) (Project 20101100727). In addition, we acknowledge the Colombian Creole Cattle breeders, including Marino Valderrama and Eduaime Cárdenas (Hartón del Valle), Juan Manuel Gonzáles and Julia Arias (BON), Rafael Torrijos and Francisco Ramón (CQT), Germán Martínez (SM), Luis Fernando Cala (ChS), Arturo Cabrera (CCC), Rodrigo Salas (ROMO), José Antonio Velásquez (VEL), and Pablo Canay and Arcesio Salamanca (CAS).

 

References

1. Alexander LJ, Stewart AF, Mackinlay AG, Kapelinskaya TV, Tkach TM, Gorodetsky SI. Isolation and characterization of the bovine Kappa–Casein gene. Eur J Biochem 1988; 178:395–401.         [ Links ]

2. Asocriollo. Creole and Colombian Breed Association. National Union of Colombian Livestock Associations. 2007; [Access date: June 2011] URL: http://www.unaga.org.co/index.htm        [ Links ]

3. Barrera G, Martínez R, Pérez JE, Herazo T. Bos taurus Mitochondrial DNA identification in Zebu cattle populations. Corpoica Journal. 2006; 7:5–11.         [ Links ]

4. Barreras A, Robinson O, Monge FJ, Vizcarra O, Navarro LH. Effects of genotypes of casein and serum proteins on milk production in Holsteins. Proceeding, Western Section. A Soc of Anim Sci 2001; [Access date: June 2011] URL: http://www.asas.org/.../Abstract%2011,%202001%20WSASAS,%20        [ Links ]

5. Barroso A, Dunner S, Cañon J. Technical note: Detection of bovine kappa–casein variants A, B, C and E by means of polymerase chain reaction single strand conformation polymorphism (PCR–SSCP). J Anim Sci 1998; 76:1535–1538.         [ Links ]

6. Beja–Pereira A, Erhardt G, Matos C, Gama L, Ferrand N. Evidence for a geographical cline of casein haplotypes in Portuguese cattle breeds. Anim Genet 2002; 33:295–300.         [ Links ]

7. Braunschweig M, Hagger C, Stranzinger G, Puhan Z. Associations between casein haplotypes and milk production traits of Swiss Brown cattle. J Dairy Sci 2000; 76:1387–1395.         [ Links ]

8. Caroli A, Chessa S, Bolla P, Budelli E, Gandini G. Genetic structure of milk protein polymorphism and effects on milk production traits in local dairy cattle. J Anim Breed Genet 2004; 121:141–142.         [ Links ]

9. Caroli AM, Chessa S, Erhardt GJ. Invited review: milk protein polymorphisms in cattle: Effect on animal breeding and human nutrition. J Dairy Sci 2009; 92:5335–5352.         [ Links ]

10. Casas I, Valderrama M. Hartón del Valle Creole cattle. Informational Bulletin. Asohartón. National University of Colombia. Palmira 1998; p.54.         [ Links ]

11. Ceriotti G, Marletta D, Caroli A, Erhardt G. Milk protein loci polymorphism in taurine (Bos taurus) and zebu (Bos indicus) populations bred in hot climate. J Anim Breed Genet 2004; 121:404–415.         [ Links ]

12. Colombian Corporation for Livestock Research– Corpoica and Asocriollo. Colombian Creole Breeds; 2006.         [ Links ]

13. Díaz HA, Álvarez LA, Muñoz JE, Posso A, Sanabria HL. Genetic variability of milk proteins (k–Casein, β–Lactoglobulin and α–Lactoalbumin) in ''Hartón Del Valle'' Creole cattle. Proceedings of the 30th International Conference on Animal Genetics. Porto Seguro, Brazil. Belo Horizonte, Brazil 2006; [Access date: June 2011] URL: http://www.isag.us/Docs/2006ISAG_Proceedings.pdf        [ Links ]

14. Excoffier L, Laval G, Schneider S. Arlequin Version 3.1: An Integrated Software Package for Population Genetics Data Analysis. Arlequin User Manual. 2006.         [ Links ]

15. FAO. Global Plan of Action for Animal Genetic Resources and the Interlaken Declaration. 2007; [Access Date: June 2011] URL: http://www.fao.org/docrep/010/a1404e/a1404e00.HTM.         [ Links ]

16. Fox PF, Brodkorb A. The casein micelle: Historical aspects, current concepts and significance. Review. Int Dairy J 2008; 18:677–684.         [ Links ]

17. Guo S, Thompson E. Performing the exact test of Hardy– Weinberg proportion for multiple alleles. Biometrics 1992; 48:361–372.         [ Links ]

18. Hallén E, Allmere T, Näslund J, Andrén A, Lundén A. Effect of genetic polymorphism of milk proteins on rheology of chymosin–induced milk gels. Int Dairy J 2007; 17:791–799.         [ Links ]

19. Hallén E, Allmere T, Lunden A, Andren A. Effect of genetic polymorphism of milk proteins on rheology of acid–induced milk gels. Int Dairy J 2009; 19:399–404.         [ Links ]

20. Harrison S, Hastings A. Genetic and evolutionary consequences of a metapopulation structure. Trends Ecol Evol 1996; 11:180– 183.         [ Links ]

21. Heck JM, Schennink A, Van Valenberg HJ, Bovenhuis H, Visker MH, Van Arendonk JA, Van Hooijdonk AC. Effects of milk protein variants on the protein composition of bovine milk. J Dairy Sci 2009; 92:1192–1202.         [ Links ]

22. Hijmans RJ, Guarino L, Bussink C, Jarvis A, O'Brien R, Mathur P, Cruz M, Barrantes I, Rojas E. DIVA–GIS, User Manual; 2005.         [ Links ]

23. Hurtado GM. Physical–Chemical characterization of Hartón del Valle Creole cattle in two different herds in Cauca Valley. Directed study. National University of Colombia, Palmira Campus 1998; 121 p.        [ Links ]

24. Ikonen T, Ojala M, Syvaoja EL. Effects of composite casein and beta–lactoglobulin genotypes on renneting properties and composition of bovine milk by assuming an animal model. Agri Food Sci 1997; 6:283–294.        [ Links ]

25. Kemenes PA, De Almeida RL, De Magalhaes RA, Packer IU,Razook AG, De Figuereido LA, Silva NA, Etchegaray MAL,Coutinho LL. k–casein; β–lactoglobulin and growth hormoneallele frequencies and genetic distance in Nelore, Gyr, Guzera, Caracu, Charolais, Canchim and Santa Gertrudis Cattle. Genet Mol Biol 1999; 22:539–541.         [ Links ]

26. Lara MAC, Gama LT, Bufarah G, Sereno JRB, Celegato EML, De Abreu UP. Genetic Polymorphism at the K–Casein Locus inPantaneiro cattle. Arch Zootec 2002; 51:99–105.         [ Links ]

27. Lirón JP, Ripoli MV, De Luca JC, Peral–Garcia PGiovambattista G. Analysis of genetic diversity and populationstructure in Argentine and Bolivian Creole cattle using five locirelated to milk production. Genet Mol Biol 2002; 24:413–419.         [ Links ]

28. López M, Hernández V, Estrada L. Correlation amonghaplotypes for k–casein and milk–production characteristics inHolstein Cattle. Colombia Journal of Biotechnology 1999; 2:16– 20.         [ Links ]

29. López A, Saldarriaga OA, Arango AE, Rugeles MT, ZuluagaFN, Olivera M, Bermúdez N, Bedoya G, Ossa JE. BlancoOrejinegro (BON) Cattle: An alternative for Colombianproduction. Colombia Journal of Livestock Science 2001;14:121–128.         [ Links ]

30. Lundén A, Nilsson M, Janson L. Marked effect of β–lactoglobulin polymorphism on the ratio of casein to totalprotein in milk. J Dairy Sci 1997; 80: 2996-3005.         [ Links ]

31. MADR–Ministry of Agriculture and Rural Development. Thestate of zoogenetic resources in Colombia. ICA, CORPOICA,UN, ACP, FENAVI, FAO, ASOCRIOLLO, UNAGA,FEDEGAN, INPA 2003.         [ Links ]

32. Martínez G. Romosinuano (Romo) Creole Cattle. ColombianLivestock Institute. ICA. AGRI 1998; 24:1–11.         [ Links ]

33. Martínez G. Seminario: Census and characterization ofColombian and Creole cattle production systems. FEDEGAN,ICA, PRONATTA and ASOBON; 1999.         [ Links ]

34. Martínez G, González HF. Sanmartinero (SM) Creole cattle andtheir productive potential. AGRI 2000; 28:7–17.         [ Links ]

35. Martínez G. National action plan for conservation,improvement, and sustainable use of Colombian animal geneticresources. FAO– MADR; 2010.         [ Links ]

36. Martinez RD, Giovambattista G, Ripoli MV, De Luca JC,Dulout FN. Patagonian Argentine Creole cattle polymorphism:comparison with North–West populations of this breed. Res VetSci 2003; 74:287–290.        [ Links ]

37. Miller SA, Dykes DD, Polesky HF. A simple salting outprocedure for extracting DNA from human nucleated cells.Nucleic Acids Res 1988; 11:1215.        [ Links ]

38. Naranjo J, Posso A, Cárdenas H, Muñoz J. Detection of kappa– casein allelic variants in Hartón del Valle Cattle. Acta Agron 2007; 56:43–47.         [ Links ]

39. Pinzón E. History of Colombian cattle. (Colombia): Livestock Bank; 1991; 8:1.        [ Links ]

40. Poli MA, Holgado F, Rabasa AE. Genotype and allelic frequencies for casein k and lactoglobulin B genes in an Argentinean Creole cattle rodeo. Third Iberoamerican Symposium on the conservation of local animal genetics and rural development. School of Veterinary Science–UDELAR 2002; p.12.         [ Links ]

41. Postiglioni AG, Rincón L, Kelly S, Llambí G, Fernández M, Angelo G, Gagliardi J, Trujillo M, De Bethencourt K, Guevara A, Castellano Y, Arruga MV. Genetic biodiversity in Uruguayan Creole cattle. Molecular marker analysis. Zootechnology Archives 2002; 51:195–202.         [ Links ]

42. Prinzenberg EM, Erhardt G. A new CSN3 allele in Bos indicus cattle is characterized by MspI PCR–RFLP. Anim Genet 1999; 30:164.         [ Links ]

43. Prinzenberg EM, Krause I, Erhardt G. SCCP analysis of the bovine CSN3 locus discriminates six alleles corresponding to known protein variants (A, B, C, E, F, G) and three new DNA polymorphism (H, I, A ). Anim Biotechnol 1999; 10:49–62.         [ Links ]

44. Saccheri I, Kuussaari M, Kankare M, Vikman P, Fortelius 1         [ Links ]

45. W, Hanski I. Inbreeding and extinction in a butterfly metapopulation. Nature 1998; 392:491–494.         [ Links ]

46. Solarte–Portilla CE, Rosero CY, Eraso JM, Zambrano GL, Cárdenas H, Burgos WO. Allelic frequencies of the Kappa Casein gene in the Holstein breed of the Nariño, Colombia high tropics. Livestock Res Rural Dev 2009; 21:1–7.         [ Links ]

47. Tsiaras AM, Bargouli GG, Banos G, Boscos CM. Effect of Kappa–Casein and Beta–Lactoglobulin loci on milk production traits and reproductive performance of Holstein cows. J Dairy Sci 2005; 88:327–334.         [ Links ]

48. Uffo O, Martín–Burriel I, Martínez S, Ronda R, Osta R, Rodellar C, Zaragoza P. Genetic characterization of six milk proteins in three Cuban cow breeds. AGRI 2006; 39:15–24.         [ Links ]

49. Van–Eenennaan A, Medrano JF. Differences in allelic protein expression in the milk of heterozygous k–casein cows. J Dairy Sci 1991; 74:1491–1496.         [ Links ]

50. Veli E, Rivas E, Rivas V, Verastegui M, Pastor S. Evaluation of variability of kappa–casein genes in Creole cattle populations in Ticllos and Huaschao, Ancash region. INIA 2004; [Access Date: June 10, 2008]. URL:http://www.inia.gob.pe/genetica/zoogeneticos/articulo%20V%20Congreso.pdf.         [ Links ]

51. Vivas NJ, Vinasco AM, Álvarez LA, Posso AM, Muñoz JE. Detection of allelic variants of kappa–casein using the PCR–SSCP technique on Holstein cattle in the Cauca Valley. Colombia Journal of Livestock Science 2008; 21:155–156.         [ Links ]

52. Wedholm A, Larsen LB, Lindmark–Ma Nsson H, Karlsson AH, Andrén A. Effect of Protein Composition on the Cheese–Making Properties of Milk from Individual Dairy Cows. J Dairy Sci 2006; 89:3296–3305.        [ Links ]

53. Weir BS, Cockerham CC. Estimating F–statistics for the analysis of populations structure. Evolution 1984; 38:1358–1370.         [ Links ]

54.Wilkins JV, Martínez F, and Rojas F. Creole Cattle. Dialogue XXXVI. Conservation and improvement of Creole cattle. IICA– PROCISUR; 1993.        [ Links ]

 

Notas

¤ To cite this article: Rosero JA, Álvarez LA, Muñoz JE, Durán CV, Rodas AG. Allelic frequency of the Kappa–Casein gene in Colombianbreeds. Rev Colomb Cienc Pecu 2012; 25:173–182.

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