Relatedness between Polymorphism Growth Hormone Gene and Growth Traits of Limousin Cross Madura Cattle - repository civitas UGM

  th Proceedings of the 15 AAAP Animal Science Congress

  26-30 November 2012, Thammasat University, Rangsit Campus, Thailand Code Title Page

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  and Growth Traits of Limousin Cross Madura Cattle T. Hartatik, S. D. Volkandari and Sumadi (Indonesia)

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  th Proceedings of the 15 AAAP Animal Science Congress

  26-30 November 2012, Thammasat University, Rangsit Campus, Thailand

Relatedness between Polymorphism Growth Hormone Gene

and Growth Traits of Limousin Cross Madura Cattle

  

T. Hartatik, S. D. Volkandari and Sumadi

  Faculty of Animal Science, Universitas Gadjah Mada, Bulaksumur, Yogyakarta, Indonesia The research was conducted to know the genotype of Limousin cross Madura cattle (Limura) and investigate the effect of polymorphism growth hormone (GH) gene for growth traits. The growth traits were examined based on quantitative data which contain birth weight and body weight at day 60, average daily gain (ADG), body length, heart girth, withers height and hip height. The sample for the experiment consist of 35 Limura calves and 10 Madura calves. Analysis of polymorphism GH gene were using Polymerase Chain Reaction-Restriction Fragment Lenght Polymorphism (PCR-RFLP) method. The PCR product of GH gene in this research (211 bp) was digested by AluI enzyme. The relatedness between genotype and growth traits were analyzed using one way ANOVA, where three genotype groups (LL_Limura, LV_Limura, and LL_Madura) as factors. The results indicated that the genotype of Madura cattle were 100% LL and the genotype of Limura cattle were LL (82.86%) and LV (17.14%). There are no significant differences of birth weight and pre-weaning body weight in three groups of genotype. However, both genotype LL and LV of Limura cattle had greater ADG, body length at birth and pre-weaning hip height gain than those of LL_Madura genotypes (p<0.05). In conclusion, the polymorphism GH gene does not affect the growth traits, but the differences of two breeds (Limura dan Madura cattle) seem influence the growth trait of the cattle.

  Key Words: Genotype, GH gene, Growth traits, Limousin cross Madura cattle (Limura)

  

INTRODUCTION

  Indonesia has very high biodiversity (mega biodiversity), one of which is genetic resources of Indonesian cattle such as local and crossbreed cattle. The most popular crossbreed cattle called by native farmer as Madrasin, and the scientist called it as Limura. Limura cattle derived from crossing between Limousin bull and Madura cow by artificial insemination which developed well in Madura Island. The growth performance of Limura cattle was greater than that of Madura cattle. Growth hormone (GH) plays a central role in the regulation of growth and metabolism in animals. Growth Hormone is a polypeptide hormone with 191 amino acid sequence which is located on bovine chromosome 19q26qter position (Hediger et al., 1990) with 5 exons and 4 introns. The fifth exon in the GH gene located at position 127 amino acid polymorphisms have been found with two alleles, namely L and V corresponding to valine and Leucine variant polypeptide at GH locus. Previous researchers reported that the genotype on GH gene associated with birth weight in dairy cows (Biswas et al., 2003) and adult weight in beef cattle (Zwierzchowski et al., 2001). The aim of this study was to analyze

  th Proceedings of the 15 AAAP Animal Science Congress

  26-30 November 2012, Thammasat University, Rangsit Campus, Thailand

  polymorphism GH gene in Limura and Madura cattle as a control. This molecular approach intends to be an introductory step for genetic characterization of these indigenous cattle (Madura) and the crossbreed (Limura). The relationship between polimorphisme GH gene and growth traits is very important to know the potential of local cattle and the offspring of crossbreed cattle.

  

MATERIALS AND METHODS

Sample and measurement of growth traits

  The sample of the research were consist of 35 Limousin cross Madura calves and 10 Madura calves. Body weight and body size was measured at the birth ( day 1) and 60 days of age (day 60). The body size was measured using a tape FH and ruler FHK brand. The body size data including: body length, heart girth, withers height and hip height. The body length (absolut) is the straight distance that measured between Tuberculum lateralis to Tuberculum ischiadicum. Heart girth is the length of the circular (the circumference) as measured on chest that right in the back of withers on the rib (costae) 3-4. Withers height is straight distance from the plane to the highest point in withers. Hip height was obtained by measuring the distance from the plane until the highest point of the hip. Data of body measurement was taken with parallelogram position of the livestock.

  Detection of the polymorphism growth hormone gene

  DNA sample was extracted from blood sample using SDS/ProteinaseK modified methode from Sambrook et al (1989). A 211 bp fragment of GH was amplified by polymerase chain reaction (PCR) which refer to Reis etal

  .(2001). The PCR reaction contains 1 μl DNA sample, 1 μl each primer (10 pmol), 10 μl PCR KIT (Fastart, Roche) and 7 μl aquabidest in total solution 20 μl. Amplification in vitro was conducted for 35 cycles using GeneAmp® PCR System 9700 (Applied

  Biosystems™). The PCR product (5μl) was digested with 1 units AluI enzyme (Fermentas) in a total volume 20 μl and incubated at 37ºCfor 3 hours. The polymorphism GH gene was analyzed by separating restriction product in vertical electrophoresis (BioRad) with 12% polyacrilamide gel (PAGE), 1X TBE buffer and running at 50 Voltage for 3 hour 30 minutes.

  Data analysis

  Genotype frequency represents the ratio of a genotype to total population (Falconer and Mackay, 1996). Analysis of the quantitative data (body weight and body size) were analyzed descriptively by calculating the means and standard deviation. The relatedness between genotype with growth traits were analyzed using one way ANOVA, where three genotype groups (LL_Limura, LV_ Limura and LL_Madura) as a factor.

  th Proceedings of the 15 AAAP Animal Science Congress

  26-30 November 2012, Thammasat University, Rangsit Campus, Thailand

RESULTS AND DISCUSSION

  Based on PCR-RFLP methods, we characterized the associations between genetic polymorphisms in GH gene and growth traits of Limura and Madura cattle. The result of genotyping GH gene of Limura cattle were 82.86% LL and 17.14% LV, whereas Madura cattle were 100% LL. The relationship between genotype and growth traits can be seen in Table 1-3. In this study, we measured and evaluated for ADG, body weight, body lenght, heart girth, withers height, and hip height at day 1 and day 60 of calves.

  Table 1 Body weight (kg) and average daily gain (kg/day) of Limura and Madura cattle Limura (LL) (n=29) Limura (LV) (n=6) Madura (LL) Parameter _{a a a} (n=10) Birth weight (day 1) 22.73 ± 4.59 21.96 ± 5.61 20.55 ± 3.43 _{a a a} Body weight day 60 55.39 ± 12.96 55.79 ± 18.65 44.39 ± 5.78 _{a a b} Average daily gain 0.54 ± 0.16 0.56 ± 0.24 0.39 ± 0.09 _a,b

  Superscript at the same row indicate significantly different (p<0,05)

  The results indicated that both genotype LL and LV of Limura cattle had greater ADG, body length at day 1 and hip height gain than those with genotypes LL of Madura (p<0.05). The two genotype within Limura breed does not show the significant different in all growth traits. Average calf birth weight of Madura cattle and crossbreed was greater than that of Bali cattle ( 16 kg for females and 17.5 kg for males). The pre-weaning ADG for Bali cattle was 0.4 kg/day (Julianto et al., 2010). Thus, Limura cattle had ADG higher than that of Bali cattle and Madura cattle. Its seem that Madura and Bali cattle had the same pre-weaning ADG.

  Table 2 Body measurement of Limura and Madura cattle at day 1 and day 60 (cm) Parameter Limura (LL) (n=29) Limura (LV) (n=6) Madura (LL) (n=10) Day 1: _{a a b}

Body length 55.56 ± 4.74 54.43 ± 3.03 49.04 ± 2.03

_{a a a}

Heart girth 67.08 ± 5.29 66.85 ± 3.31 64.85 ± 2.76

_{a a a}

Withers height 65.75 ± 5.09 66.51 ± 4.81 66.42 ± 2.96

_{a a a}

Hip height 69.36 ± 5.19 69.53 ± 4.83 69.61 ± 4.34

  Day 60: _{a a a}

Body length 72.95 ± 5.50 73.34 ± 5.66 68.28 ± 7.88

_{a a a}

Heart girth 87.27 ± 6.95 87.47 ± 7.49 81.74 ± 4.28

_{a a a}

Withers height 79.10 ± 4.90 79.52 ± 5.75 79.57 ± 4.38

_{a a a}

Hip height 84.83 ± 6.43 84.21 ± 7.66 80.14 ± 4.50

_a,b Superscript at the same row indicate significantly different (p<0,05)

  Proceedings of the 15 ^th AAAP Animal Science Congress

  In conclusion, the polymorphism GH gene does not affect the growth traits, but the differences of two breeds (Limura dan Madura cattle) seem influence the growth trait of the cattle.

  Hip height gain 0.26 ± 0.08

  a

  0.24 ± 0.12

  a

  0.17 ± 0.06

  b Superscript ^a,b at the same row indicate significantly different (p<0,05)

  

REFERENCES

Biswas T.K., T.K. Bhartacharya, A.D. Narayan, S. Badola, P. Kumar and A. Sharma. 2003.

  0.22 ± 0.05

  Growth hormone gene polymorphism and its effect on birth weight in cattle and buffalo. Asian-Aust J. Anim. Sci. 16 (4): 494-497. Falconer, D. S and Mackay, T. F. C. 1996. Quantitative Genetics. 4th ed. Longman Group Ltd., Essex, UK. Hediger, R., S.E. Johnson, W. Barendse, R.D. Drinkwater, S.S. Moore and J. Hetzel. 1995.

  Assignment of the growth hormone gene locus to 19q26-qter in cattle and to 11q25-qter in sheep by in situ hybridization. Genomics Volume 8 (1): 171 (Abstr.) Julianto, T.B., T. Panjaitan, J. Fordyce, and D. Poppi. 2010. Breeding Bos javanicus d’Alton cattle in eastern Indonesia: Cattle growth. In: Proceeding 5

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  International Seminar on Tropical Animal Production, Yogyakarta, Indonesia. pp. 474-477

  Reis, C., D. Navas., M. Pereira and A. Cravador. 2001. Growth Hormone AluI Polymorphism Analysis in Eight Portuguese Bovine Breeds. Arch. Zootec. 50 : 41-48. Sambrook J., E. F. Fritsch., and Maniatis T. 1989. Molecular Cloning, A Laboratoty Manual.

  ColdSpringHarbour Laboratory Press: Cold Spring Harbour, USA. Zwierchowski, L., J. Oprzadek, E. Dymnicki and P. Dzierzbicki. 2001. An Association of growth hormone, k- casein, β-lactoglobulin, leptin and Pit-I loci polymorphism with growth rate and carcass traits in beef cattle. Animal Science Papers and Reports 19:65-

  a

  a

  26-30 November 2012, Thammasat University, Rangsit Campus, Thailand Table 3 Body measurement gain of Limura and Madura (cm/day)

  Heart girth gain 0.34 ± 0.08

  Parameter Limura (LL) (n=29) Limura (LV) (n=6) Madura (LL) (n=10) Body length gain 0.29 ± 0.09

  a

  0.31 ± 0.09

  a

  0.32 ± 0.13

  a

  a

  0.22 ± 0.06

  0.34 ± 0.08

  a

  0.28 ± 0.05

  a

  Withers height gain 0.22 ± 0.05

  a

  78.