Blood group and protein polymorphism in the Finnish native cattle populations Juha Kantanen and Matti Ojala Kantanen, J. & Ojala, M. 1994. Blood group and protein polymorphism in the Finnish native cattle populations. Agricultural Science in Finland 3: 169-176. (Department of Animal Science, P.O. Box 28, FIN-00014 University of Helsinki, Finland. Present address: Juha Kantanen, Agricultural Research Centre of Finland, Institute of Animal Production, FIN-31600 Jokioinen,Finland.) Nine blood group loci and five polymorphic protein loci were investigated in the native East-, North- and West-Finnish cattle populations. The studied East-, North- and West-Finnish cattle populations comprised 74, 55 and 121 individuals, respec- tively. According to the average degree of heterozygosity, East-Finnish cattle had the highest genetic variation and North-Finnish cattle the lowest. Within the loci investigated, the East- and North-Finnish cattle populations, which are threatened by extinction, did not lack genetic diversity. The genetic distances between West- and North-Finnish cattle calculated by the Nei’s (1972) standard method ranged from 0.019 to 0.052 in three partly different locus groups and between East- and North-Finnish cattle from 0.034 to 0.046. The distances between East- and West- Finnish cattle were 0.030 in all cases. According to these results. East-, West- and North-Finnish cattle could be regarded as three differentnative breeds. Key words: local cattle breed, heterozygosity, genetic distance Introduction The Finnish native cattle populations were named on the basis of their geographic breeding areas. East-Finnish cattle (EFc) are usually brown-sid- ed. North-Finnish cattle (NFc) are white with brown or black spots and West-Finnish cattle (WFc) are brown. Indigenous cattle are almost exclusively polled. At the beginning of this cen- tury there were three native cattle herd book so- cieties in Finland. Since the fusion of the socie- ties in 1947, the populations were regarded as one population and breed, Finncattle. The number of Finnish native cows has de- clined drastically. Still in 1970 there were 307 600 Finncattle, but in 1980 their number had dropped to 42 800 and in 1991 down to only 7 900, which is 1.6 % of the total dairy cattle population in Finland. West-Finnish cattle are the largest group - 7 700 out of the total 7 900. The EFc and NFc populations, with 70 and 60 pure- bred cows left, respectively, can be regarded as threatened by extinction. The indigenous cattle have been replaced by Finnish Ayrshire (FAy) and Finnish Friesian (FFr). The polymorphic character of genetically de- termined red cell antigens and blood protein sys- tems, and their simple mode of inheritance make them valuable for studying the origin, structure and relationship of breeds. Blood polymorphism can also be utilized as a measure of genetic vari- ation. This study was conducted to determine the alleles or the phenogroups of the blood groups and blood protein systems in the Finnish native 169 Agricultural Science in Finland 3 (1994) https://www.c-info.fi/en/info/?token=LhMAOpkbjiTQUyh1.-nWTnZWxjdKnJx_zIuerCQ.ArL741mr_Ld3yjkCw4TB68VAlh_CZ_1i9MZE9EQc8denNGScm2h9H8fE7dV8DSOFypS7NdP2OBEAuN_WPTHUHk44INZtaTGVE1XFBw1shhW6wdgbYORGWTtZKlFRCVOUVdUYXS7fAG2qAxy1OeuOjzG2KGM2e0cB63gkp_i4N_4bYB8S6ueuapoxHpN1oWvbvR5_lLLLOhGwhY3jlxQUl5leljav3e0F0qYeTavbyfKbbn1qbkkuZpD8iwQVb_xrTn224A_PNvibPKQbqYsXNBGIUKg cattle populations, to investigate the average de- gree of heterozygosity and to calculate the genet- ic distances between East-, North- and West-Finn- ish cattle. Material and methods Blood samples were collected from a total of 250 native cattle (Table 1). The animals belonged to 43 herds located in different parts of Finland. For comparison, 50 Finnish Ayrshire (FAy) and 50 Finnish Friesian (FFr) animals were tested as well. The number of sires of the sampled animals were estimated to be between 22 and 37 in EFc, be- tween 16 and 18 in NFc and 50 in WFc. Of these, 7 EFc, 10 NFc and 48 WFc males had been used in artificial insemination (AI) and the rest of the bulls only in their birth herds. East-, North- and West-Finnish cattle AI bulls had on an average 3.7, 4.3 and 2.4 progenies, respectively. Both FAy and FFr animals had been sired by 50 AI bulls. The sires of all the studied populations were not included in the present material. Blood samples were typed for nine red cell antigen and five polymorphic protein systems (Table 2). The variants in the five blood plasma protein systems were determined by horizontal and two-dimensional polyacrylamide gel electro- phoresis. The technique employed in blood group tests and electrophoresis has been described by Braend (1959), Gahne et al. (1977) and Juneja and Gahne (1980). Table 1. Number of tested animals. POPULATIONS EFc NFc WFc FAy FFr Female 52 37 117 47 45 Male 22 18 4 3 522 18 4 3 5 Total 74 55 121 50 50 Populations: EFc = East-Finnish cattle NFc = North-Finnish cattle WFc = West-Finnish cattle FAy = Finnish Ayrshire FFr = Finnish Friesian Table 2. Symbols for the blood group and protein systems tested and, in parentheses, the factors and variants within each system. BLOOD GROUP SYSTEMS A (A.H) B(B,G,K,I 1 1,2,I2 ,0 |,0 3 ,0 11 ,P,Q,T 1,T 2,Y 1,Y2>A’2,8’, B”,D’ >E’ 2>E’ 3> GM’,J’K’,O’,P’,Y’,F’ | ) F (F,V) J(J) L (L) M (M) Z(Z) R’(R’,S’) T’(T’) BLOOD PROTEIN SYSTEMS Pa (Post-albumin F and S) Tf (Transferrin A, D and E) Ptf 1 (Post-transferrin 1 A and B) Ptf 2 (Post-transferrin 2 F and S) Pi-2 inhibitor F, I and S) The term allele is used in this report instead of a factor or a variant, except in the B blood group system where the terms B factor and B pheno- group are employed. The presence of a factor, e.g. G, means that the animal reacts positively with the reagent for G. A phenogroup stands for a combination of factors which is inherited as a block, e.g. BGOr The F and R’ blood groups and the investi- gated protein systems form so-called closed sys- tems, where an individual’s genotype can be di- rectly determined in a laboratory test. Other blood group systems are open systems where an indi- vidual’s genotype may be deduced from its par- ents’ or progeny’s phenotypes. Thus, in the closed systems the alleles are codominant, whereas dom- inance is present in the open systems. Depending on the nature of the system, the allele frequen- cies were estimated by different counting meth- ods (Falconer 1981). In the B system the fre- quencies of the phenogroups were estimated with Braend’s (1963) square root method. East- and West-Finnish cattle were divided into two groups according to the geographic location 170 Agricultural Science in Finland 3 (1994) of the herd. There were 30 EFc and 40 WFc animals in South and Central Finland (region 1) and 44 EFc and 81 WFc animals in North Fin- land (region 2). In the EFc population there were four genetically isolated herds, which had 4,6, 14 and 15 animals, a total of 39 animals. In- breeding has obviously occurred in these herds, each of which formed a closed mating unit for over five cattle generations. The existence of ge- netic equilibrium and the regional and isolational effects were studied using of the codominant sys- tems. The average degree of heterozygosity of the closed systems was estimated according to Ferguson (1980). There were no half- or fullsibs or parent-progeny pairs in the calculations. The X 2 independence test was employed to calculate the statistical significance for the differences be- tween the populations in the allele frequencies of the closed systems and for investigating the ge- netic equilibrium as well as the regional and iso- lational effects. Genetic distances among EFc, NFc and WFc and among WFc, FAy and FFr were estimated according to NeTs (1972) standard method. Three locus groups were used in the calculation. The first locus group included the F and R’ blood groups and the Pa, Tf, Ptf 1, Ptf 2 and Pi-2 pro- tein systems, the second group the A, B, J, L, M, Z and T’ blood groups and the third group the closed and the open systems together. Results Allele and genotype frequencies Statistically significant differences (p<0.001) were found between EFc, NFc and WFc in the allele frequencies of the F, Pa, Tf, Ptf 2 and Pi-2 sys- tems (Table 3). The PaF allele of postalbumin in the EFc population was more than twice or three times as numerous as in WFc or in NFc, respec- tively. The Tf° was the most frequent allele of the transferrin system in NFc and WFc, but not in EFc, in which the Tf A allele had the highest frequency. The presence of the TfE and the Ptf 2 s Table 3. Allele frequencies in East-, North- and West- Finnish cattle. CLOSED SYSTEMS Allele EFc NFc WFc Level of significance 1215574N 0.673 1 0.327 I 0.763 1 0.237 J FF 0.9661 Fv 0.034 J *** 0.004 1 0.996 I1.000 N.S. 0.107 1 0.893 I 0.073 0.927 *** 0.227 ' 0.555 0.218. *** 0.455 1 0.545 I 0.518 0.482 N.S. 0.979 1 0.021 J 0.746 0.254 *** 0.045 3 0.033 [ 0.922 J 0.009 I 0.991 J � ** OPEN SYSTEMS 0.575] 0.361 [ 0.064 J 0.603 ' 0.249 0.148, 0.384 1 0.616 j 0.086 1 0.914 ] 0.278 1 0.722 I 0.2141 0.786 I LL 0.186] L’ 0.8141 0.0561 0.944 J 0.0641 0.936 ) M M - ] M m 1.0001 0.109 0.891 0.047 I 0.953 I T’r 0.041 ] T’ 1 ' 0.959 I Zz 0.312] Z* 0.688 1 0.213 0.787 0,2141 0.786] N = number of animals EFc = East-Finnish cattle NFc = North-Finnish cattle WFc = West-Finnish cattle Level of statistical significance between the allele fre- quencies in EFc, NFc and WFc: ***=p IT 0.050 0.012 0.006 0.950 0.988 1.0 0.994 0.067 0.375 0.138 0.093 0.933 0.625 0.862 0.907 0.166 0.761 0.438 0.352 0.717 0.205 0.537 0.580 0.117 0.034 0.025 0.068 Ptf l A Ptf l B Ptf 2F Ptf 2 s Pi-2 F Pi-2' 0.633 0.466 0.425 0.469 0.367 0.534 0.575 0.531 0.983 0.966 0.962 0.988 0.017 0.034 0.038 0.012 0.133 0.068 0.050 0.043 0.167 0.102 0.037 0.031 Pi-2s 0.700 0.830 0,913 0,926 N = number of animals EFc = East-Finnish cattle WFc = West-Finnish cattle Region 1 = South and Central Finland Region 2 = North Finland Table 6. Effect of isolation on the allele frequencies of the closed systems in East-Finnish cattle. Isol. herdsAllele Al-herds N 39 35 FF 0.936 0.064 1.0 Fv R-R R.S- Pa F Pa s TfA Tfn X ft 0.057 1.0 0.943 0.244 0.257 0.756 0,743 0.474 0.572 0.462 0.371 0.064 0.057 Ptf 1A Ptf 1B Ptf 2F Ptf 2 s Pi-2 F Pi-2' 0.603 0.457 0.397 0.543 1.0 0.943 0.057 0.115 0.071 0.039 0.229 Pi-2S 0.846 0.700 N = number of animals Isol. herds = isolated herds having used bulls of their own. Al-herds = herds having used Al-bulls the exception of the F blood group there were less heterozygotic animals in the isolated herds than in other herds in all loci. Five B pheno- groups -1,, A’ 2E’ 3 G’, GO,Y2 , D’GT and V - were frequent in the genetically isolated EFc herds. Degree of heterozygosity and genetic distances The average degree of heterozygosity (H) was highest in East-Finnish cattle and lowest in North- Finnish cattle (Table 7). NFc was monomorphic in the R’ blood group and this locus was least diverse also in WFc. NFc had little variation in the Pi-2 although this locus has three alleles. With respect to the level of genetic variation in the loci of the closed systems, WFc and NFc were more similar than WFc and EFc. East- and North- Finnish cattle differed the most. West- and North-Finnish cattle were genetical- ly most similar on the basis of the genetic dis- 173 Agricultural Science in Finland 3 (1994) Table 7. Degree of heterozygosity (%) in the loci of the closed systems. Locus EFc NFc WFc N 21 15 50 F 13.19 27.82 0.00 46.08 1.9813.19R’ Pa Tf 36.27 18.00 21.12 58.04 53.13 55.04 Ptf 1 Ptf 2 Pi-2 48.99 49.78 47.12 4.68 27.82 3.92 20.146.3844.93 26.13 27.91H 31.33 N = number of animals EFc = East-Finnish cattle NFc = North-Finnish cattle WFc = West-Finnish cattle H = average degree of heterozygosity tance (D) of the closed systems (locus group 1) (D=0.019) and East- and North-Finnish cattle the most different (D=0.046) (Table 8). The genetic distance among Finnish Ayrshire and Finnish Frie- sian (D=0.020) was about equal to or smaller than the distances between the native populations. With locus groups 2 and 3, NFc and WFc were genetically more different (D= 0.052 or 0.035) than EFc and WFc. The genetic distances be- tween EFc and WFc (D=0.030) were the same in all cases. Discussion The present results of the allele frequencies of WFc were almost the same as the results of Vasenius (1965), who studied allele frequencies for the transferrin locus in 313 Finncattle, and those of Maijala and Findström (1966), who studied blood group alleles in 540 West-Finnish bulls. No fixation or loss of alleles in available loci in the West-Finnish cattle population was found when comparing the present study to the investigations made in the 1960’5. The B pheno- group Y2D’G’, typical of WFc in this study, had not been observed in other native cattle popula- tions in the Nordic countries (Maijala and Lind- ström 1966). Baker and Manwell (1980) re- ported that the Tf6 is characteristic of breeds which had been developed in harsh environments. The results in this study agree with that hypothesis, since the Tf6 was found in a high frequency in North-Finnish cattle, a population ofFinnish Lap- land. EFc and NFc were not in genetic equilibrium in all loci. The small population sizes, genetic drift, isolation and an obvious inbreeding in the Table 8. Genetic distances between the populations on the basis of three locus groups. LG 1 LG 2 LG 3 FAy FFr EFc NFc EFc NFc EFc NFc EFc WFc FAy 0.046 0.034 0.040 0.034 0.025 0.030 0.019 0.030 0.052 0.030 0.035 0.020 LG 1 = locus group 1 LG 2 = locus group 2 LG 3 = locus group 3 FAy = Finnish Ayrshire FFr = Finnish Friesian EFc = East-Finnish cattle NFc = North-Finnish cattle WFc = West-Finnish cattle 174 Agricultural Science in Finland 3 (1994) isolated EFc herds may be the major causes of a slight genetic disequilibrium. YamADA (1981) has suggested that an animal population could be subdivided into several lines for the preservation of genetic resources. This situ- ation has partly developed in East-Finnish cattle. Line subdivision was less pronounced in NFc than in EFc or WFc. There were no full- or halfsibs among the WFc data used for the heterozygosity calculation. Some of the WFc animals were, how- ever, related through their maternal or paternal grandsires. Only a few WFc bulls have so far been chosen to be employed in Al on a large scale. The magnitude of the heterozygosity observed in this study is important because it implies thatEast- and North-Finnish cattle may not, in fact, suffer from a lack of genetic variation. Locus group 1 could be regarded to give the most reliable data for the calculation of the ge- netic distances, since the allele frequencies of the open systems were estimates only of the exact frequencies. West-Finnish cattle were genetically closer to Finnish Friesian than to Finnish Ayr- shire although West-Finnish cattle and Ayrshire are both classified as belonging to the North- European cattle breedgroup according to Baker and Manwell (1980). A share of WFc cows were crossbred with Ayrshire already in the 1950’sand 1960’5, and with Friesian in the 1970’s to up- grade the native cattle by these two breeds. Be- cause of this, more genetic influence of WFc was left in FFr than in FAy. The genetic distances between the Finnish native cattle populations were nearly as great as, or greater than those between Finnish Ayrshire and Finnish Friesian. In a previ- ous study among seven Spanish native cattle breeds, the genetic distances calculated by the Nei’s (1972) standard method ranged from 0.007 to 0.180 (Gonzales et al. 1987). The magnitude of the genetic distances among the Finnish native cattle populations (0.019 - 0.046)relative to those between FAy and FFr and between Spanish na- tive cattle breeds (Gonzales et al. 1987) sug- gests that East-, North- and West-Finnish cattle are three different breeds and not only three dif- ferent colour types of the same breed. Several reasons may have caused the genetic differentiationof the Finnish native cattle breeds. Before the fusion of the three herd book societies in 1947, East-, North- and West-Finnish cattle were at least partly geographically isolated. Also the founding animals with which the breeding work of each native breed was begun may have been genetically different. The present East- and North-Finnish cattle breeds are only samples of the populations of past times. Genetic drift is obviously one of the major causes for differenti- ation. East-Finnish cattle have partly been isolat- ed, and inbreeding in the isolated herds may have increased genetic differentiation in the EFc as opposed to the other native breeds. Acknowledgements. The authors are grateful to Tirri Nii- ni, Raili Huttunen, Kaarina Pirhonen and Ilona Salminen (The Blood Group Laboratory of the Finnish Animal Breed- ing Association) for the collaboration in testing blood samples. References Baker, C. M.A. & Manwell, C. 1980. Chemical classifi- cation of cattle. 1. Breed groups. Animal Blood Groups and biochemical Genetics 11: 127-150. Braend, M. 1959.Blood groups of cattle in Norway. 144p. Skandinavisk Bladforlag, Oslo. - 1963. Estimation of gene frequencies in the B-system of cattle. Immunogen. Letter 3: 43^t8. Falconer, D.S. 1981. Introduction to quantitative genet- ics. 340 p. Longman Inc., New York. Ferguson, A. 1980. Biochemical systematics and evolu- tion. 194 p. Blakkie and Son, Glasgow. Gahne, 8., Juneja, R.K. & Grolmus, J. 1977. Horizontal polyacrylamide gradient gel electrophoresis for the si- multaneous phenotyping of transferrin, post-transfer- rin, albumin and post-albumin in the blood plasma of cattle. Animal Blood Groups and biochemical Genet- ics 8: 127-137. Gonzalez, P., Tunon, M.J. & Vallejo, M. 1987. Genetic relationship between seven Spanish native breeds of cattle. Animal Genetics 18: 249-256. Juneja, R.K. & Gahne, B. 1980. Two-dimensional gel electrophoresis of cattle plasma proteins. Genetic poly- morphism of an al-protease inhibitor. Animal Blood Groups and biochemical Genetics 11: 215- 228. 175 Agricultural Science in Finland 3 (1994) Maijala, K. & Lindström, G. 1966. Frequencies of blood group genes and factors in the Finnish cattle breeds with special regard to breed comparisons. Annales Ag- riculturae Fenniae 5: 76-93. Nei, M. 1972. Genetic distance between populations. Amer- ican Naturalist 106: 283-292. Vasenius, L. 1965. Transferrin polymorphism in Finnish Ayrshire cattle. Annales Academiae Scientiarum Fen- nicae Series A. IV. Biologica 98: 1-58. Yamada, Y. 1981. The importance of mating systems in the conservation ofanimal genetic resources. FAO An- imal Production and Health Paper 24: 268-278. Manuscript received October 1993 SELOSTUS Veriryhmä- ja veren valkuaisainepolymorfismi Suomen alkuperäisissä nautaroduissa Juha Kantanen ja Matti Ojala Helsingin yliopisto Suomalaisista alkuperäisistä nautapopulaatioista itä-ja poh- joissuomenkarja (ISK jaPSK) ovatkriittisesti uhanalaisia nautarotuja, sillä näissä populaatioissa on vain 70 ja 60 puhdasrotuista lisääntyvää naarasta. Länsisuomenkarjan (LSK) lehmiä on noin 8 000. Tutkimusaineisto koostui 74 ISK-, 121 LSK-ja 55 PSK-eläimen verinäytteistä. Suori- tettujen vertailujen vuoksi analysoitiin 50 ayrshire- ja 50 friisiläisrodun eläintä. Alkuperäisrotujen geneettistä muun- telua ja geneettisiä etäisyyksiä tutkittiin yhdeksän veri- ryhmä- ja viiden veren valkuaisainelokuksen perusteella. Veriryhmät määritettiin kansainvälistä hemolyyttistä tes- tiä käyttäen. Naudan seerumiproteiinit tutkittiin yksi- tai kaksisuuntaisella polyakryyliamidi-elektroforeesilla. Ro- tujen geneettinen muuntelu arvioitiin seitsemän lokuksen heterotsygotia-asteiden keskiarvon perusteella. Geneettis- ten etäisyyksien laskennassa käytettiin kolmea lokusryh- mää. Geneettisesti muuntelevin rotu oli ISK. Vähiten muun- telua oli PSK:lla, ISK ja PSK ovat pienestä populaatio- koostaan huolimatta geneettisesti muuntelevia populaa- tioita, kun verrataan näiden heterotsygotia-asteita LSK:n heterotsygotia-asteeseen. Suomalaisten alkuperäisrotujen geneettiset etäisyydet olivat lähes yhtä suuret tai suurem- mat kuin ayrshiren ja friisiläisen välinen geneettinen etäi- syys. Veriryhmä- ja valkuaisainelokusten polymorfian pe- rusteella voitiin todeta, että itä-, länsi- ja pohjoissuomen- karja ovatkolme erillistä nautarotua. 176 Agricultural Science in Finland 3 (1994)