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Сельскохозяйственная биология, 2011, № 2, с. 51-56.

УДК 636.2:575.17:576.08

POOL OF ERYTHROCYTIC ANTIGENS AND CHROMOSOMAL INSTABILITY IN YAKUT CATTLE

E.V. Kamaldinov, O.S. Korotkevich, V.L. Petukhov

The authors sum the data on the frequency of erythrocytic antigens and chromosomal instability in aboriginal Yakut, the Black-and-White cattle and Yakut cattle introduced to West Siberia. As a result of long keeping in other, than in Yakutia, conditions of environment the introduced cattle have specific complex of antigens with other frequency ratio. In addition, these animals have higher degree of chromosomal instability.

Keywords: Yakut cattle, erythrocyte antigenes, chromosomal instability, genofund.

 

Domestication of aboriginal cattle breeds has led to fixation of unique and economically valuable features, as well as in maintaining the constant level of mutations. The development of animal husbandry provides accelerated selection, extended areal of best breeds and endangered state of many local breeds. Unidirectional increase of livestock population by interbreeding can impoverish the national gene pool and cause loss of valuable qualities of local and plant breeds of farm animals (1, 2). Irreversible loss of such breeds contributes to disappearance of unique allelic combinations that won’t be restored even by means of genetic and cell engineering (3).
Yakut cattle (Bos taurus turano-mongolicus) has a number of valuable properties formed over thousand years in extremely low temperatures of Siberia. These are: adaptability, high vitality, rapid growth and development in early postnatal period, increased fat content in milk, low demands to feed quality, valuable properties of milk and meat, good thermoregulation, rapid replacement of fetal hemoglobin to adult hemoglobin forms (4) and tuberculosis immunity (5). In winter, hair cover of the body gets thicker by 6 times compared with summer.
Erythrocyte antigens are used as genetic markers in many widespread methods allowing to assess the degree of phylogenetic similarity or difference between populations (6), the degree of animals’ heterozygosity, the consolidation of hereditary qualities of breeds, lines and types (7, 8). Blood groups of cattle are highly polymorphic. Today, it has been determined more than 80 antigenic factors (9).
Chromosomal instability reflects spontaneous mutation process in populations and it presents in almost all individuals (10-12). The major reasons of chromosomal instability can be the violation of functioning of enzymes responsible for structural integrity of the genome (13), changes in the repair or replication of chromosomes (14) and immunosuppression (15). Despite the numerous investigations of factors causing chromosomal mutations, there’s still a number of discussed issues (16).
The purpose of this study was investigation of erythrocyte antigens and somatic chromosomal instability in aboriginal Yakut cattle, Black-and-White cattle and in Yakut cattle introduced in Western Siberia.
Technique. The observations were performed in 2006-2009 in farms of the Novosibirsk province: the experimental farm “Elbashi”(the Institute of Cytology and Genetics of the RAS Siberian branch), JSC "Neudachino", LLC "Pervomaiskii" and the pedigree farm “Tulinskoe” (the school-farm of the Novosibirsk State Agrarian University). The object of study was the introduced Yakut cattle (n = 48) and Black-and-White (n = 380) breeds. Feeding rations corresponded to physiological needs of animals.
Venous blood for the analysis (10 ml) was collected into a sterile tube containing 0,5 ml heparin from the jugular vein of clinically healthy animals previously surveyed by veterinary experts. Leukocyte layer was placed in a nutrient medium 199 or RPMI (5 ml).  Cell material was thermostated at 37 °C for 24-48 hours. The dried preparations were stained with Giemsa stain. Chromosomes were stained using the method of G-differentiation (17, 18). The smears were studied by optical microscopy (Biolam L-211 and Biolam R-16 by “LOMO”, Russia). Blood groups were identified by hemolytic tests. 46 monospecific sera were used to reveal the antigens of eight genetic systems of blood groups: A, B, C, F-V, L, M, S and Z. In 100 cells, there were recorded ones with fragments, disruptions, aberrations, altered number of chromosomes, as well as hypoploid, hyperploid, aneuploid and polyploid cells.
Coefficients of genetic distance and similarity were assessed using the method of A.S. Serebrovskii (19) based on Euclidean distance (20):

, ,
where xi — the frequency of i antigen in the first compared population; yi —the frequency of i antigen in the second compared population.

1. The frequency of erythrocyte antigens in studied cattle breeds bred in Siberia (farms of Novosibirsk province,  2006-2009)
Antigen	Breed
	Introduced Yakut (n = 48)	Yakut (cyt. from 23)	Black-and-White (n = 380)
A2	0,667	0,423	0,430
B2	0,417	0,247	0,394
G2	0,750	0,277	0,459
G3	0,750	0,230	0,471
K	0,000	–	–
I1	0,000	0,090	0,071
I2	0,000	0,090	0,027
O1	0,250	–	0,093
O2	0,250	0,583	0,207
P2	0,000	0,126	0,076
Q	0,000	0,040	0,074
T2	0,000	0,030	0,013
Y2	0,833	0,720	0,601
A2	0,083	–	–
B'	0,000	0,010	0,030
D'	0,000	0,003	0,245
E1'	0,250	–	–
E2'	0,417	0,060	0,493
G'	0,083	0,300	0,241
I'	0,083	0,203	0,161
J2'	0,000	0,297	0,079
O'	0,000	0,030	0,243
P2'	0,250	0,120	–
Q'	0,500	0,133	0,581
J'	0,000	0,193	0,022
B''	0,000	0,070	0,011
G''	0,417	0,187	0,212
C1	0,750	0,553	0,565
C2	0,833	0,890	0,596
E	0,583	0,823	0,480
R1	0,000	0,003	0,080
R2	0,583	0,434	0,303
W	0,583	0,230	0,417
X1	0,000	0,503	0,140
X2	0,833	0,670	0,597
L'	0,000	0,705	0,085
F	1,000	0,884	0,973
V	0,250	0,350	0,205
L	0,333	0,124	0,405
M	0,000	0,220	0,133
S1	0,250	0,093	0,161
H1	0,833	0,893	0,753
U	0,083	–	0,046
U'	0,250	0,133	0,059
H''	0,083	0,100	0,015
Z	0,500	0,890	0,423
Note. Dash means that the antigen wasn’t tested.

Statistical analysis was performed using a free spreadsheet Gnumeric and R language of statistical computing. The parameters of descriptive statistics for qualitative traits were determined. At zero frequency, the standard error was calculated using the method of Van der Waerden. The comparison of mean values was carried out using the method of Fisher’s angular j-transformation and Student's t-test for qualitative traits (21). Null hypothesis was rejected at the significance level of p <0,05.
Results. Yakut cattle was introduced in Western Siberia in order to preserve the gene pool of this breed. Black-and-White cattle is bred in Siberia for a long time; it is well adapted to severe local climate (22). When comparing this breed with aboriginal Yakut (23) and the introduced Yakut cattle, there was assumed revealing of possible markers of adaptive processes in populations. The detailed analysis of Yakut cattle was performed by comparing it with White Siberian (24) and Red Steppe (25) breeds by antigen frequencies and proportions.
The introduced Yakut cattle were found to have significant changes in the fund of red blood cell antigens compared with aboriginal Yakut. The introduced livestock showed almost 7 times higher frequency of E2’antigen and significantly increased frequencies of antigens G2, G3 and Q’ (respectively, in 2,7, 3,2 and 3,6 times). At the same time, there were found the reduced frequencies of antigens J2’ and O2 (genetic system B), L’ and X1 (genetic system C). Microevolution process didn’t change the frequencies of antigens T2, B’, D’, O’, C2, R1, F, V, H1 and H’’.

The comparison of relative frequencies of antigens in Black-and-White and Yakut showed no significant differences for antigens A2, B2, I’, C1, F and V regardless of location of breeding Yakut cattle. Frequencies of O2, E2’, Q’, W and Z were similar in breeds from one geographical area; the differences were observed only when compared to the aboriginal Yakut cattle. It is possible that these antigens are the adaptation markers whose frequency doesn’t depend on species.
Frequencies of antigens G2, G3, X1, L’ and Z differed in all studied breeds. Probably, these antigens are the indicators of adaptation processes in populations and serve as distinguishing characteristics of studied species.
Prolonged breeding of Yakut cattle in the south of Western Siberia has led to a significant change in structure of erythrocyte antigens. Such processes happen quite often during the artificial selection in populations (26, 27). Black-and-White cattle was bred in similar environment, so the Yakut introduced in Western Siberia and Black-and-White breeds demonstrated higher genetic similarity (0,853) than was found between the introduced and aboriginal Yakut cattle (0,748). Along with it, the low genetic distance was detected between the introduced Yakut and White Siberian cattle (0,181).
Yakut cattle differed from other breeds of Siberia by the frequencies of antigens E2’, X1, and L' (Table 2). The ratio of antigens E2’, Q’, X1, and L’ in aboriginal Yakut was 1,0:2,2:8,4:11,8, while in Black-and-White, White Siberian and Red Steppe breeds - respectively 5,8:6,8:1,7:1,0; 8,3:5,1:2,7:1,0 and 2,6:3,7:1,0:1,6. For the introduced Yakut cattle, it wasn’t possible to calculate this index owing to significant differences between frequencies of these antigens. However, introduced animals showed the frequency of antigen E2’ not distinct from other breeds, probably due to manifestations of adaptive abilities of animals. E2’ has an adaptive advantage over the rest of other antigens. At the same time, antigens whose frequency wasn’t changed after breeding the Yakut cattle in Siberia can be considered as adaptive-neutral.

The main types of chromosome abnormalities detected in the introduced Yakut and Black-and-White cattle were: polyploidy, chromosome fragmentation and disruptions, aneuploidy; the latter occurred much more frequently than other anomalies (Table 3).
The degree of chromosomal instability in Yakut cattle was generally higher than in Black-and-White. Thus, the frequency of polyploidy in Yakut exceeded that in Black-and-White in 4,1 times, the rate of chromosomal aberrations – in 3,0 times. The high frequency of hypoploid cells in Black-and-White cattle can be explained by nondisjunction of chromosomes during mitosis due to certain unknown factors usually associated with destructive influence of some organic compounds on microtubules of mitotic spindle.

Chromosomal instability can be used to assess animals’ interior and the degree of environmental influence. High frequency of chromosomal instability in the Yakut breed can reflect the adaptive mechanism providing the maintenance of homeostasis in extreme conditions without losses in reproduction and productivity. It has been reported about increased ploidy of plants grown in the northern climate (28). In this study, a similar trend was observed.
Thus, there were established significant changes in the fund of erythrocyte antigens of the Yakut cattle bred in different ecological zones. The introduced Yakut cattle after the long-time breeding in conditions other than Yakutia was found to develop a specific complex of several antigens having a distinct ratio of frequencies. The introduced Yakut cattle showed the increased rate of various chromosomal abnormalities. These features can be considered as breed-specific adaptive mechanisms developed in severe environmental conditions. Intense breeding work increases the pressure on local species and reduces biodiversity. Finally, this can lead to the dominance of most common local breeds and reduced variability within breeds. Highly productive breeds, as a rule, are quite demanding to growing conditions. Today, such practice can cause adverse results under global climatic changes and rapidly mutating pathogens. The authors believe that this threat can be avoided by development of programs for conservation of gene pools of rare and endangered species.

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Novosibirsk State Agrarian University, Novosibirsk 630039, Russia, e-mail: kamevar@gmail.com

Received April 8, 2010

All-Russian Institute for Selection and Seed-Breeding of Vegetables LIK Cosmetics Institute of Agricultural Biotechnology Joint Stock Company "GosNIISyntezBelok"