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GENERAL DESIGN OF A STUDY ON MARKERS OF DISEASE RESISTANCE IN CATTLE

X XX X X XLie, 0., H. Solbu, R.L. Spooner, xxxxH.J. Larsen & xxx xv M. Syed-

NORWAY AND SCOTLAND

1. INTRODUCTIONSelection for general as well as for specific disease resi­stance in livestock has been subject to comprehensive studies and discussions especially during the last years. Also pos­sible application of disease associated immune traits in breeding programmes has been proposed (for review see Pavel 1975, Spooner et al. 1975, Gahne 1978, Govora & Spencer 1978, Almlid 1981, Krausslich et al. 1981, Lie & Solbu 1981, Oosterlee 1981). The general conclusions made by these authors are that present knowledge within the fields of immunogenetics and animal breeding combined with available computer processed disease records offer considerable sources for the planning of indirect selection schemes. On the other hand, still great research effort is required, especially for the development of a reliable balanced index of generalised resistance.The object of this study was to evaluate geveral immune traits as indicators of resistance to bovine mastitis.General design and principal preliminary results are pre­sented .2. MATERIAL AND METHODS2.1. Source of material 2ilili__Exgerimental_animalsiFrom a performance testing station of the Norwegian Red Cattle Association 130 young bulls from 4 to 12 months OF age were available. The material consisted of 14 half sib groups (sire families).

National Veterinary Institute, P.O.B. 8156, Dep. Oslo 1. Norway.Dept of Animal Breeding, Agricultural University of Norway, 1432 As-NLF,Dept, of Iinrwnology, Animal Breeding Research Organisation, Edinburgh EH*

3 JQ ScotlandDept, of Microbiology and Immunology, Veterinary College of Norway

P.O.B. 8146, Dep, Oslo 1, Norway.Dept, of Animal Husbandry and Genetics, Veterinary College of Norway

P.O.B. 8146, Dep. Oslo 1, Norway

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2l1^2^_Freeze_stored_serum_sam£lesiSerum samples from 329 other bulls at the same station were also available. These samples had been collected during a period of two years. The animals were all at approximately the same age at the time of sampling (279±8 days). This material comprised 2 1 sire families.2^1 ^ ^ _2 isease_recordsiData on mastitis frequency in half sisters of almost all the 130 young bulls were available from the Norwegian so-called health card system. This is a disease recording scheme as described by Solbu (1978) which implies computer processed data on veterinary treatments. The health card system com­prises clinical records of two thirds of the Norwegian cow population. Twelve of the fourteen sires had been progeny tested for clinical mastitis based on data from 150-200 first lactating cows per bull.2,2. Immunoassays2^2_. 1 _._§2 yine_lymghocyte_antigens__(6bLA)_,iThe BoLA types were determined in all 130 young bulls after which the likely BoLA genoype of each sire was inferred.The testing, which is an antibody/complement dependent cyto­toxic reaction, was run as described by Spooner et al. (1978). The workshop agreed specificities Spooner et al. (1979),Sales & Curie-Cohen (1-982) , which have been found to be pro­ducts of one single locus (Oliver et al. 1981) are designated W and represent the bovine Major Histocompatibility Complex (HHC) or BoLA. Other bovine lymphocyte specificities are here designated ED (abbr.for Edinburgh specificity).2^2^2_Immune_resgonsiveness^Antibody and cell mediated immune (CMI) responses were measu­red after the injection of two different antigens, of which one was multideterminant (HSA) and the other one a uniform synthetic polypeptide (TG-ALX). The antigens were admini­strated twice with an interval of one month. Both primary and secondary responses were recorded. Also CMI responses to different mitogens were determined. The antibody titers were measured employing single radial immunodiffusion and passive hemagglutination (Larsen, unpublished). The CMI acti­vity was determined according to a whole blood technique as described by Larsen (1979), slightly modified in this study.

ZiZih__§§EffiJLiX§25XSJ§_22£i¥i£¥i_ii2?i!!LZM levels were measured both in the experimental material of 130 bulls and in the 329 freeze stored serum samples. The assay was run according to a procedure previously described (Lie 1980) .

XMiles Laboratories Ltd. P.O.Box 37, Stoke Poges, S12 4LY, England

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2.2^ 4^ __§erum_hemol^tic_comglement__(HC)__ l e v e lTotal serum HC activity was determined in the 130 bulls according to Kakoma & Kinyanjui (1974) with some modifica­tion (Lie et al. unpublished data).2.3. Statistical methodThe major statistical analysis was performed employing the method of least squares.3. RESULTS AND DISCUSSION3.1. Genetic influence on immune traits37T7T7__I nunune_ responsiveness^Several sire families differed significantly (p < 0.05) in respect of both antibody and CMI responses. On the other hand, the sire effect as a whole was not quite significant with regard to these traits (0.05 < p < 0.1). In previous expe­riments with HSA on an even larger material the significant genetic influence was restricted to the primary response only (Lie 1979). Therefore, these observations suggest that the genetic influence on immune responsiveness in cattle is realtively low and for the purpose of heritability (h2) cal­culation, a larger body of material with more sires included is needed.3 i . l i .2 i__L Z M _ a c tiv ity iThe effect of sires on LZM activity was found to be highly significant (p < 0.01). The main contribution to this effect was caused by a sire family with a mean LZM level about twice that of the total material. LZM was observed to segregate into distinct groups of high and low level phenotypes when thirty offspring of the above sire were studied. The groups did not differ significantly from 50:50 proportions which suggested a simple Mendelian inheritance of this enzyme. More­over, no sire effect could be demonstrated when the effect of an apparent major gene for LZM activitv was removed, by the exclusion of all high level animals from the material (Lie & Solbu, 1982).3 . 1 . 3 . _H C _levels_.Highly significant (p < 0.01) sire effect with corresponding h in the range of 0.69 to 1.0 has been estimated for the total serum HC. This is in accordance with previous studies on complement levels in monozygous cattle twins (Sellei 1975). Also the striking breed differences in HC demonstrated by Kakoma & Kinyanjui (1974) and the strong MHC associattion with HC found in this study (see below ) support the above heritability estimate.3.2. Association of BoLA with quantitative immune traitsAs can be seen from Table 1, association of BoLA with all immune traits studied has been detected. The most striking findings are the strong influence of BoLA on HC and antibody

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r e s p o n s e t o TG-AL. A ls o MHC a s s o c i a t e d s p e c i f i c CMI r e s p o n ­s e s t o HSA an d TG-AL w e r e r e v e a l e d . C o r r e s p o n d in g f i n d i n g i n o t h e r l i v e s t o c k s p e c i e s a r e t h e MHC c o n t r o l l e d a n t i TG-AL r e s p o n s e fo u n d i n c h ic k e n (G u n th er e t a l . 1 9 7 4 ) an d t h e p i g MHC i n f l u e n c e o n HC d e m o n s tr a te d b y V aim an e t a l . ( 1 9 7 8 ) .The m it o g e n in d u c e d r e s p o n s e s a r e i n g e n e r a l r e g a r d e d a s r e f l e c t i n g t h e immune a c t i v i t y o f d i f f e r e n t s u b p o p u la t io n s o f ly m p h o c y t e s w h ic h may p o s s e s s d i f f e r e n t MHC a s s o c i a t e d a n t i g e n s . T he BoLA a n d s p e c i f i c immune r e s p o n s e a s s o c i a t e d m it o g e n r e s p o n s e s d e m o n s tr a te d i n T a b le 1 a r e t h e r e f o r e t o som e d e g r e e e x p e c t e d .

T able 1 . A s s o c i a t i o n o f som e s e r o l o g i c a l l y d e f i n e d b o v in e lymphocyte a n t i g e n s ( s p e c i f i c i t i e s ) w ith ' d i f f e r e n t immune t r a i s a s s t u d i e d i n y o u n g b u l l s . A rrow ; Up o r down d i r e c t e d a rro w i n d i c a t e t h a t i n d i v i d u a l s p o s s e s s i n g t h e s p e c i f i c i t y i n q u e s t i o n h a v e s i g n i f i c a n t l y (p < 0 .0 5 ) h ig h e r o r lo w e r , r e s p e c t i v e l y , immune a c t i v i t y th a n i n d v i d u a l s l a c k i n g t h e s p e c i f i c i t y . A rrow b e tw e e n b r a c k e t s c o r r e s p o n d t o a l e v e l o f s i g n i f i c a n c e o f 0 .0 5 < p < 0 . 0 8 .

B o v in elynphocytes p e c i f ic t y

Non s p e c i f i c imnune t r a i t s :

S p e c if ic antibody response to ;

S p e c if ic CMI response to :

U n sp ec ific (Mitogen induced)

HC LZM HSA TGAL HSA TGAL PHA PWM ConA PPDW 2" 6 4 ~ T ~ t

---------” 6 .1 t---------R-g-------- (A l___________ + ___________ 1 ___ 1 ____

---------W~ T T ~ 4 t T "" i £ ( t ) 4_____" 20 H Z

ED 28 " 4 4 4---------* ~ 1 T ~ 4

" 85 t T t t ( V" 91 t ~ T ~" 94 tM 99 t 4

3 . 3 . A s s o c i a t i o n o f BoLA w it h m a s t i t i s .

C om p arin g t h e BoLA t y p e w i t h t h e p r o g e n y t e s t r e s u l t f o r m a s t i t i s w i t h i n e a c h s i r e , s i g n i f i c a n t d i f f e r e n c e s b e tw e e n BoLA t y p e s i n s u s c e p t i b i l i t y t o t h i s d i s e a s e w e r e dem on­s t r a t e d . (p < 0 . 0 5 ) .

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4 . CeWERAL CONCLUSIONS

T h is s t u d y s u g g e s t s t h a t t h e g e n e t i c i n f l u e n c e i s s t r o n g e r a n d /o r t h a t t h e i n h e r i t a n c e i s m ore s im p l e f o r n a t u r a l o r n o n - s p e c i f i c immune m ech a n ism s ( ly s o z y m e an d c o m p le m e n t) th a n i s t h e c a s e w i t h t h e s p e c i f i c o n e s (im m une r e s p o n s i v e ­n e s s ) .M oreo v er t h e l i k e l y e x i s t e n c e i n c a t t l e o f MHC a s s o c i a t e d r e g u l a t i o n o f immune r e s p o n s i v e n e s s , an d c o m p le m e n t s h o u ld b e a d m it t e d .F i n a l l y , t h e r e i s r e a s o n t o b e l i e v e t h a t r e s i s t a n c e t o m a s t i t i s t o a c e r t a i n d e g r e e i s i n h e r i t e d t o g e t h e r w i t h an a n im a l ' s BoLA t y p e .

SUMMARY

The g e n e r a l d e s i g n o f an e x p e r im e n t t o s t u d y t h e r e l a t i o n ­s h ip o f g e n e t i c m a rk ers w i t h d i s e a s e . i s d e s c r i b e d . A num ber o f immune t r a i t s i n c l u d i n g s p e c i f i c r e s p o n s e s ( t o HSA an d TGAL) and u n s p e c i f i c o n e s ( t o m i t o g e n s ) t o g e t h e r w it h l e v e l s o f ly s o z y m e and c o m p le m e n t h a v e b e e n d e te r m in e d i n 130 y o u n g b u l l s . I n a d d i t i o n t h e BoLA g e n o t y p e s o f t h e a n im a ls w e r e d e t e r m in e d . R e l a t i o n s h i p s b e tw e e n t h e t r a i t s h a v e b e e n s t u d i e d a n d i t h e i r a s s o c i a t i o n w i t h d i s e a s e i n h a l f s i s t e r s o f t h e you n g b u l l s h a s b e e n c a l c u l a t e d .

KURZFASSUNG

Es w urde d e r g e n e r e l l e A u fb au e i n e s E x p e r im e n te s zum S tu d iu m der B e z ie h u n g z w is c h e n g e n e t i s c h e n M a r k e r - E ig e n s c h a f t e n und K r a n k h e i t s a n f a l l i g k e i t b e s c h r i e b e n . E in e A n z a h l v e r s c h i e d e - n e r im m u n o lo g is c h e r M erkm ale w ie s p e z i f i s c h e (HSA gnd TGAL) und u n s p e z i f i s c h e (M ito g e n ) Im m u n a n tw o rt, L ysozym und K o m p le- m ent w u rd en b e im 130 J u n g -B u lle n u n t e r s u c h t . A uch d e r BoLA- G en o ty p d e r B u l le n w urde g e t e s t e t . D er Zusam m enhang d i e s e r M erkm ale w urde e r m i t t e l t , und d i e A s s o z i a t i o n z u r K r a n k h e i t s ­a n f a l l i g k e i t d e r H a lb -S c h w e s te r d e r J u n g - B u l le n w urde b e s t im m t .

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