Br. vet.
J.
(1975)
13 1 ,
292
DEGENERATIVE MYOPATHY WITH MYOGLOBINURIA IN YEARLING CATTLE W. M. ALLEN, * R. BRADLEY, t SYLVIA W. H. PARR,t K. SWANNACK,+ C. R. Q. AND A. MACPHEE §
By
BERRETT, t BARTON§
*Institute for Research on Animal Diseases, Compton, Newbury, Berkshire, RG r6 oNN. tCentral Veterinary Laboratory, Wrybridge, Surrey. §South Molton, Devon +Bridget's E.H.F., Winchester, Hants. SUMMARY
The clinical, biochemical and pathological observations made during an outbreak of acute myopathy in yearling cattle which followed turnout to spring pasture are described. The affected cattle were between 13 and 20 months old and the weight range was 225-320 kg. The diet fed during the winter was vitamin E and selenium deficient. In one case the vitamin deficiency was exacerbated by preservation of the cereal portion of the diet with propionic acid. Other predisposing factors were the unaccustomed exercise following winter housing and the onset of inclement weather after turnout. The condition is compared with enzootic muscular dystrophy of young calves, and it is concluded that the condition is an atypical manifestation of the calf disease but affecting older animals. New husbandry practices, particularly the storage of high moisture grain, seem likely to increase the frequency of the condition. INTRODUCTION
A paralytic muscle disease affecting yearling cattle following turnout in the spring was described by Doig (1970) and Christl (1971). It resembled the skeletal muscle form of nutritional or enzootic muscular dystropy of calves, except that the affected cattle were considerably older. In the latter outbreak described by Christl (197 I) there was a concurrent severe myoglobinuria. During the spring of 1972, Barton & Allen (1973) observed a condition closely resembling that described by Christl (197 I) and which resolved following supportive and empirical therapy. The treatment included vitamin E and selenium administration. The other vitamin E responsive myopathy in cattle, enzootic or nutritional muscular dystrophy of calves has been extensively reviewed (Blaxter & McGill, 1955; Hadlow, 1962; Hulland, 1970). This condition is a nutritional myodegeneration and is commonly called white muscle disease. The sections of
DEGENERATIVE MYOPATHY
293
diseased bovine muscle received at the Central Veterinary Laboratory during the previous 14 years were reviewed and 24 per cent showed histological lesions indistinguishable from those observed in white muscle disease. The majority of affected animals were between 3 and 6 months of age. Further investigations to establish the aetiology and pathogenesis of the condition described by Barton & Allen (1973) were undertaken during spring 1973. This paper describes the epidemiological, clinical, pathological and biochemical investigations and results. Further details of the muscle histology and ultrastructure and the vitamin E and selenium status have been or will be published elsewhere (Allen et al., 1974). As well as observations on the Devonshire farm originally associated with the condition (farm I), a further outbreak was investigated on a neighbouring farm (farm II). A group of cattle was also observed at an experimental husbandry farm, where the condition had not occurred previously, to act as control.
MATERIALS AND METHODS
Animals At farm I, where disease occurred during spring 1972, 25 heifers were housed from November 1972 to April 1973 in three covered pens approximately 30 ft X 25 ft; two pens housed 17 cattle which were 16-18 months old at turnout. These animals will be referred to as group Ia. A further eight animals housed in the third pen were 13-16 months old and will be referred to as group lb. A further six younger animals were housed in similar accommodation, their ages were between 6-14 months at turnout and represented group Ie. On farm II, 17 cattle of various breeds, Friesian, Hereford, HerefordFriesian crosses and Charolais crosses, were housed over the winter in a large covered yard, and these constituted group II. The four animals killed for pathological examination all came from this group (numbers 1-4). Their age at turnout was between 13 and 20 months. At the Experimental Husbandry Farm, 40 yearling heifers, housed in semicovered yards from November-April, were between 15 and 20 months old at turnout. These animals constituted group III. Group III animals were weighed regularly, and the weights of the other groups were estimated from tape measurements round the anterior thorax. Diet The heifers in groups la, Ib, and Ie were fed on a diet of home-grown hay and home-grown propionic-acid-conserved barley with proprietary balancer included at 6·25 per cent as in 1972 (Barton & Allen, 1973). Gr oup II animals received home-grown straw and propionic-acid-treated hay and conventionally stored barley together with a barley balancer. The group III cattle received home-grown straw and silage and silo-stored moist barley with a proprietary barley balancer included at 20 per cent.
294
BRITISH VETERINARY JOURNAL, 13 1, 3
Feed analysis The vitamin E content of feedingstuffs was estimated as follows: the unsaponifiable matter from an acetone extract of the sample obtained as described in the report of the vitamin E panel of the S.A.C. (I959), was chromatographed on a column of deactivated alumina (Christie, Dean & Milburn, I973) to separate the vitamin E into ex- and non ex-fractions. The ex fraction was chromatographed on silica gel G thin-layer plates, developed with di-isopropylether in petroleum ether (Whittle & Pennock, I967) to separate ex-tocopherol and IX-tocotrienol. Each zone was eluted with ethanol, and the concentration of ex-tocopherol and ex-tocotrienol was determined by reaction with ferric chloride and bathophenanthroline (Tsen, I96I). The selenium content was estimated following wet oxidation of a sample dried at 60°-70°C, by measurement of the fluorescence of the cyclohexane extract of 2,3 diaminonapthaleneJselenious acid complex. The peroxide values of the fat were determined iodometrically following extraction from the grain with cold chloroform. Feed intake An approximate measure of daily feed intake for each group was obtained by either weighing feedstuffs on one single day or from total consumption over a given period. Details of the estimated feed intake for all groups, Ia-c, II and III, are included in Table I. Blood samples In groups Ia-c and group III (30 of the 40 animals) blood samples were collected on at least two occasions before and on two occasions after turnout. The animals in group II were sampled only after turnout, coincident with and subsequent to the appearance of clinical disease symptoms. Approximately 40 ml of whole blood was collected into heparinized vacutainers. Plasma was collected after centrifugation, and the concentrations of glucose, potassium, sodium, magnesium, calcium, chloride, urea and ex-ami nonitrogen and the activities of creatine kinase (CK), aldolase (ALD) and lactate dehydrogenase (LDH) were measured either using test kits* or by the routine methods used at the Central Veterinary Laboratory. The distribution of LDH activity among the various isoenzyme bands were determined using a method described by Dietz & Lubrano (I967). From a small number of animals in groups I and II, including the four animals which were killed, blood samples were collected into EDTA anticoagulant and thin blood smears were made for examination for protozoa. Total red blood cells, white blood cells and differential counts, together with packed cell volume and haemoglobin concentrations, were measured.
* The Boehringer Corporation Ltd, London, or with CK a Calbiochem test kit containing cysteine as activator.
TABLE I EST IMATED TOTAL DAILY FEED, VITAMIN E AND SELENIUM INTAKE
Vitamin E
Feedstuff
Selenium
Group Ia
b
Bodywt" (kg)
Feed
Daily intake (kg fresh wt")
310
Barley Hay
0"6 7"3
250
220
II
III
275
300
Barley Hay Barley Hay
1"3 4"5
1"8 2"7
Concentration vitamin E (mg/kg dry matter (a.-tocopherol + a.-tocotrienol) ) 3"5 9"4
3"5 9"4
3"5 9"4
Barley Straw (barl ey)
1"8 3"2
Barley Straw (barley) Silage
3"2 3"2
15"7 8"7
2"7
?
13"0 8"0
Daily intake vitamin E (mg)
Intake/kg bodywt"
Concentration (mg/kg dry matter)
Daily intake (mg)
1"7 61"2 62"9
0"04 0"04
0" 0 19 0"26 3 0"282
3"64 38 "10 41"75
0"04 0"04
5"04 22"7 27"74
0"04 0"04
19"2 23"0 -4 2 "2
0"04 0"04
4 2 "8 23"6
0"03 0"03
?
?
>66"4
0"04 2 0"162 0 "204 0"05 8 0"097 0 "145 0"059 0"115 0"174
Vitamin E (mg/kg body wt")
Selenium (mg/kg bodywt")
0
ttl Q
t':l 0"20
0"0009
Z
ttl :;d ;l>
>-l .....
0"17
0"0007
-
-l
:r:
>
0"22
> 0"0006
0"082 0"086
? > 0"168
IV
J.
(1975)
13 1 ,
292
DEGENERATIVE MYOPATHY WITH MYOGLOBINURIA IN YEARLING CATTLE W. M. ALLEN, * R. BRADLEY, t SYLVIA W. H. PARR,t K. SWANNACK,+ C. R. Q. AND A. MACPHEE §
By
BERRETT, t BARTON§
*Institute for Research on Animal Diseases, Compton, Newbury, Berkshire, RG r6 oNN. tCentral Veterinary Laboratory, Wrybridge, Surrey. §South Molton, Devon +Bridget's E.H.F., Winchester, Hants. SUMMARY
The clinical, biochemical and pathological observations made during an outbreak of acute myopathy in yearling cattle which followed turnout to spring pasture are described. The affected cattle were between 13 and 20 months old and the weight range was 225-320 kg. The diet fed during the winter was vitamin E and selenium deficient. In one case the vitamin deficiency was exacerbated by preservation of the cereal portion of the diet with propionic acid. Other predisposing factors were the unaccustomed exercise following winter housing and the onset of inclement weather after turnout. The condition is compared with enzootic muscular dystrophy of young calves, and it is concluded that the condition is an atypical manifestation of the calf disease but affecting older animals. New husbandry practices, particularly the storage of high moisture grain, seem likely to increase the frequency of the condition. INTRODUCTION
A paralytic muscle disease affecting yearling cattle following turnout in the spring was described by Doig (1970) and Christl (1971). It resembled the skeletal muscle form of nutritional or enzootic muscular dystropy of calves, except that the affected cattle were considerably older. In the latter outbreak described by Christl (197 I) there was a concurrent severe myoglobinuria. During the spring of 1972, Barton & Allen (1973) observed a condition closely resembling that described by Christl (197 I) and which resolved following supportive and empirical therapy. The treatment included vitamin E and selenium administration. The other vitamin E responsive myopathy in cattle, enzootic or nutritional muscular dystrophy of calves has been extensively reviewed (Blaxter & McGill, 1955; Hadlow, 1962; Hulland, 1970). This condition is a nutritional myodegeneration and is commonly called white muscle disease. The sections of
DEGENERATIVE MYOPATHY
293
diseased bovine muscle received at the Central Veterinary Laboratory during the previous 14 years were reviewed and 24 per cent showed histological lesions indistinguishable from those observed in white muscle disease. The majority of affected animals were between 3 and 6 months of age. Further investigations to establish the aetiology and pathogenesis of the condition described by Barton & Allen (1973) were undertaken during spring 1973. This paper describes the epidemiological, clinical, pathological and biochemical investigations and results. Further details of the muscle histology and ultrastructure and the vitamin E and selenium status have been or will be published elsewhere (Allen et al., 1974). As well as observations on the Devonshire farm originally associated with the condition (farm I), a further outbreak was investigated on a neighbouring farm (farm II). A group of cattle was also observed at an experimental husbandry farm, where the condition had not occurred previously, to act as control.
MATERIALS AND METHODS
Animals At farm I, where disease occurred during spring 1972, 25 heifers were housed from November 1972 to April 1973 in three covered pens approximately 30 ft X 25 ft; two pens housed 17 cattle which were 16-18 months old at turnout. These animals will be referred to as group Ia. A further eight animals housed in the third pen were 13-16 months old and will be referred to as group lb. A further six younger animals were housed in similar accommodation, their ages were between 6-14 months at turnout and represented group Ie. On farm II, 17 cattle of various breeds, Friesian, Hereford, HerefordFriesian crosses and Charolais crosses, were housed over the winter in a large covered yard, and these constituted group II. The four animals killed for pathological examination all came from this group (numbers 1-4). Their age at turnout was between 13 and 20 months. At the Experimental Husbandry Farm, 40 yearling heifers, housed in semicovered yards from November-April, were between 15 and 20 months old at turnout. These animals constituted group III. Group III animals were weighed regularly, and the weights of the other groups were estimated from tape measurements round the anterior thorax. Diet The heifers in groups la, Ib, and Ie were fed on a diet of home-grown hay and home-grown propionic-acid-conserved barley with proprietary balancer included at 6·25 per cent as in 1972 (Barton & Allen, 1973). Gr oup II animals received home-grown straw and propionic-acid-treated hay and conventionally stored barley together with a barley balancer. The group III cattle received home-grown straw and silage and silo-stored moist barley with a proprietary barley balancer included at 20 per cent.
294
BRITISH VETERINARY JOURNAL, 13 1, 3
Feed analysis The vitamin E content of feedingstuffs was estimated as follows: the unsaponifiable matter from an acetone extract of the sample obtained as described in the report of the vitamin E panel of the S.A.C. (I959), was chromatographed on a column of deactivated alumina (Christie, Dean & Milburn, I973) to separate the vitamin E into ex- and non ex-fractions. The ex fraction was chromatographed on silica gel G thin-layer plates, developed with di-isopropylether in petroleum ether (Whittle & Pennock, I967) to separate ex-tocopherol and IX-tocotrienol. Each zone was eluted with ethanol, and the concentration of ex-tocopherol and ex-tocotrienol was determined by reaction with ferric chloride and bathophenanthroline (Tsen, I96I). The selenium content was estimated following wet oxidation of a sample dried at 60°-70°C, by measurement of the fluorescence of the cyclohexane extract of 2,3 diaminonapthaleneJselenious acid complex. The peroxide values of the fat were determined iodometrically following extraction from the grain with cold chloroform. Feed intake An approximate measure of daily feed intake for each group was obtained by either weighing feedstuffs on one single day or from total consumption over a given period. Details of the estimated feed intake for all groups, Ia-c, II and III, are included in Table I. Blood samples In groups Ia-c and group III (30 of the 40 animals) blood samples were collected on at least two occasions before and on two occasions after turnout. The animals in group II were sampled only after turnout, coincident with and subsequent to the appearance of clinical disease symptoms. Approximately 40 ml of whole blood was collected into heparinized vacutainers. Plasma was collected after centrifugation, and the concentrations of glucose, potassium, sodium, magnesium, calcium, chloride, urea and ex-ami nonitrogen and the activities of creatine kinase (CK), aldolase (ALD) and lactate dehydrogenase (LDH) were measured either using test kits* or by the routine methods used at the Central Veterinary Laboratory. The distribution of LDH activity among the various isoenzyme bands were determined using a method described by Dietz & Lubrano (I967). From a small number of animals in groups I and II, including the four animals which were killed, blood samples were collected into EDTA anticoagulant and thin blood smears were made for examination for protozoa. Total red blood cells, white blood cells and differential counts, together with packed cell volume and haemoglobin concentrations, were measured.
* The Boehringer Corporation Ltd, London, or with CK a Calbiochem test kit containing cysteine as activator.
TABLE I EST IMATED TOTAL DAILY FEED, VITAMIN E AND SELENIUM INTAKE
Vitamin E
Feedstuff
Selenium
Group Ia
b
Bodywt" (kg)
Feed
Daily intake (kg fresh wt")
310
Barley Hay
0"6 7"3
250
220
II
III
275
300
Barley Hay Barley Hay
1"3 4"5
1"8 2"7
Concentration vitamin E (mg/kg dry matter (a.-tocopherol + a.-tocotrienol) ) 3"5 9"4
3"5 9"4
3"5 9"4
Barley Straw (barl ey)
1"8 3"2
Barley Straw (barley) Silage
3"2 3"2
15"7 8"7
2"7
?
13"0 8"0
Daily intake vitamin E (mg)
Intake/kg bodywt"
Concentration (mg/kg dry matter)
Daily intake (mg)
1"7 61"2 62"9
0"04 0"04
0" 0 19 0"26 3 0"282
3"64 38 "10 41"75
0"04 0"04
5"04 22"7 27"74
0"04 0"04
19"2 23"0 -4 2 "2
0"04 0"04
4 2 "8 23"6
0"03 0"03
?
?
>66"4
0"04 2 0"162 0 "204 0"05 8 0"097 0 "145 0"059 0"115 0"174
Vitamin E (mg/kg body wt")
Selenium (mg/kg bodywt")
0
ttl Q
t':l 0"20
0"0009
Z
ttl :;d ;l>
>-l .....
0"17
0"0007
-
-l
:r:
>
0"22
> 0"0006
0"082 0"086
? > 0"168
IV
