Chapter 11.

Trace elements and vitamins; deficiency and

excess

Introduction | Biochemistry | Clinical signs of deficiency | Sub-clinical deficiency | Clinical

pathology in the investigation of deficiencies | Interpretation of results | Diagnosis of clinical
syndromes | Response trials | Prevention and treatment | Poisoning | Recommended reading

Return to Sheep Health & Production Index

Introduction

Trace elements are those minerals required by animals in amounts measured in micrograms
and milligrams per kilogram of dry matter ingested. They include iron, manganese, zinc,
copper, iodine, cobalt, molybdenum and selenium. In Australia and New Zealand, deficiency
syndromes of grazing sheep are well recognised for copper, iodine, cobalt and selenium.
Poisoning is chiefly restricted to copper and, less commonly, selenium, although molybdenum
excess can induce a syndrome of copper deficiency. Vitamin nutrition of sheep is occasionally
deficient; inadequate intake of vitamins A, D and E being responsible for the most common
deficiency conditions. Vitamin B12 deficiency occurs when cobalt intake is inadequate.
The major deficiencies considered here are those of copper, selenium, cobalt and vitamin E.
The grouping together of these common trace element and vitamin deficiencies is not
intended to suggest that the deficiency syndromes have in common any pathogenesis or
clinical features, or that they are likely to occur together. What they do have in common is a
tendency to cause sub-clinical and insidious losses of productivity in wool-growing sheep,
particularly young sheep, and a number of strategies for prevention or correction of
deficiency; such as intra-ruminal pellets, concentrates added to anthelmintic preparations and
paddock 'licks' or 'blocks'. The notion that trace element deficiencies cause general 'failure to
thrive' or 'illthrift' in young sheep despite apparently adequate pasture feed also leads
producers to request their veterinary practitioners to demonstrate a deficiency with broad
ranging diagnostic tests, often with limited justification other than a vague history of poor
performance.
The correction of trace element deficiencies does at times result in spectacular improvement
in the production of grazing animals and widespread, but less obvious sub-clinical responses.
They remain, however, relatively unimportant problems compared to parasitism and nutritional
deficiencies of energy and protein.

Biochemistry

Copper
Copper is required for body, bone and wool growth, for pigmentation, myelination of nerve
fibres and leucocyte function. It is involved in a number of enzymes which catalyse oxidase-
type reactions in both plants and animals. Ceruloplasmin is a copper containing compound
which is important in the mobilization of iron for haemoglobin synthesis from intestinal
mucosal cells, liver parenchymal cells and reticulo-endothelial cells. The formation of Fe(III)-
transferrin is dependent on ceruloplasmin. Copper is essential for the enzymes amine and
lysyl oxidase, which contribute to the cross-linking of bone collagen which, in turn, affects
collagen solubility and bone strength. Copper is essential for the enzyme cytochrome oxidase
which is involved in cellular respiration and phospholipid synthesis; and essential for the
polyphenyl oxidases which catalyse conversion of tyrosin to melanin[1].

Selenium and vitamin E

The best known biological activity of selenium is in the seleno-enzyme glutathione peroxidase
(GSHPx). Vitamin E and GSHPx have similar and complementary physiological roles in
protecting cells from damage caused by endogenous peroxides. These products, which are
formed as part of the essential synthesis of prostenoids from polyunsaturated fatty acids
(PUFA)[2], can damage lipid-rich cell membranes and intracellular organelles leading to
cellular degeneration and necrosis. GSHPx acts to destroy peroxides before they attack cell
membranes, while vitamin E acts within the membrane to attract unsaturated fatty acid
molecules and form loose chemical complexes until they are metabolised during cell
respiration, thus preventing the formation of fatty acid hydroperoxides[3]. The peroxides can
also denature cellular proteins and while GSHPx is protective, Vitamin E is not. This may
explain why selenium therapy of deficient animals is able to prevent some forms of muscular
degeneration which cannot be prevented by administration of vitamin E[4]. Similarly, vitamin E
may have a unique preventitive role in clinical syndromes associated with peroxidation in
tissues which are inherently low in GSHPx, regardless of the adequacy of selenium nutrition.
The similarities in the myopathies associated with selenium and vitamin E deficiency originally
led to the conclusion that the two nutrients had interchangeable roles. This is now known not
to be true; selenium will not protect sheep from weaner nutritional myopathy but vitamin E will;
vitamin E will not protect sheep from white muscle disease, but selenium will.
Unsaturated fatty acids, particularly if polyunsaturated, form both a part of the energy supply
and an essential part of tissue membranes. The unsaturated double bonds of membrane
PUFAs are inherently unstable and are readily attacked by peroxides and other forms of
active oxygen. It follows that the greater the level and input of PUFA to the cells, the greater
the amount of antioxidants needed to prevent the reaction(3). Animals with high PUFA diets
particularly require satisfactory nutrition with selenium and vitamin E to avoid the risk of
myopathy.
In species other than sheep, selenium has a role in resistance to disease. In sheep there is
evidence that selenium deficient animals are no less able to produce antibodies against
bacterial antigens[5] and no less able to prevent the establishment of internal parasites[6]
than sheep with adequate selenium intake.

Vitamin B12
This vitamin is synthesised by rumen microbes from dietary cobalt. Propionate, derived from
the fermentation of plant cellulose in the rumen, is the major source of energy in the ruminant.
It is converted to succinate to provide energy. The conversion of methylmalonyl-CoA to
succinyl-CoA is dependent on methylmalonyl-CoA mutase, a Vitamin B12-containing
mitochondrial enzyme involved in the major pathway through which propionate and several
amino acids are metabolised. Deficiency of vitamin B12 leads to an accumulation of
methylmalonic acid in the liver and a high level of propionate in the blood. The increased
impairment of propionate metabolism is accompanied by a progressive loss of appetite.

Iodine, molybdenum, iron, manganese and zinc

Iodine is essential for thyroid hormone production, molybdenum is required for several
metalloenzymes, iron is required for haemoglobin synthesis, manganese for carbohydrate
and lipid metabolism and zinc is a component of many enzymes.

Clinical signs of deficiency

Copper

Changes in wool growth and character

The loss of wool crimp and increased lustre of wool, producing 'steely wool', is one of the first
clinical signs of copper deficiency to develop in sheep. The wool has crimps at 3 to 4 times
the normal width and greatly reduced tensile strength. In black-woolled sheep, failure of
pigment-ation during growth of the follicle leads to the formation of bands of grey or white
wool in the staple; reflecting periods during which copper nutrition was inadequate. There
may be several narrow bands interspersed with normally pigmented wool indicating that, over
a period of months the animal dipped in and out of 'normality' for a few days or weeks at a
time. Excess dietary molybdenum or sulphate, in particular, leads to copper-responsive
aberrations in wool growth.

Enzootic ataxia

Enzootic ataxia occurs in lambs born of copper deficient ewes; the seasonal variation in
copper availability usually leads to the restriction of this syndrome to lambs born in late winter
and spring. Lambs are affected at birth or develop signs within 1 or 2 months of birth. Affected
animals have posterior paresis which is particularly apparent when the lambs are driven. They
appear weak and incoordinated in the hind limbs, knuckling over on the fetlocks and swaying
from side to side as they attempt to rise or stay upright. The disease is similar to 'swayback' of
lambs, recorded in Great Britain and also associated with copper deficiency.
Histopathologically, the diseases differ, possibly reflecting a different severity of copper
deficiency during gestation.

Bone disorders
Osteoporosis in lambs due to copper deficiency results in bone fragility and an increased
prevalence of fractures of long bones and rib bones.

Selenium

In young sheep, two selenium-responsive conditions are recognised; one a myopathy of

lambs, white muscle disease (WMD), the other a syndrome of lowered productivity varying in
severity from sub-clinical depression of wool production to a clinical condition of poor growth
and increased mortality rates - so called selenium-responsive unthriftiness (SRU). In adult
sheep, a selenium-responsive infertility of ewes has been reported. While the diseases are
clearly associated with selenium deficient areas, they have been difficult to reproduce
experimentally or to predict on the basis of dietary selenium or animal tissue selenium levels.
The possibility exists that other factors are involved in the expression of disease.

White muscle disease (WMD)

This syndrome is also called nutritional or enzootic muscular dystrophy. The myopathy of
WMD can affect heart muscle or different skeletal muscle groups; clinical signs varying with
the muscle groups involved. Muscles of the upper limbs are frequently involved causing
affected lambs to walk stiffly, if they can rise at all. Cardiac myopathy can cause sudden
death. WMD has been classified as congenital[7] or delayed, and acute or subacute(4). In the
congenital form, affected lambs are either born dead or die suddenly after exertion, such as
that required for suckling, within a few days of birth. Lesions are usually confined to the
myocardium. The delayed and sub-acute forms occur predominantly in lambs from 3 to 6
weeks of age, sometimes up to 3 months of age. Affected lambs have a stiff, stilted gait
(leading to an alternate name stiff lamb disease) and an arched back. They prefer to remain
in sternal recumbency then become prostrate and die within a few days. The myopathy
involves several or many skeletal muscle groups.

Selenium responsive unthriftiness (SRU)

A selenium-responsive syndrome of reduced weight gain and increased mortality rates,

particularly in sheep 3 to 18 months of age, has been recorded in NZ and Australia. Until
effective preventive measures became widely used, SRU was considered the most
widespread and economically important of all the selenium-responsive diseases of New
Zealand livestock. In sheep, it varies from a subclinical inability to maintain expected growth
rate to a clinical unthriftiness which may, in some seasons, lead to a heavy mortality[8].
Affected animals lose condition, become weak, develop dry, open fleeces and some have
diarrhoea[9]. There is usually a history of WMD and/or selenium-responsive infertility on the
same property.
SRU is less commonly observed in Australia but there are regions where the disease is
endemic. In a response trial in the Strathbogie Ranges of central Victoria, treatment with
selenium (0.1mg/kg per os) of lambs at marking and weaning improved weaning weight by 2
kg (10%) and reduced mortality rate from 17.5% to nil. Despite the selenium responses, no
WMD occurred during the trial but it had occurred on the same property in the previous year.
Fleece weight responses were also large. In other regions of Victoria, SRU is generally
considered unlikely because pasture selenium levels are rarely sufficiently low (< 0.02mg/kg)
for more than a few weeks or months in spring[10].

Selenium responsive infertility

In New Zealand, selenium treatment has resulted in marked improvements in fertility in ewes
in regions where WMD and SRU occur[11]. The losses appeared to be associated with higher
embryonic mortality in untreated ewes (24% to 26%) than in treated ewes (2% to 5%)
detected at slaughter 4 to 8 weeks after mating.
In Australia, there are only a few reports of selenium responses in fertility of ewes. Godwin et
al[12] reported increased percentages of fertile ewes and increased percentages of lambs
marked per ewe mated with selenium treatment of ewes on Kangaroo Island (SA) on a
property where WMD had occurred. Wilkins and Kilgour[13] recorded improvements in fertility
in 12 of 14 flocks on 8 properties in the northern Tablelands of NSW following treatment with
5mg selenium orally 3 weeks before joining. In that trial, 9% of treated ewes and 16% of
untreated ewes failed to lamb. Of the ewes mated in the first 6 weeks, 46% of the untreated
ewes and 5% of the treated ewes did not return to service yet failed to lamb. This result
supported the observation from the NZ study reported above and other studies in the
Northern Tablelands of NSW[14] that the failure is one of embryonic loss.
Most attempts to obtain fertility responses to selenium in Australia have failed. For example,
another study in the northern Tablelands of NSW[15] found that selenium supplementation
had no effect on weaning rate at a low stocking rate. There was a positive effect at a high
stocking rate but it was due to higher lamb survival rates, rather than improved fertility.

Vitamin E

Vitamin E deficiency can lead to weaner nutritional myopathy (WNM) which has clinical and
necropsy signs similar to the myopathy associated with selenium deficiency but occurs
despite an apparently adequate selenium status[16]. It has been mainly reported in weaner
sheep grazing dry cereal stubble paddocks in south western Western Australia. Deaths occur
after mustering and yarding or spontaneously in the paddock. Clinical signs included
weakness and a staggering gait, frothing from the mouth and nostrils, inability to stand and
paddling in lateral recumbency. It can occur in sheep which are in satisfactory condition.
Lesions involve skeletal and cardiac musculature[17], particularly muscle groups containing
Type II fibres[18].

Cobalt

Ill-thrift

Sheep which have an inadequate dietary source of cobalt, unless supplemented with
parenteral vitamin B12, develop a syndrome of reduced appetite and weight loss, even in the
presence of otherwise sufficient pasture feed. The wasting is accompanied by lethargy, ocular
discharge, scaly ears and a mild anaemia. Before cobalt deficiency was recognised, there
were areas of New Zealand, the south coast of Western Australia and South Australia and
other regions in other countries, where sheep and cattle could not be grazed without
developing enzootic marasmus, also called bush sickness or coast disease in South Australia.
Affected ruminants became weak and emaciated, progressively anaemic and died. Oral, but
not parenteral, treatment with cobalt was ultimately found to prevent the disease, although
copper was also necessary for complete treatment and prevention of coast disease(1).

White liver disease

Ovine white liver disease occurs in New Zealand[19], Victoria[20], Western Australia[21] and
Tasmania[22]. The disease appears to be a vitamin B12 deficiency complicated by liver
damage. It principally affects lambs 2 to 6 months of age causing severe ill-thrift, hepatopathy,
depression, serous ocular discharge, crusty lesions on the ears probably as a result of
photosensitization, high morbidity with mortality rates of 10% to 15% or, rarely, over 80%(21).
Affected sheep lose weight rapidly. Clinical signs are present for up to 14 days before death.
Vitamin B12 or cobalt therapy leads to rapid improvement except in advanced cases. At
necropsy, the liver is pale and, in some cases, almost white, and very swollen. Lipidosis is
usually present.

Iodine

The presence of goitre - enlarged thyroid glands - in newborn animals, often with high
mortalities, is the characteristic sign of iodine deficiency. The enlargement, however, is often
hard to detect and a careful dissection and weighing of the glands is necessary. Affected
lambs lack vigour and have less than a normal covering of wool. They die readily of cold.

Molybdenum

A clinical deficiency of molybdenum has not been reported in sheep. Toxicity can occur in the
form of a molybdenum-induced copper deficiency.

Zinc

Zinc deficiency may lead to sub-normal growth and fertility, crusty proliferations, cracking of
the skin and loss of hair on the muzzle, vulva, anus, tail-head, ears, back of hindlegs, knee
folds, flank and neck, and a loss of appetite.

Manganese and Iron

Manganese deficiency is rare. Dietary iron deficiency does not occur in grazing ruminants.

Sub-clinical deficiency
Copper

Bodyweight and wool growth depression are likely to occur at marginal levels of copper
deficiency before clinical signs are seen. Reduced growth rate and anaemia is particularly
associated with copper deficiency secondary to high intakes of dietary molybdenum and
sulphur.

Selenium

SRU is effectively the severe and clinically apparent manifestation of a spectrum of disease
which includes, as a mild form, sub-clinical losses of production. The most significant sub-
clinical selenium response is an increase in fleece weight. In an experiment in the northern
Tablelands over 4 years from 1984 to 1987, selenium-supplemented ewes, which had mean
blood selenium concentrations over 0.05μg/ml, had higher liveweights (1 to 2kg higher)
through most of the experiment, higher fleece weights in 3 years (3.8% to 7.5% higher) with
corresponding increases in fibre diameter (0.5μm higher) than unsupplemented ewes (mean
blood selenium concentration around 0.02μg/ml)[23]. Despite these responses, no clinical
signs of deficiency, including depressed fertility, occurred in the ewes and the incidence of
WMD in lambs was very low. Lambs of untreated ewes had lower liveweights at birth and at
weaning, more so at a high stocking rate (3.1kg) than at a low SR (1.7kg) and more
pronounced in lambs of Merino ewes than in lambs of crossbred ewes[24].
In general, responses to selenium are most likely to occur in young animals. Fleece weight
responses of the order of 3% to 8% in WA[25], 4% to 12% in northern NSW[26], 9% and 17%
in the Southern Tablelands of NSW[27] and 14% in Victoria have been reported for sheep
supplemented in their first year of life on commercial farms. Liveweight responses also
occurred in many cases but these are generally of lesser economic significance, unless
severe enough to be associated with SRU. A feature of district trials which include a number
of farms in each district is that responses are not usually detected on all farms, despite blood
selenium concentrations which suggest responses might occur.

Vitamin E

A sub-clinical myopathy in sheep on low vitamin E diets has been identified(18). Lesions in the
muscles are detectable at autopsy but no production responses to treatment have been
reported.

Cobalt

Large body weight and wool growth responses have followed supplementation of lambs with
low vitamin B12 status but which are not showing clinical signs of deficiency.

Zinc

Sub-normal growth and fertility can occur.

Clinical pathology in the investigation of deficiencies

Trace element nutrition, particularly dietary deficiencies, can be investigated by the collection
of a varying range of tissue and soil samples. The choice of the most appropriate samples
depends on the circumstances prompting the investigation and the time of the year it is
performed.

Reasons for using clinical pathology

• When specific clinical signs are evident, such as an outbreak of long bone fractures
or myopathy
• When 'ill-thrift' is suspected or when sheep (particularly young sheep) fail to grow and
produce wool as well as expected
• For routine monitoring, to determine trace element nutritional status of a flock of
sheep; this can detect deficiencies which are completely sub-clinical or which only
occur in occasional years

Seasonal variations in trace element availability

As pasture dries off in late spring, a number of nutritional changes occur, summarised in Table
11-1.

Table 11-1 : Changes in dietary nutrients from spring to summer

Nutritional elements which increase inNutritional elements which decline in

availability after spring availability after spring
Copper Vitamin E
Selenium Crude protein
Cobalt Digestible energy
Iodine Phosphorus
Sulphur

In general, winter to early spring is a good time for routine monitoring or investigations to
determine the probability of occurrence of a significant deficiency of the four trace elements in
the left column of Table 11-1. Certain additional factors, however, should be borne in mind
when planning an investigation.

Copper

Copper nutrition depends on the herbage copper concentration, the amount of herbage
available and the availability of copper from the herbage. The concentration in herbage is
likely to be lowest in winter and it varies between seasons in response to variations in rainfall
and temperature. The amount of herbage available is usually low in winter. The relative
availability of copper from herbage is lower in winter than in summer. For these reasons,
copper deficiency occurs seasonally in southern Australia in the winter period and resolves
itself by summer.
High dietary intakes of Mo, S, Zn, Fe, Cd, Ca and soil all decrease the availability of dietary
copper. Molybdenum application to pastures can exacerbate a copper deficiency. Liming of
pastures can decrease copper availability to plants and animals and increase the availability
of molybdenum. Some of the clinical syndromes associated with low copper intake occur only
when dietary molybdenum levels are high. Sulphur has a highly complex interaction with
molybdenum and a direct effect on copper absorption[28]. Increased dietary sulphur (either
organic sulphur or inorganic sulphate) depresses the absorption of copper and depresses it
more when molybdenum intake is higher. Increasing molybdenum intake has a depressing
effect on copper absorption, an effect which itself is sulphur dependent.

Selenium

Selenium nutrition varies throughout the season with the lowest levels in spring(10) and rising
levels in autumn. Blood selenium concentrations are also inversely related to the previous 12
month rainfall so deficiency syndromes are more likely in wet years. Selenium levels in
pasture are lower in high rainfall areas, possibly as a consequence of leaching, and in
association with particular soil types which tend to be low in minerals, such as granitic soils,
basalt soils and sandy soils. Pasture improvement, particularly the introduction of clovers and
subsequent soil acidification, is an additional risk factor for the depression of pasture
selenium concentrations. Superphosphate application is usually a precursor or concurrent
activity to pasture improvement but may of itself exacerbate marginal selenium availability to
plants by raising soil sulphate levels with consequent decline in the uptake by plants of
selenite salts(9). High stocking rates (leading to lower pasture availability) has also been
associated with decreased blood selenium concentrations.
In improved pastures, clover growth is usually greatest in spring. Clover is similar or lower
than grasses in selenium concentration, is high in PUFA (polyunsaturated fatty acids) and
promotes rapid growth of lambs. Tests on animal tissues performed in late winter will enable
early warning of the risk of selenium-responsive diseases. Tests performed in spring require
interpretation in light of the seasonal variations in selenium nutrition.

Vitamin E

Deficiencies of vitamin E occur over summer and autumn in winter rainfall zones when sheep
have no green feed in their diet. Most fresh green forages contain about 50mg α-tocopherol
per kg DM. Losses occur in sun-curing and hays generally contain 10mg or less per kg.
Cereal grains mostly contain about 8mg/kg DM(3). There is little point in testing sheep which
have a significant intake of green feed.

Cobalt

The seasonal variations in cobalt nutrition are significant. The concentration of cobalt in
pastures and of B12 in plasma is lowest in spring. There are also large fluctuations in the
availability of cobalt between years. Seasons with lush pasture growth favour the
development of cobalt deficiency - perhaps because the sheep ingest less soil. Soil provides
a more concentrated source of cobalt to the ruminant than does pasture[29]. As many cases
of cobalt deficiency syndromes are recognised in spring and summer, early spring is often a
good time to test.

Iodine

There is a marked seasonal variation in intake of iodine. It increases during November (in
Victoria), peaks in summer, declines rapidly within days after autumn rain. Dietary intake
continues to decline during winter. In Victoria, only lambs or kids born between August and
October are susceptible (generally). Seasonal conditions which favour iodine deficiency are
those with early rains, good pasture growth in May and June and rainfall in May, June and
July over 80 mm/month.
Selection of animals for clinical pathology

Copper

Generally, animals of any age can be tested but animals which have grazed dry pasture (over
a summer) are less likely to be deficient than young animals born in late autumn, winter or
spring.

Cobalt

Plasma B12 levels reflect current cobalt nutrition rather than liver stores. Consequently, pre-
ruminant lambs and lambs with recently developed rumens will have low plasma B12 levels
even if liver stores are adequate. By 4 months of age, testing of plasma B12 levels should
indicate cobalt availability and hence whether deficiency is likely to exist or to develop.

Selenium

The animals at risk should be tested. For routine monitoring, lambs 3 to 6 months of age are
suitable.

Selection of tissues for clinical pathology

Copper

When copper absorption exceeds tissue requirements, it accumulates in the liver. When
intake is below requirements, liver copper levels fall. The declining levels increasingly limit
ceruloplasmin synthesis and, eventually, plasma ceruloplasmin concentration declines. Tissue
levels of copper-containing enzymes may, however, remain at functional levels for some
weeks after ceruloplasmin levels start to decline. Measurement of liver copper levels will both
confirm an existing deficiency and enable a prediction of impending deficiency syndromes but
collection of liver samples is not always as convenient as collection of blood samples.
Both plasma copper and plasma ceruloplasmin levels can be measured. Ceruloplasmin is a
copper-protein complex which contains 70% to 90% of plasma copper. Care should be taken
when collecting samples to prevent the formation of a clot - ceruloplasmin levels are variably
lower in serum than in plasma due to sequestration in the clot.
75% of the copper in erythrocytes is in copper-superoxide dismutase (CuSOD). CuSOD
appears to be synthesised at erythrocyte initiation and therefore reflects copper nutrition 2 to
3 months earlier than the time of testing. Compared to plasma ceruloplasmin, CuSOD levels
may give a better indication of the effect that copper deficiency is having at tissue level.
Provided pasture samples are also assayed for molybdenum and sulphate, these can provide
additional useful information about the pathogenesis of a copper deficiency and the likelihood
of recurrence. Soil copper levels, however, are poorly correlated with animal copper nutrition.

Cobalt

Plasma or serum B12 levels are both satisfactory tests of cobalt nutrition. (Note that, for cattle,
liver B12 is preferred because the relationship between cobalt nutrition and plasma levels is
less precise than in sheep.) Soil levels of cobalt provide little or no information about the
likelihood of deficiency in animals.

Selenium

Both blood GSHPx and blood or plasma selenium concentrations give useful indications of
selenium nutrition. Over 95% of blood enzyme activity is associated with erythrocytes and
whole blood samples must be collected with anticoagulant. Selenium is incorporated into
erythrocyte GSHPx at the time of erythropoiesis so measurements of erythrocyte selenium or
GSHPx reflect selenium nutrition at a time up to 2 months in the past. Consequently, plasma
selenium levels are preferable when one wishes to estimate current selenium nutrition.
Plasma concentrations of creatine kinase (CK) and aspartate amino transferase (AST,
formerly SGOT) have been frequently used to assist with the diagnosis of WMD. The
comments below in relation to the diagnosis of WNM with muscle enzymes are probably also
applicable to diagnosis of WMD.
Liver can also be used for testing selenium status. Soil testing is too insensitive to be useful.
Pasture testing is reliable but will not necessarily give any additional information. SRU is likely
to occur when pasture selenium concentrations fall below 0.02mg/kg and are not likely if
levels are above 0.03mg/kg[30]. Responses in fertility cited previously(13) were obtained in
northern NSW where, on one property, pasture selenium levels in March were 0.015mg/kg.

Vitamin E

Both plasma and liver α-tocopherol concentrations provide good indication of vitamin E
nutritional status. Plasma levels of muscle enzymes are frequently used to assist in the
diagnosis of nutritional myopathies. Plasma CK in particular is used in all species because the
enzyme is normally distributed only in muscle. Plasma levels of this enzyme fluctuate widely
in sheep with sub-clinical WNM and the enzyme has a half-life of only a few hours in sheep
and cattle. Consequently, measurement of plasma levels of alanine amino transferase (ALT)
is recommended, alone or in conjunction with CK, in diagnosis of WNM. AST and lactate
dehydrogenase (LDH) are also sensitive indicators of muscle damage but are not specific;
they are also raised whn liver damage is present[31][32].

Interpretation of results

'Normal' levels for trace element concentration in particular animal and plant tissue are
published in a number of sources and it is the usual practice of most laboratories to provide
'normal' and 'marginal' ranges. Rather than remembering particular cut-off levels, one should
try to appreciate that there are generally a large number of factors which influence the
response or lack of response to supplementation with particular trace elements. The results of
tissue testing give useful assistance in determining the risk and likely cost of any clinical or
sub-clinical deficiency.

Copper

• Are levels about to rise without treatment?

• Is deficiency due to a primary copper deficiency or secondary to excess molybdenum
etc? If primary, is problem one of low soil copper or failure by plants to pick it up?
• Is deficiency severe? Will it occur again? If yes, how often?

Cobalt

• Are levels low because lambs are too young, or because cobalt nutrition is low? B 12
injection will not lead to extended maintenance of plasma levels
• Are cobalt pellets in the sheep but not working?
• Are levels low enough to warrant treatment every year or should further testing be
advised?

Selenium

Paynter et al[33] described a relationship between blood GSHPx levels and liveweight
responses to selenium (Appendix B). These authors suggested that GSHPx levels may have
a better predictive value than blood selenium concentrations. As liveweight may be less
responsive than fleece weight, values in the 'marginal' range between 'deficient' and 'normal'
(see Appendix B) will also justify treatment in many situations.

Diagnosis of clinical syndromes

The differential diagnosis of ill-thrift is discussed in the section on weaner management and
that of infertility in the section on reproduction in ewes.

Myopathy
The myopathy associated with vitamin E can be differentiated from that of selenium deficiency
on the history of access to green feed and with the aid of clinical pathology, particularly
estimations of plasma selenium, GSHPx and vitamin E. Lupinosis, although primarily a
hepatotoxicity, has also been associated with a myopathy in sheep in Western Australia
(lupinosis associated myopathy, LAM)[34]. LAM is not preventable with selenium or vitamin E
although it has been proposed that the pathophysiology of the muscle damage is similar to
that of WMD and WNM and at least partly associated with the high PUFA and low vitamin E
and selenium levels in lupin crop residues and lupin grain, as well as the effects of the
phomopsin[35]. LAM has a history of recent exposure to lupin stubbles and jaundice is usually
present in at least some of the flock with similar exposure to phomopsin. At autopsy, varying
degrees of liver damage are detectable either grossly or histologically in sheep with LAM.
Sudden death, while a feature of outbreaks of WMD/WNM, are not so common with LAM(34).
The estimation of muscle and liver enzymes and blood biochemistry can also assist
differentiate between syndromes of myopathy with liver damage and those without. In
lupinosis, liver damage leads to elevation of plasma gamma-glutamyl transpeptidase (GGT)
and bilirubin, both conjugated and unconjugated. In myopathies without liver damage, GGT
and bilirubin are normal, CK and ALT are elevated. Plasma AST is raised with both muscle
and liver damage[36].
Sheep with congenital progressive ovine muscular dystrophy also develop a stiff hind-limb
gait. Affected sheep fail to thrive and their locomotory deficit worsens, so that they are culled
or die of other causes before 2 years of age[37]. The condition is a true dystrophy rather than
a degeneration as in WMD, WNM and LAM and a distinctive feature at post-mortem is the
involvement of the vastus intermedius muscle, which is not involved in the myopathy of
vitamin E deficiency. The disease is rare.
Exertional rhabdomyolysis has also been reported in sheep, following prolonged energetic
chasing or mustering with dogs[38]. Chapman[39] also includes monensin toxicity, poisoning
with Cassia spp and Ixidaena spp, ischaemic myopathy, trauma, snake bite, clostridial
myositis and cereal grain associated myopathy.
Posterior paresis and recumbency due to enzootic ataxia and from spinal cord abscess both
involve neurological deficits of limb placement not seen in skeletal myopathies. Arthritis will
also cause apparent paresis and restricted gait in lambs and weaners.

Response trials

In some ways, a response trial is the only 'proof' that a deficiency exists. There are a number
of problems, however, associated with using response trials for diagnosis of deficiency. Firstly,
failure to demonstrate a response in 1 year does not mean that a response will not occur in
other years. Secondly, trials must be set up and conducted carefully to give meaningful
results and this adds to the cost of diagnosis. Reference levels have been calculated from
trial work so one has to ask if a response trial is in fact an exercise in rediscovering the wheel.

Prevention and treatment

Mineral supplements are frequently provided to sheep in proprietary and home-made licks
and blocks. Blocks are solid 'bricks', approximately 30 cm cubes, of salt, molasses or
composite material in which trace and macro nutrients are mixed. Blocks are placed in
paddocks where sheep graze with the intention that they will lick, nibble or chew the block
and ingest useful quantities of nutrients missing from the pasture feed. Licks are powder
mixtures prepared with the same intention but are 'fed' in drums or troughs or tractor tyres cut
in half to form circular troughs. These preparations are probably ineffective in the face of
significant dietary deficiencies. Commercial blocks produced for widespread sale must
contain very low levels of copper and selenium, for example, because increased consumption
of these elements by sheep in some areas could lead to poisoning. Those products designed
for specific areas and specific degrees of deficiency are, however, potentially useful. A
limitation of their use is the variable intake of sheep within a flock. The subject is further
discussed by McDonald[40]. Other, more direct methods of prophylaxis are discussed below.

Copper

Copper in soil is not particularly subject to leaching losses and if adequate amounts of copper
have been applied to soils (2 kg Cu/ha) there is evidence that the soil will provide adequate
copper for pasture and sheep production for at least 23 years[41]. Where copper
concentrations in pasture approach 10mg/kg and copper deficiency still occurs, application of
additional copper to the soil is not the most appropriate way to increase copper availability to
the grazing animals. In such cases the animals should be treated directly with copper[42].
Oral dosing of copper as oxidised copper wire particles is the most effective method for
prophylaxis of copper deficiency in sheep. The particles, administered in a gelatin capsule
('Cuprax', Pitman-Moore), are retained in the abomasum and release copper slowly through
acid solubilization of the copper. 2.5g cupric oxide raise liver copper levels for 10 weeks and
develop sufficient stores to maintain adequate copper status for a further 20 weeks[43].
Treatment should be timed to ensure satisfactory copper nutrition during the expected period
of low dietary copper (usually winter).
Copper is also administered as an oral drench, either alone or mixed with anthelmintics.
Treatment gives only short-term prophylaxis - perhaps as little as 2 weeks and there is a
danger of poisoning. Addition to water supplies is practised in some areas.

Selenium

Selenium can be supplied to animals in a number of forms. Oral drenches, as sodium selenite
concentrate for addition to anthelmintic drenches or for dilution with water and drenching, are
effective short-term (up to 3 months) therapeutic and preventive treatments. Injectable
selenium, usually included with clostridial vaccine is frequently used to protect lambs at
marking. For adult sheep, intra-ruminal selenium pellets (5% elemental selenium, 95% iron)
are effective in raising and maintaining tissue levels of selenium-containing enzymes,
including GSHPx[44]. Pellets will provide adequate selenium nutrition for 3 to 4 years[45]
although they may need to be accompanied by a steel grinder to prevent coating of the pellet.
Selenium can be applied to deficient pastures as encapsulated selenium (Selcote, Mintech
NZ Ltd). When added to phosphatic fertilizers and applied at the rate of 10g/hectare of
selenium, it is a safe and effective method of supplementation which lasts for over 12
months[46].

Vitamin E

The oral route is probably preferable to parenteral routes of administration of vitamin E,

particularly when treating clinically affected animals. Intramuscular injection of vitamin E in an
oily base has been reported to give longer periods of elevated plasma α-tocopherol in most
field situations than oral administration, suggesting that IM injections are preferable to oral
routes for prophylaxis[47]. Intramuscular injections, however, may cause a local myopathy
and the vitamin become sequestered in regional lymph nodes for extended periods[48].
120 mg/kg liveweight of dl α-tocopherol acetate given orally will adequately raise vitamin E
liver reserves for 2 months but sub-cutaneous treatment will not.

Cobalt

The sub-cutaneous injection of vitamin B12 is recommended for the immediate treatment of
deficient animals and for short-term (up to 3 months) prevention of deficiency. It is the
preferred method of prophylaxis for pre-ruminant lambs and is often administered at marking
time, when lambs are 1 to 8 weeks of age. Intra-ruminal cobalt pellets ('bullets') are the most
efficient method of long-term prevention in endemically deficient areas. They provide
adequate cobalt nutrition for several years, although a small percentage may be regurgitated
and lost. They are usually administered to weaners over 6 months of age or hoggets,
because pre-ruminant (under 10 weeks of age) sheep do not benefit from cobalt and because
larger sheep are easier to dose with pellets. In flocks grazing calcareous soils the pellets
become coated with a calcium phosphate deposit which reduces the rate of cobalt dissolution.
The co-administration of a metallic 'grinder' prevents coating. Oral drenches of cobalt salts
are used but give only short-term protection. Salt licks and mineral blocks may be effective
provided sheep ingest at least 0.05mg of cobalt per day. Addition of cobalt to pastures with
fertilizers and foliar sprays can be useful, particularly when cobalt is being administered as an
aid to preventing phalaris staggers.
Ewes grazing cobalt-deficient pastures should receive cobalt bullets at least 8 weeks before
lambing to ensure adequate colostral vitamin B12 and to provide adequate foetal liver reserves
of the vitamin. Non-colostral milk contains only low levels of vitamin B12[49].

Poisoning

Poisoning with copper is a significant problem in many areas of Australia and supplemental
treatment should not be given without consideration of the risk of poisoning. The subject is
discussed further under diseases of the liver.
Selenium can also be toxic and poisoning can occur naturally (NIA), by treating animals which
already have high selenium stores or by over-dosing. Doses of 15 mg have been fatal in
lambs of 10 kg liveweight. Young lambs may be particularly susceptible to selenium poisoning
due to incomplete rumen development[50]. Signs of acute selenium toxicity include blindness,
abdominal pain, excessive salivation, paralysis and death after 1 to 7 days. In a number of
field trials, high doses of selenium ( > 0.1 mg/kg liveweight) have resulted in negative
responses while, in the same trials, doses of 0.1 mg/kg or less resulted in positive
responses(9,33>).
Allen et al[51] identified 3 therapeutic uses of zinc which could lead to intoxication of sheep
with this element. These are the use of zinc against facial eczema, against lupinosis and in
the treatment of footrot. They reported the death of 19 of 100 treated weaners, 14 within 24
hours of oral treatment with 3g of zinc. At necropsy, there was marked necrosis and a lime
green discolouration of the mucosa of the abomasum and duodenum.

Recommended reading

Hosking, WJ, Caple IW, Halpin CG, Brown AJ, Paynter DI, Conley DN, North-Coombes PL
(1986) Trace elements for pastures and animals in Victoria, Victorian Government Printer, PO
Box 203 North Melbourne 3051
Caple IW (1990) Trace element deficiencies In Sheep Medicine, University of Sydney Post-
graduate Committee in Veterinary Science, Proceedings No 141, p 367
Caple IW (1990) Myopathies: Selenium and Vitamin E In Sheep Medicine, University of
Sydney Post-graduate Committee in Veterinary Science, Proceedings No 141, p 359
Campbell EA (1983) Nutritional Deficiencies and Diseases of Livestock, Australian Agricultural
Health and Quarantine Service/Australian Government Publishing Service, Canberra (Animal
Health in Australia vol 3)

[1] Underwood EJ (1977) Selenium In Trace elements in human and animal nutrition, 4th
edition, Academic Press, New York, p 302
[2] Rice D and Kennedy S (1988) Vitamin E: function and effects of deficiency Br vet J 144 p
482
[3] Putnam ME and Comben N (1987) Vitamin E Vet Rec 121 p 541
[4] Radostits OM, Blood DC and Gay CG (1994) Veterinary Medicine 8th edition, Bailliere
Tindall, New York, p 1415
[5] Ellis TM, Masters HG, Hustas L, Sutherland SS and Evans R (1990) The effect of
selenium supplementation on antibody response to bacterial antigens in Merino sheep with a
low selenium status Aust vet J 67 p 226
[6] Jelinek PD, Ellis T, Wroth RH, Sutherland SS, Masters HG and Petterson DS (1988) The
effect of selenium supplementation on immunity, and the establishment of an experimental
Haemonchus contortus infection, in weaner Merino sheep fed a low selenium diet Aust vet J
65 p 214
[7] Grant AB, Drake C and Hartley WJ (1960) Further observations on white muscle disease
in lambs NZ vet J 8 p 1
[8] Andrews ED, Hartley WJ and Grant AB (1968) Selenium-responsive diseases of animals
in New Zealand NZ vet J 16 p 3
[9] McDonald JW (1975) Selenium responsive unthriftiness of young Merino sheep in central
Victoria Aust vet J 51 p 433
[10] Caple IW, Andrewartha KA, Edwards SJA and Halpin CG (1980) An examination of the
selenium nutrition of sheep in Victoria Aust vet J 56 p 160
[11] Hartley WJ (1963) Selenium and ewe fertility Proc NZ Soc Anim Prod 23 p 20
[12] Godwin KO, Kuchel RE and Buckley RA (1970) The effect of selenium on infertility in
ewes grazing improved pastures Aust J exp Agric Anim Husb 10 p 672
[13. Wilkins JF and Kilgour RJ (1982) Production responses in selenium supplemented sheep
in northern New South Wales 1. Infertility in ewes and associated production Aust J Exp Agric
Anim Husb 22 p 18
[14] Piper LR, Bindon BM, Wilkins JF, Cox, RJ, Curtis YM and Cheers MA (1980) The effect
of selenium treatment on the fertility of Merino sheep Proc Aust Soc Anim Prod 13 p 241
[15] Langlands JP, Donald GE, Bowles JE and Smith AJ (1991) Subclinical selenium
insufficiency 2. The response in reproductive performance of grazing ewes supplemented
with selenium Aust J exp Agric 31 p 33
[16] Steele P, Peet RL, Skirrow S, Hopkinson W and Masters HG (1980) Low alpha-
tocopherol levels in livers of weaner sheep with nutritional myopathy Aust vet J 56 p 529
[17] Allen JG and Steele P (1980) Distribution of lesions in ovine weaner nutritional myopathy
in Western Australia (Aust vet J 56 p 560
[18] Allen JG, Steele P, Masters HG and D'Antuonon MF (1986) A study of nutritional
myopathy in weaner sheep Aust vet J 63 p 8
[19] Sutherland RJ, Cordes DO and Carthew GC (1979) Ovine white liver disease - an
hepatic dysfunction associated with vitamin B12 deficiency NZ vet J 27 p 227
[20] Mitchel PJ, McOrist S, Thomas KW and McCausland IP (1982) White liver disease of
sheep Aust vet J 58 p 181
[21] Richards RB and Harrison MR (1981) White liver disease in lambs Aust vet J 57 p 565
[22] Mason RW and McKay R (1983) Ovine white liver disease Aust vet J 60 p 219
[23] Langlands JP, Donald GE, Bowles JE and Smith AJ (1991) Subclinical selenium
insufficiency 1. Selenium status and the response in liveweight and wool production of
grazing ewes supplemented with selenium Aust J exp Agric 31 p 25
[24] Langlands JP, Donald GE, Bowles JE and Smith AJ (1991) Subclinical selenium
insufficiency 3. The selenium status and productivity of lambs born to ewes supplemented
with selenium Aust J exp Agric 31 p 37
[25] Gabbedy BJ (1971) Effect of selenium on wool production, body weight and mortality of
young sheep in Western Australia Aust vet J 47 p 318
[26] Wilkins JF and Kilgour RJ (1982) Production responses in selenium supplemented sheep
in northern New South Wales 2. Liveweight gain, wool production and reproductive
performance in young Merino ewes given selenium and copper supplements Aust J Exp Agric
Anim Husb 22 p 24
[27] Dove H, Axelsen A and Watt R (1986) Selenium responses in grazing ewes and their
lambs Proc Aust Soc Anim Prod 16 p 187
[28] ARC (Agricultural Research Council) (1980) Trace elements In The Nutrient
Requirements of Ruminant Livestock. Commonwealth Agricultural Bureaux, Farnham Royal,
Slough, England.
[29] Hosking, WJ, Caple IW, Halpin CG, Brown AJ, Paynter DI, Conley DN, North-Coombes
PL (1986) Trace elements for pastures and animals in Victoria, Victorian Government Printer,
Melbourne
[30] Andrews ED, Hartley WJ and Grant AB (1968) Selenium responsive diseases of animals
in New Zealand NZ vet J 16 p 3
[31] Fry JM, Allen JG, Speijers EJ and Roberts WD (1994) Muscle enzymes in the diagnosis
of ovine weaner nutritional myopathy Aust vet J 71 p 146
[32] Smith GM, Fry JM, Allen JG and Costa ND (994) Plasma indicators of muscle damage in
a model of nutritional myopathy in weaner sheep Aust vet J 71 p 12
[33] Paynter DI, Anderson JW and McDonald JW (1979) Glutathione peroxidase and
selenium in sheep II The relationship between glutathione peroxidase and selenium-
responsive unthriftiness in Merino lambs Aust J Agric Res 30 p 703
[34] Allen JG (1978) The emergence of a lupinosis-associated myopathy in sheep in Western
Australia Aust vet J 54 p 548
[35] Allen JG, Steele P, Masters HG and Lambe WJ (1992) A lupinosis-associated myopathy
in sheep and the effectiveness of treatments to prevent it Aust vet J 69 p 75
[36] Allen JG and Randell AG (1993) The clinical biochemistry of experimentally produced
lupinosis in the sheep Aust vet J 70 p 283
[37] Dent AC and Richards RB (1979) Congenital progressive ovine muscular dystrophy in
Western Australia Aust vet J 55 p 297
[38] Peet RL, Dickson J, Masters H and Johnston J (1980) Exertional rhabdomyolosis in
sheep Aust vet J 56 p 155
[39] Chapman HM (1990) Myopathies that do not respond to selenium In Sheep Medicine,
University of Sydney Post-graduate Committee in Veterinary Science, Proceedings No 141,
p 483
[40] McDonald JW (1983) Mineral licks In Sheep Production and Preventive Medicine,
University of Sydney Post-graduate Committee in Veterinary Science, Proceedings No 67, p
283
[41] Hannam RJ, Judson GJ, Reuter DJ, McLaren LD and McFarlane JD (1982) Current
requirements of copper for pasture and sheep on sandy soils in the upper south-east of South
Australia Aust J exp Agric Anim Husb 22 p 324
[42] McFarlane JD, Judson GJ and Gouzos J (1990) Copper deficiency in ruminants in the
south east of South Australia Aust J exp Agriculture 30 p 187
[43] Judson GJ, Brown TH, Gray D, Dewey DW, Edwards JB and McFarlane JD (1982)
Oxidised copper wire particles for copper therapy in sheep Aust J Agric Res 33 p 1073
[44] Paynter DI (1979) Glutathione peroxidase and selenium in sheep. I Effect of intraruminal
selenium pellets on tissue glutathione peroxidase activities Aust J Agric Res 30 p 695
[45] Judson GJ, Ellis NJS, Kempe BR and Shallow M (1991) Long-acting selenium
treatments for sheep Aust vet J 68 p 263
[46] Halpin CG, Hanrahan P and McDonald JW (1987) Selenium fertilizer for the control of
selenium deficiency in grazing sheep In Temperate Pastures: Their production, use and
management edited by JL Wheeler, CJ Pearson and GE Robards, Australian Wool
Corporation-CSIRO, Melbourne, p 386
[47] Doncon GH and Steele P (1988) Plasma and liver concentrations of α-tocopherol in
weaner sheep after vitamin E supplementation Aust vet J 65 p 210
[48] Dickson J, Hopkins DL and Doncon GH (1986) Muscle damage associated with
injections of vitamin E in sheep Aust Vet J 63 p 231
[49] Caple IW and McDonald JW (1983) Trace element nutrition In Sheep Production and
Preventive Medicine, University of Sydney Post-graduate Committee in Veterinary Science,
Proceedings No 67, p 236
[50] Lambourne DA and Mason RW (1969) Mortality in lambs following overdosing with
sodium selenite Aust Vet J 45 208
[51] Allen JG, Morcombe PW, Masters HG, Petterson DS and Robertson TA (1986) Acute
zinc toxicity in sheep Aust vet J 63 p 93