Vet Pathol 44:342–354 (2007)

Chondrodysplastic Calves in Northeast Victoria

P. J. MCLAREN, J. G. CAVE, E. M. PARKER, AND R. F. SLOCOMBE
Gribbles Veterinary Pathology, Clayton, Victoria, Australia (PJM); Department of Primary Industries,
Wodonga, Victoria, Australia (JGC); Department of Primary Industries, Leongatha, Victoria, Australia
(EMP); and University of Melbourne Veterinary Clinical Centre, Werribee, Victoria, Australia (RFS)

Abstract. Outbreaks of chondrodysplasia in calves occur sporadically every 10–15 years, particularly
following prolonged drought conditions, throughout Northeastern Victoria and the Southern Table-
lands of New South Wales, Australia. An outbreak spanning 2 calving seasons (2003–2004) involving
numerous losses through stillbirth, perinatal loss, and poor growth was investigated. Investigations of 4
representative cases are presented here with a definition of the gross and histopathologic defects and an
overview of epidemiologic data gathered from affected farms. Calves showed variable disproportionate
dwarfism without arthrogryposis. Long bones were shortened and showed axial rotation. Articular
surfaces were distorted with misshapen weight-bearing surfaces associated with variable thickness of
articular cartilage. Physes were distorted and variable in thickness with occasional foci of complete
closure. The major histologic abnormality in the physes was disorderly development of the zones of
cartilage hypertrophy, with reduced number and irregular arrangement of hypertrophic chondrocytes;
similar less severe changes were present in the zones of cartilage proliferation. Histochemical staining of
the cartilage matrix was variable in intensity, and there was evidence of abnormal resorption of cartilage
matrix at the level of the primary spongiosa. Osteoid formation and subsequent bone remodelling
seemed unaffected, and diaphyseal cortical bone appeared normal at the gross and light microscopic
level. No infectious agents were identified, and other known causes for chondrodysplasia in calves were
excluded. The most likely cause for the syndrome was considered to be congenital manganese deficiency.
Further surveys of tissue and blood manganese levels from cows and calves with and without clinical
signs from the region are planned.

Key words: Bovine; chondrodysplasia; deficiency; disproportionate; drought; dwarf; manganese;

skeleton.

Sporadic congenital deformities are expected in
the bovine population. Outbreaks of higher in-
cidence of congenital disease such as that described
here may be associated with considerable losses of
animals and consequent financial loss. Across
Northeastern Victoria and the Southern Tablelands
of New South Wales (NSW), Australia, outbreaks
of chondrodysplastic disease have been reported,
particularly in years following prolonged drought
conditions. Outbreaks occurring in 2 consecutive
spring calving seasons (July to November, 2003
and 2004) and involving numerous losses through
stillbirth, perinatal loss, and poor growth were
investigated to exclude potential infectious disease
and to define the gross and histopathologic
defects. The cause of the syndrome is still un-
resolved, although congenital manganese deficien-
cy is suspected; despite marked improvement in the
climatic conditions in this area, a lesser number of
farms saw deformities among calves in a third
consecutive calving season (2005). Investigations
are ongoing.
342
Congenital disease may result from genetic
abnormality, infectious disease, or other in utero
insults. ‘‘Chondrodysplasia’’ is a general term for
abnormalities of cartilage, usually caused by de-
fective nutrition or metabolism of cartilage, which
results in disproportionate dwarfism.19
Genetic chondrodysplastic syndromes are re-
ported in Holstein, Dexter, and several other
breeds of cattle. Beef breeds selected for short
stocky phenotype have an increased prevalence of
brachycephalic calves, with longitudinal compres-
sion of the vertebrae and shortening of the distal
long bones, believed to be hypochondroplasia
rather than a dysfunction of endochondral ossifi-
cation.19
There are many examples of toxic and nutrition-
al causes of musculoskeletal deformity. Hypervita-
minosis A causes vertebral abnormalities and may
be compounded by protein-energy malnutrition.5
‘‘Crooked calves’’ are caused by ingestion of
certain Lupinus sp. containing toxic concentrations
of the quinolizidine alkaloid, anagyrine, by the
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria
dam, particularly between 40 and 70 days of
gestation.23 This leads to arthrogryposis, spinal
deformities, and disordered long bone growth,
possibly due to reduced intrauterine motility either
through tonic contraction of the uterus or sup-
pressed motility and apparent sedation of the
fetus,3 leading to a functional arthrogryposis
without identifiable histologic or histochemical
lesions in the bone or muscle.1 Many other plant-
associated toxins are implicated in skeletal disease,
for example teratogenic piperidine alkaloids in
Astragalus sp., Oxytropis sp., and Nicotiana glauca,
which may act in a similar manner.3 Fungal toxins
have been presumptively associated with outbreaks
of congenital spinal stenosis and disproportionate
dwarfism, leading to myelomalacia and posterior
paralysis in 3 outbreaks in Western Canada, where
heavy growth of Penicillium sp. and Fusarium sp. in
wet straw bales was also causally associated with
pruritus and mortality in dams.20 ‘‘Acorn calves,’’
described in western regions of North America and
also recorded in New Zealand and South Africa,
were associated in the original reports with in-
gestion of acorns.2,19 Affected calves showed joint
laxity, varus, and valgus deformities, and dispro-
portionate dwarfism. More recent work has sug-
gested that some of these cases may be consistent
with a deficiency in dietary intake of manganese, as
supported by small feeding trials in cattle.6,7,10,22
Infectious causes of skeletal deformities seen in
cattle in Australia include bovine viral diarrhea
virus (BVDV), Akabane and Aino viruses. These
tend to cause arthrogrypotic deformity of the
skeleton rather than chondrodysplasia.
Consideration and, to a large part, elimination of
these differential diagnoses combined with evalua-
tion of the anatomic pathology of several cases led
investigators to propose that deficiency of manga-
nese could be responsible for the abnormalities
in this outbreak. Manganese deficiency was sus-
pected, although not conclusively proven, in several
previous field outbreaks of skeletal disease in
calves.24,28
This article presents the gross and histopatho-
logic features of 2 outbreaks of congenital disease
in calves, defines the disease, and discusses the
potential relationship with manganese deficiency.
Methods
Veterinary field staff and epidemiologists per-
formed clinical investigations with visits to numer-
ous affected farms and epidemiologic data collec-
tion through analysis of questionnaires completed
by farmers. The case definition formulated among
the investigative group was as follows: ‘‘Birth of
343
abnormally short, stunted, or deformed calves with
a spectrum of axial and appendicular skeletal
defects predominantly of reduced long bone/di-
aphysis length, angular rotations, irregular growth
plates, and articular cartilages, but without signif-
icant abnormality of cortical bone, born to dams
without clinical signs in the area of Northeast
Victoria and Southwest New South Wales.’’
Portions of vertebrae and long bones from calf
No. 1, a stillborn calf from a farm in Northern
Victoria, were received fixed in 10% neutral
buffered formalin (NBF) and examined grossly
and histopathologically. Calf No. 2, one live 21-
day-old calf from a second farm, and calves Nos. 3
and 4, entire stillborn calves from a third farm,
underwent full postmortem and histologic exami-
nation using standard methods. Carcasses of calves
Nos. 2–4 were weighed, and skeletal components of
calves Nos. 3 and 4 were recorded using standard
measuring tapes. Radiographic analysis of intact
demuscled limb bones and sections through verte-
brae were prepared using a band saw, after fixation
but before decalcification, on calves Nos. 3 and 4
(not presented).
Five-millimeter sections were taken from a wide
range of axial and appendicular skeletal sites and
fixed in 10% NBF for 24–48 hours before de-
calcification, using a standard aqueous solution of
18% formic acid and 3.5% sodium formate for
approximately 1 week at room temperature with
continuous slow agitation. Following standard
paraffin embedding, 4-mm sections were cut and
stained with hematoxylin and eosin (HE). Histo-
chemical stains used to evaluate cartilage and bone
included Giemsa and Acid Giemsa, periodic acid–
Schiff (PAS), Alcian blue, Alcian blue–PAS,
toluidine blue, safranin O/fast green,15 and Ver-
hoeff elastic stain.
For comparison of bone measurements and
histopathology, 20 femora of grossly normal young
calves randomly collected by staff at an abattoir in
an unaffected region were weighed and measured,
and histologic sections were prepared from the
distal physis of each. Age and sex matching was not
possible in this investigation.
Fresh frozen liver was collected from calves
Nos. 3 and 4. Glutathione peroxidase was analyzed
using an enzymatic method according to the
Standing Committee on Agriculture, Common-
wealth Scientific and Industrial Research Organ,
Australia, 1993. Vitamin B12 was analyzed spec-
trophotometrically with competitive binding assay,
and copper was analyzed by atomic absorption
spectrophotometry. Manganese concentration was
determined following wet combustion in perchlo-
344 McLaren, Cave, Parker, and Slocombe Vet Pathol 44:3, 2007

ric/nitric acid solution and dilution in dilute
hydrochloric acid to correct volume by flame
atomic absorption spectrophotometry at 279.5 nm
on an AAnalayst 300 Spectrometer (PerkinElmer,
Waltham, MA). The reference ranges available for
concentration of each of these elements are based
on adult cattle, and no reference range for neonates
was available at the start of the study.
Agar gel immunodiffusion was performed on
serum samples from calves Nos. 2–4 and several
dams and live calves from affected farms to detect
BVDV and Akabane virus antigen. Virus neutral-
ization was performed to detect Aino virus antigen.
The National Arbovirus Monitoring Program of
Animal Health Australia was consulted to assess
presence of insect vectors and serologic evidence of
arboviruses in the region during the periods of
gestation (www.namp.com.au/reports/report_0203.
pdf, 2005).
Results
Many farms across Northeastern Victoria expe-
rienced births of deformed calves during the spring
(July to November) of 2003 and 2004, common
calving periods for this region. Although some
farms reported births of marginally increased
numbers of mildly chondrodysplastic calves in
preceding years, exceeding the expected number
of sporadic cases, the outbreaks in 2003–2004 were
widespread with significant portions (#90%) of
cohort groups affected. The spectrum of severity
ranged from stillbirths and perinatal deaths (due to
inability to stand and respiratory compromise) to
calves deformed but able to suckle and follow the
dam. Less-affected calves were vigorous and
seemed to show some clinical reduction of defor-
mities over time. Total reported losses due to high
morbidity and mortality numbered into hundreds
of calves; given economic conditions in this region
that had suffered severe drought for several years,
unreported losses were expected to approximate
1,000 calves.
Dams of all age groups produced deformed
calves. Affected calves appeared to be clustered in
management cohort groups rather than showing
a pattern of spread expected with an infectious
agent. The groups of dams giving birth to affected
calves had predominantly been at pasture on hilly
granitic country that had been extremely dry over
the majority of each gestation period, although not
all dams were in poor body condition. No toxic
plants were found on repeated examination of the
grazing areas or reported during careful question-
ing of farmers. Some farms had been blanketed in
heavy bushfire smoke during the first gestation
period (2003), but there did not appear to be an
association between density of smoke and the
affected cohort groups.
Common supplemental food, medication, or
management procedures, and genetic links were
not identified during clinical investigation and
epidemiologic study. The calves affected were
a range of beef breeds and crosses, including
Murray Grey, Hereford, and Angus. Numerous
sires were involved, and cows giving birth to
deformed calves had previously produced normal
calves and, in many cases, went on to produce
normal calves in the following season. One mildly
affected heifer calf was observed to produce
a normal calf at her first calving.
Gross pathology
External examination of the calves revealed
similar deformities in all calves presented intact.
All had low body weight (Table 1) and short
stature, with moderate-to-marked limb-trunk dis-

Table 1. Selected individual and average measurements of skeletal elements of calves Nos. 2–4.

No. 2 No. 3 No. 4 Average

Body weight (kg) 25 30 25 26.7
Head length (cm) 23.5 25 20.5 23
Poll–anus length (cm) –* 81.5 76 78.8
Right Left
Metacarpal diameter (cm) 1.9 1.6 – 1.7
Metacarpal length (cm) 11.8 11 11 – 11.3
Metacarpal total length/diaphyseal diameter 6.4 6.9 – – 6.5
Ulna length (cm) 18 16.7 15 – 16.6
Humerus length (cm) 13 11 11.5 – 11.8
Femur length (cm) 15.7 14 14.5 – 14.7
Tibia length (cm) 15.5 14.5 14 – 14.7
Metatarsal length (cm) – 13 11.3 – 12.2
* – 5 not available.
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria 345

Fig. 1. Carcass, calf No. 4. Disproportionate dwarf-

ism, kyphosis, domed skull, and supinatory rotation of
distal limbs are seen.

proportion (Fig. 1). The heads appeared slightly

domed in calves Nos. 2–4 (Fig. 1). There was
variable brachygnathism, and calves Nos. 3 and 4
showed protrusion of the tongue with oversize
relative to the oral cavity.
Limb deformities varied in severity, with the live
21-day-old calf (calf No. 2) showing a bilateral
valgus deformity of the forelimbs, over-extension
of the fetlocks, disproportionate shortening
(length-to-epiphyseal diameter) predominately of
proximal limb bones, and reduced range of
movement of most joints. Calves Nos. 3 and 4,
periparturient mortalities, showed more severe
deformities with moderate-to-marked dispropor-
tionate shortening of proximal limb bones and
supinatory rotation of the distal forelimbs and, to
a lesser extent, the hind limbs (Fig. 2). This
appeared to result from distortion of the diaphyseal
shafts of the long bones rather than from abnormal
direction of joint movement. The joints did not
show significant laxity or fixation (arthrogryposis),
and muscle mass appeared normal relative to the
size of the calves.
The vertebral columns showed variable kyphosis
and lordosis. The atlas was irregularly distorted
with increased concavity of the cranial and caudal
articular surfaces. Growth plates of the vertebrae
appeared distorted and showed variable thickness.
Irregular interlacing trabeculae of cartilage extend-
ing between the cranial and caudal vertebral
growth plates traversing the vertebral bodies were
noted in all calves with variable frequency and
extent along the vertebral column (Fig. 3). The
significance of these changes is uncertain since the
authors have not identified studies characterizing
morphology of the bovine neonate spinal column.
The shape of several of the vertebral bodies
appeared distorted in calves Nos. 2–4, with cranial
Fig. 2. Midsagittal section of forelimb; calf No. 4.
Distortion of proximal limb bones and irregular growth
plates are noted.
concavity and caudal convexity of articular sur-
faces and loss of the central waist normally seen in
the vertebral body (Fig. 3). Compression of the
spinal cord was not evident except in calf No. 1,
where the spinal canal was virtually occluded in the
single cervical vertebra (atlas) presented for exam-
ination. Changes similar to those of the vertebrae
were also identified in basicranial bones; distortion
of basicranial bones appeared to lead to shortening
of the ventral cranial vault and thus skull doming.
Calf No. 4 showed a depression fracture of the
cranial vault, fracture and displacement of the
basicranial bones with hemorrhage before death,
considered consistent with trauma shortly after
birth (e.g., trampling by the dam).
Examination of the limbs revealed longitudinal
shortening of the long bones, most marked in the
proximal limbs, with the distal limbs below the
metacarpus and the metatarsus relatively less
affected. The scapulae were normal in shape and
346 McLaren, Cave, Parker, and Slocombe Vet Pathol 44:3, 2007

Fig. 3. Midsagittal section of cervicothoracic ver-

tebral column; calf No. 4. D-shaped distorted vertebral
bodies are seen.

proportionate to body size. On longitudinal sec-

tion, numerous long bones revealed undulating
irregularities of the physes. These were thin and
irregularly wavy, with several stepped graduations
in thickness across the growth plate (Figs. 4, 5). In
calves Nos. 2–4, variably prominent growth-arrest
lines were visible in the metaphyses and epiphyses
(Fig. 5).

Fig. 5. Midsagittal section of humerus; calf No. 3.

Marked longitudinal shortening and growth-arrest lines
in the metaphyses and proximal epiphysis (arrow)
are depicted.

The humeri showed marked bilaterally symmet-

Fig. 4. Midsagittal section of hindlimb; calf No. 4.
Note prominent distortion with irregularity of growth
plates of the femur and tibia.
ric diaphyseal shortening with mild outward
(supinatory) rotation of the distal epiphysis. The
elbows showed prominent lateral ridges and lateral
displacement of the axial ridges of the radial
articular surfaces, the radius and ulna were bowed,
and the distal articulation was markedly irregular
with numerous articular facets corresponding with
the markedly curving distorted proximal carpal
bones. All carpal bones were abnormal, misshapen,
and difficult to classify (Fig. 6).
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria
Fig. 6. Carpal joint; calf No. 3. The carpus is
opened to reveal markedly distorted and irregular
articular facets leading to abnormally reduced range of
movement without arthrogryposis.
Hind limbs were similarly deformed, with the
femoral and tibial bones showing markedly de-
creased diaphyseal length and proportionally en-
larged epiphyses similar to that of the forelimbs.
The femora showed shortening and marked distal
outward rotational deformity, with flattened troch-
lear ridges leading to laterally displaced patellae in
the severely affected calf No. 2 and laterally placed
patellae remaining within the trochlear groove in
calf No. 4. The distal femoral articular surfaces
were asymmetrically distorted. The tibial crest was
spirally rotated from the proximal surface, with
medial elevation of the tibial plateau. The diaphysis
of the tibia was medially bowed, and the distal
epiphysis was rotated inwardly (pronated), partial-
ly reversing the effect of femoral supination on the
limb as a whole in calves Nos. 3 and 4.
The articular cartilages of all cases showed
irregular thickness, with some areas of erosion
and prominent subchondral vessels. There was
variable multifocal lifting and separation of the
articular cartilages in calf No. 2, with suppurative
polyarthritis and osteomyelitis of the proximal
tibia.
Only skeletal elements were examined in calf
No. 1; abnormalities of nonskeletal elements were
limited to hypoplasia and lateral flattening of the
tracheal cartilage rings and extensive primary
atelectasis, with only small areas of inflation of
the lungs in calves Nos. 3 and 4.
Morphometric analysis
Measurements of skeletal components of calves
Nos. 3 and 4 are presented (Table 1) and compared
347
Table 2. Average length of 20 femora sourced from
normal calves at an abattoir in a nonaffected region
of Victoria.
Femoral Length
No. of Calves Mean Range
20 20.6 18.7–23.0
with measurements of the femora of young but not
age-matched calves from an abattoir (Table 2).
Previously measured skeletal elements of calves are
compared (Table 3).
The Student’s t-test was used to compare the
mean length of the femora from affected animals
(group A) with that of the femora from normal
animals (group N). There was a difference between
the mean length of the femora from group A
(14.7 cm) and the mean length of femora from
group N (20.6 cm) (P , .001). The assumption of
independence of observations may not be satisfied
because femora were randomly selected after
boning and may represent femora from between
10 and 20 animals in the N group. Poll–anus length
(81.5 cm) and eye–anus length (90 cm) for calf
No. 3 (closest to the smallest body weight group in
the referenced data) are less than those of average
calves in this source (90.3 and 100.8 cm, respec-
tively).18,25 However, variables including breed, sex
and regional variations in average neonatal calf
weight limit interpretation.
Histopathologic findings
The cases all showed widespread physeal and
metaphyseal abnormalities of varying degrees of
severity. At the physis, the zone of cartilage
hypertrophy was irregularly reduced, consisting of
disorderly columns varying markedly in number
and alignment along the growth plate, often with
only 3–6 cells (Fig. 7), compared with neat col-
umns of 10–20 cells in control animals (Fig. 8). The
zones of resting and proliferating cartilage ap-
peared relatively normal, although small multifocal
nodules of increased numbers of proliferative
chondrocytes were seen in each case. Multifocal
cystic spaces were present both within the zone of
hypertrophy and interspersed between the degen-
erate/calcifying chondrocytes of the zone of miner-
alization, suggestive of failure of formation of
matrix or premature loss of chondrocytes and
matrix. These cysts showed basophilic granular-to-
fibrillar staining with toluidine blue and PAS,
suggesting abnormal cartilage matrix filling the
spaces (Fig. 9). A high rate of osteoclastic activity
was indicated in these areas by the presence of
348 McLaren, Cave, Parker, and Slocombe
Table 3. Selected average skeletal measurements of normal neonatal beef calves.18,25
Measurement
Body weight (kg)*
Head length (cm)*
Poll–anus length (cm)*
Metacarpal total length/
diaphyseal diameter{
* 5 from Mee 199518
{ 5 from Tyler et al. 196125
abundant osteoclasts. Trabeculae within the pri-
mary spongiosa were short and irregular; a low
proportion showed evidence of fracture close to the
physis, with osteoclasts present at the blunt distal
ends of trabeculae (Fig. 7).
The abnormalities of the cartilage matrix were
most clearly demonstrated by variable granular
Alcian blue staining, compared with that of
controls (Figs. 10, 11).
The epiphyseal surface of the physis of the long
bones examined in calf No. 1 showed development
of a band of mature bone horizontal and adjacent
to the physis (terminal plate), indicating closure of
the growth plate (Fig. 12). The fused epiphyseal
and articular cartilages were apparently less affect-
ed. Horizontal metaphyseal trabeculae of bone
(growth-arrest lines suggesting periods of reduced
dam nutritional status) were found running parallel
to and at a short distance from the physes of some
bones in all cases (Fig. 5).
Fig. 7. Metaphyseal aspect of physis, femur; calf
No. 2. Reduced number and irregular arrangement of
hypertrophic chondrocytes, presence of osteoclasts at
abnormally truncated primary spongiosa, and irregular
resorption of cartilage are seen. HE. Bar 5 100 mm.
Vet Pathol 44:3, 2007
Range
31–33 41–43 53–63
Mean (SD)
25.9 (0.89) 27.4 (0.85) 29.7 (1.46)
90.3 (2.16) 94.3 (2.81) 107.1 (3.5)
Mean (range)
6.25 (4.7–7.5)
Sections of many long bones examined from all
cases showed variable osteopenia, with a reduced
quantity of trabecular bone in the epiphysis and
medulla. Cortical lamellar bone was normal in
histologic appearance as was the perichondrial
collar, a lamellar plate of bone surrounding the
physis.
Sections of vertebrae examined from each animal
showed irregular outlines of the cranial and caudal
physes due to variation in depth of both cranial
and caudal aspects of the zones of hypertrophy and
mineralization (Fig. 13).
Sections of mandible demonstrated decreased
medullary trabecular bone, but lamellar cortical
bone was within normal limits. The bones lining
the nasal cavity and turbinates were composed of
fine irregular spicules of bone showing high
cellularity, numerous irregular growth-arrest lines,
and irregular scalloped surfaces lined with reduced
osteoblast populations.
In calf No. 1, the atlas showed a pathologic in
utero fracture of a focus of ectopic cartilage,
disorderly proliferation of new bone, and cartilage
Fig. 8. Metaphyseal aspect of physis, femur; neo-
natal calf control. Neatly aligned columns of hypertro-
phic chondrocytes and long primary spongiosa are
compared with Fig. 7. HE. Bar 5 100 mm.
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria
Fig. 9. Metaphyseal aspect of physis, femur; calf
No. 2. Irregular mineralization of truncated spongiosa,
cystic space filled with faintly granular mucinous
material, and irregular granularity of chondrocyte
cytoplasm are shown. Periodic acid–Schiff. Bar 5
100 mm.
with altered staining characteristics and increased
cellularity and proliferation of the periosteum. This
caused distortion and compression of the spinal
canal and cord.
The proximal tibial physis of calf No. 2 had
bacterial osteomyelitis. A necrotic cartilage seques-
trum was surrounded by clusters of degenerate
neutrophils grouped around colonies of small
coccobacilli and associated with dense infiltrates
of lymphocytes, plasma cells, and macrophages and
a thick rim of fibroblastic connective tissue
bridging the physis, thus causing pathologic
fracture. In this case, lymph nodes taken from
Fig. 10. Physis, femur; calf No. 4. Variably dense
and granular staining of physis is seen. Alcian blue. Bar
5 100 mm.
349
Fig. 11. Physis, femur; neonatal calf control. Com-
pare regularity of staining and physeal structure with
Fig. 10. Alcian blue. Bar 5 100 mm.
mesenteric and prescapular regions all showed
a mild hypertrophy of follicles, consistent with
excessive immune stimulation; however, viscera
and neurologic tissues showed no significant
changes. Microbial cultures were not performed.
Mineral analysis
In calves Nos. 3 and 4, portions of fresh frozen
liver were submitted for mineral analysis (Table 4).
The manganese concentrations detected in these
unsuckled neonates were below the normal refer-
ence range; however, since this range was de-
termined for adult cattle, further analysis is re-
quired to assess the range for neonatal calves
before reliable interpretation can be made.
Fig. 12. Epiphyseal aspect of physis, femur; calf
No. 1. Formation of a terminal plate of osteoid
indicates closure of the epiphyseal aspect of the physis.
HE. Bar 5 100 mm.
350 McLaren, Cave, Parker, and Slocombe
Fig. 13. Vertebral end plate physis; calf No. 2.
Irregularity of the physis and cartilage cores are present
within the vertebral body. HE. Bar 5 1,000 mm.
Virology
Serologic analysis for BVDV, Akabane and Aino
viruses performed on these calves was negative as
was that from numerous other animals from herds
affected during the outbreak (data not presented).
The National Arbovirus Monitoring Program did
not report any seropositive animal or insect vector
detection in the affected regions during the
gestation periods.
Discussion
These cases showed sufficient gross and histo-
logic similarity and distinction from other reported
chondrodysplastic syndromes to constitute a recog-
nizable clinical and pathologic entity. Disorder of
the zone of cartilage hypertrophy with abnormal
growth and resorption of cartilage, particularly at
the physes of long bones and vertebrae, the
basicranial and turbinate bones, but excluding
facial and mandibular bones, was observed. Gross
and histologic features suggest a chondrodysplastic
disorder confined to bones of mesodermal and
sclerotomal origin and not affecting bones derived
Table 4. Liver manganese, copper, cobalt, and selenium analysis from calves Nos. 3 and 4.
No. Manganese* Copper{
3 22 1.48
4 24 1.37
Reference 46–109 0.08–2.09
* mmol/kg wet weight.
{ mmol/kg wet weight.
{ U/g wet weight.
Vet Pathol 44:3, 2007
from the neural crest. The presence of abnormal-
ities of the physis not affecting the perichondrial
collar and diaphyseal shaft suggests the disorder is
limited to bone formed through interstitial growth
from cartilage and by endochondral ossification
but not by appositional growth, consistent with
chondrodysplasia.
The common causes of chondrodysplasia in-
cluding genetic defects, most toxic and nutritional
causes, and infectious agents were excluded se-
quentially by epidemiologic and pathologic in-
vestigation.
In the affected herds in this outbreak, no familial
link could be established, and the range of breeds
and crosses affected was felt to preclude any
possibility of a genetic etiology.
Vitamin A supplementation was rarely per-
formed; therefore, hypervitaminosis A was exclud-
ed. Many farms with diverse management practices
were affected; therefore, disease related to man-
agement procedures or therapeutic product admin-
istration was considered unlikely.
Pastures had been bare for much of the gestation
period in these outbreaks. While there was no
known access to any toxic plants, under conditions
of undernutrition, cattle will graze plants normally
rejected as unpalatable. The presence of toxic
plants during the lengthy period of development
and growth of bone and cartilage (from 40 days
onward), and not during investigation several
months later, cannot be excluded. The skeletal
changes within individual affected calves and the
range of severity seen among cohort groups suggest
a prolonged period of exposure extending through-
out the period of skeletal mineralization and
growth, increasing the likelihood of detection of
toxic plants. In this region of Australia, lupins are
not common, and oak trees are rare; aflatoxicosis
was unlikely since there was little rain and silage
feeding was uncommon. No epidemiologic associ-
ation could be made between bushfire smoke
intensity and the congenital abnormalities.
Infection with teratogenic viruses was unlikely
since serology was negative and the lesions were
Cobalt (as vitamin B12){ Selenium (as glutathione peroxidase){
330 8.8
305 6.9
200–1,500 3.0–25.0
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria
dissimilar to those expected with infectious defor-
mities. Viral culture and immunoglobulin G
analysis was performed in other cases without
evidence of viral disease or in utero immune
challenge (data not presented). Viral agents known
to cause congenital defects (e.g., BVDV, Akabane
and Aino viruses) tend to cause neurologic defects
and arthrogryposis rather than deformity at the
growth plate. No histologic lesions of the nervous
system were detected. Nationwide surveillance
programs showed that insect vectors responsible
for transmission of arboviral disease were excluded
from these areas throughout the period of gesta-
tion; thus, Akabane and Aino viruses were
considered unlikely.
Anecdotal and unpublished data on previous
Australian outbreaks similar to this case have
suggested a possible association with long periods
of drought, deformed calves tending to present
following prolonged dry conditions or in the
seasons following early onset of more normal rain
patterns, particularly in the southeastern regions of
NSW. Extensive investigations have always failed
to ascertain a definite cause (A. Philbey, T. Ross,
personal communication).
Sequential elimination of reported causes of
deformities in this investigation led the authors to
suspect involvement of manganese deficiency in the
pathogenesis. Histopathologic descriptions of ex-
perimental manganese deficiency–induced deformi-
ties have been sparse to date, although the gross
pathology described is similar to that in affected
cases presented here. In a field investigation of
suspected manganese deficiency, narrowed irregu-
lar growth plates with reduced zone of cartilage
hypertrophy were described in three 3-month-old
calves, but the authors did not describe histologic
changes in neonates.24 Growth-plate irregularity
with reduced and disorderly proliferating chondro-
cytes and absence of hypertrophic chondrocytes,
with residual necrotic remnants among fibrillar
matrix, was described in another field outbreak
putatively associated with manganese deficiency.28
Respiratory difficulty has been a common factor in
this and in other investigations18,24 and may
represent abnormalities of the tracheal cartilage
rings leading to reduced luminal diameter, dis-
torted nasal passages, or a combination of these
and other factors. Calves not so severely affected as
to die in the perinatal period in outbreaks of
postulated manganese deficiency have reportedly
shown limited-to-good clinical improvement with
time, if assistance is given to feed and stand, with
reduction in deformation but residual stunting, as
noted in this outbreak.28
351
Evaluation of manganese concentration
Normal reference ranges for manganese concen-
tration are not available in neonates and work to
formulate these is still in progress, hampered by the
difficulty in obtaining fresh tissues from normal
calves in the affected areas. This has limited the
interpretation of manganese concentrations in
affected calves.
Previous soil and pasture mineral analyses suffer
similar difficulty in interpretation of results due to
regional variation in soil composition and numer-
ous other compounding factors in manganese
absorption (e.g., variable manganese uptake by
plants, combination of pasture species, and in-
terference from other elements).
Our review of the literature suggests liver
concentration of manganese is most appropriate
for analysis in these neonatal calves, although the
element can be routinely detected in bone ash,
blood, and hair samples.8,9,14 Liver samples are
easily obtained by farmers or veterinarians. They
are simple to process, and liver concentrations of
manganese are relatively high compared with
manganese concentrations in other tissues, which
may allow more accurate analysis. However, some
authors suggest that liver manganese concentration
is less well correlated to deficiency states than
manganese concentrations in other tissues, including
plasma and heart muscle, at least in growing
animals.26 Reports of alteration in hair content of
manganese with environmental factors unrelated
to total body concentration and with hair pigmenta-
tion have been identified, so this simple test was not
used.9
Manganese in skeletal disease
Manganese is an essential trace element involved
as a cofactor in several critical enzymatic pathways
of glycosaminoglycan and collagen synthesis and as
a component of metalloenzymes, including super-
oxide dismutase (SOD), among other functions.
Deficiency of SOD may be implicated in damage to
membranes of mitochondria, endoplasmic reticu-
lum, and golgi bodies and thus lead to loss of
integrity, potentially leading to inability to synthe-
size mucopolysaccharides.14 Manganese is also
essential in incorporating carbohydrates into mu-
copolysaccharides at the epiphyseal plate through
manganese-dependant activation of some glycosyl-
transferases, which transport trisaccharides to link
polysaccharide to protein in bone.8,16 Unlike many
other metal-activated enzymes, similar metals such
as magnesium or iron are less effective than
manganese at activation of these particular en-
zymes.27
352 McLaren, Cave, Parker, and Slocombe
The literature on manganese metabolism and
deficiency has been thoroughly reviewed.9,14,26,27
Deficiency of manganese has been associated
experimentally and clinically with reduced repro-
ductive performance7,22 and skeletal abnormali-
ties7,10,22,24,28 in calves and laboratory animals and
with ataxia associated with abnormal development
of the otoliths in laboratory rodents but not cattle.13
Manganese bioavailability
Concentrations of manganese in plants vary.
Grasses are richer than legumes, and straw and
corn silage have particularly low concentrations.9
Uptake of bivalent manganese by plants is more
efficient than that of the trivalent or tetravalent
forms present in highest concentrations in soil. Low
pH soils have a relatively high proportion of
bivalent manganese.
Discussion with nutritionists in this investigation
led to a suggestion that rumen pH may have
a similar effect on bioavailability of manganese. At
the time of writing, investigations were being
undertaken to review the current knowledge of
this interaction and determine whether manipula-
tion of rumen pH rather than direct supple-
mentation of manganese may aid in improving
manganese absorption (T. Parker, personal com-
munication).
Overall bioavailability from ingested plant mat-
ter is low, with ,1% intestinal absorption in cattle.
It appears from some studies that high dietary
levels of iron, copper, zinc, sulphur, and, to a lesser
extent, calcium and phosphorus lead to further
reduction in manganese bioavailability.8,9 Although
the soil in the affected area is high in granite, which
usually has a high level of manganese, the bio-
availability of the manganese may be reduced by
high dietary intake of other elements in soil such as
copper, cobalt, calcium, phosphorus, and iron,
which show significant interference with manga-
nese.26 Under severe drought conditions, cows may
ingest a high volume of soil and dust, leading to
altered dietary mineral balance.
Manganese has been shown in rats and sheep to
be preferentially sequestered by the placenta in
conditions of manganese deficiency of the dam, but
this sequestration is insufficient to return the fetal
manganese concentration to normal.11 Unlike
neonates, fetuses are not able to preferentially
concentrate manganese in the liver.14 It is believed
that manganese concentrations in the liver of
neonatal presuckled calves represent the manga-
nese status of the dam.26
Retention of absorbed manganese varies with
age; calves may apparently alter the percentage of
Vet Pathol 44:3, 2007
dietary manganese retained after intestinal absorp-
tion according to the concentration of the element
in the food and in response to deficiency.12
Manganese concentration in tissues has been
shown to change rapidly after birth and onset of
suckling in calves if there is high manganese
concentration in the diet, as indicated by radioac-
tive isotope studies of manganese metabolism.4
Young calves are able to rapidly incorporate
supplemental manganese from oral and intrave-
nous administration into tissues, and the percent-
age retained varies according to the dietary and
whole body concentrations of the element at the
time of administration of manganese.4 Once
suckling commences, it is probable that the
manganese concentration in liver rises rapidly and
more markedly than in other tissues. This is
attributed to the liver’s role in manganese homeo-
stasis through storage in mitochondria and excre-
tion in bile. Higher manganese concentrations were
found more often in suckled affected calves of a few
days old (data not presented) than in neonatal
unsuckled affected calves in this outbreak. Results
suggested that within a few days, the liver
concentration of manganese in affected calves
approached normal levels despite the low concen-
tration of manganese found in milk.
The percentage of dietary manganese absorbed
in adults is relatively constant. Manganese is
largely removed from first-pass portal circulation
by the liver, and peripheral blood has a very low
concentration of manganese bound to a-2-macro-
globulin.
The apparent difference in manganese absorp-
tion may reflect the incorporation of the majority
of dietary manganese into ruminal micro-organ-
isms in adults, as evidenced by more efficient
cellulose digestion by micro-organisms in the
presence of supplemental manganese.17 The reason
adult cows are unable to enhance retention of
manganese if deficient is unclear.
Feeding of silage alone through winter in
northern latitudes has been identified as a risk
factor for a condition known as congenital
joint laxity and dwarfism, which has some similar-
ities to experimental manganese deficiency.21 It is
postulated by some authors that this may relate in
part to manganese deficiency indirectly through
reduced bioavailability of manganese present in
silage.10
In this outbreak, drought conditions may have
led to rumen content with a high percentage of dry
matter and thus a relatively high pH. Under these
conditions, it is postulated that manganese is
present within the rumen mostly as poorly bio-
Vet Pathol 44:3, 2007 Chondrodysplastic Calves in Northeast Victoria
available forms and thus absorption may be
reduced. One potential avenue of investigation at
present is analysis of the effects of rumen pH on
manganese absorption; alterations of rumen pH
may be more readily achieved through the use of
supplemental grain feeding or dietary cation-
anion–deficit rations without the need to supple-
ment manganese directly.
Conclusion
The outbreak was characterized by dispropor-
tionate dwarfism, with reduced long bone/diaphysis
length secondary to abnormality of the hyaline
cartilage of physes and epiphyses evident on gross
and histologic examination. Changes affected all
bones formed by endochondral ossification; thus,
the vertebral column was shortened and the skull
domed due to reduced basicranial bone length.
Tracheal rings were distorted, indicating a general-
ized abnormality of hyaline cartilage.
Manganese deficiency remains the main postu-
lated cause at the time of writing. Confirmation of
manganese deficiency in calves is difficult and time-
consuming. Further studies are ongoing, and
veterinary nutritional specialists are assessing
advice to be given to farmers in the area. Ideally,
supplementation will be planned to provide a
controlled experiment of the effect of dietary
manganese on the birth of deformed calves in this
region, provided sufficient resources are available
to gather the epidemiologic data and for manga-
nese concentration analysis and other ongoing
exclusions. One review has recommended the
addition of 0.5 g/day per head of manganese as
sulphate, the most soluble form, or as oxide,
carbonate, or methionine salts to cattle.8 Avoid-
ance of management procedures that increase the
pH of soil such as liming may also aid in disease
prevention.26
Acknowledgements
We thank Professor K. V. F. Jubb for his support and
advice regarding pathologic investigations of submitted
cases and Associate Professor John Glastonbury for
critical review of the manuscript. We acknowledge Josie
Wilson, Dennis Miller, Paul Benham, Garry Anderson,
and Dayle Tyrrell of the Faculty of Veterinary Science,
the University of Melbourne, for assistance with
preparation of histology sections, postmortem examina-
tions, photography, statistical analysis, and radiologic
interpretation, respectively. The many Department of
Primary Industries Victoria field staff, Stephen J.
Whittaker of the Rural Lands Protection Board, and
Alex Stephens and Luzia Rast of NSW Department of
Primary Industries in particular must be acknowledged
for their field investigation work. Tony Parker is
353
acknowledged for his investigations into the effect of
rumen pH on bioavailability of manganese, and his
assistance in planning therapeutic strategies. Staff at
Primary Industries Research Victoria for further path-
ologic investigations, including mineral analysis and
virology studies; Gribbles Veterinary Pathology for
serological and mineral analysis; and Regional Labora-
tory Services Benalla for mineral analysis are gratefully
acknowledged.
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