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peer reviewed

Use of a lateral tibial head buttress plate in the repair of an

open comminuted supracondylar femoral fracture in a German
Shepherd dog
Mark R. Glyde1, David A. Lidbetter2 and Wing T. Wong2
1
2

Department of Small Animal Surgery, Faculty of Veterinary Medicine, University College Dublin, Belfield, Dublin 4, Ireland.
Veterinary Clinic and Hospital, University of Melbourne, Werribee, Victoria 3030, Australia.

An 8.5-month-old neutered female German Shepherd dog with a grade 1 open left
supracondylar femoral fracture was repaired with the application of an AO/ASIF
right lateral tibial head buttress (LTHB) plate applied to the lateral surface of the
femur. The unique design of this plate enabled placement of five screws in the
distal fracture fragment, thereby achieving excellent stability without compromising
stifle joint mobility. The fracture healed without long-term complications.
The LTHB plate proved to be a simple and effective method for supracondylar
femoral fracture repair and overcame the limitations of standard bone plates in
treating this type of fracture. Further studies are warranted to evaluate the use of
modified bone plates in the repair of supracondylar femoral fractures in dogs.

Keywords
Dog,
Supracondylar femoral fracture,
Lateral tibial head buttress plate.

Irish Veterinary Journal

Volume 56: 87 - 93, 2003

Introduction
Fracture of the femur is the most commonly occurring fracture
type in dogs and cats. Diaphyseal fractures of the femur are
most common, followed by distal physeal fractures in immature
animals. Supracondylar femoral (SCF) fractures affecting the
distal metaphyseal region in adult dogs and cats are uncommon
(Kolata and Johnson, 1975; Braden et al., 1990).
Surgical repair options for fractures occurring through the distal
femoral physis in immature dogs and cats have been described
(Parker and Bloomberg, 1984; Whitney and Schrader, 1987;
Brinker et al., 1990a; Sukhiani and Holmberg, 1997). The use
of crossed pins is the most commonly used technique and has
been shown to be biomechanically superior in distal physeal
fractures in immature dogs (Parker and Bloomberg, 1984;
Whitney and Schrader, 1987; Sukhiani and Holmberg, 1997).
Corresponding author:
Mark Glyde
Department of Small Animal Surgery
Faculty of Veterinary Medicine
University College Dublin
Belfield, Dublin 4, Ireland
Tel: +353 1 716 6058
Fax: +353 1 716 6061
E-mail: mark.glyde@ucd.ie

Distal femoral physeal fractures in immature animals are

relatively simple to repair and they heal quickly compared to
SCF fractures in adult animals. They are usually low-energy
fractures with limited soft tissue damage and minimal
comminution. They are inherently stable after anatomic
alignment and fracture repair due to interdigitation of the Wshaped physis with the femoral metaphysis (Brinker et al.,
1990a; Braden, 1993; Lidbetter and Glyde, 2000).
SCF fractures in adult dogs are more difficult to repair and
provide a substantial challenge to the veterinary surgeon for
several reasons. They are often comminuted and may have
significant soft tissue damage as a result of the higher-energy
force needed to fracture the bone of the adult animal compared
to that required to cause physeal separation in the immature
animal. The short length of the distal fracture segment, the
large medullary canal and the caudal bow of the distal femur
make application of effective implants difficult (Milton, 1993;
Lidbetter and Glyde, 2000). When axial loads are applied on
weight-bearing, the eccentric position of the femoral condyle
produces large bending moments that must be counteracted by
implants which are typically seated in a small distal bone
fragment. The caudal bow of the distal femur is more
pronounced in chondrodystrophoid breeds (Lewis et al., 1993;

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FIGURE 1a: Pre-operative lateral

radiograph of the left femur.

FIGURE 1b: Pre-operative

craniocaudal radiograph of the left
femur.

Milton, 1993). The location of the fracture adjacent to a highmotion joint such as the stifle complicates implant placement.
Implants must avoid the intercondylar fossa, cruciate ligament
origins and articular surfaces yet permit free movement of the
patella mechanism without causing patella subluxation
(Lidbetter and Glyde, 2000).
Many SCF fracture fixation methods have been described and
include the use of crossed pins, Rush pins, dynamic
intramedullary (IM) crossed pins (modified Rush pins), single
IM pins, two or more IM pins, paired convergent pins, external
skeletal fixators, standard bone plates, customised hook plates,
reconstruction plates and lag screws (Milton et al., 1980;
Franczusk et al., 1986; Whitney and Schrader, 1987; Brinker et
al., 1990a; Klause et al., 1990; Lewis et al., 1993; Milton,
1993; Robins et al., 1993; Lidbetter and Glyde, 2000). The use
of a lateral tibial head buttress (LTHB) plate in the repair of a
pathologic femoral fracture has been reported (Dueland and
VanEnkevort, 1995). This case report aims to detail the use of
an AO/ASIF LTHB plate in an open SCF fracture in a German
Shepherd dog.

Case report
An 8.5 month-old neutered female German Shepherd dog
weighing 35kg was presented for an acute traumatic nonweight-bearing left pelvic limb lameness. Physical examination
was otherwise normal with the exception of crepitus and
instability in the distal left femur consistent with a femoral
fracture. Neurological abnormalities were not apparent.

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FIGURE 2a: LTHB plate adjacent

to the aluminium bending template
prior to contouring.

FIGURE 2b: LTHB plate adjacent

to the aluminium bending template
after contouring.

Analgesics were administered (buprenorphine 10m g/kg

subcutaneously and carprofen 4mg/kg intravenously).
Lateral and craniocaudal radiographs of the left femur (Figures
1a and 1b) revealed an oblique comminuted distal femoral
fracture with a butterfly fragment from the cranial aspect of the
distal segment extending into the supracondylar area.
Longitudinal fissures were apparent in both the proximal and
distal fragments extending 1.0 to 1.5cm from the fracture. The
distal femoral physis had closed. Abnormalities were not
apparent on thoracic radiographs.
Premedication with acetylpromazine (0.05mg/kg) and atropine
(20m g/kg) was administered subcutaneously. General
anaesthesia was induced with thiopentone (12mg/kg)
administered intravenously to effect and maintained with 1.5%
halothane in oxygen (1.0L/min) and nitrous oxide (2.0
L/min). Cefazolin (22mg/kg) was administered intravenously
at induction and repeated two hours later. Intraoperative
analgesia comprised buprenorphine (15m g/kg) and carprofen
(4mg/kg) intravenously at induction. Lactated Ringers
solution was administered intravenously at 10ml/kg/hour for
the duration of the surgery and then reduced to
2.5ml/kg/hour for 12 hours postoperatively.
During preparation of the limb for surgery a small full-thickness
skin wound measuring approximately 10mm by 5mm was
apparent on the lateral aspect of the femur at the level of the
fracture consistent with a grade 1 open fracture (Gustilo et al.,

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FIGURE 3: Craniocaudal
radiograph of the intact
contralateral femur.

FIGURE 4: Intraoperative
photograph of the LTHB plate
prior to closure. The closed
arrows indicate the proximal and
distal margins of the butterfly
fragment on the cranial femur. The
open arrow points to the lateral
ridge of the femoral trochlea.

1990; Brinker et al., 1990b; Anson, 1993).

A lateral approach to the femur was combined with a lateral
approach to the stifle joint (Piermattei, 1993). The surgical site
was lavaged with 1 litre of warmed sterile 0.9% NaCl. There was
no evidence of gross contamination. The butterfly fragment had
a tenuous soft tissue attachment to the torn femoropatellar joint
capsule reflection distally and laterally. The attachment was
maintained and the fragment was replaced in the fracture defect
but not fixed in place; otherwise it was not manipulated.
A 166mm seven-shaft-hole 4.5mm right LTHB plate was
contoured using a standard aluminium plate template applied to
the fracture site following anatomic reduction (Figures 2a and
2b) and a craniocaudal radiograph of the intact contralateral
femur, which showed a mild varus deviation of the distal femur
(Figure 3).
The plate extended distally to the lateral epicondylar area of the
femur avoiding the articular cartilage. Nine 4.5mm cortical
screws were used. Four screws were placed in the proximal
fragment and five in the distal fragment. Two screw holes over
the fracture site were not filled (Figure 4). The intercondylar
fossa was visualised to facilitate placement of the two most distal
screws. The wound was lavaged with 0.9% NaCl and was closed
routinely. On recovery, buprenorphine (10m g/kg) was
administered intravenously as required. Postoperative
radiographs revealed good fracture alignment (Figures 5a and
5b).
The dog was weight-bearing on the leg the following day and

FIGURE 5a: Lateral postoperative radiograph of the

repaired left femur.

FIGURE 5b: Craniocaudal postoperative radiograph of the

repaired left femur.

was discharged into the owners care on amoxycillin/clavulanic

acid (500mg twice daily per os for seven days) and carprofen
(75mg twice daily per os for seven days, then 75mg once daily
per os for a further seven days).
Re-examination at ten days postoperatively was routine and, as
expected, the dog was progressively increasing weight-bearing
on the fractured leg. Seventeen days postoperatively the dog
presented to the referring veterinarian with an acute worsening
of the lameness. Physical examination by the referring
veterinarian revealed a moderate weight-bearing lameness,
pyrexia and pain localised to the left femur. A presumptive
diagnosis of osteomyelitis or soft tissue infection was made and
clindamycin (300mg per os twice daily) was prescribed. Three
days later the dog was examined and found to have a mild
weight-bearing lameness, without pyrexia. Radiographs were
obtained of the left femur. These revealed good fracture
alignment with no change in the position of the implants. There
was reasonable evidence of callus formation although a piece of
the caudomedial cortex showed little periosteal reaction. A fine
needle aspirate was taken from the fracture site. Cytology
revealed primarily red blood cells and only a small number of
nucleated cells; bacteria were not noted. Spindle cells were
prominent and moderately pleomorphic. The cytologic findings
were suggestive of normal osteoblastic and fibroblastic
proliferation as seen in callus formation. Antibiotics were
discontinued. Four days later the surgical site was explored.
Gross evidence of infection was not apparent. The butterfly
fragment had not moved, although it did not appear to have

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FIGURE 6a: Lateral radiograph of

the left femur showing fracture
union 16 weeks postoperatively.

FIGURE 6b: Craniocaudal

radiograph of the left femur
showing fracture union 16 weeks
postoperatively.

any soft tissue attachment. The fragment was removed and

submitted for aerobic and anaerobic culture. The site was
lavaged with two litres of warmed sterile 0.9% NaCl and closed
routinely. Then, cefazolin (22mg/kg) was administered
intravenously and clindamycin (300mg per os twice daily) was
continued postoperatively pending the results of the cultures.
Anaesthetic and analgesic protocols were as described for the
first surgery.
There was no bacterial isolate on either the aerobic or anaerobic
cultures of the bone fragment. Clindamycin (300mg per os twice
daily) was continued for a total of 14 days post-operatively.
On physical examination eight weeks postoperatively the patient
was clinically normal without lameness, with normal range of
movement of the stifle joint and slight palpable crepitus of soft
tissue over the lateral aspect of the supracondylar area. Standard
radiographs of the femur showed good remodeling of the callus.
At 16 weeks postoperatively, the patient was walking normally,
without pain on manipulation of the limb, and she had a full
range of stifle joint motion without any crepitus. Radiographs
confirmed that the fracture had healed without complications
associated with the implant (Figures 6a and 6b). There was no
radiographic evidence of degenerative change within the stifle
joint.
Twelve months after the surgery, the owner reported that the
patient was not lame. A clinical examination was normal with
the exception of slight periarticular fibrosis over the lateral
aspect of the stifle joint of the fractured limb.

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FIGURE 7: Seven and nine shafthole left LTHB plates.

Discussion
LTHB plates are designed for use in humans with proximal
tibial fractures but they have been used for repair of
supracondylar femoral fractures in humans (Schatzker and Tile,
1996) and for repair of a pathologic distal femoral fracture in a
dog (Dueland and VanEnkevort, 1995). LTHB plates have the
same shaft thickness as the AO/ASIF 3.5 broad plate; however,
the shaft is 2mm wider (14mm). They have five to 13 shaft
holes and range in length from 130 to 274mm. One end of the
plate is curved either right or left with an abaxial angle of 13
degrees and widens to a maximum width of 26mm. The
thickness of this widened end of the plate gradually reduces to
1.57mm centrally. The shaft screw holes are of the dynamic
compression design. The widened section of the plate has three
round holes and one oval hole most adjacent to the shaft
(Figure 7). All of the screw holes, including the shaft holes, will
fit either 4.5mm or 6.5mm screws (Dueland and VanEnkevort,
1995). The designation of the plates as either right or left is in
reference to their application to proximal tibial fractures in
humans.
Repair of distal femoral and SCF fractures in adult dogs presents
significant biomechanical and technical challenges. Achieving
effective stability and implant security while avoiding both intraarticular structures and interference with the patellar mechanism
is difficult (Milton, 1993; Lidbetter and Glyde, 2000).
Little information exists on the relative biomechanical merits of
the different fixation methods for SCF fractures (Sukhiani and
Holmberg, 1997). Crossed pins, paired convergent pins and

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dynamic pins had statistically similar strength in a biomechanical

study of pinning techniques used to stabilise distal femoral
physeal fractures. Crossed pins were significantly stronger than
single IM pins (Sukhiani and Holmberg, 1997). While the use
of crossed pins is indicated in the repair of the majority of distal
femoral physeal fractures, their use in SCF fractures in adult
animals is limited to simple transverse or short-oblique fractures
in cats and small dogs (Sukhiani and Holmberg, 1997;
Lidbetter and Glyde, 2000). Crossed pins, paired convergent
pins, Rush pins and dynamic IM pins are contraindicated in the
repair of comminuted SCF fractures and in large breed dogs.
Chondrodystrophoid breeds are unsuitable for cross pinning as
the marked caudal bow of the distal femur limits bone stock for
pin placement and, consequently, probably cannot provide
sufficient stability (Lidbetter and Glyde, 2000).
Intramedullary pinning techniques described in SCF fractures
employ either a single pin or two or more pins (Milton et al.,
1980; Parker and Bloomberg, 1984; Whitney and Schrader,
1987; Klause et al., 1990). Single IM pins are ineffective in
stabilising comminuted supracondylar fractures (Klause et al.,
1990) and provide poor rotational stability in transverse
fractures (DeYoung and Probst, 1993). Adequate stabilisation is
difficult to achieve due to the poor distal cancellous bone stock
for pin seating and consequent poor frictional resistance
(Milton, 1993). Pin seating can be improved by over-reducing
the distal fragment allowing pin penetration further into the
condyle; however, the resulting malalignment may decrease
stability (Milton, 1993). This problem is exacerbated in
chondrodystrophoid breeds (Lewis et al., 1993).
The location of the body wall limits femoral external skeletal
fixator (ESF) constructs to type 1 frames; however, the stiffness
of these can be increased with the addition of a second or third
frame connected on the medial side to a full condylar pin (Aron,
1996; Lidbetter and Glyde, 2000). The use of modified
unilateral ESFs in four comminuted SCF fractures has been
reported. Complications occurred in two of these cases, one of
which required revision with a bone plate to achieve fracture
union (Klause et al., 1990). With more recent developments in
ESF equipment and technique, modified type 1 ESFs are
considered to be suitable for use in most SCF fractures though
some morbidity is expected consequent to pin placement
adjacent to a high motion joint such as the stifle (Aron, 1996;
Lidbetter and Glyde, 2000).
The anatomy of the distal femur makes it difficult to apply
standard dynamic compression (DCP) plates to SCF fractures.
DCP plate application commonly impinges on the lateral
trochlear ridge and parapatellar fibrocartilage. This may interfere
with the patella and quadriceps mechanism, causing lateral
patella subluxation and difficulty in closing the joint capsule.
The use of DCP plates also often compromises the principles of
bone plating (Lidbetter and Glyde, 2000). A minimum of four
and preferably six cortices on either side of a fracture stabilised
with a bone plate are necessary to achieve stability. Eight

cortices are recommended in large breed dogs (Brinker et al.,

1990c). In most supracondylar fractures usually only three to
five cortices can be achieved with DCP plates (Klause et al.,
1990).
Customised hook plates were reported to achieve better stability
than standard bone plates without interfering with stifle joint
function in the repair of SCF fractures (Robins et al., 1993).
Customised hook plates are modified DCP plates that allow
increased cortical purchase over a smaller bone stock length
when compared to DCP plates. They are technically more
difficult to apply than DCP plates and are not commercially
available (Lidbetter and Glyde, 2000).
Reconstruction plates are useful in distal femoral and SCF
fractures in chondrodystrophoid breeds and small dogs and cats
with comminuted fractures. They are well suited to SCF
fractures as they can be contoured in three planes to fit the
caudal bow of the distal femur. This increases implant security
by increasing the number of screws engaged in the distal
fragment without impinging on the lateral trochlear ridge and
patella mechanism. They are ideal for chondrodystrophoid
breeds and are considered to be the fixation method of choice
in these animals. They should be reinforced with a Kirschner
wire if used in comminuted fractures and are not recommended
for use in large-breed dogs (Lewis et al., 1993; Lidbetter and
Glyde, 2000).
In this case a LTHB plate was chosen for repair in consideration
of the type of fracture, body weight and temperament of the
animal, availability of implants and surgeon preference. Preoperative planning suggested that ten cortices could be achieved
with minimal interference with the patella mechanism and
lateral trochlear ridge. Other suitable options considered in this
case were a modified type 1 ESF, which was not used in
consideration of patient temperament and anticipated
morbidity, and a modified hook plate, which would have
achieved possibly eight cortices though with slightly more
interference with the patella and more technical difficulty than
the LTBH plate. Pinning techniques and the use of DCP or
reconstruction plates were considered to be unsuitable in this
case due to the comminution of the fracture and the body
weight of the dog.
Application of the LTHB plate to the femur is relatively simple
and follows standard principles for internal fixation (Brinker et
al., 1990c). Care should be taken prior to placement of the two
most distal screws through the epicondylar region of the femur
to ensure screw placement does not penetrate the intercondylar
fossa. Visualisation of the femoro-tibial joint space prior to
placement of these screws and consideration of the anatomy of
the distal femur facilitated correct screw placement.
While placement of this plate is intra-articular it does not
impinge on the abaxial surface of the lateral trochlea ridge to
the degree of a standard plate. In addition, the LTHB plate
allows placement of more screws in the distal fragment than a
standard bone plate, thereby achieving superior stability without

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impinging on the articular surfaces of the femur. Even in more

distal fractures it would be anticipated that, due to the unique
design of this plate, a minimum of eight cortices would be
achieved. This overcomes the difficulty of a short, caudally
bowed distal fragment in SCF fractures.
To the authors knowledge, biomechanical studies of the use of
LTHB plates in the distal femur have not been published.
Application of a LTHB plate to the femur of a dog requires
bending the wide thinner end of the plate against the contour
with which it was manufactured. It could be expected that this
reverse contouring would have an effect on plate strength;
however, the biomechanical significance of this is not known. A
limitation to the use of these plates in veterinary surgery is that
they are essentially suitable for application only to large dogs
greater than 30 to 35 kg in weight.
Autogenous cancellous bone grafts accelerate bone healing via
the processes of osteogenesis, osteoinduction and
osteoconduction. They are recommended in the surgical
management of comminuted fractures, fractures in mature dogs,
non-union and delayed union fractures, arthrodeses and for
filling bone defects (Fitch et al., 1997). Autogenous cancellous
bone grafts were not used in this case. In the original fracture
repair a bone graft was not placed in view of the limited
comminution and the age of the dog. At the subsequent
exploratory surgery placement of a bone graft to fill the defect
created by the removal of the butterfly fragment might have
aided subsequent bone healing.
The decision to re-explore the fracture for lavage and sampling
for bacterial culture was made in consideration of a possible
diagnosis of osteomyelitis and concern over the possibility of
sequestration of the butterfly fragment. This was based on the
history of a grade 1 open fracture and the clinical signs despite
the absence of radiographic or cytological evidence of
osteomyelitis. Radiographic evidence of osteomyelitis is usually
not apparent for more than two weeks (Anderson, 1998; Hulse
and Johnson, 1997). Needle biopsy techniques are less reliable
in obtaining the causative organisms in osteomyelitis than
culture of tissue obtained during surgical debridement (Perry et
al., 1991). Aerobic and anaerobic bacterial cultures of the
fracture site and of the removed butterfly fragment did not
produce a growth. Whether this was a result of the prior
antimicrobial therapy or whether infection was not present is
not known. Discontinuation of antibiotics for 24 to 72 hours
prior to bacterial culture is recommended (Smith, 1993;
Anderson, 1998). Antimicrobial therapy had been discontinued
96 hours prior to samples being obtained for culture.
The recommendations for the use of antibiotics after surgery for
open fractures are controversial (May, 1998). The prophylactic
use of bactericidal antibiotics such as amoxycillin/clavulanic acid
or cephalosporins for five days is currently recommended if the
original bacterial culture is negative. Further antibiotics are
administered only if bacterial culture is positive or if
osteomyelitis develops subsequently (May, 1998).

92

The use of bactericidal antibiotics is recommended for three to

four weeks in acute osteomyelitis and for four to six weeks or
more in chronic osteomyelitis (Anderson, 1998; Hulse and
Johnson, 1997). Clindamycin was used in this case for 14 days
following the second surgery despite the negative bacterial
cultures. Clindamycin is a lincosamide antibiotic with efficacy
against Gram-positive cocci, including penicillin-resistant
staphylococci, and many anaerobes. It may be bacteriostatic or
bactericidal depending on the organism and drug concentration
(Tennant, 1999).

Conclusion
A LTHB plate was used successfully to repair a comminuted
grade 1 open SCF fracture in a German Shepherd dog. The
unique plate design overcame the limitations of a short, caudally
bowed distal fragment that typically makes fixation of these
fractures difficult. Effective fracture fixation was achieved
without compromising the principles of internal fixation or the
articular surfaces of the stifle joint.
LTHB plates should be considered for repair of SCF fractures in
large breeds of dogs. Further studies are warranted to evaluate
the biomechanical effect of plate contouring necessary in
application of these plates and to assess larger case numbers and
long-term results of LTHB plate repair of SCF fractures in dogs.

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Surgery. Edited by D. Slatter. Second edition, pp 1685-1694.
Philadelphia: Saunders.
Sukhiani, H.R. and Holmberg, D.L. (1997). Ex vivo biomechanical
comparison of pin fixation techniques for canine distal femoral
physeal fractures. Veterinary Surgery 26: 398-407.
Tennant, B. (1999). BSAVA Formulary. Third edition, pp 60-61.
Cheltenham: BSAVA.
Whitney, W.O. and Schrader, S.C. (1987). Dynamic intramedullary
crosspinning technique for repair of distal femoral fractures in dogs
and cats: 71 cases (1981-1985). Journal of the American Veterinary
Medical Association 191: 1133-1138.

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Occurrence and characterization of thermophilic Campylobacter

spp. in a poultry processing plant in Northern Ireland
John E. Moore1, Tom S. Wilson1 David R.A. Wareing2 and Philip G. Murphy1
1
2

Northern Ireland Public Health Laboratory, Department of Bacteriology, Belfast City Hospital, Belfast, Northern Ireland, BT9 7AD.
Preston Public Health Laboratory, Royal Preston Hospital, Sharoe Green Lane, Preston, Lancashire, United Kingdom.

Although there have been numerous studies investigating the prevalence of

campylobacters in cooked foods, there are limited published data on the occurrence of
these organisms in raw poultry in Ireland. Thermophilic campylobacters were detected in
254/275 (92.4%) batches of specimens of neck skin examined from raw processed chicken
at slaughter, where one batch represented 10 individual samples taken from 10
consecutive birds, thus giving a total population of 2,750 birds examined. C. jejuni, C. coli
and C. lari accounted for 78%, 20% and 2% of isolates, respectively. Quantitatively, there
was a mean count of 202 cfu campylobacters/g neck skin and counts ranged from 32 to
480 cfu campylobacter/g. Subspecies characterization of a subset of total isolates
recovered (i.e., approximately every sixth isolate; 17%) showed a large degree of
phenotypic diversity, with several biotypes and phage-types being identified. All isolates
were typed successfully by biotyping, but approximately only half of both C. jejuni and C.
coli isolates were typed successfully by phage-typing. Neither species shared a common
biotype; however, phagetypes 90, 91 and 121 were common to both C. jejuni and C. coli.
In conclusion, this study demonstrated the high prevalence of a phenotypically diverse
population of thermophilic campylobacters in poultry from Northern Ireland; however,
their association with human disease remains unclear and an epidemiological study is
required to address this potentially important relationship. Therefore, it is important that
intervention controls are established at farm-level to minimize colonisation of the
gastrointestinal tract of the bird and that emphasis continues to be placed on adoption of
HACCP practices on the farm, during processing, at sale and distribution, and in the
commercial or domestic kitchen.

Keywords
Poultry,
Biotype,
Campylobacter jejuni,
Phagetype,
Phenotype.

Irish Veterinary Journal

Volume 56: 95 - 98, 2003

Introduction
The past three decades have witnessed the rise of Campylobacter
enteritis in man from virtual obscurity to notoriety, with present
isolation rates superseding those of other enteric pathogens such
Correspondence:
Dr John E. Moore
Northern Ireland Public Health Laboratory,
Department of Bacteriology,
Belfast City Hospital, Belfast, N. Ireland, BT9 7AD.
Tel:
+44 (28) 9026 3554
Fax: +44 (28) 9026 3991
E-mail: jemoore@niphl.dnet.co.uk

as Salmonella spp. and Shigella spp. in most developed

countries. Unlike the salmonellae and other enteric pathogens,
the majority (ca. 99%) of clinical reports concerning
Campylobacter are sporadic and Campylobacter enteritis
outbreaks are rare. The lack of well-developed typing schemes
has hindered the epidemiological investigations seeking natural
reservoirs of the organism and modes of transmission from these
sources to man. Only about 15% of clinical isolates are identified
to species level, thus making epidemiological investigations
extremely difficult to perform. Since the development of more
sophisticated isolation techniques, the true disease potential of
these organisms has become apparent and today
campylobacteriosis is regarded as a zoonosis, which is capable of

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being transmitted to man by a wide range of domestic animals.

The laboratory isolation of these organisms has become routine
from clinical specimens as well as from environmental specimens
and, although relatively complicated to perform, routine
isolation has been carried out with success for this past 20 years
or so.
Campylobacter spp. are the most common cause of acute
gastroenteritis in Northern Ireland (N.I.), as well as in Britain
(Anon., 2002a). In N.I. there has been a gradual increase in the
number of reported cases since 1981 (Anon., 2002a).
Thermophilic campylobacters, i.e., those Campylobacter spp.
which are able to proliferate at 42C, particularly C. jejuni, C.
coli and C. lari, are of particular interest to the food industry, as
these campylobacters form the natural microflora of the
gastrointestinal tract of several domestic and pet animals,
including poultry (Shane, 1992; Whyte et al., 2001), cows
(Giacoboni et al., 1993), pigs (Stich-Groh, 1982; Moore and
Madden (1998) and frequently occur in the environment,
including untreated water sources (Aho et al., 1989; Taylor et
al., 1983) and other sources related to animal production
(Abeyta et al., 1993). Although campylobacters are the most
common cause of acute human enteritis, their routes of
infection and transmission to man are still not fully understood.
In addition, poultry have been shown to be a major reservoir of
these organisms; however, there have been limited reports in the
literature decribing the situation in Ireland. Consequently, this
study was carried out in order to establish the level of
contamination of commercially available raw poultry from a
single processor in Northern Ireland and to subsequently
examine the phenotypic diversity of recovered isolates, as well as
to establish a collection of archived isolates for future
epidemiological studies.

Materials and methods

Collection of chicken specimens
Freshly killed birds (n=2,750) originating from 106 growers
were examined over a three-month period (May July) at a
single processing plant in N.I., following evisceration and
dressing but prior to packaging. Approximately 2.5g neck skin
was taken from each bird and combined into a composite 25g
specimen from 10 consecutive birds from the same
grower/production batch to give a total of 275 specimens
examined. All specimens were transported to the laboratory at
4C and were examined within 17h following collection.

Isolation and characterization of Campylobacter spp.

Campylobacter spp. were isolated according to the method of
Moore and Madden (1998). Briefly, 25g of neck skin was
homogenized in 250ml of Exeter selective enrichment broth
(37C; 48h) in a shaking incubator (25rpm)(Innova 4000, New
Brunswick Scientific, Canada). The Exeter broth consisted of
nutrient broth no. 2 (1L) (Oxoid CM67), lysed horse blood
(50ml) (Beckton Dickson, England), sodium pyruvate

96

(200mg/L) (BDH Ltd, England), sodium metabisulphite

(200mg/L), ferrous sulphate (200mg/L), trimethoprim
(10mg/L)(Sigma Chemicals Ltd, England), rifampicin
(10mg/L), cefoperazone (15mg/L), polymyxin B sulphate
(4mg/L) and amphotericin (2mg/L) (E.R. Squibb Ltd,
England). After 48h incubation, 20ml of broth was streaked
onto Exeter selective agar (as broth with 15g/L Agar no. 1).
Plates were incubated microaerophilically (42C; 72h) in
anaerobic jars with a micraerophilic gas generating kit (Oxoid
BR56) and examined for the presence of typical Campylobacter
colonies.
All presumptive positive colonies were confirmed and speciated
in accordance with the following tests as described in Abeyta et
al. (1993): Gram stain, motility/morphology test, growth in
air, growth at 25C, utilization of glucose, catalase activity,
oxidase activity, haemolytic activity, susceptibility to nalidixic
acid and cephalothin, hippurase activity, hydrolysis of indoxyl
acetate and urease activity.

Quantitative enumeration of campylobacters on neck skin

Quantitative counts were performed on approximately 4%
(11/275) specimens examined by adding 10g neck skin to
90ml 0.1% [w/v] peptone saline (Oxoid). Further serial
dilutions were prepared in 0.1% peptone saline to 10-9 and
100m l of several dilutions were plated onto Exeter agar and
incubated for 72h, as described above. Resulting dilutions
showing between 30 and 300 colonies were enumerated and
the count calculated and defined as colony forming units (cfu)
campylobacters/g neck skin.

Sub-species characterization of isolates

Approximately every sixth Campylobacter-positive isolate
recovered from raw poultry (n=40) was characterized at the subspecies level by both Preston bio-(resisto)-typing (Bolton et al.,
1984) and phage-typing (Salama et al., 1990) techniques.

Results and discussion

Thermophilic campylobacters were detected in the batched
specimens examined, giving a prevalence value of 254/275
(92.4%) for the load processed. C. jejuni, C. coli and C. lari
accounted for 78%, 20% and 2% isolates, respectively. The
sensitivity of the enrichment method could not detect lower
than 10 cfu/g in 24h from a background flora of log10 9.06
cfu/g. Quantitatively, there was a mean count of 202 cfu
campylobacters/g neck skin and counts ranged from 32 to 480
cfu campylobacter/g. Subspecies characterization of a subset of
total isolates recovered (i.e., approximately every sixth isolate;
17%) showed a large degree of phenotypic diversity (Table 1),
with several biotypes and phage-types being described. All
isolates were typed successfully by biotyping, but approximately
only half of both C. jejuni and C. coli isolates were typed
successfully by phage-typing. Neither species shared a common

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TABLE 1: Distribution of Preston biotypes and phagetypes amongst isolates of Campylobacter jejuni and C. coli recovered from
raw chicken in a poultry processing plant in Northern Ireland

TOTAL

C. jejuni isolates
Preston
No of isolates
Phagetype (with phage type)

No. of isolates
(with biotype)

6000
6002
6004
6006
6010
6014
6015
6016
6052
6100
6146
6152
6304
6321
6346
6356
6404
6410
6450
6452
6604
6704
6710
6714
24

2
1
2
2
1
2
1
1
1
1
1
1
1
2
1
1
2
1
1
1
1
1
1
1
30

52
69A
90
90A
91
121
125
Non-typable

30

10

10

biotypes

isolates

phagetypes

isolates

biotypes

isolates

phagetypes

isolates

2
1
2
3
4
2
1
15

biotype; however, phagetypes 90, 91 and 121 were common to

both C. jejuni and C. coli.
Poultry is now well-established as a major reservoir of
thermophilic campylobacters (Shane, 1992), which have the
potential of being transmitted to humans via cross
contamination of ready-to-eat foods with raw poultry
(particularly drip) or via undercooking of contaminated
carcasses or products. Although thermophilic campylobacters
are not a health problem for poultry (Shane, 1992), their high
prevalence in commercial broiler flocks is a major problem for
human medicine. In this study, a high proportion of specimens
were positive for campylobacters. Although these positive
specimens originated from 102 out of 106 growers, it is difficult
to ascertain the true numbers of positives coming from
individual growers, as sampling of carcasses was performed at
the final stages of processing, which allowed for several
opportunities during processing for cross-contamination of
campylobacters from positive carcasses to campylobacter-free

Preston
biotype

No. of isolates
(with biotype)

2014
2040
2110
2454
2510
2514
2600
2640
2644

1
1
2
1
1
1
1
1
1

C. coli isolates
Preston
No of isolates
Phagetype (with phage type)

Preston
biotype

90
91
125
Non-typable

3
2
1
4

birds. Therefore, regardless of several effective intervention

measures that may be established in the broiler house, including
improved biosecurity, reduced thinning of flocks, etc., it is
important that controls are established in the processing plant
to minimize cross-contamination between positive and negative
flocks, otherwise the value of such intervention measures are
lost.
Such controls may include increased chlorination of the rinse
tank water, appropriate process flow and production staging,
whereby negatively tested flocks are processed before positive
flocks. For further information on additional HACCP controls,
please see Anon. (2002b).
In conclusion, this study demonstrated the high prevalence of a
phenotypically diverse population of thermophilic
campylobacters in poultry from Northern Ireland; however,
their association with human disease remains unclear and an
epidemiological study is required to address this potentially
important relationship. Therefore, it is important that

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intervention controls are established at farm-level to minimize

colonisation of the gastrointestinal tract of the bird and that
emphasis continues to be placed on adoption of HACCP
practices and that food handlers are sufficiently trained in the
safe handling of foodstuffs, in order to exclude these organisms
from cooked foods intended for consumption without any
further heating.

Acknowledgements
The authors thank Mr Clive Leckey and Mr Neville Heaney and
staff of the Food Hygiene Laboratory, Northern Ireland Public
Health Laboratory, for laboratory assistance and Dr Claire
Hughes, for help with collection of the specimens. This work
was funded by the Department of Health and Social Services
(NI).

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