Journal of Applied Animal Research

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Review: castration – animal welfare considerations

Gabriela A. Marquette, Stephanie Ronan & Bernadette Earley

To cite this article: Gabriela A. Marquette, Stephanie Ronan & Bernadette Earley (2023)
Review: castration – animal welfare considerations, Journal of Applied Animal Research, 51:1,
703-718, DOI: 10.1080/09712119.2023.2273270

To link to this article: https://doi.org/10.1080/09712119.2023.2273270

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UK Limited, trading as Taylor & Francis
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JOURNAL OF APPLIED ANIMAL RESEARCH
2023, VOL. 51, NO. 1, 703–718
https://doi.org/10.1080/09712119.2023.2273270

Review: castration – animal welfare considerations

Gabriela A. Marquettea, Stephanie Ronanb and Bernadette Earleya
a
Animal and Bioscience Research Department, Animal & Grassland Research and Innovation Centre (AGRIC), Teagasc, Dunsany, Ireland; bDepartment
of Agriculture, Food and the Marine, Agriculture House, Dublin 2, Ireland

ABSTRACT ARTICLE HISTORY

The castration of male cattle is an integral part of routine farm management. The nature and duration of Received 25 January 2023
an animal’s response to castration are dependent on a number of factors, including the method Accepted 17 October 2023
employed, the age of animals, the post-castration management, and whether or not pain relief is
KEYWORDS
provided with the procedure. Scientific assessments of the impact of castration on cattle welfare, Castration; cattle; age; breed;
including pain and injury, stress, inflammation, immune, and production, are the subject of this castration methods;
review. The objectives of this review are to describe (1) the different methods of castration, (2) the Burdizzo; surgical; rubber
pain responses associated with each of those methods, and (3) how age and pain mitigation ring; banding; pain
strategies affect those responses. Research studies are presented that have addressed the challenges mitigation
imposed by castration procedures on the welfare of cattle based on two main biological events: (1)
the changes in biological functions required to cope with the procedure, and (2) the biological
consequences to the animals. Indices of animal well-being are described that have objectively
demonstrated: (1) the degree of noxiousness that an animal experiences following castration and the
success of the coping mechanisms, and (2) the benefit of using pain management in modulating
these responses.

1. Introduction that tissue damage has occurred, is occurring or is likely to

1.1. Why castrate animals
Bulls are castrated to prevent sexual behaviour, reduce aggres-
sion, and increase animal handling safety (Warriss 1984; Ten-
nessen et al. 1985). In addition, steers (which are castrated
bovine animals from 1 d of age or more) have a reduced risk
of dark-cutting meat at slaughter (Tarrant 1981; Warriss 1984)
and can be comingled with heifers without the risk of
unwanted matings in the herd. Almost 8 million tonnes of
bovine meat are produced in the European Union annually
(Hocquette et al. 2018). A high production of beef from steers
is observed in the UK (50.9%) (Rutherford et al. 2021) and
Ireland (84%) (McGee et al. 2022). The scientific community
and producers worldwide (Spooner et al. 2012; Moggy et al.,
2017) agree that castration is a painful procedure and the use
of an anaesthetic in combination with an analgesic as a
measure to alleviate pain during castration is recommended
(Coetzee 2013b; Ede et al. 2022).
1.2. Pain
Pain in animals can be defined as ‘an aversive sensory and
emotional experience; it changes the animal’s physiology and
behaviour to reduce or avoid damage, to reduce the likelihood
of recurrence and to promote recovery’ (Molony and Kent
1997). Thus, pain is an important stimulus to alert an animal
occur, which allows immediate escape, withdrawal or other
avoidance behaviour (Martin et al. 2022). Painful experiences
also help animals to learn to avoid any similar pain-causing cir-
cumstances. Perceived pain varies according to the character-
istics of the noxious stimulus (e.g. location, duration,
intensity) and according to other factors (e.g. experience,
emotional state, individual variation) that modify the way in
which the central nervous system processes the perception of
pain (Mellor et al., 2000). The evaluation of pain in animals is
only possible using indicators that can be detected by external
observers. Most of these indicators are based on physiological
or behavioural responses (Reviewed by Prunier et al. 2013;
Coetzee 2013b; Tschoner 2021).
Physiological changes associated with pain occur mainly
through two related pathways. Firstly, pain is a powerful stressor
that directly stimulates the release of hormones from the hypo-
thalamic–pituitary–adrenal axis (HPA) and sympathetic-adrenal-
medullary axis (SAM). Secondly, tissue damage activates the
immune system and the release of numerous inflammatory
mediators (e.g. cytokines) which may also activate the adrenal
axis (Prunier et al. 2013). Hence, physiological indicators of pain
involve hormones from the adrenal and sympathetic axes, their
metabolic and physiological consequences, plasma markers of
an inflammatory state and mediators involved in the physiologi-
cal mechanisms of pain. At present, a number of tools for the rec-
ognition of pain are being used in research studies, including use

CONTACT Bernadette Earley bernadette.earley@teagasc.ie Animal and Bioscience Research Department, Animal & Grassland Research and Innovation Centre
(AGRIC), Teagasc, Grange, Dunsany, Co. Meath, C15 PW93, Ireland
© 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, dis-
tribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a
repository by the author(s) or with their consent.
704 G. A. MARQUETTE ET AL.

Table 1. Legislation concerning the use of anaesthesia for castration in calves.

Country Methods allowed Anaesthesia required Reference
Ireland Surgical, Burdizzo, rubber ring/band Surgical or Burdizzo: over 6 months of age DAFM (S.I. No. 107, 2014) (Animal Health
Rubber ring/band: over 7 days of age and Welfare Act 2014).
United Kingdom Surgical, Burdizzo, rubber ring/band Rubber ring/band: over 7 days of age (DEFRA 2013)
Other methods: over 2 months of age
Germany Not specifieda All methods: over 4 weeks of age (Daanje 2013)
Belgium Surgical, Haemostatic clamp At any age (Daanje 2013)
Switzerland Surgical, Burdizzoa At any age (The Swiss Federal Council 2008)
Australia Surgical, Burdizzo, rubber ring/band All methods: over 6 months of age (Animal Health Australia 2016)
New Zealand Surgical, Burdizzo, rubber ring/band Band: at any age (Reddy 2018)
Other methods: over 6 months of age
Canada Surgical, Burdizzo, rubber ring/band All methods: Beef: over 6 months of age; Dairy: at any age (National Farm Animal Care Council, 2013)
a
Rubber ring/band are not allowed.

of algometry to measure mechanical nociceptive thresholds,
accelerometers and pedometers to measure activity, the use of
infrared thermography, and the assessment of heart rate and
heart rate variability (Tschoner 2021).
1.3. Discrepancies in regulations between countries
While the use of anaesthetics is mandatory during castration of
calves at any age in Belgium and Switzerland, calves can be
castrated without use of anaesthetic up to a certain age in
some countries (Table 1). Castration is a painful procedure,
and pain relief is recommended when castrating calves at all
ages (Stafford and Mellor 2005; Coetzee 2013a; 2013b;
Canozzi et al. 2017; Mijares et al. 2022). For cattle there are rec-
ommendations from the Council of Europe (CoE 1988) that
allow husbandry management procedures such as castration,
freeze branding, dehorning, disbudding, ear notching, nose
ringing, vasectomies and chipping (Spoolder et al. 2016). The
principle is that ‘mutilations should be avoided’. While there
are derogations to this principle, mitigation measures are
always recommended (based on age of the animal, provision
of pain relief, presence of a veterinarian, etc). Indeed, the legis-
lation concerning the use of anaesthesia for castration in cattle
varies considerably among different countries and depends on
the method used and age of the animal (Table 1).
2. Castration methods
Castration procedures are generally divided into two categories:
surgical and bloodless (Burdizzo and rubber rings/banding).
2.1. Surgical castration
Surgical castration mainly involves removal of the testes by
splitting or removing the distal one third of the scrotum and
removing the testes by severing the spermatic cord in a
manner that minimizes bleeding, usually with an emasculator
or Henderson castrating tool (Jennings 1984). One other
method is the incision of the lateral scrotal walls with a
scalpel or a Newberry knife to expose the testicles. Using the
Newberry knife, both lateral walls and the median septum are
simultaneously incised, thus facilitating drainage from the
wound (Fubini and Ducharme 2016). In addition, the Newberry
knife allows the scrotal skin to be split without danger of other
tissues being cut. After the incision, the testes and spermatic
vasculature are pulled and exposed (surgical pull) to allow com-
plete removal of the testicles.
2.2. Bloodless castration
Bloodless castration is generally performed by using an emas-
culatome (i.e. Burdizzo) or elastic band.
2.2.1. Burdizzo castration (emasculatome)
When using a Burdizzo, the scrotum remains intact while the
spermatic cord of each testicle (within the scrotum) is placed
in the jaws of the tool and crushed. The resulting damage
stops blood flow to the testes with eventual testicular atrophy
within the scrotum. The Burdizzo is applied twice (the second
crush is repeated distally with one cm space between the two)
to each spermatic cord along the neck of the scrotum (Ting
et al. 2003a). The jaws of the instrument are closed for approxi-
mately 10 s in a single (Robertson et al. 1994) or two crushes
(Fisher et al. 1996, 1997; Ting et al. 2003a) to ensure that both
blood supply and nerves to the testes are irreversibly destroyed.
2.2.2. Rubber ring and banding
Banding involves using an elastrator to place a heavy elastic
band around the neck of the scrotum with both testes inside.
The band disrupts blood flow to the testes and scrotum,
which atrophy over a long period of time and slough off.
Rubber ring and elastic band castration are considered blood-
less castration since there is no incision of the scrotum
(Becker et al. 2012). Small rubber rings (rubber ring castration)
are used for calves less than one month (mo-) of age. For older
calves heavy wall latex bands are used along with a grommet to
securely fasten the mechanically tightened tubing at the appro-
priate tension (Pang et al. 2009a; 2009b). Animals generally
receive tetanus prophylaxis to minimize the risk of tetanus
(O’Connor et al. 1993). The effect of banding and Burdizzo cas-
tration of continental × beef bulls (12.0 ± 0.2 mo-old; (mean ±
SE); weight 341 ± 3.0 kg (mean ± SE) was investigated by
Pang et al. (2009a). The authors found that banding compared
to Burdizzo castration caused more inflammatory associated
gene expression changes in the epididymis and scrotum. In a
further study, Pang et al. (2009b) investigated measures of neu-
trophil function in response to banding or Burdizzo castration
of bulls. Neutrophil functioning in terms of phagocytosis and
respiratory burst and serum IL-8 concentration were not
affected by banding, Burdizzo, and cortisol infusion. These
findings indicate that non-surgical castration is unlikely to

induce a severe acute systemic inflammatory response in terms
of neutrophil function.
2.3. Other methods of castration
2.3.1. Immunocastration (ImC)
Immunocastration (ImC) uses immunological methods to
destroy the hormone balance of the hypothalamic–pituitary–
gonadal (HPG) axis (also known as the reproductive axis).
Using this approach, the hypothalamic gonadotropin-releasing
hormone (GnRH) is reduced, which cause a decrease in luteiniz-
ing hormone (LH) and follicle-stimulating hormone (FSH),
thereby reducing the level of gonadal hormone and eventually
leading to gonadal atrophy and functional inhibition (Adams
et al. 1993; Mazon et al. 2019). Studies conducted in Mexico
(Pérez-Linares et al. 2017), Canada (Marti et al. 2015), and
Brazil (Amatayakul-Chantler et al. 2013; Miguel et al. 2014)
found that vaccinating cattle against gonadotropin-releasing
hormone (GnRH) produces effects similar to traditional cas-
tration methods. However, Mazon et al. (2019) reported that
ImC decreases the tenderness of fresh meat and negatively
affects the overall liking and tenderness sensory ratings. A dis-
avantage of the ImC method is that the effect of immunocastra-
tion is of short duration (4 months) and requires repeated
immunization to maintain effective suppression of testosterone
concentrations. A commercial vaccine is available for use in
cattle in some countries, but not in Europe.
2.3.2. Chemical castration
Chemical castration involves the injection of lactic acid, directly
into the testes, causing oedema and sloughing (Capucille et al.
2002). This method is reported to cause less pain and compli-
cations compared to physical methods of castration.
However, it is recommended only for calves less than 70 kg
(Skarda 1986). In addition, this method does not always
provide a successful castration (Hill et al. 2010) and
therefore chemical castration is not considered a useful tech-
nique for castrating calves (Coventry et al. 1989). Furthermore,
Fordyce et al. (1989) found that scrotal necrosis occurred in
25% of chemically-castrated calves which may have been
associated with leakage of the chemical from the testes
under the high pressure of injection. The authors reported
that healing time for chemical castrates was twice that for sur-
gical castrates and also found that 18% of chemically-castrated
calves retained one testis.
3. Castration induced pain
The age of an animal is a relevant factor which contributes to
individual pain differences (Tucker, 2018). In most research
studies, castration procedures are considered least stressful
on calves when performed at the earliest age possible.
Studies investigating castration-induced pain have been per-
formed in calves at different ages (see reviews by Bretschneider
2005; Coetzee et al. 2012; Coetzee 2013a; 2013b; Canozzi et al.
2017). These studies suggest that castration at a younger age is
less painful, however, it is difficult to draw a general conclusion
since there are many differences between studies, such as
breed of calves, environment (presence of the dam or not),
JOURNAL OF APPLIED ANIMAL RESEARCH 705
and plane of nutrition. Age could be an important factor
influencing the changes in stress response between studies.
Indeed, Stafford and Mellor (2005) in their review on castration
stress stated that it is complex and needs to be studied rigor-
ously to determine effect of breed and age of the animals,
the methods used and how animals are reared before
castration.
The pain experience of individuals differs according to their
sensitivity to noxious stimuli, susceptibility to developing neu-
ropathic pain after injury, and their analgesic response to
pharmacological therapy (Mogil 2012). A better understanding
of the basis of these differences in cattle may assist with our
ability to recognize and manage pain effectively through all
its stages.
3.1. Cortisol concentrations and inflammatory
responses
Castration is considered both a stressor and a painful experi-
ence for animals (Fisher et al. 2001; Bretschneider et al., 2005;
Stafford and Mellor, 2011). Animal welfare concerns thus
warrant that castration procedures are performed in a way
that causes the least stress and pain. One of the defining fea-
tures of the stress response to castration in male cattle is the
secretion of excess glucocorticoids (cortisol), and a biological
consequence associated with this reaction is the temporal sup-
pression of the adaptive immunity. A high peak of cortisol after
a known stressor may be a sign of a well-functioning HPA axis
with an established glucocorticoid cycle (Moberg and Mench,
2000; Blecha, 2000; Mitra et al., 2009; Hulbert and Moisá
2016). Castration by Burdizzo and surgical methods has been
shown to result in a rapid and large increase in plasma cortisol
concentrations (Fisher et al. 1996). It would appear that a sig-
nificant part of the castration-induced cortisol rise is caused
by pain as the provision of local anaesthesia reduced the corti-
sol surge for both burdizzo and surgical methods, while local
anaesthesia alone had no effect on plasma cortisol (Fisher
et al. 1996). Pain is difficult to quantify in animals; however,
plasma concentration of the stress hormone cortisol is often
used as an indicator of castration induced stress related pain.
King et al. (1991) investigated the plasma cortisol concentration
changes in beef bull calves castrated at 2.5 or 5.5 mo-old by
either surgery or Burdizzo. In their study, uncastrated calves
were restrained in the same manner as castrated calves for 2
min, which was the average length of time taken for each cas-
tration. The authors concluded that the cortisol response to sur-
gical or Burdizzo castration was reduced by castrating calves at
2.5 mo-old compared with 5.5 mo-old, but no direct compari-
son was made between age groups due to the study design.
Interestingly, uncastrated 2.5 mo-old calves had a greater
increase in cortisol concentration after sham handling and
restraint compared to the 2.5 and 5.5 mo-old Burdizzo
castrated calves, indicating that restraint is also a stressor. In
their study, all calves were separated from their dams for
sham handling, castration and blood collection. It is likely
that the abrupt separation of the calves followed by restraint
resulted in an increased cortisol response in all calves regard-
less of treatment (King et al. 1991). A similar reduction in corti-
sol response in young dairy calves was found in other studies.
706 G. A. MARQUETTE ET AL.
Robertson et al. (1994) evaluated plasma cortisol responses of
dairy calves castrated by Burdizzo, surgery or rubber ring cas-
tration at 6, 21 and 42 days old. At each age, the peak of cortisol
concentration of Burdizzo and surgical groups increased
sharply after castration (Robertson et al. 1994). Similarly,
another study using Holstein bull calves castrated by Burdizzo
at 1.5, 2.5, 3.5, 4.5 and 5.5 mo-old found that the plasma cortisol
area under the curve (AUC) from 0 to 3 h after castration was
significantly reduced in 1.5 and 4.5 mo-old calves by 47% and
34%, respectively, with intermediate reductions in 2.5 and 3.5
mo-old calves (27 and 23%, respectively), compared to 5.5
mo-old castrates. However, no significant changes were
observed on area under the curve (AUC) from 3 to 12 h after
castration between age groups (Ting et al. 2005). In a study
by Dockweiler et al. (2013), plasma cortisol concentration of
Holstein bull calves was increased in 6 mo-old calves relative
to their 2-mo-old counterparts, regardless of treatment
applied which included sham handling of uncastrated calves,
and surgical (cut and clamp; cut and pull) and non-surgical
(band) castration procedures. The authors suggested that
younger calves may be less acutely stressed by handling and
castration procedures than older calves.
However, further studies found no difference in the cortisol
response of calves castrated at different ages. Petherick et al.
(2015) investigated the effect of band and surgical castration
on plasma cortisol concentration of Belmont Red calves at 3
and 6 mo-old. In their study, plasma cortisol concentrations
increased immediately after castration but was not different
between age treatments 30 min post-castration; only 6-mo-
old calves castrated by band showed increased plasma cortisol
concentrations above baseline 2 h post-castration. Similarly,
Meléndez et al. (2017b) evaluated effect of band and knife cas-
tration on chronic indicators of pain in 1 week old, 2 and 4 mo-
old Angus crossbred calves. They found no differences in sali-
vary cortisol concentration at 60, 120 min, or 7 d after castration
in 1 week-old calves. Additionally, the baseline salivary cortisol
concentration for 1 week-old calves was greater than for 2 and
4 mo-old calves supporting the suggestion that calf-dam separ-
ation and handling of 1 week-old calves are cumulative stres-
sors and that a high intensity of circulating glucocorticoids
was present before castration (Meléndez et al. 2017a).
However, the comparison between ages was not possible as
the studies were conducted at different time of the year
using different restraint conditions due to calf size. Further-
more, Sutherland et al. (2013) investigated behavioural and
physiological changes following surgical castration, dehorning,
or both of 3 mo-old Holstein calves (101.5 ± 1.45 kg; mean, SE)
and found that the responses were cumulative when the two
procedure were carried out simultaneously. Therefore, combin-
ing castration and dehorning procedures will exacerbate
responses to the procedures and should be avoided in view
of the negative welfare consequences.
3.2. Behaviour
Several postural and behavioural indicators of pain have been
reported in animals, such as avoidance and defensive beha-
viours, vocalisations, behaviours directed towards the painful
areas. The pain of castration occurs first as acute, short-term
pain associated with the actual castration procedure. Chronic
pain is the longer-lasting pain that occurs in the days following
castration until the injury is healed, associated with inflam-
mation and neural injury.
Boesch et al. (2008) examined the short-term pain response
of 30 calves less than 1-week of age to Burdizzo castration with
and without local anaesthesia. Twenty minutes before cas-
tration, 10 ml lidocaine (n = 10), bupivacaine (n = 10) or saline
solution (n = 10) was injected into each spermatic cord and dis-
tributed subcutaneously in the scrotal neck. The behaviour of
the calves and response to local palpation was assessed on
the day before and after castration during 8 h periods.
Plasma cortisol levels were determined on the day of castration.
The struggling behaviour exhibited by the calves during cas-
tration and their response to local palpation of the spermatic
cords indicated that the procedure was painful. Nevertheless,
postures and behavioural elements thought to be associated
with pain were not readily apparent after castration in the
majority of calves. The pain caused by injection of the anaes-
thetic was considered minimal. Local anaesthesia reduced the
immediate pain response during castration as evidenced by
less struggling during castration and lower cortisol levels.
However, some calves struggled during the procedure and
had distinct pain-indicating signs afterwards. There was no
apparent difference between the analgesia provided by lido-
caine and bupivacaine.
Molony et al. (1995) concluded that burdizzo castration,
when performed correctly, was probably less stressful to
young calves than surgical castration, and that castration
using a rubber ring induced abnormal behaviour for up to 42
d, possibly associated with chronic pain. Nogues et al. (2021)
compared two castration methods; surgical (n = 10 calves)
and rubber ring (n = 11). Pre-weaned dairy calves were
castrated at 28 days of age using multimodal pain control.
During the 8 week period post-castration, wound healing,
local inflammation, body weight, milk and calf starter intake,
lying time, and wound-directed behaviour were recorded. Sur-
gical wounds were fully healed on average 4 weeks after the
procedure, but only 1 calf in the rubber ring treatment fully
healed within the 8-week study period. Inflammation was
greater after rubber ring castration; skin temperature in the
area around the lesion was higher (+1.7 ± 0.35 °C; mean ± sd)
than for the surgical treatment. Compared with surgically
castrated calves, those castrated by rubber ring gained less
weight over the study period (on average 11.9 ± 5.1 kg less),
a difference due in part to lower intake of calf starter (on
average 1.8 ± 0.6 kg less). Calves in the rubber ring treatment
spent less time lying down (on average 4.2 ± 1.2% fewer
scans per day) and licked their lesions more frequently (on
average 16.0 ± 3.3 more licks per day). The authors concluded
that the rubber ring calves experienced more pain in the
weeks following the procedure and thus recommend that sur-
gical castration be favoured for pre-weaning dairy calves.
Thuer et al. (2007) investigated the behavioural and cortisol
responses of calves as indicators of pain to assess short- and
long-term effects of bloodless castration methods with and
without local anaesthesia. Seventy calves, aged 21–28 days
old, were control handled (n = 20) or castrated using the Bur-
dizzo (n = 25) or rubber ring technique (n = 25). Either 10 ml

lidocaine or NaCl was distributed in both spermatic cords and
the scrotal neck. The plasma cortisol response was recorded
for 72 h, and behavioural and clinical traits monitored over a
three month period. Local anaesthesia reduced the level of
indicators of acute pain after both the Burdizzo and rubber
ring techniques. It did not, however, result in a totally painless
castration. As there was evidence of chronic pain lasting for
several weeks after rubber ring castration, the Burdizzo
method was considered to be preferable to the rubber ring
technique based on the animal’s behavioural and physiological
responses. Indeed, rubber ring castrated calves responded up
to 4 weeks longer than Burdizzo calves on local palpation
and continued to show an increase in proportion of abnormal
postures after the first week following castration.
Lambertz et al. (2014) evaluated the effect of castration on
the stress response assessed by behavioural observations as
well as blood traits and performance of calves weaned at the
same time of castration or 4 weeks afterwards. The treatments
consisted of bulls that were (1) castrated and concurrently
weaned in week 0 after housing; (2) castrated in week 0 and
weaned in week 4; (3) bulls weaned in week 0; and (4) bulls
weaned in week 4. Weaning was found to have a greater
effect on the number of vocalizations, standing/walking and
lying behaviour and ADG compared with Burdizzo castration.
The authors concluded that, with undertaking the procedures
separately, concurrent castration and weaning did not affect
behaviour and haematological parameters or impaired animal
performance.
Petherick et al. (2015) investigated effect of band and surgi-
cal castration on behaviour of tropical breed calves (Belmont
Red) castrated at 3 and 6 mo-old, by band or surgery. In their
study, some behavioural indicators of restlessness/activity
were influenced by calf age, being greater in 3-mo-old than
6-mo-old calves on the day of castration. These greater levels
of restlessness in the younger compared with older calves
may have been due to a greater motivation for calves to re-
establish contact with their mothers, which is a response fre-
quently reported in studies where the calf and dam are separ-
ated for some time. In the same study, during the 4 to 7 h
period post castration, by which time calves had been separ-
ated from their mothers for at least 4 or 5 h, there were
greater levels of vocalization by the control (not castrated)
calves compared with the castrated calves (Petherick et al.
2015). Although vocalization behaviour can be used to assess
pain, in this case, calves may be experiencing the least pain
and showing more normal behaviours, by trying to establish
contact with their mothers. In fact, these authors concluded
that in experimental situations where unweaned calves are
temporarily separated from their mothers for data collection,
pain-related behavioural responses may be attenuated due to
calves being motivated to re-establish contact with their
dams, and this in turn may switch their attention from pain
(Petherick et al. 2015).
Two studies evaluated the effects of band and knife cas-
tration on acute (Meléndez et al. 2017b) and chronic (Marti
et al. 2017a, 2017b) indicators of pain, such as behaviour, in 1
week old, 2 mo-old and 4-mo-old Angus crossbred suckler
calves. Investigating acute signs of pain in 1 week old calves,
knife castrated calves showed a greater number of pain-
JOURNAL OF APPLIED ANIMAL RESEARCH 707
related indicators on the day of castration (stride length and
tail flicking), whereas calves castrated by elastic bands exhib-
ited pain-related behaviours on days 2 and 3 (lying duration
and standing and lying bouts) The authors commented that
the lack of differences between tail flicking in banded and
control calves may be due to high individual variation for tail
flicking compared to the rest of the behaviours. In 2 mo-old
calves, knife castrated calves had greater standing percentage,
and walking duration, as well as decreased eating and lying
behaviours on the day of castration. In addition, greater stand-
ing and lower lying percentages were observed during the first
7 days after castration in knife castrated calves, suggesting that
knife castration is more painful than band castration in 2 mo-
old calves. In 4 mo-old calves, band castrated calves presented
greater restless behaviour 2 to 3 days after castration. However,
knife castrated calves had greater frequency of leg movement,
and vocalizations at the time of castration, shorter stride length
immediately after castration, a greater number of tail flicks 2–4
h after castration, and greater standing activity for the first 5 d
after castration. This indicates that behaviour responses associ-
ated with pain lasted for a longer period after knife castration.
Using the same animals as the previous study, the same
authors found no differences in standing or lying, or duration
of lying bouts among treatments, and no chronic effect of cas-
tration method on stride length, eating time, or behaviours
related to pain (tail flicks, foot stamping, head turning, or
lesion licking) throughout the trial in 1-week and 2-mo-old
calves (Marti et al. 2017a, 2017b). However, in 4 mo-old
calves, lying time after castration differed between treatments;
band castrated calves spent more time lying compared to knife
and control calves (Marti et al. 2017a, 2017b). In those studies,
cow-calf pairs were kept together in the same pen (Marti et al.
2017a, 2017b; Meléndez et al. 2017b). Throughout the day,
pain-related behavioural responses can be reduced by the
calf shifting it’s attention elsewhere (mediated by conscious
awareness of pain) (Gentle 2001). Consequently, the presence
of the dam with the calf can shift their attention from the
pain and change the expression pattern of pain-related
behaviours.
3.3. Wound healing and sensitivity
Petherick et al. (2015) investigated the impact of band and sur-
gical castration on wound healing of tropical breed (Belmont
Red) calves castrated at 3 or 6 mo-old, by band or surgery.
The scrotal wounds of calves castrated at 3-mo-old took
longer to heal compared to calves castrated at 6-mo-old. In
contrast, Norring et al. (2017) found no difference in wound
healing after surgical castration of beef calves at 3 d or 2.4
mo-old calves. In their study, younger calves reacted to
lighter pressure of Von Frey monofilaments (48% more sensi-
tive) compared to castrates at 2.4 mo-old especially in the
first stages of the healing process, and there were other signs
indicative of inflammation processes in this region at this
time (Norring et al. 2017). The incisions of younger calves
healed more quickly than older ones (fully healed, median
(95% confidence interval); 39 (32–61) v. 61 (61–77) days),
however, they had relatively increased swelling in the days
after castration.
708 G. A. MARQUETTE ET AL.
3.4. Scrotal temperature and circumference
The presence of heat and oedema following tissue trauma, such
as castration, are used as signs of inflammation. Ting et al. (2005)
investigated the changes in scrotal temperature and circumfer-
ence following Burdizzo castration of calves at different ages.
In their study, the increase in scrotal circumferences of calves
castrated at ages between 2.5 and 4.5 mo was not different
from the increase in scrotal circumference in the 5.5 mo-old
calves. However, the 1.5 mo-old castrates had little change in
scrotal circumference on the first day following castration com-
pared with all other castrates, and less change on days 7, 21 and
28 post-castration compared with either 4.5 or 5.5 mo-old cas-
trates. Since the younger calves had the least genital tissue
development, castration at younger ages could be done to mini-
mize tissue inflammation, as suggested by Ting et al. (2005).
There is evidence to show that surgically castrated older calves
take longer to heal their wounds and to resolve swelling com-
pared with younger calves. Healing after surgical castration
can take between 10 days (Molony et al. 1995) and 4–6 weeks
(Fisher et al. 2001; Stafford et al. 2002; Mintline et al. 2014;
Marti et al. 2017a, 2017b).
3.5. Thermal nociception threshold
The physiological development of the calf and responses to
stressful stimuli at a young age was also evaluated. Ting et al.
(2010) assessed the thermal nociception threshold of calves
castrated at different ages (1.5, 2.5, 3.5, 4.5 and 5.5 mo-old) to
a heat spot laser which was directed to the lower leg of the
calves after Burdizzo castration. The thermal nociception was
not significantly increased following Burdizzo castration at
any age. However, the study showed other age-related differ-
ences between calves. The skin temperature of the hind legs
of calves 1.5 mo-old were lower before and after castration
compared to calves 2.5, 3.5, 4.5 and 5.5 mo-old. The reaction
time to the heat spot laser increased in all calves after cas-
tration. However, calves 1.5 mo-old, which also had a lower
skin temperature, tolerated the heat spot laser for a longer dur-
ation than the older calves. The lower skin temperature and
more tolerance to the heat spot laser suggests that younger
calves are less capable of showing behavioural signs of pain
(Ting et al. 2010).
Variations in skin temperature are likely due to changes in
tissue perfusion, metabolism, and blood flow in the superficial
veins, and sympathetic nerve activity. However, the question
remains to be addressed concerning the physiological develop-
mental stage of calves at 1.5 mo of age versus older calves (2–6
mo-old) and their responses to castration procedures, since the
laser-based thermal nociceptive assay used in the study by Ting
et al. (2010) was influenced by the initial skin temperature and
the age of calves. This finding suggests that the assessment of
pain should be multimodal and based on physiological and
behavioural indicators.
3.6. Electroencephalogram responses
Dockweiler et al. (2013) investigated the age-related differ-
ences in pain response of Holstein bull calves subjected to
surgical and band castration at 8 weeks and 6-mo of age.
Desynchronized electroencephalogram (EEG) and electroder-
mal activity readings (both indicative of pain response) were
greater in 6-mo-old compared to 8-week-old calves after cas-
tration. However, the absence of desynchronization across cas-
tration methods in young calves does not imply that they do
not require analgesia at times of castration. The cortical func-
tion in these young calves are not developed enough to
show the same EEG responses observed in older calves.
However, this does not necessarily indicate they are not experi-
encing pain. This study illustrates the importance of measuring
different variables, whenever it is possible. Lehmann et al.
(2017) investigated the mitigating effects of administration of
local anaesthetic or systemic meloxicam on the EEG responses
during surgical castration of bull calves. The EEG changes indi-
cated nociceptive responses in all three groups during surgical
castration, being greater in calves receiving local anaesthetic
compared to calves that received no preoperative analgesia
and those that received preoperative meloxicam. Lidocaine
and meloxicam administered prior to castration attenuated
these responses in bull calves.
Bergamasco et al. (2021) investigated effect of unmitigated
surgical castration on the EEG responses of male Holstein calves
in three age categories [<6 weeks, 3, 6 mo of age; 10 calves per
age group]. Calves were subjected to a simulated castration
session (sham) followed 24 h later by surgical castration
without analgesia. The EEG total power decreased, and
median frequency increased relative to sham in 6 week and 3
mo-old calves only following treatment. The authors reported
variation in EEG responses following unmitigated surgical cas-
tration in calves and suggested that the responses could be
age specific.
3.7. Substance P (SP)
There is some evidence to suggest that SP might be a suitable
biomarker for nociception in cattle, but results of research are
heterogenous. Bergamasco et al. (2021) also investigated
effect of unmitigated surgical castration on the plasma sub-
stance P concentrations in calves of different ages under the
same experimental conditions, as described above. Substance
P concentrations decreased in the castrated treatment com-
pared to the sham treatment at the later times; 6 week old
calves showed lower substance P concentration at castration
relative to sham. The authors reported variation in SP concen-
trations following unmitigated surgical castration in calves
which may be age specific. In a recent systematic review,
Tschoner and Feist (2022) concluded that SP concentrations
of calves and adult cattle differ throughout studies and that
further research is warranted to investigate factors others
than nociception which might influence the SP concentrations
in the circulation. The authors stated that although studies
showed that SP concentrations differed significantly by age,
with 6-months-old calves showing higher concentrations
than 8-week-old calves (Dockweiler et al. 2013), there was no
consistency among age groups of animals included in
studies. Even within the same gender and age group, high
between- and within-calf variations were found (Coetzee
et al. 2008).

3.8. Acute phase proteins – haptoglobin and fibrinogen
Acute phase proteins (APPs) are induced primarily in the liver
and often described as positive (up-regulated) or negative
(down-regulated) acute phase proteins in response to the chal-
lenge (Murata et al. 2004). The former have important roles in
the inflammatory response, whereas the latter are important
carrier proteins such as albumin, corticosteroid binding
protein, and transferrin (metal-binding protein). Examples of
positive APPs include haptoglobin, fibrinogen, serum amyloid
A, C-reactive protein, and α−1-glycoprotein.
While some studies have found an increase in plasma hapto-
globin concentration after a stressor (Murata and Miyamoto,
1993; Morrow-Tesch and Whitehead, 1998), others have failed
to demonstrate any effect of stress on the haptoglobin
response, even in the presence of increased levels of other
acute phase proteins such as serum amyloid A (SAA) and fibri-
nogen (Alsemgeest et al. 1995; 1996; Hickey et al. 2003). It
therefore appears that the haptoglobin response to stress is
complex. A reduction in cortisol and APP production following
castration with analgesia would indicate that the ‘noxiousness’
associated with the stimulus has been effectively alleviated,
and hence impact on the welfare of castrated animals would
be minimized (Earley and Crowe 2002).
3.9. Effect of castration on animal performance
There is a belief that delaying castration could extend the pro-
duction advantages of keeping animals as bulls until weaning
or beyond puberty. However, a number of studies have
shown that there is no advantage in delaying castration of
bulls from 5 to 7 mo of age up to 17 mo in terms of liveweight,
growth rate, or carcass weight at slaughter (Keane 1999; Knight
et al. 1999a, 1999b). Keane (1999) reported that Burdizzo cas-
tration of spring-born calves in their first autumn (complete
castration) at 5–6 mo-old did not significantly affect the
overall 347 d liveweight gain compared with: (1) delayed unilat-
eral castration – the right testicle removed in autumn and left
testicle the following spring with ∼178 d apart; or (2) split cas-
tration in spring with about one month interval between
removal of each testicle. Furthermore, in that study, no inter-
action between castration treatment and breed type (Friesian
versus Charolais × Friesian) was found. Knight et al. (1999a,
1999b) reported that the age at surgical castration (7–15
versus 17 mo-old) of post-pubertal bulls had no effect on
either liveweight or carcass weight when the animals were
slaughtered at 22 mo-old.
Pang et al. (2008) assessed the effect of banding or Burdizzo
castration, performed on farms, on plasma testosterone, acute-
phase proteins, scrotal circumferences, growth, and well-being
of bulls. The 243 Continental bulls (12 mo-old; 399.2 ± 5.72 kg;
mean ± SE) used in the study were on three different commer-
cial farms. The bulls were allocated at random, after stratifica-
tion on weight within breed type, to one of three treatment
groups: banding castration, Burdizzo castration and controls
(intact). The authors reported that banding and Burdizzo cas-
tration significantly reduced plasma testosterone concen-
tration; reduced average daily weight gain mainly during the
first 2 weeks, which was not compensated during the
JOURNAL OF APPLIED ANIMAL RESEARCH 709
subsequent 16 weeks; increased withdrawal of stored energy
and increased plasma protein concentration. Burdizzo
showed an advantage over banding in growth during days
15–28 following castration. Theurer et al. (2019) investigated
the distal splitting of the scrotum at time of banding of bulls,
ranging in weight from 306 to 552 kg (average 436 kg), and
reported improved average daily gain, healing time, and
increased ribeye area compared to leaving the scrotum intact.
Castration has been shown to elicit a reduction in growth to
varying degrees. Fisher et al. (1996) reported that surgical cas-
tration of 5.5 mo-old calves resulted in lower 35 d growth rates
compared with Burdizzo castration, and the depressive effect
occurred mainly during the first week after castration.
However, the animals used in their study were maintained in
individual tie stalls which may influence their growth compared
with animals raised in the feedlot or pasture environment. King
et al. (1991) found no effect of either surgical or Burdizzo cas-
tration on the liveweight or daily gain of 2.5 mo-old calves com-
pared with bulls over a three-month period after castration. In
contrast, Fenton et al. (1958) found no difference in either sur-
gical, Burdizzo, or elastrator castration procedures in 7-week-
old calves in terms of liveweight gain five weeks after cas-
tration, but the control calves had higher gains than castrates.
However, the authors reported a significant retardation of
growth for the elastrator group on the 28th day after castration
due to chronic pain and sepsis (based on visual assessment and
palpation of the scrotum) proximal to the ring. This is sup-
ported by the findings of Molony et al. (1995) who showed
trends for lower 36-d growth rates in rubber ring, and com-
bined rubber ring plus Burdizzo castrated 1-week-old calves
compared with intact, surgical or Burdizzo castrated calves.
Bretschneider (2005) reviewed the effects of age at cas-
tration on performance of beef cattle and found that castration
at birth or close to birth drastically reduced weight loss associ-
ated with castration. Later, studies designed to compare the
impact of age at castration on performance agreed with his
findings. Norring et al. (2017) found that beef calves surgically
castrated at 2.4 mo had a reduction of 57% on ADG (average
daily gain) from 1 to 77 days after castration compared to
calves castrated at 3-d-old (Table 2). In agreement with the pre-
vious study, the study by Petherick et al. (2015) found that
surgery or rubber ring castrated calves at 3 mo had superior
weight gains compared with calves castrated 6 mo-old calves.
Meléndez et al. (2017a) found no difference on ADG from d
−1–7 of calves castrated at 1-week or 2-mo-old, by either
band or surgically, compared to intact calves within respective
age group. However, in the same study, calves surgically
castrated at 4-mo-old had a significant lower ADG, from d
−1–7, compared to intact calves at same age.
Conversely, other studies found no difference in perform-
ance of calves castrated at different ages. Ting et al. (2005)
found no difference in growth trends up to 42 days after Bur-
dizzo castration of Holstein-Friesian calves castrated at 1.5,
2.5, 3.5, 4.5, and 5.5 mo-old. In another study, Micol et al.
(2009) evaluated the effect of age (2 or 10 mo) at castration
by rubber band on performance of Charolais steers and their
muscle characteristics and meat quality traits. They found no
difference in liveweights and average daily gains according to
castration age. In addition, meat quality traits of tenderness,
 710
Table 2. Summary of the scientific literature examining the effect of age at castration (cx) on welfare outcomes.
Sedative/Anaesthetic/ Castration Age range Percent Significance
Author Population Analgesic Methods (treatments) Comparison group Outcome parameter Change (%) (P value)
Ting et al. (2005) Holstein-Friesian None Burdizzo 1.5 mo cx (n = 5.5 mo castrated Plasma cortisol (Cmax) −54.05 <0.05

G. A. MARQUETTE ET AL.
Control (sham) 10/trt) calves (n = 10) AUC (–3 h) −46.55 <0.05
AUC (3–12 h) −10.18 NS
Haptoglobin (day 3) −75.64 <0.05
Fibrinogen (day 3) −29.66 <0.05
WBC (day 1)a +38.54 <0.05
WBC (day 2)
Growth rate (day −7–35) −52.98 NS
2.5 mo cx Plasma cortisol (Cmax) −28.82 <0.05
AUC (0–3 h) −26.72 NS
AUC (3–12 h) +1.85 NS
Haptoglobin (day 3) −45.72 <0.05
Fibrinogen (day 3) −19.49 <0.05
WBC (day 1) +25.68 <0.05
WBC (day 2)
Growth rate (day −7 to 35) −31.78 NS
3.5 mo cx Plasma cortisol (Cmax) −26.12 <0.05
AUC (0–3 h) −23.27 NS
AUC (3–12 h) +15.74 NS
Haptoglobin (day 3) −23.93 NS
Fibrinogen (day 3) −16.95 <0.05
WBC (day 1) +20.00 <0.05
WBC (day 2)
4.5 mo cx Plasma cortisol (Cmax) −25.22 <0.05
AUC (0–3 h) −34.48 <0.05
AUC (3–12 h) +22.22 NS
Haptoglobin (day 3) −16.66 NS
Fibrinogen (day 3) −8.47 NS
WBC (day 1) +17.60 <0.05
WBC (day 2)
Growth rate (day −7–35) −10.59 NS
Ting et al. (2005) Holstein-Friesian None Burdizzo 5.5 mo sham 5.5 mo castrated Plasma cortisol (Cmax) −77.47 <0.05
Control (sham) calves (n = 10) AUC (0–3 h) −69.83 <0.05
AUC (3–12 h) −33.33 NS
Haptoglobin (day 3) −82.90 <0.05
Fibrinogen (day 3) −43.22 <0.05
WBC (day 1) −3.6 NS
WBC (day 2)
Growth rate (day −7–35) −8.00 NS
Ting et al. (2010) Holstein-Friesian None Burdizzo 1.5 mo cx (n = Each group baseline Skin Temperature
Control (sham) 10/trt) (−72 h) 12 h post castration +30.14 <0.05
24 h post castration +19.62 <0.05
48 h post castration +15.79 <0.05
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +15.84 NS
24 h post castration +20.79 NS
48 h post castration +36.63 NS
2.5 mo cx Skin Temperature
12 h post castration +7.39 <0.05
24 h post castration −1.05 NS
48 h post castration 0 NS

(Continued)
Table 2. Continued.
Sedative/Anaesthetic/ Castration Age range Percent Significance
Author Population Analgesic Methods (treatments) Comparison group Outcome parameter Change (%) (P value)
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +47.44 NS
24 h post castration +17.95 NS
48 h post castration +43.59 NS
Ting et al. (2010) Holstein-Friesian None Burdizzo Control 3.5 mo cx Each group baseline Skin Temperature
(sham) (−72 h) 12 h post castration +1.32 NS
24 h post castration −3.97 NS
48 h post castration −11.92 <0.05
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +47.89 NS
24 h post castration +76.06 NS
48 h post castration +28.17 NS
4.5 mo cx Skin Temperature
12 h post castration +21.08 NS
24 h post castration −4.15 NS
48 h post castration −3.51 NS
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +41.89 NS
24 h post castration +56.76 NS
48 h post castration +55.41 NS
5.5 mo cx Skin Temperature
12 h post castration +1.60 NS
24 h post castration −4.79 P < 0.05
48 h post castration −7.09 P < 0.05
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +9.52 NS
24 h post castration +27.38 NS
48 h post castration +46.43 NS
Ting et al. (2010) Holstein-Friesian None Burdizzo 5.5 mo sham Each group baseline Skin Temperature

JOURNAL OF APPLIED ANIMAL RESEARCH

Control (sham) (−72 h) 12 h post castration +1.60 NS
24 h post castration −1.88 NS
48 h post castration +0.63 NS
Thermal Nociception Threshold
(latency to leg withdrawal)
12 h post castration +37.50 NS
24 h post castration +10.94 NS
48 h post castration +23.44 NS
Norring et al. (2017) Angus-Hereford cross- Lignocaine 3% ring Surgical 2.4 mo (n = 15) Surgical castrated 3 d ADG (1–77 d post castration) −57.14 P < 0.05
bred bull calves block (all calves) old (n = 16)
Skin temperature
(1–77 d post castration) +2.00 P < 0.05
Time to resolve swelling −60.00 P < 0.05
Sensitivity to wound palpation
(1–77 d post castration) +48.08 P < 0.05
Abbreviations: Trt treatment; AUC, area under the curve; WBC, white blood cells; ADG, average daily gain in body weight.
a
WBC percent changes were calculated between day 1 and day 2 of the same treatment group.

711
712 G. A. MARQUETTE ET AL.

juiciness and flavour were equivalent for the two age groups of 4. Attributed to age at castration
steers (Micol et al. 2009). These studies suggest that the effect
A misconception that castration is less stressful for younger
of the age at castration on performance of the calves also
animals likely arose as older animals have a greater peak of
depends on the castration method with surgical castration
plasma cortisol after castration, and calves did not gain as
having a greater impact on performance mainly in older calves.
much weight after castration as older animals (Bretschneider
2005; Stafford and Mellor, 2011). In fact, Murray and Leslie
3.10. Effect of castration on animal health (2013) suggest that pain may be even greater among neonatal
animals compared with mature animals since their nervous
To our knowledge, detailed epidemiological studies on the
system and HPA axis are not developed. Kampen et al. (2006)
effects of castration per se on health are unavailable,
reported that immune parameters in young calves differ from
however, earlier work by Addis and colleagues in the mid 70s
what is found in older calves and adult animals, therefore
indicated that castration of calves causes adverse effects on
these age-related factors should be considered when assessing
animal health, particularly when it was performed at any time
immunological responses in young calves to castration. Robert-
near weaning or shipment (cited in Zweiacher et al. 1979).
son et al. (1994) examined the effect of rubber ring, Burdizzo
This supports the theoretical framework developed by
and surgical castration in Ayrshire bull calves at different ages
Moberg (2000) who postulated that exposure to multiple stres-
(6, 21 and 42 days old) on behavioural responses. In their
sors leads to an adverse ‘cumulative’ effect on the health and
study, the younger calves showed less tail wagging and foot
welfare of an animal. Zweiacher et al. (1979), in two separate
stamping and more head turning and spent more time lying
studies, found that calves (mean bodyweight; 180 kg) pur-
normally than the older calves (21 and 42 days old). The total
chased as steers had less health problems than those pur-
time spent in abnormal postures was increased following cas-
chased as bulls and subsequently castrated by surgical
tration by all methods, and the 21 and 42 d old calves spent
methods on arrival at a feedlot (mean percentage of animals
less time eating and suckling than non-castrated controls, indi-
requiring treatment for sickness = 17.5% for steers [n = 97]
cating that the pain induced by castration temporarily domi-
and 38.4% for surgical castrates [n = 104]). These health
nated their behaviours. The time spent in abnormal standing
effects were not related to the time (castration on arrival
postures increased in Burdizzo and surgical castrates and
versus after 1 week or after 2 weeks) or methods of castration
lasted for 24 min in 6 d old calves and up to 120 min in 21
(surgery with either use of elastrator band for ligation, or crimp-
and 42 d old calves, indicating that older calves were experien-
ing of testicular cords with an emasculator to prevent the blood
cing discomfort for a longer time.
loss). However, the death losses were greatest in bulls castrated
Effect of age at castration on physiological, immunological
on the day of arrival at the feedlot compared with bulls
stress indices, and behavioural responses using 60 Holstein-
castrated one week, or two weeks after arrival (mean percen-
Friesian bull calves was examined by Ting et al. 2005, 2010.
tage death loss was 8.5% for bulls castrated on arrival [n =
Calves were sourced such that they were in one of five age
35], 0.0% for bulls castrated after one week [n = 35], and 3%
groups for Burdizzo castration on day 0 (n = 10 per treatment),
for bulls castrated after two weeks [n = 34]). These animals
1.5, 2.5, 3.5, 4.5, and 5.5 mo of age, or were sham castrated at
were also branded, dehorned, wormed and vaccinated on
5.5 months of age (Ting et al. 2005, 2010). Castration was
arrival at the feedlot. In their second study, surgical castration
shown to be stressful across all ages between 1.5 and 5.5
occurred on the day of arrival and prophylactic treatment
mo-old, as indicated by the increased (∼180 to >300% mean
with oxytetracycline for the first three days appeared to
increase) integrated plasma cortisol response for the first 3 h
reduce, but not prevent the higher occurrence of sickness in
after treatment relative to the intact controls. However by redu-
the castrates (mean percentage of animals requiring treatment
cing the age at castration, the integrated cortisol response was
for sickness was 2.6% for steers [n = 38] and 13.0% for surgical
markedly reduced (by nearly half) in the 1.5 mo-old and by one-
castrates [n = 85]; Zweiacher et al. [1979]). These findings are
third in 4.5 mo-old castrates, but the ∼20% reduction observed
supported by Berry et al. (2001) who found that calves (mean
in the 2.5 and 3.5 mo-old castrates were not significant. The
bodyweight, 166 kg) purchased as bulls and subsequently
peak cortisol responses to castration were reduced by castrat-
castrated by surgical method the day after arrival at the
ing calves at younger ages. However, there was no evidence
feedlot had greater incidences of sickness (requiring treatment
to suggest that the welfare (including performance and
for respiratory disease) than calves purchased as steers.
immune responses) of calves was adversely affected by Bur-
However, in another study, these authors did not find any
dizzo castration from 1.5 to 5.5 months of age (without use
differences in terms of health response between the steers,
of local anaeshetic) (Ting et al. 2005). This is an important con-
banded calves or surgically castrated calves on arrival. The
sideration; the cortisol response was short-lived, there was no
main problems associated with the studies on the effects of
adverse effect on immune variables and performance at the
castration on animal health reported so far are that there
age range studied.
were numerous other confounding factors involved (e.g. trans-
Ting et al. (2005) also reported that plasma concentration of
port, branding and dehorning). Thus more detailed and specific
the acute phase proteins (APPs), haptoglobin and fibrinogen,
epidemiological studies may be warranted in order to draw a
were reduced by castrating calves at younger ages. Plasma
more definitive conclusion. Furthermore, Sutherland et al.
haptoglobin concentration on the third day after castration
(2013) investigated behavioural and physiological changes
was markedly reduced by castrating calves at 1.5 and 2.5 mo-
induced by surgical castration, dehorning, or both and found
old (reduction of 75 and 45%, respectively); but no significant
that the responses were cumulative when carried out together.

changes were found in haptoglobin concentration of calves
castrated at 3.5 and 4.5 mo-old, compared to calves castrated
at 5.5 mo-old (Ting et al. 2005). In the same study, plasma fibri-
nogen concentration was reduced on the third day after cas-
tration at 1.5, 2.5 and 3.5 mo-old (reduction of 29, 19 and
17%, respectively) compared to 5.5 mo-old castrates (Ting
et al. 2005). This indicated that the inflammation caused by cas-
tration was reduced by castrating calves at a younger age.
Marquette et al. (2021b) examined effect of age at castration
on stress indicators and performance of 40 crossbred suckler
beef calves. Calves were assigned to two age groups and two
castration treatments; calves 2.5 or 5.0 mo-old (mean body
weight (SD) = 120.8 (29.3), 218.1 (30.8) kg, respectively) were
sham (control) or Burdizzo castrated in a 2 × 2 factorial
design. Following castration, peak plasma cortisol concen-
trations were greater in 2.5 and 5.0 mo-old calves compared
with corresponding controls, while peak cortisol concen-
trations in control animals were greater in 5 mo-old compared
with 2.5 mo-old calves. The integrated cortisol responses for
the first 4 h after castration were not different between 2.5
and 5.0 mo-old calves. However, the integrated cortisol
response was greater in 5.0 mo-old calves from 4 to 9 h post
treatment. The increase in scrotal circumference after castration
was greater in 5.0 mo-old calves, and abnormal postures were
observed more often in 5 mo-old castrates. There was no effect
of castration on haematology profiles, haptoglobin, metab-
olites, body temperature and growth performance.
Following castration, calves spent more time standing
during the first hour, regardless of the treatment. Duration
spent lying was only affected by time which was decreased
in the first hour compared with the second and third hour
block after treatment. There was no difference in time spent
standing in abnormal position between 2.5 or 5.0 mo-old
control calves for any sampling time. However, calves castrated
at 5.0 mo-old spent more time standing in abnormal position
(928 s) compared with calves castrated at 2.5 mo-old (358 s),
in the first hour post-castration. Castrated calves exhibited
more foot stamping compared with control calves, regardless
of the age at castration. Before treatment (−24 h), the mean
latitudinal scrotal circumferences was greater for 5.0 mo-old
calves compared with 2.5 mo-old calves. Both latitudinal and
longitudinal scrotal circumferences were increased in castrated
calves at 24 and 48 h after treatment compared with their
respective baseline. Calves castrated at 5.0 mo-old had a
greater increase in latitudinal scrotal circumference at 24 and
48 h compared with 2.5 mo-old castrates. No difference was
observed in longitudinal scrotal circumference between 2.5
and 5.0 mo-old castrated calves.
The increased plasma cortisol concentration following cas-
tration observed in this study suggests that Burdizzo castration
is stressful for both 2.5 and 5.0 mo-old suckler beef calves.
However, an increase in abnormal postures, scrotal swelling
and a prolonged plasma cortisol response observed in older
calves suggest that these calves suffered more stress induced
by handling and castration. Therefore, castration of younger
suckler calves is preferable from an animal welfare point of
view. The age-related differences in stress response found in
this study need to be considered when recommending the
use of anaesthesia and analgesia during castration, especially
JOURNAL OF APPLIED ANIMAL RESEARCH 713
for older calves. The timing of pain relief (analgesia) adminis-
tration should be planned to effectively reduce the stress
response according to the age of the calf. The practical impli-
cations of the study are that if calves are to be castrated
without analgesia or anaesthesia, then it would be preferable
to do this at 2.5 mo-old rather than later, in order to minimize
the stress associated with Burdizzo castration.
In cattle, most of the immune system maturity is seen by 5–8
months of age. For example, T cells (CD4+, CD8 + and TCRγδ+
cells) do not reach peak levels until the animal is 8 months of
age (Cortese, 2009). Research has demonstrated that from
birth, there is a decrease in immune responses until day 3 in
calves, when they are at their lowest levels (Cortese, 2009). By
day 5, these responses are back to the level of immune
responses seen on the day of birth. Procedures like disbudding,
castration and movement need to be considered as stressors
that have the potential to decrease immune system function
temporarily in younger calves.
4.1. Attributed to pain mitigation strategy
The use of analgesia after castration of calves differs with age
and breed. A Canadian survey reported that 6.9% of beef
calves and 18.7% of dairy calves under 6 mo of age and
19.9% of beef calves and 33.2% of dairy calves over 6 mo of
age, received some form of anaesthesia at the time of cas-
tration (Hewson et al. 2007), but very few received post-operat-
ive analgesia. In a 2010 survey of veterinarians in the United
States, Coetzee et al. (2010) reported that approximately 20%
of respondents used some form of pain relief for surgical cas-
tration in cattle. Similarly, Fajt et al. (2011) reported that
approximately 30% of respondents provided some analgesic
drugs when castrating calves. A UK survey of veterinarians by
Remnant et al. (2017) found that 67% of respondents used
local anesthesia in calves undergoing castration.
Research studies have demonstrated that local anaesthesia
alone or combined with systemic analgesic drug administration
prior to castration mitigates physiological, and neuroendocrine
changes usually associated with pain and distress (Fisher et al.
1996; Earley and Crowe 2002; Ting et al. 2003a, 2003b). Calves
administered ketoprofen, a non-steroidal anti-inflammatory
drug (NSAID), prior to castration exhibited increased feeding
and rumination activities and fewer pain-associated behav-
ioural responses than those castrated without ketoprofen
(Ting et al. 2003a). Local anaesthesia prolonged the increase
in acute-phase proteins suggesting that LA administration
may further exacerbate inflammatory reactions associated
with castration, including pain responsiveness. Ketoprofen
was more effective than lignocaine or epidural in decreasing
cortisol and partially reversed the reduction in average daily
gain following castration. The use of ketoprofen or epidural
was more effective than local anaesthesia in decreasing pain-
associated behavioural responses observed during the first 6
h after treatment. Systemic analgesia with ketoprofen, was
more effective in reducing inflammatory responses associated
with castration than local or epidural anaesthesia. However,
the authors did not investigate the pain caused by the epidural
injection in uncastrated bull calves.
714 G. A. MARQUETTE ET AL.
The effect of carprofen administration before banding or
Burdizzo castration of bulls on plasma cortisol, in vitro inter-
feron-gamma production, acute-phase proteins, feed intake,
and growth was investigated by Pang et al. (2006). Overall,
the integrated cortisol response was greater in the castrates
than in control, whereas carprofen treatments tended to
reduce this response compared with banding and Burdizzo cas-
tration without carprofen treatment, respectively. The authors
concluded that carprofen (1.4 mg/kg of BW) tended to
reduce the integrated cortisol response, and it reduced cortisol
secretion in banded animals between 6 and 12 h post-cas-
tration. There was an increased acute-phase protein production
following castration; this response was effectively reduced by
the administration of carprofen before castration.
Pang et al. (2011) investigated the effect of Banding or Bur-
dizzo castration of bulls on the gene expression profile of a
range of peripheral leukocyte inflammatory cytokines (IL-1, IL-
6, IL-8, IL-10, interferon-γ and tumor necrosis factor-α) and
determined the response to administration of carprofen
before castration. Thirty Holstein-Friesian bulls (5.5 months
old; 191 ± 3.7 kg, mean ± SE) were blocked by weight and ran-
domly assigned to one of five treatments: (1) Untreated control
(Con); (2) Banding castration at 0 min; (3) Banding following an
i.v. injection of 1.4 mg/kg BW of carprofen (C) at −20 min; (4)
Burdizzo castration at 0 min; or (5) Burdizzo following 1.4
mg/kg BW of carprofen at −20 min. Generally, there were no
differences among treatment groups in haematological vari-
ables following castration. Cortisol concentrations were
unchanged throughout the experimental period in Con bulls.
Burdizzo animals had greater cortisol concentrations than
banded and controls at 6 h post treatment. Of particular inter-
est is IL-6, an inflammatory cytokine, which plays a critical role
in the development of pathological pain. At 24 h post-cas-
tration, Pang et al. (2011) found the relative quantity of IL-6
mRNA was greater in Burdizzo calves than banded calves at
24 h post-castration. This finding is in agreement with their pre-
vious study (Pang et al. 2006) which found that banding was
associated with a greater cortisol secretion than Burdizzo at
12 h post-castration (Pang et al. 2006) and that of Molony
et al. (1995) that banding caused more chronic inflammation
than Burdizzo.
Others NSAID’s have been studied, for example, nalbuphine
hydrochloride (Coetzee et al. 2012), flunixin meglumine (Nordi
et al. 2019), and meloxicam (Roberts et al. 2015; Vindevoghel
et al. 2018; Melendez et al., 2017a, 2018a, 2018b, 2019). Stilwell
et al. (2008) showed that epidural anaesthesia with lidocaine
will reduce temporarily the pain caused by Burdizzo castration.
Inflammation, pain-related behaviours and high levels of corti-
sol were still present at greater than 48 h after clamp castration
and were only abolished after the use of a long-acting NSAID
(Pang et al. 2006; Stilwell et al. 2008). Several studies have
assessed the efficacy of different analgesic protocols. Thuer
et al. (2007) showed that a local anaesthetic, injected into the
spermatic cord and subcutaneously at the neck of the
scrotum, reduced acute pain during and immediately after
calves were castrated by use of a castration clamp. Therefore,
providing calves with pain relief, in the form of local anesthetic
and an NSAID, can markedly reduce both the behavioural and
physiological response to castration. As evidenced, pain
management for castration is achieved by pharmacological
interventions. However, factors affecting the development of
pain and inflammation, such as the age of animals, the tech-
niques for castration and associated severity of injury, and
the post-operative animal management could be be optimized
in future studies to reduce the adverse effects of castration.
5. Conclusion
Depending on the techniques used (Burdizzo, surgical, banding
(typically using either a rubber ring or band for constriction)),
and the age of animals, castration of male cattle causes
varying degrees of acute distress by inference from increased
plasma cortisol concentrations, acute-phase protein pro-
duction, leukocytosis with lymphopenia, temporal suppression
of immune function, increased scrotal swelling, depressed
temperature gradient between the core body and scrotal
skin, and increased expression of pain-associated behaviours.
In this review, studies also suggest that effect of calf age at cas-
tration on cortisol response depends on the calf breed types
(beef versus dairy). Reduced cortisol responses were found in
studies with dairy breed calves while studies with beef calves
found no difference among age groups. The findings of these
studies could be attributed to the different husbandry manage-
ment of dairy and beef calves. Beef calves are typically separ-
ated from the dams for castration, which could increase the
stress response, regardless of the age of the calf. Early separ-
ation of the dairy calves from their dams occurs within a few
hours of birth which makes it easier to measure the effect of
castration alone on those calves.
The pain, stress and inflammatory responses to tissue injury
elicited by castration procedures constitute an acute challenge
in cattle of all ages. Therefore, it is desirable to devise measures
to alleviate the adverse effects of castration through optimizing
pain management. While there is clear evidence from research
studies that multimodal analgesic protocols are associated with
a significant decrease in plasma cortisol concentration after
castration, the practical implications of these interventions at
farm level require further investigation. In future studies it
would be of interest to develop a comprehensive view of the
dynamic inter- and intracellular cytokine fluxes and associated
changes in cell signaling pathways following castration, in
order to elucidate basic mechanisms involved in the modu-
lation of pain including ‘inflammatory-anti-inflammatory’ cas-
cades. The subjective emotional experience of castration pain
is probably one of the most significant determinants of acute
distress in cattle. However, suitable models for measurement
of the emotional distress associated with the perception of cas-
tration pain in cattle have yet to be fully developed. Further-
more, the behaviour-based approach is likely to be the most
practical, non-invasive method for the assessment of castration
stress and pain in cattle.
Studies investigating the effect of age at castration on stress
response are important to draw recommendations on what is
the optimum age to castrate calves. In addition, understanding
how calves castrated at different ages respond to pain is also
helpful to recommend the optimum timing of pain relief.
When considering how age at castration affects calf welfare,
the consensus is that the younger the calf is at time of

castration, the less impact castration may have on its welfare.
Research has indicated that the pain associated with castration
of younger calves is at a lower level and has less of a negative
impact on the calf in terms of welfare, health and performance.
In practice, there is no advantage of banding over burdizzo
castration in adult animals. The administration of anti-tetanus
vaccination and antibiotics at castration add to the financial
cost. There are also some contradictory reports on growth sup-
pression associated with castration stress, which may be due to
the ages / weights of the animals studied. It appears that
banding or burdizzo castration had less impact on growth in
5.5 months old than in 14 months old castrates. A combination
of banding and burdizzo techniques has been evaluated in
young lambs and indicates that there is a benefit in terms of
reduced stress responses. However, only one study in the litera-
ture (Molony et al., 1995) has attempted to address this as an
alternative technique for castration of calves and this was
done at a very young age (5–7 days). Further evaluation of com-
bined burdizzo / banding approaches to castration is warranted
in cattle.
Regarding the impact of castration on immune function and
inflammation, transcriptomics and proteomics could be
employed to demonstrate an integrated and thorough profile
of leukocyte (and /or other cells) functioning (probably in
terms of gene expression) during and post castration.
Castration has been shown to elicit physiological stress,
inflammatory reactions, pain-associated behaviour, suppres-
sion of immune function, and a reduction in growth to
varying degrees. The scientific assessment of animal health
and welfare has been significantly advancing through multidis-
ciplinary approaches including physiology, behaviour studies,
immunology, qualitative and quantitative molecular biology,
and functional genomics. This should allow the impact of cas-
tration on cattle welfare to be further assessed using biological
indices of welfare.
Disclosure statement
No potential conflict of interest was reported by the author(s).
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