Applied Animal Behaviour Science xxx (xxxx) xxxx

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Applied Animal Behaviour Science

journal homepage: www.elsevier.com/locate/applanim

Effect of analgesic strategies on pain behaviour associated with combined

ring castration and hot iron tail docking in Merino lambs
Nikki J. Kellsa,*, Ngaio J. Beausoleila, A. Jonathan R Godfreyb, Katherine E Littlewooda,
R. Neil Warda, Craig B. Johnsona
a
Animal Welfare Science & Bioethics Centre, School of Veterinary Science, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand
b
School of Fundamental Sciences, Massey University, Private Bag 11-222, Palmerston North 4442, New Zealand

A R T I C LE I N FO A B S T R A C T

Keywords: Most countries don’t require analgesia to be provided to lambs < 6 months of age undergoing castration and tail-
Analgesia docking. However, increasing concern for the welfare of farmed livestock means that analgesia is likely to be
Behaviour mandatory at all ages in the future. We investigated whether prior administration of local anaesthesia (LA),
Castration alone or in combination with a systemic non-steroidal anti-inflammatory drug (meloxicam), reduced beha-
Merino lambs
vioural indicators of pain in 4–6-week-old New Zealand Merino lambs following ring castration and hot iron
Pain
docking. Ninety singleton male lambs were randomly allocated to one of six treatment groups: sham castration
Tail docking
and docking (C/D) (SHAM); C/D without analgesia (CON); C/D with LA (LA); C/D with systemic meloxicam
(MEL); C/D with LA and meloxicam administered separately (LA + M); or C/D with LA and meloxicam mixed
and administered together (LAM). Pain behaviours were scored continuously during three 30-minute periods
prior to (period A), shortly after (15–45 min; Period B), and 4 h (Period C) after treatment. Significant treatment
x period interactions were identified for proportions of time spent normal standing/walking (P < 0.001), ab-
normal/standing walking (P = 0.002) and twisted lying (P < 0.001), and the frequency of foot kicks/stamps
(P < 0.001), turns to hindquarters (P = 0.025), restlessness (P < 0.001), hindlimb stretches (P < 0.001) and
tail lifts (P < 0.001). CON lambs spent more time standing/walking abnormally and twisted lying and less time
standing/walking normally in Period B, relative to Period A and relative to SHAM lambs. CON lambs also
exhibited higher frequencies of foot stamp/kicks, restlessness, turns to hindquarters, hindlimb stretches and tail
lifts than SHAM lambs in Period B. All behaviours returned to pre-treatment values in Period C. Prior admin-
istration of LA, alone or with meloxicam, reduced the time spent twisted lying and the frequency of hindlimb
stretching and tail lifting in Period B but did not influence other pain-related behaviours. Meloxicam alone had
no effect on pain behaviour in any period. Due to the short duration of behavioural responses and the limited
analgesic effects observed, the efficacy of the novel mixture of LA and meloxicam relative to LA and meloxicam
administered separately could not be established. There appeared to be no deleterious effects to combining the
two agents for quicker administration. In conclusion, prior administration of LA reduced the acute pain asso-
ciated with ring castration and hot iron docking to some degree. Further research is required to determine an
optimal analgesic strategy for reducing pain associated with ring castration and hot iron docking in Merino
lambs.

1. Introduction
Castration and tail docking are routinely performed in most sheep
production systems, to prevent undesirable behaviour and unwanted
breeding, and to improve ease of handling and processing (Sutherland,
2011) and reduce the risk of flystrike (French et al., 1994), respectively.
There is substantial evidence that both of these procedures, applied
separately or in combination, elicit acute pain and/or other negative
affective experiences (e.g. Mellor and Stafford, 2000; Grant, 2004;
Melches et al., 2007).
It has previously been demonstrated that different methods used for
castrating and/or tail docking lambs elicit highly specific behavioural
responses (Lester et al., 1996; Grant, 2004). The specific combination of
rubber ring castration and hot iron tail docking is the method most
commonly used for Merino lambs in New Zealand (Kevin Stafford,
Personal communication). To date, only two studies have reported the

⁎
Corresponding author.
E-mail address: n.j.kells@massey.ac.nz (N.J. Kells).

https://doi.org/10.1016/j.applanim.2019.104914
Received 4 February 2019; Received in revised form 30 August 2019; Accepted 7 November 2019
0168-1591/ © 2019 Elsevier B.V. All rights reserved.

Please cite this article as: Nikki J. Kells, et al., Applied Animal Behaviour Science, https://doi.org/10.1016/j.applanim.2019.104914
N.J. Kells, et al.
behavioural responses of lambs to the specific combination of ring
castration and hot iron docking. The first, involving 4–5 week old New
Zealand Romney lambs, looked only at lamb postures (Lester et al.,
1996), whilst the second, involving 5 week old Australian Merino
lambs, evaluated both posture and the frequencies of various putative
pain behaviours (Paull et al., 2009). Although a number of studies have
investigated the behavioural responses of lambs of different breeds and
ages to ring castration or hot iron docking alone (e.g. Lester et al., 1996;
Sutherland et al., 1999; Stewart et al., 2014), differences in genetics,
age and management are likely to have influenced the observed beha-
viours. Thus, there is a need to characterise the complete behavioural
responses of New Zealand Merino lambs raised under New Zealand
management conditions to the specific combination of ring castration
and hot iron docking.
Castration and docking of lambs are typically carried out in the first
weeks of life, without the provision of analgesia. Internationally, in-
creasing concern for the welfare of farmed animals (Cornish et al.,
2016; Coleman, 2017) has seen the development of regulations gov-
erning the performance of painful husbandry procedures such as cas-
tration and tail docking. For example, in New Zealand and Australia it is
mandatory to provide pain relief when castrating or tail docking lambs
older than 6 months of age (Anonymous, 2014, 2018). However,
younger lambs also exhibit signs of pain in response to castration and/
or tail docking (Molony et al., 1993; Kent et al., 1998), and the pro-
vision of analgesia at any age may become mandatory in the future. In
addition, evidence is emerging of the production benefits, such as
greater average daily weight gain, of providing analgesia for painful
husbandry procedures (Bates et al., 2015, 2016). Thus, there is a need
to develop practical and effective methods for managing pain in live-
stock during routine husbandry procedures.
The objectives of the current study were threefold: i) to characterise
the behavioural responses of New Zealand Merino lambs to the specific
combination of ring castration and hot iron docking; ii) to evaluate the
effect of previously validated analgesic strategies on the behavioural
responses of Merino lambs to this combination of procedures; and iii) to
evaluate the effects of a novel combined local anaesthetic (LA)/non-
steroidal anti-inflammatory drug (NSAID) formulation on the beha-
vioural responses of Merino lambs to ring castration and hot iron
docking. We hypothesised that provision of LA, alone or in combination
with a NSAID, would reduce the expression of pain-related behaviour in
the 60 min immediately following castration and tail docking, whereas
prior administration of a NSAID would reduce pain-related behaviour in
the period 4–5 h after castration and tail docking. Further, we hy-
pothesised that the administration of a novel combined LA/NSAID
formulation would be at least as effective in reducing pain-related be-
haviour in the 60 min after processing as LA and NSAID delivered se-
parately.
2. Materials and methods
The study was carried out on a commercial high-country farm in the
Ashburton Lakes district of New Zealand over three consecutive days
during early summer (Dec) 2017. All procedures were approved by the
Massey University Animal Ethics Committee (protocol 17/85) and were
conducted in accordance with the Massey University Code of Ethical
Conduct for the Use of Live Animals in Research, Testing and Teaching.
2.1. Animals, facilities and procedures
Ninety entire male singleton Merino lambs aged 4–6-weeks,
weighing 15.4 ± 0.23 kg (mean ± SE) were used in the study. On the
first day, the whole ewe-lamb mob was brought down from a hill
paddock at approximately 0630 h (a distance of approximately 1.5 km).
The mob was allowed time to settle and for the ewes and lambs to pair
up before sorting. The mob was subsequently held in a paddock ad-
jacent to the yards and sheds and was mustered into the yards at
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Applied Animal Behaviour Science xxx (xxxx) xxxx
approximately 0700 h on days two and three.
Each day, thirty singleton male lambs were drafted off the mob,
weighed and randomly assigned to one of six treatment groups in a pre-
determined sequence based on random-number generation in Microsoft
Excel. After weighing, each lamb was marked on the head and rump
with coloured stock marker according to treatment group. The lambs
selected for that day were allocated to one of five pens in the adjacent
shearing shed, such that each pen contained one lamb from each
treatment group (n = 6 per pen). After allocation to their pens, the
lambs were left to settle for a minimum of 30 min.
On each day, pens were processed sequentially, with treatments
applied to all lambs in one pen and video data collection begun before
moving to the next. Within each pen, each lamb was caught and held by
a researcher. Analgesia was administered by a veterinarian, after which
treatment (castration and docking or sham) was performed by a single
farm staff member. In each pen, the order in which lambs were caught
determined the treatment order for that pen.
The indoor pens were approximately 2.5 × 4 m, with slatted
wooden floors and were located in a single row along one side of the
shed. Alloy gates (Prattley Industries Ltd, Temuka, NZ) were used to
partition the existing pens into equal areas; these permitted visual and
olfactory contact between adjacent pens.
Lambs were inside the shed for a total of approximately six hours on
their day of testing. No feed or water was provided in the pens. After
data collection (and treatment of sham lambs – see below), lambs were
returned to their dams in the adjacent paddock. Prior to being reunited
with the mob, all lambs were ear tagged, vaccinated against scabby
mouth (contagious ecthyma or pustular dermatitis) and given a B12
injection by farm staff as per normal farm practice. At the end of day
three, the mob was returned to the original paddock.
2.2. Treatments
Lambs were randomly assigned to one of the following six treatment
groups (n = 15 per treatment):
1 Castration and tail docking (CON). Lambs were picked up and held
on their back with their rump resting on the side of the pen.
Castration was performed by the application of a latex rubber ring
proximal to the testes using an elastrator device. Tail docking was
then carried out using a portable gas-heated docking iron to remove
the distal approximately 4/5 of the tail; enough of the tail was left to
cover the anus. After castration and docking, the lamb was returned
to the pen.
2 Sham castration and tail docking (SHAM). Lambs were handled and
restrained in the same manner and for the same duration as ca-
strated and tail docked lambs. The testes and tail were briefly and
gently manipulated to simulate application of the ring and hot iron,
before returning the lamb to the pen.
3 Local anaesthesia plus castration and tail docking (LA). Immediately
prior to castration and docking, 5 mL of lignocaine solution (1 mL of
2 % lignocaine (Lopaine, Ethical Agents Ltd, Auckland, NZ) diluted
to 5 mL in saline) was injected subcutaneously into the scrotal neck,
spermatic cords and tail using a 25-gauge needle. A 1 mL volume
was injected into the anteriomedial surface and 0.5 mL into each
lateral surface of the scrotum and massaged to disperse. A further
0.5 mL was injected into each spermatic cord, followed by 1 mL into
either side of the base of the tail. Castration and tail docking were
then carried out as for CON.
4 Meloxicam plus castration and tail docking (MEL). Immediately
prior to treatment, 1 mL of 2 % meloxicam (Metacam 20, Boerhinger
Ingelheim, Auckland, NZ) was injected intramuscularly into the
neck using a 20-gauge needle. Castration and tail docking were then
carried out as described for CON.
5 Local anaesthesia and meloxicam (administered separately) plus
castration and tail docking (LA + M). Immediately prior to
N.J. Kells, et al.
treatment, 5 mL of lignocaine solution was administered as de-
scribed for LA and 1 mL of meloxicam was administered as de-
scribed for MEL. Castration and tail docking were then carried out as
described for CON.
6 Local anaesthesia and meloxicam (administered in combination)
plus castration and tail docking (LAM). A mixture of 20 mg lig-
nocaine and 20 mg of meloxicam was prepared by adding 1 mL of 2
% lignocaine and 1 mL of 2 % meloxicam to a 5 mL syringe and
diluting to 5 mL with saline. Immediately prior to treatment, 5 mL of
the mixture was administered subcutaneously as described for LA
above. Castration and tail docking were then carried out as de-
scribed for CON.
At the end of data collection on each day (approximately 5 h after
castration and docking), lambs in the SHAM group were castrated and
tail docked after receiving LA and systemic meloxicam (as per LA + M
above). At this time lambs in the CON group were given systemic me-
loxicam (as per MEL above) to reduce any residual inflammatory pain.
2.3. Behaviour observations
Ten HD video cameras (6 Sony Handycam DCR-SR20 and 4 Sony
Handycam DCR-SR85 (Sony Corporation, Tokyo, Japan) were used to
record the behaviour of lambs. Two cameras were mounted on the
rafters, one at either end of each pen.
For each pen, pre-treatment behaviour (period A) was continuously
recorded for 30 min following the settling period. All lambs in the pen
were then treated. After the last lamb in the pen was treated, 60 min of
post-treatment behaviour was recorded (Period B). This was repeated
sequentially for each of the five pens. At the end of the 60-minute post-
treatment period, the cameras were turned off and the lambs were left
undisturbed in the shed for a period of three hours. At 4 h post-treat-
ment the cameras were activated in each pen sequentially to con-
tinuously record behaviour for a further 60 min (Period C).
Lamb behaviour was scored from the video recordings by a single
trained observer. Thirty consecutive minutes of video were scored from
each observation period for each lamb, as shown in Fig. 1. The first
15 min post-treatment were excluded to avoid disturbances due to
treatment of lambs in the adjacent pen. Thus, period B comprised
15–45 min post-treatment. Likewise, in period C the first 15 min of re-
cording were excluded to avoid any disturbance caused by activation of
cameras in the adjacent pen. Therefore, period C comprised 4.25–4.75 h
post-treatment.
Continuous behavioural observations were scored using
Behavioural Observation Research Interactive Software (BORIS) ver-
sion 5.0.1 (Friard and Gamba, 2016). The observer was blind to treat-
ment group; however, lambs in the SHAM group were unavoidably
distinguishable by the presence of intact tails. Behaviour scoring
training was carried out over a period of approximately four weeks.
Initial training on recognition of specific behaviours and using BORIS
comprised sample scoring of six 10-minute video clips (two each from
periods A, B and C). The first 10-minutes of each period was selected for
training, as these periods represented the widest range of behaviours.
Fig. 1. Schematic indicating the time periods for which lamb behaviour was scored from video recordings. A =30 min immediately prior to treatment;
B = 15–45 min post-treatment; C = 255–285 min post-treatment.
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Applied Animal Behaviour Science xxx (xxxx) xxxx
Following initial training, and once the scorer was confident in iden-
tifying behaviours, two 30-minute videos (one each from periods A and
B in a single pen) were scored in duplicate and the intra class correla-
tion coefficient calculated for each behaviour. Actual scoring was only
undertaken once consistency in scoring had been demonstrated (Intra-
class correlation coefficients of 0.75 or greater) for all behaviours.
The state and event behaviours scored in this study were selected
based on their previous validation as pain indicators in lambs under-
going painful husbandry procedures (Lester et al., 1996; Graham et al.,
1997; Dinniss et al., 1999; Thornton and Waterman-Pearson, 1999,
2002; Stewart et al., 2014; Marini et al., 2017). State behaviours ana-
lysed included proportions of time spent in various ‘normal’ or ‘ab-
normal’ postures or movements (see Table 1). The event variables
‘stamp’ and ‘kick’ were combined into a new variable ‘foot’ due to poor
intra-rater reliability when these were assessed separately.
2.4. Statistical analyses
To evaluate rater reliability, intra-class correlation coefficients (ICC)
were calculated for scored lamb behaviours using the Mixed procedure
in SAS version 9.4 (SAS Institute Inc., Cary, NC, USA), based on the
method described in Winer (1971).
Measurements of lamb behaviour fell into two main categories;
proportions of total time, expressed as percentages (state behaviours),
and counts of specific behaviours (event behaviours). For state beha-
viours, some of the observed proportions summed to 100 % (i.e. normal
and abnormal states) and thus were mutually interdependent. To ac-
count for this, we modelled each of the proportions as if they were
independent, but assessed the outcome of the models in combination.
All proportion data were arcsine square root transformed to allow a
standard linear model to be fitted. For results depicted in graphs, fitted
values from the models have been back-transformed onto the original
proportion scale.
Following data collection, a very high number of zero counts were
observed for several event behaviours, limiting the available options for
subsequent statistical analyses. Some event behaviours were expressed
by relatively few lambs (i.e. there were many 0 values) but some in-
dividual lambs exhibited high counts of the behaviour. Therefore, sta-
tistical models that could account for both the range of counts and the
high number of zero values were needed. These data were analysed
using generalised linear models for Poisson count data with log link
function, zero inflated mixture models with the logit link function for
proportion of zeroes observed and the log link for the counts.
In addition, some event behaviours could only be performed when
standing or lying. Thus, the proportion of time spent in the appropriate
state (standing or lying) was used as a covariate in the generalised
linear models, but was omitted from the zero inflated models as it
added no statistical value. Manual effort was required to explore the
range of zero inflated models that were best suited to each count
variable, while a stepwise routine was used to determine the best
generalised linear model. In all cases, the Akaike information criterion
was used to compare all models, with final model selection based on
human selection.
N.J. Kells, et al. Applied Animal Behaviour Science xxx (xxxx) xxxx

Table 1
Ethogram used for behavioural observations of 4–6-week-old Merino lambs (n = 90) that were either sham castrated and tail docked, or castrated and tail docked
with no analgesia, local anaesthesia (LA) alone, meloxicam alone, LA and meloxicam given separately, or LA and meloxicam given as a mixture (n = 15 per
treatment).
Category Behaviour Description

State Normal standing or walking Standing or walking with no apparent abnormalities

Abnormal standing or walking Statue standing (immobile standing with obvious withdrawal from interaction with pen mates and outside stimuli); or Standing with
arched/hunched back; or Unsteady standing or walking (swaying, staggering, stiff or otherwise abnormal gait); or Backward walking
(walking backward, leading with hind limbs, all legs must move backward); or Stretched standing (standing with both hind feet
planted behind base of tail); or Standing with tail lifted away from anus
Normal lying Sternal recumbency with hind limbs tucked under or near body and head up or down
Twisted lying Sternal recumbency with one or both hind limbs partly or fully extended. The hind legs were considered extended if one leg was fully
visible and the underlying leg was visible from the hoof to the hock.
Lateral lying Lying on one side with shoulder on ground with head up or down
Event Kick A kicking movement of the hind limb up towards the belly when standing or lying: circular type motion
Foot stamp A fast, forceful movement of the forefoot or hind foot onto the ground: vertical motion
Turn to hindquarters Turn head to look at, sniff/bite or lick scrotal or tail area. Head greater than 90 degrees to the shoulder, neck curved toward rear.
Restlessness Number of times lamb stood up or lay down with one count each time it stood up and one count each time in lay down (including up
to or down from knees)
Body/neck stretch Neck or forelimbs and torso extended rostrally for a brief period (< 2 seconds) while lying or standing
Hindlimb stretch Extension of hind limbs away from body while lying down
Tail lift Tail lifted ≥90 degrees away from the anus
Tail wag Movement of tail from side to side. One count for each occurrence, regardless of duration. A new count began when the behaviour
was absent for > 2 seconds
Abdominal arch Tucked abdomen, arched back
Head butt Butting head at pen walls or another lamb
Mouthing Nibbling/chewing/licking other lambs, objects (excluding licking scrotal area). One count for each occurrence, regardless of
duration. A new count began when the behaviour was absent for > 2 seconds
Hop Both back hooves leave the ground simultaneously (within one second of each other) in a leaping
jumping upward and forward action
Shake Brief, vigorous shaking of body (voluntary)
Ease quarters Weight was shifted slowly between hindquarters, without walking
Roll One roll counted when lamb rolled from lying on one side of the body to the other, or when lamb rolled from side of the body to the
back and returned to the same side

The event behaviours foot stamp/kick, restlessness and abdominal
arch were analysed using generalised linear models with log link
function. For all other event behaviours, the high number of zero counts
observed warranted the use of zero inflated models.
Although significant interaction effects between day and some other
variables were identified (see Supplementary Table 1), these are not
presented for two reasons. Firstly, these were generally trivial in nature
and secondly, day was modelled as a randomised block effect in the
zero inflated models, as the tools to analyse day as a random effect in
these models were not available. Therefore, these interactions were not
considered meaningful in the context of this study.
All behaviour analyses were performed in R version 3.5.2 (R Core
Team, 2018), with zero inflated models fitted using the pscl add-on
package (Jackman, 2017). Unless specified otherwise, default treatment
contrast methods were used.
3. Results
The event behaviours hop and roll were never observed, whilst ease
quarters was only observed in two individuals on one occasion each.
These behaviours were therefore excluded from further analyses.
The behavioural responses of Merino lambs to ring castration and
hot iron tailing were relatively short-lived, with most having resolved
(returned to pre-treatment level) by 5 h. The exceptions to this were
reduced time spent lying normally and elevated time spent standing/
walking abnormally; both changes persisted at 4–5 h after treatment
(Fig. 2).
3.1. Characterisation of behavioural responses to combined ring castration
and hot iron docking
Significant treatment x period interactions were identified for the
state behaviours normal standing/walking, abnormal standing/walking
and twisted lying (Table 2). Significant treatment x period interaction
effects were observed for all event behaviours except abdominal arch
and shake (Table 3). Graphs of back-transformed mean frequency of
behaviour in each observation period for each treatment group, aver-
aged over the three days, are presented in Figs. 3–7.
In the 15–45 min after treatment (period B), sham lambs spent less
time lying normally and more time standing/walking normally than
before treatment (Fig. 2a, c). Lambs castrated and docked without an-
algesia (CON) also spent less time lying normally in the 5 h following
treatment. However, they spent less time in period B standing/walking
normally and more time standing/walking abnormally and twisted
lying than they had before treatment and compared to sham-treated
lambs in that period (Fig. 2a, b, d). Whilst time spent standing/walking
normally and twisted lying had returned to pre-treatment values 4–5 h
after treatment in CON lambs, time spent standing/walking abnormally
remained elevated relative to pre-treatment.
Sham-treated lambs exhibited no changes in the frequency of
stamping/kicking (foot events) or turning towards their hindquarters
(Fig. 3a,b), getting up and lying down (restlessness; Fig. 4), or
stretching their neck/body or hindlimbs (Fig. 5a,b) over the 5 -h ob-
servation period. Sham lambs showed a small progressive increase in
the frequency of tail wagging and lifting over the 5 h after treatment but
these behaviours occurred infrequently in this group (Fig. 6a,b). In
contrast, in the 15−45 min following castration and docking, CON
lambs exhibited marked increases in the number of stamps/kicks, turns
to hindquarters, restlessness, hindlimb stretches and tail lifts and a
small increase in the frequency of tail wagging. Five hours after treat-
ment, all counts had returned to pre-treatment values. The frequency of
neck/body stretching did not change in either group.
Mouthing occurred infrequently and the small changes in incidence
observed for both CON and SHAM lambs over the 5 -h observation
period were unlikely to be significant (Fig. 7a). The incidence of head
butting differed markedly among treatment groups in the pre-treatment
period (Fig. 7b). A general pattern of reduced frequency of head butting
in period B, returning to pre-treatment levels in period C, was observed

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N.J. Kells, et al. Applied Animal Behaviour Science xxx (xxxx) xxxx

Fig. 2. Proportion of time spent a) normal standing/walking, b) abnormal standing/walking, c) normal lying, and d) twisted lying by Merino lambs (n = 90) in each
of three thirty-minute observation periods (illustrated in Fig. 1): prior to treatment (A), 15–45 min after treatment (B) and 4.25–4.75 h after treatment (C). Lambs
were allocated to one of six treatment groups (n = 15 per group): sham castration and tail docking (SHAM), castration and tail docking without analgesia (CON),
castration and tail docking after administration of local anaesthesia alone (LA), castration and tail docking after administration of meloxicam alone (M), castration
and tail docking after administration of LA + meloxicam given separately (LA + M), or castration and tail docking after administration of LA + meloxicam ad-
ministered as a mixture (LAM).

for both CON and SHAM lambs.
3.2. Evaluation of the efficacy of previously validated analgesic strategies
The effects of prior administration of local anaesthetic (LA) and
meloxicam (M), alone or in combination (LA + M), on the behaviour of
lambs subjected to combined ring castration and hot iron tail docking
are depicted in Figs. 2–7.
Over the five hours after castration and docking, lambs in all three
analgesic groups exhibited a similar pattern of behavioural responses to
lambs which received no analgesia (CON). Specifically, in period B,
lambs castrated and docked after administration of LA, M, or LA + M
showed a similar reduction in the time spent standing/walking nor-
mally and a similar increase in the time spent standing/walking ab-
normally as was seen in CON lambs (Fig. 2). Time spent standing/
walking abnormally remained elevated above pre-treatment levels in
Period C in all castrated/docked lambs, regardless of analgesia ad-
ministration. Provision of local anaesthetic and/or meloxicam had no
significant effect on the number of stamps/kicks or restlessness in ca-
strated and docked lambs in period B (Fig. 3a and 4). Similarly, there
was no reduction in the number of turns toward the hindquarters in this
period. In fact, there was some indication that prior administration of
local anaesthesia (LA or LA + M) increased the performance of this
behaviour and the higher incidence persisted in LA + M lambs at 5 h
post-treatment (Fig. 3b). Likewise, lambs in the LA and LA + M treat-
ments exhibited a marked increase in tail wags in period B, whereas the
M group was similar to the control and sham treatments (Fig. 6a).
There was some evidence of an effect of analgesia on the expression
of twisted lying. While all castrated and docked lambs showed a tran-
sient increase in the time spent in this posture (period B), the

Table 2
Summary of analyses of proportion of time lambs spent in various state behaviours following castration and tail docking with different analgesic strategies. For each
variable, F statistics and p-values are shown for the main effects of day, treatment and time period (before, immediately after and 4 -hs after treatment) and the
interaction between treatment and period. Significant p-values (< 0.05) are bolded.
Standing/Walking Lying

Effect Normal Abnormal Normal Twisted Lateral

F P F P F P F P F P

Day 1.201 0.3026 3.841 0.0227 0.655 0.5204 2.165 0.0752 2.332 0.0995
Treatment 13.258 0.0000 4.214 0.0011 1.572 0.1648 11.123 0.0000 0.603 0.6978
Period 30.247 0.0000 112.75 0.0000 18.679 0.0000 60.276 0.0000 6.763 0.0014
Treatment*Period 4.772 0.0000 2.798 0.0027 1.169 0.3120 3.358 0.0004 0.844 0.1822

5
N.J. Kells, et al. Applied Animal Behaviour Science xxx (xxxx) xxxx

Table 3
Summary of analyses of event behaviours following castration and tail docking with different analgesic strategies. For each variable, analysis of deviance test
statistics and p-values are shown for all main effects and treatment by period interactions (where included in the final Poisson models). NA = effect not included in
the final model. Results for all interactions included in the final models are provided in Supplementary information 1.
Variable Day Treatment Period Treatment*Period

Dev P Dev P Dev P Dev P

Foot 8.9 0.0155 885.0 < 0.0001 6630 < 0.0001 533.2 < 0.0001
Turn to hindquarters 4.36 0.1131 63.45 < 0.0001 194.6 < 0.0001 20.44 0.0254
Restlessness 16.3 0.0003 786.5 < 0.0001 4693 < 0.0001 263.7 < 0.0001
Stretch neck/body 9.00 0.0111 57.09 < 0.0001 56.26 < 0.0001 55.05 < 0.0001
Stretch hindlimb 6.40 0.0407 326.74 < 0.0001 1748.5 < 0.0001 53.06 < 0.0001
Tail wag 27.11 < 0.0001 119.33 < 0.0001 389.49 < 0.0001 133.56 < 0.0001
Tail lift 129.45 < 0.0001 26.28 < 0.0001 308.86 < 0.0001 113.06 < 0.0001
Abdominal arch NA NA 19.20 0.0018 3.15 0.2075 NA NA
Mouthing 40.76 < 0.0001 34.68 < 0.0001 81.18 < 0.0001 42.31 < 0.0001
Shake 8.59 0.0136 17.71 0.0033 41.93 < 0.0001 8.07 0.6217
Head butt 80.89 < 0.0001 156.21 < 0.0001 63.74 < 0.0001 72.84 < 0.0001

Fig. 4. Mean number of restlessness events (transitions from standing to lying

or vice versa) performed by Merino lambs (n = 90) in each of three thirty-
minute observation periods (illustrated in Fig. 1): prior to treatment (A),
15–45 min after treatment (B) and 4.25–4.75 h after treatment (C). Lambs were
allocated to one of six treatment groups (n = 15 per group): sham castration
and tail docking (SHAM), castration and tail docking without analgesia (CON),
castration and tail docking after administration of local anaesthesia alone (LA),
castration and tail docking after administration of meloxicam alone (M), cas-
tration and tail docking after administration of LA + meloxicam given sepa-
rately (LA + M), or castration and tail docking after administration of
LA + meloxicam administered as a mixture (LAM).

Lambs receiving meloxicam alone responded similarly to CON lambs

Fig. 3. Mean number of a) foot events (stamps and kicks) and b) turns to
hindquarters performed by Merino lambs (n = 90) in each of three thirty-
minute observation periods (illustrated in Fig. 1): prior to treatment (A),
15–45 min after treatment (B) and 4.25–4.75 h after treatment (C). Lambs were
allocated to one of six treatment groups (n = 15 per group): sham castration
and tail docking (SHAM), castration and tail docking without analgesia (CON),
castration and tail docking after administration of local anaesthesia alone (LA),
castration and tail docking after administration of meloxicam alone (M), cas-
tration and tail docking after administration of LA + meloxicam given sepa-
rately (LA + M), or castration and tail docking after administration of
LA + meloxicam administered as a mixture (LAM).
magnitude of the increase was smaller in lambs that had received local
anaesthetic (alone or with meloxicam: LA, LA + M groups) than for
those that received meloxicam (M) or no analgesia (CON) (Fig. 2d).
for all state behaviours over the 5 h after treatment.
Provision of local anaesthesia also reduced the performance of
hindlimb stretching and tail lifting behaviours in the 15–45-minutes
post-treatment (relative to CON). Although both control lambs and
those that received LA, LA + M or M exhibited an increase in hindlimb
stretching compared with sham lambs, the magnitude of this increase
was smaller in the LA + M and LA groups (Fig. 5b). Likewise, while
both control and analgesia lambs exhibited an increase in tail lifting in
period B, this was smaller in the LA + M and LA groups (Fig. 6b).
Lambs that received LA + M showed an increased frequency of
head/neck stretches in Period B which was not observed in any other
treatment group (Fig. 5a). This behaviour occurred in 3/15 lambs, with
very the high counts observed for two individuals (21 and 23 events in
30 min) likely contributing strongly to the observed effect.

6
N.J. Kells, et al.
Fig. 5. Mean number of a) neck or body stretches and b) hind limb stretches
performed by Merino lambs (n = 90) in each of three thirty-minute observation
periods (illustrated in Fig. 1): prior to treatment (A), 15–45 min after treatment
(B) and 4.25–4.75 h after treatment (C). Lambs were allocated to one of six
treatment groups (n = 15 per group): sham castration and tail docking (SHAM),
castration and tail docking without analgesia (CON), castration and tail docking
after administration of local anaesthesia alone (LA), castration and tail docking
after administration of meloxicam alone (M), castration and tail docking after
administration of LA + meloxicam given separately (LA + M), or castration
and tail docking after administration of LA + meloxicam administered as a
mixture (LAM).
3.3. Evaluation of a novel combination of local anaesthetic and meloxicam
The reduction in twisted lying seen in lambs that received local
anaesthetic and meloxicam separately was also evident in lambs that
were given the novel combined formulation; however, the effect was
less pronounced (Fig. 2b). There were few differences in the perfor-
mance of event behaviours when LA and meloxicam were administered
separately or together.
Separate injections of LA and meloxicam (LA + M) significantly
reduced the rise in the frequency of hindlimb stretching in period B
while LAM had no such effect (Fig. 5b). In contrast, LAM abolished the
small increase in tail wagging observed in CON lambs while lambs re-
ceiving LA + M showed a much larger increase in this behaviour
shortly after treatment (Fig. 6a). Additionally, LAM completely ame-
liorated the post-treatment rise in tail lifting observed in CON lambs,
while this effect was present but less pronounced for LA + M.
4. Discussion
The present study sought to characterise the behavioural responses
7
Applied Animal Behaviour Science xxx (xxxx) xxxx
Fig. 6. Mean number of a) tail wags and b) tail lifts performed by Merino lambs
(n = 90 in each of three thirty-minute observation periods (illustrated in
Fig. 1): prior to treatment (A), 15–45 min after treatment (B) and 4.25–4.75 h
after treatment (C). Lambs were allocated to one of six treatment groups
(n = 15 per group): sham castration and tail docking (SHAM), castration and
tail docking without analgesia (CON), castration and tail docking after ad-
ministration of local anaesthesia alone (LA), castration and tail docking after
administration of meloxicam alone (M), castration and tail docking after ad-
ministration of LA + meloxicam given separately (LA + M), or castration and
tail docking after administration of LA + meloxicam administered as a mixture
(LAM).
of New Zealand Merino lambs to combined ring castration and hot iron
docking performed at 4–6 weeks of age, to evaluate the efficacy of
various combinations of local anaesthesia and/or the NSAID meloxicam
in ameliorating pain-related behaviour, and to determine whether
combining local anaesthetic and meloxicam in the same injection al-
tered their analgesic efficacy.
4.1. Behavioural responses to ring castration and hot iron docking
A suite of behaviours consistent with those identified in previous
studies of ring castration and hot iron tail docking have been char-
acterised in 4-6-week old New Zealand Merino lambs. These include
reduced time spent in normal postures and movements and more time
spent standing/walking and lying abnormally. In addition, active be-
haviours such as foot stamping/kicking, restlessness, turning towards
the hindquarters, hindlimb stretching and tail lifting appear to be good
indicators of pain in the 60 min following this combination of proce-
dures.
The observed reduction in time spent in ‘normal’ postures is con-
sistent with that reported in 4-6-week-old Romney (Lester et al., 1996)
and Australian Merino (Paull et al., 2009) lambs to the same
N.J. Kells, et al.
Fig. 7. Mean number of a) mouthing events (licking, chewing, nibbling) and b)
head butts performed by Merino lambs (n = 90) in each of three thirty-minute
observation periods: prior to treatment (A), 15–45 min after treatment (B) and
4.25–4.75 h after treatment (C). Lambs were allocated to one of six treatment
groups (n = 15 per group): sham castration and tail docking (SHAM), castration
and tail docking without analgesia (CON), castration and tail docking after
administration of local anaesthesia alone (LA), castration and tail docking after
administration of meloxicam alone (M), castration and tail docking after ad-
ministration of LA + meloxicam given separately (LA + M), or castration and
tail docking after administration of LA + meloxicam administered as a mixture
(LAM).
combination of ring castration and hot iron docking. Paull et al. (2009)
also reported increased restlessness and foot stamping or kicking in
Australian Merino lambs. Increased time spent twisted lying, hindlimb
stretching, turning toward hindquarters or tail lifting have not pre-
viously been reported.
To our knowledge, hindlimb stretching or tail lifting events have not
previously been evaluated as pain indicators in lambs. Although full or
partial hindlimb extension in ventral recumbency has been in-
corporated in descriptions of abnormal posture following castration or
tail docking (e.g. Graham et al., 1997; Dinniss et al., 1999; Molony
et al., 2002; Thornton and Waterman-Pearson, 2002; Grant, 2004),
discrete hindlimb stretch/extension events have not been reported. Tail
stretching, defined as ‘holding the tail in an unusual stretched posture
in the absence of urination/defecation’ was similarly incorporated into
an ‘abnormal posture’ index in castrated 10–24-week-old lambs
(Melches et al., 2007), but was not been reported as a discrete event.
Based on the results of the present study, i.e. the absence of these be-
haviours in sham handled lambs and the high incidence in control
lambs, it appears these may be sensitive indicators of pain in lambs
following combined ring castration and hot iron docking.
In line with prior studies evaluating lamb responses to this
8
Applied Animal Behaviour Science xxx (xxxx) xxxx
combination of ring castration and hot iron docking, the expression of
pain behaviour was relatively short lived. For example, Paull et al.
(2009) reported elevated expression of abnormal behaviour 0−4 hours
after treatment but not 4−8 hours after. Likewise, Lester et al. (1996)
reported low incidence of restlessness after the first 60 min and re-
solution of abnormal walking/standing within 2 h. Thus, it appears that
the behavioural expression of acute pain associated with this specific
combination of husbandry procedures in NZ Merino lambs is generally
consistent in terms of duration and character with those reported in
other breeds, or managed under different conditions.
4.2. Effects of analgesic strategies on behavioural responses to castration
and docking
Overall, only limited evidence of effective analgesia was found in
this study. Local anaesthetic ring block of the tail and scrotum applied
immediately prior to treatment significantly reduced the proportion of
time spent twisted lying and the incidence of hindlimb stretching and
tail lifting, but had little effect on the proportion of time spent abnormal
standing/walking, or the incidence of restlessness. Twisted lying, hin-
dlimb stretching and tail lifting were reduced to an even greater degree
when LA was accompanied by a separate injection of meloxicam
(LA + M). Previous studies have also demonstrated significant reduc-
tions in twisted lying (Kent et al., 1998; Molony et al., 2002) or ab-
normal lying (incorporating twisted lying) (Stewart et al., 2014) in
lambs given LA prior to ring castration.
Prior studies of lambs castrated with rubber rings also reported no
effect of LA on the proportion of time standing or walking abnormally
(Molony et al., 1997; Kent et al., 1998; Dinniss et al., 1999). In contrast
to the present study, Dinniss et al. (1999) reported a reduction in
restlessness in ring castrated lambs that were given 2 % lignocaine in
either the scrotal neck (2 mL) or spermatic cords (0.5 mL in each cord)
15 min prior to ring application. Likewise, Stewart et al. (2014) found
that injection of 6 ml of 2 % lignocaine (2 mL into each testis and 2 mL
into the scrotal neck) immediately prior to ring castration effected a
reduction in postural changes (akin to restlessness). No such reduction
in restlessness was observed in the present study where lambs received
approximately 0.6 mL of 2 % lignocaine as a scrotal block immediately
prior to rubber ring castration. This may reflect differences in the site,
volume, or timing of anaesthetic injection between studies.
Prior administration of meloxicam alone had no effect on the ex-
pression of pain-related behaviour in the 60 min after treatment. This
was expected, given that NSAIDs primarily act against inflammatory
pain, which manifests secondary to tissue damage. Because behavioural
responses to treatment had largely resolved by 4–5 h after treatment,
when meloxicam was expected to exert its main effects, there was no
real possibility to gauge any effects. Paull et al. (2012) similarly re-
ported no reduction in ‘active’ pain behaviours (including restlessness,
foot stamps or kicks and wound licking) in Merino lambs that were
given meloxicam prior to ring castration. The authors did, however,
observe an effect of meloxicam on postural behaviours over the 12 h
after ring castration. The reason for these discrepancies may be due to
the sampling regime or the dose and route of meloxicam administra-
tion. Paull et al. (2012) measured state behaviours by scan sampling
every 15 min for 12 h. As such, they would have captured any changes
that occurred between 1 and 4 h that were not assessed in the current
study. Furthermore, the total dose of meloxicam was considerably
higher than that used in the present study (100 mg versus 20 mg) and
was injected subcutaneously around the scrotum prior to ring applica-
tion, whereas in the present study meloxicam was injected in-
tramuscularly in the neck.
In the current study, lambs that received LA (LA, LA + M, LAM)
exhibited more tail wagging and turns toward the hindquarters than did
control lambs. It is possible that these responses, particularly the tail
wagging, relate to the sensation of numbness induced by LA; however,
this has not been substantiated. In addition, lambs given LA + M
N.J. Kells, et al.
showed a marked increase in the frequency of neck/body stretching
after castration and docking. This behaviour was very rarely expressed
overall and the finding likely reflects the very high counts observed in
two individuals, rather than a biologically relevant treatment effect.
4.3. Comparison of efficacy of separate versus combined local anaesthetic
and meloxicam administration
Due to the short duration of pain responses and the limited of evi-
dence of effective analgesia, it was difficult to compare the efficacy of
the novel combination of LA and meloxicam with administration of the
two agents separately. There was some suggestion that separate ad-
ministration might have been more effective than the mixture in re-
ducing behavioural responses to castration and docking. For instance,
the analgesic effect of separate injections of LA and M on time spent
twisted lying was more pronounced than the effect of the combined
injection (LAM). Likewise, separate injections significantly reduced the
post-treatment rise in hindlimb stretching while LAM had no such ef-
fect.
In contrast, LAM completely abolished the post-treatment rise in tail
lifting observed in control lambs, while this effect was present but less
pronounced for LA + M. Thus, the comparative analgesic effect of the
two approaches to administration is still equivocal. However, there
appear to be no deleterious effects to combining the two agents for
quicker administration.
5. Conclusions
Behavioural responses of Merino lambs subjected to ring castration
and hot iron docking were characterised by increases in foot stamping/
kicking, restlessness, turning toward hindquarters, hindlimb stretching
and tail lifting, in addition to changes in the proportions of time spent
in normal and abnormal postures. Overall, there was some indication
that local anaesthetic ameliorated the acute pain of castration and
docking to a degree and that concurrent administration of meloxicam
improved this effect, while meloxicam alone had no effect on pain-re-
lated behaviour over the 5 -h observation period. Pain behaviour was
short-lived, having resolved by 5 h post treatment. The use of a novel
combination of local anaesthetic and meloxicam delivered in a single
infection produced similar analgesic effects to the two drugs adminis-
tered separately. Further research is required to determine the most
appropriate combination of drugs and/or route of administration to
alleviate pain in Merino lambs undergoing painful husbandry proce-
dures.
Acknowledgements
This research was funded by “W3 - Wool Unleashed", a Primary
Growth Partnership programme between The New Zealand Merino
Company Ltd and the Ministry for Primary Industries. The authors
gratefully acknowledge the cooperation of the owners and manager of
the property where data collection took place, and Boehringer
Ingelheim for supplying the Metacam for the trial.
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
online version, at doi:https://doi.org/10.1016/j.applanim.2019.
104914.
Applied Animal Behaviour Science xxx (xxxx) xxxx
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