DOI: 10.

1638/2017-0194 Journal of Zoo and Wildlife Medicine 51(3): 524–536, 2020

Copyright 2020 by American Association of Zoo Veterinarians

ANESTHESIA AND SURGERY PROTOCOLS FOR INTRA-

ABDOMINAL TRANSMITTER PLACEMENT IN FOUR SPECIES OF
WILD ARMADILLO

Danilo Kluyber, MsC, Rodrigo Pinho Gomez Lopez, DVM, Gabriel Massocato, BSc, Nina Attias, PhD,
and Arnaud Léonard Jean Desbiez, PhD

Abstract: The anatomy and physiology of armadillos make anesthetic procedures and the placement of
telemetry devices challenging. From June 2011 to November 2019, a total of 73 free-living armadillos were
captured and anesthetized in the Pantanal, Brazil. Giant (Priodontes maximus), six-banded (Euphractus sexcinctus),
southern naked-tailed (Cabassous unicinctus), and nine-banded (Dasypus novemcinctus) armadillos were divided into
two groups. In group 1, 30 armadillos were anesthetized for collection of biological samples, body measurements,
and placement of a microchip tag. Anesthetic combination BDM was applied: butorphanol tartrate, detomidine
hydrochloride, and midazolam hydrochloride, each at 0.1 mg/kg. In group 2, 43 armadillos received ketamine
hydrochloride at 10 mg/kg 20 min after BDM injection, and intra-abdominal radio transmitters were surgically
implanted. The transmitter was inserted freely into the abdominal cavity. Vital signs were monitored during
anesthesia every 10 min and varied within species and between groups. Rectal temperature varied from 33.1 6
1.36 to 35.34 6 1.218C, heart rate (beats/min) from 19 6 2.14 to 84.71 6 9.25, respiratory rate (breaths/min) from
11 6 4.16 to 31 6 2.82, and oxygen saturation values (SPO2%) from 84.17% 6 2.39 to 98% 6 1.20. Both groups
received the antagonist combination NYF: naloxone hydrochloride (0.02 mg/kg), yohimbine hydrochloride (0.125
mg/kg), and flumazenil (0.01 mg/kg). Recovery varied according to intravenous or intramuscular injection from 2
6 4 to 8.08 6 2.93 min respectively. BDM protocol was considered satisfactory and provided enough time to
complete the procedures (60 6 85 to 133.20 6 9.12 min) according to the species and group. Ketamine added to
the BDM provided enough time and a surgical plane of anesthesia (97 6 22 to 137 6 39.5 min). The surgical
procedure technique chosen did not appear to have a negative impact on armadillos studied. Implantable
transmitters provide a cost-effective method for long-term monitoring of wild individuals.

INTRODUCTION fall off in a few days or weeks, due to the

Armadillos belong to the Cingulata order and
the family Dasypodidae, with nine genera and 20
species that range from the tiny pink-fairy arma-
dillo (90 g) (Chlamyphorus truncatus) to the giant
armadillo (60 kg) (Priodontes maximus).28,29,31 Com-
pared with other mammals, armadillos are chal-
lenging to study due to their low metabolism, low
body temperature, and primitive anatomy. Their
anatomy prevents placement of conventional
telemetry devices (e.g., collars) and external
devices fitted on the carapace usually break or
From the Naples Zoo at Caribbean Gardens, 1590
Goodlette-Frank Road, Naples, FL 34102, USA (Kluyb-
er); from the Instituto Promar, Estrada Lagoa do
Cassange, 34-Marau 45520-000, Bahia, Brasil. (Lopez);
from the Instituto de Conservação de Animais Silvestres-
ICAS, Rua Afonso Lino Barbosa, 142 Chácara Cachoeira,
Cep: 79040290, Campo Grande, Mato Grosso do Sul,
Brasil (Massocato, Attias, Desbiez). Present address
(Attias): Programa de Pós-Graduação em Biologia Animal,
Universidade Federal do Mato Grosso do Sul, Cidade
universitária, 79070-900, Campo Grande, Mato Grosso do
Sul, Brasil. Correspondence should be directed to Danilo
Kluyber (dkluyber@live.com).
armadillos’ fossorial behavior.6,13,24
Implantable transmitters can be a solution to
monitor animals for which the anatomy hinders
the attachment of external tracking devices.6
Nevertheless, few studies using implantable trans-
mitters have been performed with armadil-
los.6,12,13,24 A study using implantable transmitters
in nine-banded armadillo (Dasypus novemcinctus)
highlighted the difficulty of finding and tracking
animals with implants, but reported no deaths as a
consequence of implantable transmitters.13 Re-
cently, wild six-banded armadillos (Euphractus
sexcinctus) were also successfully implanted.6 In
contrast, another study reported that two nine-
banded armadillos died due to the implant12 and a
study that applied implantable transmitter tech-
nique in giant armadillos considered it challeng-
ing due to species behavior and ecological
characteristics.24
Although previous authors6,13,24 consider intra-
abdominal radio transmitters as one of the only
reliable methods for long-term monitoring of
armadillos, they also highlight their concerns
about performing an invasive surgery in the field,
with limited ability to provide ideal antiseptic
conditions, the possibilities of device failure, and

514

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 KLUYBER ET AL—ANESTHESIA AND SURGERY IN ARMADILLOS
deaths as a consequence of surgery and implants.
Hence, one of the requirements for the use of
intra-abdominal transmitters is the identification
of safe and effective anesthetic protocols to
perform implantable transmitter surgery in the
field. Anesthetic protocols evaluations have been
described for armadillos, including both inject-
able and gaseous anesthetics.5,6,8,10,13,19,20,26 Howev-
er, to our knowledge, few studies were done
developing anesthetic protocol focusing on wild
armadillos.5,6,9,10,13,21
Armadillo’s low body temperature, low metab-
olism, anatomy, and the ability to perform apnea,
are the main characteristics that make anesthesia
challenging for this group. Although individuals
of the super order xenarthra (armadillos, anteat-
ers, and sloths) are considered, in general, more
resistant to short episodes of hypoxia,19,20,30 their
particular anatomy makes it impossible to intu-
bate most of the species, and a tracheostomy is
required during emergency.2 Hence, some anes-
thetic protocols that commonly cause hypoxia or
respiratory arrest are considered impractical to
perform under field conditions.
Inhalation anesthesia is highly recommended
for several armadillo species and has been previ-
ously reported in nine-banded armadillos and
giant armadillos.8,13,26 However, even with portable
equipment, this is not easy to perform in field
studies and costs may be preclusive.
Ketamine has been widely used as a dissociative
agent alone or in combination with a-2 agonists,
opioids, or benzodiazepines.5,8,10,13,19,20,26 Studies
report that this agent provides wide safety mar-
gins, good immobilization, and increased time of
unconsciousness when combined with other
agents.8,13,20,26 Ketamine provides a short and
smooth induction, however it does not produce
good muscle relaxation, especially for xenar-
thrans.8,10,16 Furthermore, there is a lack of infor-
mation about its analgesic effects in armadillos.
Opioids and a-2 agonists, combined with
ketamine have been used as anesthetic protocols
in armadillos.13 This combination is also recom-
mended for induction, reducing the dose and
volume of dissociative agents, and hence their
side effects. Most authors also report reversal
characteristic to be a major advantage of the use
of ketamine, providing rapid recovery in the
field.5,6,13
The combination of butorphanol, ketamine,
and medetomidine and the antagonist agents
naloxone, yohimbine, and atipamezole, were suc-
cessfully used for implantation of devices in nine-
banded armadillos.13 A similar anesthetic proto-
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515
col (butorphanol, xylazine, and midazolam) was
used for three captive giant armadillos and fast
recovery was observed, while muscle relaxation
and some short-term analgesic effects were also
provided.8
The combination of ketamine, xylazine, and
midazolam for six-banded armadillos, was de-
scribed as an effective protocol combination for
implantable transmitter surgery, providing ade-
quate time and good immobilization for the
procedure.6 The same authors claimed that this
combination provided a safety margin, can be
partially reversed with antagonists, and has the
added benefits of low volume.6 However, little is
known about the effects of these anesthetics for
other species of armadillos. In this study, two new
anesthetic protocols were developed based on the
previously described protocols. The first anes-
thetic protocol was developed for chemical im-
mobilization and provides sufficient time for
collection of biological samples, body measure-
ments, and microchip placement. The second
protocol was developed to provide a surgical
plane of anesthesia and sufficient time for surgical
implantation of an intra-abdominal radio trans-
mitter. The aims of this study were to: 1) describe
and evaluate the safety and efficiency of two
anesthetic protocols, used for two different pur-
poses and 2) describe and evaluate the surgical
methods to implant intra-abdominal transmitters
in four species of wild armadillos.
MATERIALS AND METHODS
All procedures were authorized by the Brazilian
Ministry of the Environment (MMA), Instituto
¸
Chico Mendes de Conservacão da Biodiversi-
dade–ICMBio, license number 27587-1, issued
on 18 April 2011.
Ninety-five percent of the Pantanal ecosystem
consists of private lands, and this study was
conducted at the Baı́a das Pedras lodge and ranch
(where the Giant Armadillo Conservation Program
[GACP] is based), located in the Nhecolândia
Pantanal subregion, municipality of Aquidauana,
Mato Grosso do Sul state, Brazil (21 K 0627555
UTM 7870292).
Four species of wild armadillos have been
included for the long-term ecology and health
assessment initiatives developed by the GACP,
and 73 armadillos were anesthetized for this
purpose between June 2011 and November
2019. Overall, a total of 33 giant armadillos
(GA; 15 males, 18 females), 26 six-banded
armadillos (SBA; 10 males, 16 females), 2 nine-
banded armadillos (NBA; 1 male, 1 female) and
 516 JOURNAL OF ZOO AND WILDLIFE MEDICINE
12 southern naked-tailed armadillos (Cabassous
unicinctus) (SNT; 4 males, 8 females) were studied.
Small armadillos like SBA and SNT were
captured walking in an open area during the
hottest period of the day, using long-handled dip
nets. The same method was used for NBA, but
researchers observed from the vehicle, and cap-
tures occurred at night. The capture method for
GA was based on tracking footprints or traces (i.e.
recently predated termites). When a burrow was
found with a GA inside (entrance completely
sealed with sand), a jequi-type trap (iron funnel
trap) was placed in front of the burrow entrance,
given that GA burrows have only a single
entrance-exit. Once captured, armadillos were
placed in iron-reinforced wooden crates until
procedures began.
After capture, nocturnal species (GA and NB)
were kept inside cages overnight and procedures
began in the morning at the capture location.
Species considered more active during the day,
like SBA and SNT, were kept inside cages (for
approximately 1 hr), until equipment and tents
were set up for the anesthesia.
At the end of the procedures, all animals were
kept in crates until complete recovery and were
released at the same capture location. For 12 to 22
days (10 days 6 2) after the procedure, each
individual was monitored daily by the GACP
team, using a telemetry receiver (model TR-4) and
a VHF antenna (model RA-23K; Telonics, Inc,
Mesa, AZ 85204-6699, USA). Telemetry was used
to find the animals inside burrows (not active). To
ensure animal well-being, once the armadillo was
encountered, camera traps (model WR6 1080P
HD; Reconyx, Holmen, WI 54636, USA) were
placed in front of the burrow to observe individ-
ual general health condition (e.g. weight loss),
skin suture healing, and behavior. Individuals
were recognized in photos by skin or carapace
scars and the different forms and colors of
carapace scales.7 After this first period of inten-
sive monitoring, armadillos were monitored
monthly through telemetry and camera traps,
until the end of the intra-abdominal transmitter’s
operational life.
Anesthesia
Adult armadillos were divided into two groups
according to the procedure performed and anes-
thetic protocol used. Individuals of group 1 were
anesthetized for collection of biological samples
(blood, feces, swabs, ectoparasites, ear biopsy)
and body measurements, and were tagged with
subcutaneous microchips (size 2.1 3 12 mm;
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Anilhas Caprit Allflex, São Paulo, SP, 04726-
010, Brazil). Individuals of group 2 underwent the
same initial procedures, but intra-abdominal
radio transmitters were surgically implanted (Te-
lonics, Inc).
For chemical immobilization, individuals were
manually handled by team members and received
intramuscular (IM) injections into their hind
limbs. Group 1 consisted of 30 armadillos: 6 GA
(4 males, 2 females), 22 SBA (6 males, 16
females), and 2 SNT (2 females) and received
the following anesthetics: butorphanol tartrate
(Zoetis Brasil, São Paulo, SP, 04711-130, Brazil;
0.1 mg/kg), detomidine hydrochloride (União
Quı́mica Farmacêutica Nacional S/A, São Paulo,
SP, 05676-120, Brazil; 0.1 mg/kg), and midazolam
hydrochloride (Cristália Produtos Quı́micos
Farmacêuticos Ltda, Itapira, SP, 13974908, Bra-
zil; 0.1 mg/kg), from here on referred to as BDM.
Group 2 included 43 armadillos: 27 GA (11
males, 16 females), 4 SBA (4 males), 10 SNT (4
males, 6 females), and 2 NB (1 male, 1 female).
This group received BDM, and ketamine hydro-
chloride (União Quı́mica Farmacêutica Nacional;
10 mg/kg) was given IM (hind limbs), approxi-
mately 20 min after initial induction.
At the beginning of the anesthetic procedure
(20 min), each armadillo was weighed using a
portable scale (model A04, Xtrad, São Paulo, SP,
01023-050, Brazil). While animals were washed
and cleaned, physical examinations were per-
formed including general appearance, body con-
dition, hydration status, mucous membrane color,
capillary refill time, respiratory auscultation, and
presence of scars or wounds. During anesthesia
maintenance, physical examinations were per-
formed to evaluate health status and detect
pregnancy (palpation), and only adult individuals
in good body condition with no detected health
abnormalities received implants. The animals
received 0.9% sodium chloride (Eurofarma, Ri-
beirão Preto, SP, 14095-000, Brazil) given subcu-
taneously, with a dose of 20 ml for SNT, 60 ml for
SBA and NB, and 250 ml for GA.
Vital signs were monitored from the moment
animals reached an anesthetic plane and were
recorded every 10 min during anesthetic mainte-
nance. Heart rate (beats/min) was measured using
a stethoscope Littmannt Classic III (3M, Sumaré,
SP, 13181-900, Brazil), respiratory rate (beats/
min) was obtained by observing respiratory
movements. Relative oxyhemoglobin saturation
(SpO2%) was measured with a pulse oximeter
(sensor attached to tongue or ear), model BN-
Oxy9Vet Plus (Bionet, Tustin, CA 92780, USA)
 KLUYBER ET AL—ANESTHESIA AND SURGERY IN ARMADILLOS
and temperature (8C) was recorded with a digital
thermometer (Incoterm, Porto Alegre, RS,
91751-000, Brazil).
After the procedures, anesthesia was reversed in
both groups using a NYF combination that was
composed of naloxone hydrochloride (Cristália
Produtos Quı́micos Farmacêuticos Ltda; 0.02
mg/kg), yohimbine hydrochloride (Drogavet, São
Paulo, SP, 01234-000, Brazil; 0.125 mg/kg) and
flumazenil (União Quı́mica Farmacêutica Nacio-
nal S/A; 0.01 mg/kg) as antagonists of butorpha-
nol tartrate, detomidine hydrochloride, and
midazolam respectively. Group 1 received NYF
by intravenous (IV) injection (saphenous vein),
and group 2 received NYF by IM injection (hind
limbs), to provide a slower, calmer, and smoother
recovery after the surgical procedure (Table 1).
All individuals were released in the same place
or burrow from which they had been captured and
remained inside burrows for 24 to 120 hr (average
of 48 hr) before leaving for the first time. An
analysis of variance (ANOVA) was used to asses if
the induction time varied among species and a
Tukey post hoc test was used to verify which pairs
of species differed statistically. A Wilcoxon test
for nonparametric data15 was used to evaluate if
body mass, vital signs, and anesthetic parameters
such as induction time (min), mean total anes-
thetic maintenance time (min), and mean com-
plete recovery time after antagonists were applied
(min), differed for each species between the
groups subjected to each anesthetic protocol.
Due to sample size restrictions, comparison
between groups was performed only for GA and
SBA. Graphical representation of the measured
parameters for both groups of each species are
given by boxplots in Supplementary Material 1.
Surgical procedure (implantable transmitter)
Transmitters were disinfected in a sterile plastic
bag with 2% glutaraldehyde solution (Ibasa, Porto
Alegre, RS, 90220-030, Brazil) prior to surgery
(24 hr). Implants were washed twice using 0.9%
sterile physiological saline (Eurofarma) and then
immersed in another sterile plastic bag containing
the same solution (0.9% sterile physiological
saline) to completely remove glutaraldehyde res-
idues or smell.
After being washed, implants were kept in
sterilized envelopes until being implanted. Im-
plants (Telonics, Inc) consisted of a VHF micro-
processor powered by a lithium battery, covered
in wax and dual water barrier. GA received the
IMP 310 model (9.4 3 2.3 cm, 42 g) operational
life (OL) 36 mo, SBA and NB received the IMP
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200 model (6.1 3 2.3 cm, 25 g) with OL of 9 mo,
and SNT received the IMP 130 model (5.3 3 1.9
cm, 21 g) with OL of 5 mo.
Armadillos were washed with warm water and
chlorhexidine (2%) (Indústria Farmacêutica Rio-
quı́mica, S.J. Rio Preto, SP, 15057-441, Brazil).
SNT, SBA, and NB individuals had their abdom-
inal fur shaved. A plastic tent was set up to protect
individuals from the sunlight or rain, and the back
of the truck was covered with a plastic canvas and
used as a table for the procedure.
Prior to the surgical procedure, intradermal
local anesthesia—lidocaine hydrochloride, 2 mg/
kg (Eurofarma)—was infiltrated into the infra-
umbilical region. Ketamine hydrochloride (10
mg/kg) was given IM as a supplementary anes-
thetic immediately after the animals had been
completely washed.
Armadillos were placed in dorsal recumbency
and the abdominal skin was prepared for surgery
using 70% alcohol, 2% povidone iodine (Indústria
Farmacêutica Rioquı́mica), and a surgical drape
(27 3 27 feet) with 3 3 3-foot fenestration
(Veterinary Specialty Products, Shawnee, KS
66214, USA) was applied.
A longitudinal, supra-umbilical skin, subcutane-
ous tissue incision was performed. Muscles were
divided through the linea alba and a peritoneal
incision made. The transmitter was inserted into
the abdominal cavity, where it free-floated.
For GA, simple interrupted suture was applied
for the first layer (peritoneal membrane) using
Vicryl 2 (Johnson & Johnson, Nova Odessa, SP,
13460-000, Brazil). For SNT, SBA, and NBA the
same type of suture was performed, using Vicryl 0
(Johnson & Johnson). The muscle and subcuta-
neous layers were sutured using cruciate pattern,
simple interrupted with Vicryl 2 (Johnson &
Johnson) for GA, and Vicryl 0 (Johnson &
Johnson) for SNT, SBA, and NBA.
Surgical glue—Vetbond tissue adhesive (3M)—
was applied and hydrocolloid adhesive (Convatec,
São Paulo, SP, 04717-004, Brazil) used to tempo-
rarily protect the surgical wound. Meloxicam
(Ourofino Saúde Animal, Cravinhos, SP, 14140-
000, Brazil; 0.1 mg/kg) was injected subcutane-
ously and enrofloxacin (Bayer, São Paulo, SP,
04779-900, Brazil; 8.0 mg/kg) was administered
through deep IM injection.
RESULTS
Anesthetic protocol
The median values of body mass, vital signs,
and duration of each phase of the procedures for
 518

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Table 1. Vital parameters (median 6 SD) of 73 wild armadillos during anesthetic stages of two different drug association protocols. Armadillos in group 1 (G1; n ¼
30) were anesthetized using a combination of butorphanol 0.1 þ detomidine 0.1 þ midazolam 0.1 mg/kg to allow the collection of biological samples and body
measurements. Armadillos in group 2 (G2; n ¼ 43) were anesthetized using the same combination of drugs added to ketamine 10 mg/kg to allow the surgical
implantation of intra-abdominal radio transmitters in addition to the procedures described for group 1. Reversal for both groups was done through a combination
of naloxone 0.02 þ yohimbine 0.125 þ flumazenil 0.01 mg/kg. These protocols were used in wild individuals of four armadillo species: Giant armadillos (GA,
Priodontes maximus); six-banded armadillos (SBA, Euphractus sexcinctus); southern naked-tailed armadillo (SNT, Cabassous unicinctus); and nine-banded armadillos
(NBA, Dasypus novemcinctus). Anesthetic stages are characterized by median and SD values of: induction time, total anesthetic maintenance time, and complete
recovery time after antagonists were applied.

Anesthesia Wilcoxon test; Wilcoxon test;

stage Parameters G1 G2 P-value G1 G2 P-value G1 G2 G2

GA, n ¼ 6 GA, n ¼ 27 GA SBA, n ¼ 22 SBA, n ¼ 4 SBA SNT, n ¼ 2 SNT, n ¼ 10 NBA, n ¼ 2

(4 males, (11 males, (6 males, (4 males) (2 females) (4 males, (1 male,
2 females) 16 females) 16 females) 6 females) 1 female)
Induction Body mass (kg) 30.6 6 5.95 30 6 4.52 W ¼ 108; 4.08 6 0.87 4.25 6 0.5 W ¼ 31; 1.4 6 0.28 1.8 6 0.24 4.82 6 0.4
P ¼ 0.47 P ¼ 0.7
Induction time 5.5 6 2.5 5.5 6 3 W ¼ 98; 3 6 2.5 3.5 6 2 W ¼ 28; 2.5 6 1 2 6 1.5 3.8 6 0.8
(min) P ¼ 0.77 P ¼ 0.31
Anesthesia Heart rate 37.36 6 18.64 38.45 6 5.10 W ¼ 111.5; 74.9 6 16.47 74.53 6 2.49 W ¼ 44.5; 81.8 6 1.13 84.71 6 9.25 19 6 2.14
(beats/min) P ¼ 0.38 P ¼ 0.75
Respiratory 11 6 4.16 12 6 1.67 W ¼ 75.5; 22.5 6 7.69 21.81 6 3.58 W ¼ 53; 31 6 2.82 23 6 9.34 20 6 3.88
rate (breaths/ P ¼ 0.51 P ¼ 0.43
min)
Oximetry (% 91.16 6 3.79 93.1 6 2.65 W ¼ 82; 95.2 6 14.09 92.03 6 11.1 W ¼ 41; 86.35 6 2.05 84.17 6 2.39 98 6 1.20
SpO2) P ¼ 0.71 P ¼ 0.97
Rectal 34 6 2.54 33.1 6 1.36 W ¼ 104.5; 35.19 6 1.31 35.34 6 1.21 W ¼ 39; 33.4 6 0.71 34.83 6 1.45 34.0 6 1.21
temperature P 0.85
JOURNAL OF ZOO AND WILDLIFE MEDICINE

P ¼ 0.56 ¼
(8C)
Total 104 6 59.5 137 6 39.5 W ¼ 62.5; 78 6 25 97 6 22 W ¼ 18.5; 60 6 85 98 6 44 133.20 6 9.12
anesthetic P ¼ 0.22 P ¼ 0.09
maintenance
time (min)
Recovery Complete 467 6 6 8.5 W ¼ 85.5; 264 3.5 6 8.5 W ¼ 37.5; 3 6 0.5 3 6 67 8.08 6 2.93
recovery time P ¼ 0.82 P ¼ 0.76
(min)

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 KLUYBER ET AL—ANESTHESIA AND SURGERY IN ARMADILLOS 519

Figure 1. Duration of each anesthetic phase for the studied species of wild armadillo. Anesthetic phases are
represented by: induction time (min), total anesthetic maintenance time (hr), and complete recovery time after
antagonists were applied (min). Seventy-three wild armadillos were anesthetized: 33 giant armadillos (GA,
Priodontes maximus); 26 six-banded armadillos (SBA, Euphractus sexcinctus); 12 southern naked-tailed armadillo
(SNT, Cabassous unicinctus); and 2 nine-banded armadillos (NBA, Dasypus novemcinctus).

each group and species are presented in Table 1
and Fig. 1. These measures are represented by
median and SD values. Body mass did not differ
between groups of GA or SBA, allowing the
comparison of vital signs and anesthetic stage
duration between groups. None of the vital signs
monitored (i.e. heart rates, respiratory rates,
relative oxyhemoglobin saturation [SpO2], and
rectal temperature) considerably differed between
groups for any of the studied species (Fig. 2–5).
Induction time differed among species (F(3,68) ¼
9.02, P , 0.01). According to the post hoc test,

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 520 JOURNAL OF ZOO AND WILDLIFE MEDICINE

Figure 2. Vital parameters of 33 wild giant armadillos (Priodontes maximus) anesthetized using two different drug
association protocols. Armadillos in group 1 (n ¼ 7) were anesthetized using a combination of butorphanol 0.1 þ
detomidine 0.1 þ midazolam 0.1 mg/kg to allow the collection of biological samples and body measurements.
Armadillos in group 2 (n ¼ 26) were anesthetized using the same combination of drugs added to ketamine 10
mg/kg to allow the surgical implantation of intra-abdominal radio transmitters in addition to the procedures
described for group 1. Reversal for both groups was done through a combination of naloxone 0.02 þ yohimbine
0.125 þ flumazenil 0.01 mg/kg.

induction for GA was longer than for SBA (P ,
0.01) and SNT (P , 0.01) (Fig. 1). Nevertheless,
there was no significant difference in induction
time between groups for each species (Table 1).
For both groups, the BMD combination resulted
in a quick and smooth induction for all armadillo
species (median ¼ 4 6 3 min) and supplementary
doses were not necessary for animals to reach an
anesthetic plane.
Prolonged apnea (6 7 min) only occurred in one
GA from group 1. Chest massage was given and
respiratory movements returned to normal; intu-
bation was not necessary. During this prolonged
apnea episode, cardiac rate and relative oxyhe-
moglobin saturation (SpO2) remained stable and
no clinical signs of hypoxia were observed.
Total anesthetic maintenance time varied be-
tween species (F(3,68) ¼ 7.86, P , 0.01). GA had
longer immobilization periods than SBA (P ,
0.01) and SNT (P ¼ 0.04) (Fig. 1). Only three
individuals from group 1 (1 GA and 2 SBA)
showed voluntary reflexes, moving their limbs, but
after the procedure had finished, 6 96 min for GA
and 54 min for SBA, and before antagonists were
given. Nevertheless, procedures were completed
for group 1 in 104 6 59.5 min for GA, 78 6 25 min
for SBA, and 60 6 85 min for SNT. For group 2,
137 6 39.5 min for GA, 97 6 22 min for SBA, 98
6 44 min for SNT, and 133.20 6 9.12 min for
NBA. No supplementary drugs were needed for
all procedures performed in this study.
Although there were no significant differences
in recovery time (Fig. 1), individuals of group 1 of
all species, which had their anesthesia reversed
through IV (saphenous vein) NYF, presented less
variation in recovery time (Table 1) and recovered
faster than individuals from group 2, which
received an IM injection of NYF in the hind

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 KLUYBER ET AL—ANESTHESIA AND SURGERY IN ARMADILLOS 521

Figure 3. Vital parameters of 26 wild six-banded armadillos (Euphractus sexcinctus) anesthetized using two
different drug association protocols. Armadillos in group 1 (n ¼ 22) were anesthetized using a combination of
butorphanol 0.1 þ detomidine 0.1 þ midazolam 0.1 mg/kg to allow the collection of biological samples and body
measurements. Armadillos in group 2 (n ¼ 4) were anesthetized using the same combination of drugs added to
ketamine 10 mg/kg to allow the surgical implantation of intra-abdominal radio transmitters in addition to the
procedures described for group 1. Reversal for both groups was done through a combination of naloxone 0.02 þ
yohimbine 0.125 þ flumazenil 0.01 mg/kg.

limbs. Armadillos showed signs of complete
recovery up until the moment of release. No
animals died during capture or due to anesthetic
procedures.
Surgical recovery
None of the armadillos died due to a field
surgical procedure or due to a complication of
intra-abdominal implantation. In five of the GA,
devices failed (suddenly ceased sending signals) at
a time earlier than the manufacturer’s determined
operational battery life. Implantable devices were
not removed from the animals and monitoring
was interrupted once the battery failed.
DISCUSSION
The combination of butorphanol, detomidine,
and midazolam resulted in quick, smooth, and
adequate induction in all armadillos from both
groups. The initial induction period required
careful supervision due to transitory apnea for
the first 20 min. In group 2, ketamine was only
given after the first 20 min of anesthesia (time
needed for preparation and cleansing of skin prior
to the surgical procedure), and did not affect
induction time.
Nevertheless, vital parameters normalized and
remained constant throughout the procedure after
this period (20 min). Oximetry values for both
groups were recorded between 84.17 6 2.39 and
98 6 1.20%. Similar low oximetry values, were
reported for the long-nosed armadillo (Dasypus
kappleri) and nine-banded armadillo.10 The au-
thors also reported that no animals showed
tachycardia, cyanosis, or any other clinical signs
in this study. Transitory apnea should not be
worrisome for armadillos because, compared with

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 522 JOURNAL OF ZOO AND WILDLIFE MEDICINE

Figure 4. Vital parameters of 12 wild southern naked-tailed armadillo (Cabassous unicinctus) anesthetized using
two different drug association protocols. Armadillos in group 1 (n ¼ 2) were anesthetized using a combination of
butorphanol 0.1 þ detomidine 0.1 þ midazolam 0.1 mg/kg to allow the collection of biological samples and body
measurements. Armadillos in group 2 (n ¼ 10) were anesthetized using the same combination of drugs added to
ketamine 10 mg/kg to allow the surgical implantation of intra-abdominal radio transmitters in addition to the
procedures described for group 1. Reversal for both groups was done through a combination of naloxone 0.02 þ
yohimbine 0.125 þ flumazenil 0.01 mg/kg.

other mammal species, armadillos are able to hold
their breath and tolerate hypoxemia.5 Neverthe-
less, none of the individuals studied showed
clinical signs of hypoxemia throughout anesthetic
procedure.
Opioids have been seldom used in xenarthran
studies.4,8,13,26 Most of the opioid’s agents have
been used as pre-anesthetic, combined or not with
benzodiazepines like diazepam, but its use is
considered not practical in field situations.1,4,8,13,22
Butorphanol is the synthetic opioid that has been
chosen for this study and has been previously
recommended for armadillos.8,13 Usually com-
bined with other anesthetic drugs, butorphanol
reduces the volume or doses of other drugs and is
applied for pain control after surgical proce-
dures.13 A rapid recovery was also reported in
giant anteaters22 and nine-banded armadillos,
after antagonist was applied.13
Medetomidine is a selective a-2 agonist drug
that can provide analgesia, sedation, and sympa-
tholysis with opioid-sparing effects.1,25 Considering
all a-2 agonist drugs, this agent is a better option
for armadillos and xenarthrans in general.19,30
Xylazine, medetomidine, or dexmedetomidine
have been used for giant anteaters and armadillos
to promote good muscle relaxion; quick, smooth
induction; and recovery.8,10,11,13,14,17,18,19,21,22 This
agent promotes better muscle relaxation, shorter
analgesic, and less adverse effects. It also lowers
the dose of ketamine needed compared with
protocols with xylazine.10,13,19,25
Although a-2 agonists are reported to cause
regurgitation and abortion,3,11 during this study,
pregnancy was detected by palpation and baby’s
heart auscultation (stethoscope) was performed in
two armadillos’ females (1 GA and 1 SNT). Those
females were tracked by the team and babies were

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 KLUYBER ET AL—ANESTHESIA AND SURGERY IN ARMADILLOS 523

Figure 5. Vital parameters of 2 wild nine-banded armadillo (Dasypus novemcinctus) anesthetized using the drug
association protocols of group 2. Armadillos in group 2 (n ¼ 2) were anesthetized using a combination of
butorphanol 0.1 þ detomidine 0.1 þ midazolam 0.1 mg/kg to allow the collection of biological samples and body
measurements, and ketamine 10 mg/kg to allow the surgical implantation of intra-abdominal radio transmitters.
Reversal was done through a combination of naloxone 0.02 þ yohimbine 0.125 þ flumazenil 0.01 mg/kg.

seen with the adult females after the anesthetic
procedure, ruling out the possibility of an abor-
tion. However, detomidine was the a-2 agonist
agent chosen for this study due to its cost-
effectiveness and availability.
Due to its cost-effectiveness, the use of disso-
ciative agents like ketamine were commonly
applied alone for edentates,3 but this is not
recommended anymore.11 Used alone, ketamine
usually requires high doses and may cause
adverse effects like respiratory depression, cata-
tonia, and rough recovery, especially in the
field.10,13 The other most common drug widely
applied for armadillos and combined with keta-
mine are a-2 agonists, like xylazine.5,6,13,30
The advantages of these drugs are their specific
antagonism, their ability to synergize ketamine’s
dissociative effects, and the need for lower
ketamine doses. This combination is also recom-
mended for providing excellent muscle relaxation,
faster recovery, high safety margin, and its low
cost.1,13,20,22 However, severe cardiovascular abnor-
malities were reported when these agents were
used in giant anteaters.30 Similar results were
observed in a study with D. novemcinctus and D.
kappleri.10
As a-2 agonists, many protocols were developed
combining dissociative drugs with benzodiaze-
pines, like ketamine and midazolam or tiletamine
and midazolam,5,6,8,13 for short procedures,19,20 and
especially due to the wide safety margin and
excellent muscle relation,1,25 for xenar-
thrans.17,21,22,27,30 Midazolam is also recommended
to avoid the risk of regurgitation, abortion, respi-
ratory arrest, and hypotension caused by xyla-
zine.11,13,22,30
Armadillos are considered imperfect homeo-
thermic species16 and have low body temperatures
compared with other mammals (30 to 368C). No
signs of hypothermia or hyperthermia were ob-

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 524 JOURNAL OF ZOO AND WILDLIFE MEDICINE
served during capture or chemical immobilization
procedures. Body temperatures recorded were
similar to previous studies involving different
armadillo species like D. novemcinctus, D. kappleri,
E. sexcinctus.5,6,8,10,13
The safety of the BDM protocol is enhanced by
its reversibility through respective antagonists,
which can be performed at any time throughout
the procedure. No adverse effects or emergency
situation was faced, and no need of specialized
equipment during any of the procedures was
required. This provides a key contingency plan
in case of emergency or adverse collateral effects.
In addition, no armadillos from group 2, which
received ketamine hydrochloride as a supplemen-
tary drug, showed significant changes in vital
signs, nor were undesirable side effects that could
compromise efficiency and safety during the
procedure or anesthetic recovery recorded.
Choosing these anesthetic protocols was justi-
fied due to their high safety margin, low level of
side effects, and ability to provide adequate
immobilization, muscle relaxation, and uncon-
sciousness. Supplementing the BDM combination
with ketamine provided an ideal anesthetic plane
for surgical implantation and enough time to
collect biological samples and make body mea-
surements. Further studies are recommended to
investigate potential effects of these agents on
armadillos. The large therapeutic window and low
cost of this combination per procedure and/or
individual, makes this method attractive for field
conditions, when long induction and recovery
times are undesirable and/or resources are limit-
ed.
This study highlights that transmitter implan-
tation is practically irreversible, and armadillos
will carry implants throughout their whole lives.
Furthermore, the impact of transmitters on ar-
madillos’ health was not evaluated after the
batteries stopped working or devices failed.
Although the implants appear to have minimal
impacts on animal health and behavior, the risks
of surgical procedure, device and battery failure,
or injuries caused by surgical implants, suggest
that this procedure should be carefully evaluated
as an alternative for radiotracking armadil-
los.6,12,13,23,24 Based on the long period in which
animals remained in burrows after surgery, the
transmitter implantation surgery procedure
should only be performed on healthy adult
animals within the average weight range for the
species (Table 1). This will help ensure that
surgical recovery will not hinder their health.
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CONCLUSIONS
These protocols can be used for short and
noninvasive procedures and/or to implant intra-
abdominal transmitters. Both can be a low-cost
alternative especially in countries where other
anesthetic agents are expensive, restricted, or not
available.
The advantage of antagonizing the BDM anes-
thetics combination is that it significantly reduces
the undesirable effects of long recoveries. The low
adverse effects and high safety margin provided
by both protocols avoid the chances of emergency
situations, which are especially delicate in field
conditions.
The intra-abdominal implant did not appear to
have a negative impact on the armadillos studied.
Nevertheless, further studies with long-term
monitoring of armadillos are required to assess
the true impacts of intra-abdominal implants.
Despite the complexities of using implantable
transmitters, including technological limitations,
high costs, and the conditions of surgical proce-
dures in the field, this type of device is currently
the most efficient and recommended method for
long-term monitoring of wild armadillos. These
devices are efficient for long term-monitoring of
wild armadillos if implantation follows strict
protocol, safety, and antisepsis criteria,13 and
includes careful evaluation.
Acknowledgments: We would like to thank Dr.
Yamil Di Blanco and the giant armadillo
project team. This study would not have been
possible without all the Giant Armadillo Con-
servation Program supporters: https://www.
icasconservation.org.br/partners.
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Accepted for publication 16 March 2020

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