TERATOLOGY 61:203–210 (2000)
Comparison of Cleft Palate Induction by
Nicotiana glauca in Goats and Sheep
K.E. PANTER,1* J. WEINZWEIG,2 D.R. GARDNER,1 B.L. STEGELMEIER,1 AND L.F. JAMES1
1UnitedStates Department of Agriculture, Agricultural Research Service,
Poisonous Plant Research Laboratory, Logan, Utah 84341
2Department of Plastic Surgery, Brown University, Providence, Rhode Island 02903
ABSTRACT The induction of cleft palate by
Nicotiana glauca (wild tree tobacco) during the first
trimester of pregnancy was compared between Spanish-
type goats and crossbred western-type sheep. Cleft
palate was induced in 100% of the embryonic/fetal
goats when their pregnant mothers were gavaged with
N. glauca plant material or with anabasine-rich extracts
from the latter, during gestation days 32–41. Seventy-
five percent of newborn goats had cleft palate after
maternal dosing with N. glauca during gestation days
35–41, while no cleft palates were induced when
dosing periods included days 36–40, 37–39, or day 38
only. The induced cleft palates were bilateral, involving
the entire secondary palates with complete detachment
of the vomer. Eleven percent of the newborn goats from
does gavaged during gestation days 32–41 had extra-
cranial abnormalities, most often contractures of the
metacarpal joints. Most of these contractures resolved
spontaneously by 4–6 weeks postpartum. One new-
born kid also had an asymmetric skull due to apparent
fetal positioning. No cleft palates were induced in lambs
whose mothers were gavaged with N. glauca plant or
anabasine-rich extracts during gestation days 34–41,
35–40, 35–41, 36–41, 35–51, or 37–50. Only one of
five lambs born to three ewes gavaged with N. glauca
plant material during gestation days 34–55 had a cleft
palate, but all five of these lambs had moderate to
severe contractures in the metacarpal joints. The slight
to moderate contracture defects resolved spontane-
ously by 4–6 weeks postpartum, but the severe contrac-
tures resolved only partially. Embryonic/fetal death and
resorption (determined by ultrasound) occurred in 25%
of pregnant goats fed N. glauca compared to only 4%
of pregnant sheep. Nicotiana glauca plant material
contained the teratogenic alkaloid anabasine at 0.175%
to 0.23%, dry weight, demonstrating that Spanish-type
goats are susceptible to cleft palate induction by the natural
toxin anabasine, while crossbred western-type sheep are
resistant. However, clinical signs of toxicity were equally
severe in goats and sheep, even though maternal alkaloid
tolerance was generally lower in sheep. We postulate that
an alkaloid-induced reduction in fetal movement during the
period of normal palate closure is the cause of the cleft
palate and multiple flexion contractures. Teratology 61:203–
210, 2000. Published 2000 Wiley-Liss, Inc.†
Published 2000 WILEY-LISS, INC. † This
article is a US
government work and, as such, is in the public domain of the
United States of America.
INTRODUCTION
Plant-induced cleft palate in cattle was first docu-
mented more than 30 years ago, when musculoskeletal
defects known as ‘‘crooked calf disease’’ were attributed
to maternal ingestion of Lupinus spp. (Palotay, ’59;
Wagnon, ’60; Binns and James, ’61; Shupe et al., ’67a,b,
’68). Lupine-induced cleft palate and skeletal malforma-
tions continue to cause heavy losses in the cattle
industry (Panter et al., ’97). The animals with cleft
palate often have other significant craniofacial devia-
tions, such as asymmetry of the skull, maxillary hypo-
plasia, brachygnathia, or malocclusion. Arthrogryposis,
scoliosis, torticollis, kyphosis, and flexure and exten-
sion contractures of the carpal, metacarpal, tarsal, and
metatarsal joints are also common. These anomalies
usually occur after extended periods of fetal exposure.
The incidence, type, and severity of malformations in
cattle depend on the alkaloid composition of the plant,
the stage and length of gestation when ingestion occurs,
and the amount of the teratogen ingested (Shupe et al.,
’67a,b; Panter et al., ’98b).
Soon after lupines were identified as the cause of
crooked calf disease, the ␣-pyridone quinolizidine alka-
loid anagyrine was identified as a principal lupine
teratogen (Keeler, ’76). While many species of lupine
contain anagyrine (Davis and Stout, ’86), others like
Lupinus formosus and L. arbustus contain other terato-
gens, such as the piperidine alkaloid ammodendrine
(Keeler and Panter, ’89; Panter et al., ’98b). Currently,
one quinolizidine alkaloid, anagyrine, and five piperi-
dine alkaloids (coniine, N-methyl coniine and ␥-conice-
ine from Conium maculatum, anabasine from Nicoti-
ana spp. and ammodendrine from Lupinus formosus
and L. arbustus) have been demonstrated to be terato-
genic (Keeler, ’76; Keeler and Balls, ’78; Keeler et al.,
’81; Panter et al., ’98a). Structure/activity experiments
of these piperidine alkaloids suggest that the 1,2-
dehydropiperidine alkaloids (␥-coniceine, anabaseine,
and N-acetyl hystrine) exhibit the most potent activity,
*Correspondence to: Kip E. Panter, United States Department of
Agriculture, Agricultural Research Service, Poisonous Plant Research
Laboratory, 1150 E. 1400 N., Logan, UT 84341.
E-mail: kpanter@cc.usu.edu
Received 25 May 1999; Accepted 20 September 1999
204 PANTER ET AL.
followed by the piperidine alkaloids (coniine, anaba-
sine, and ammodendrine), followed by the least active
N-methyl derivatives [N-methyl coniine, N-methyl
anabasine, and N-methyl ammodendrine (Panter et al.,
’98b, Panter et al., ’99)]. Conium maculatum (poison-
hemlock), Nicotiana tabacum (burley tobacco), and N.
glauca (wild tree tobacco) have been shown to cause
similar MCC-type skeletal malformations and cleft
palate in cattle, sheep, goats, and pigs (Keeler and
Balls, ’78; Keeler et al., ’81, ’84; Keeler and Crowe, ’84,
’84; Keeler, ’79; Panter et al., ’85a,b, ’88, ’90; Panter and
Keeler, ’92).
Anabasine from Nicotiana tabacum was identified as
a teratogen after an epidemic of deformed newborn pigs
occurred in Missouri and Kentucky during the late
1960s (Crowe, ’69). It was determined that the practice
of feeding tobacco stalks to pregnant sows was respon-
sible for the epidemic. These stalks contained high
levels of the simple piperidine alkaloid teratogen, anaba-
sine [2-(3-pyridyl)piperidine], which was structurally
similar to other known piperidine alkaloid teratogens
from Conium maculatum (Keeler and Balls, ’78; Keeler
et al., ’81). Nicotiana glauca was subsequently identi-
fied as a rich source of anabasine and was used for
extraction and isolation of the compound for confirma-
tion studies. Subsequently, a goat model was developed
to study the mechanism of action of lupine, poison-
hemlock, and tobacco-induced malformations in live-
stock (Panter et al., ’90, Panter and Keeler, ’92).
An induced congenital goat model of cleft palate
repair in utero was recently characterized; this model
resembles closely cleft palate in humans (Weinzweig et
al., ’99a,b). Previous research on cleft palate has fo-
cused on the development of palate defects that had
been surgically created in the sheep (Hedrick et al., ’96).
Other models designed to study fetal wound repair
have been developed in the mouse, rat, rabbit, guinea
pig, swine, opossum, and monkey (Adzick and Long-
aker, ’91). The smaller, short-gestation models, such as
the mouse, rat, guinea pig, and rabbit, only allowed
fetal manipulation later in gestation, when the post-
operative intrauterine period is short, and after the
privileged period of fetal scarless healing had passed.
By contrast, larger, long-gestation animals, such as
the monkey, sheep, and goat, provide for easier
and more extensive manipulation during the period of
early gestation (⬍day 100). Early surgical intervention
has proven far superior to postnatal repair as fetal
repair is often scarless. The earlier procedure also
provides a longer postoperative intrauterine period for
monitoring healing and development. This is especially
true of craniofacial defects, as early surgical repair
maximizes the advantage of the privileged fetal environ-
ment for scarless healing and minimizes disfiguring lip
and palate scarring in the newborn (Longaker and
Adzick, ’91).
Nicotiana glauca-induced cleft palate and musculo-
skeletal contracture malformations in the goat have
proved an excellent model with which to study the
mechanism of action of similar lupine-induced malfor-
mations in cattle (Panter et al., ’90; Panter and Keeler,
’92). The model has shown that alkaloid-induced reduc-
tion in fetal movement over specific periods of gestation
is directly responsible for cleft palate and musculoskel-
etal malformations. Similar models have established
the specific cleft palate induction periods in pigs (gesta-
tion days 30–45; Panter et al., ’85b), goats (35–41 days
of gestation; Panter and Keeler, ’92) and cattle (40–50
days gestation; Panter et al., ’98a).
The lengths of gestation in goats (caprine) and sheep
(ovine) at full term are similar. The average gestational
age when normal parturition occurs for the caprine
species is 150 days (0 ⫽ day of breeding) with a range of
146–155, while the average in the ovine species is 151
days with a range of 143–155, depending on the breed
(Morrow, ’86). Normal palate closure occurs near day 38
of gestation (0 ⫽ day of breeding) in the sheep (Evans
and Sack, ’73). This coincides with the late embryo (day
32), early fetal (day 40) stage of gestation (Evans and
Sack, ’73).
Consequently, N. glauca and its anabasine-rich ex-
tracts are now used in conjunction with the goat model
to study the mechanism of action of plant-induced cleft
palate and multiple congenital contractures, pathogen-
esis of cleft palate development and the potential for
fetal intervention. Specifically, this study was done: (1)
to compare the incidence of fetal cleft palate induction
between goats and sheep, and (2) to define more pre-
cisely the period of gestation for maximal induction of
cleft palate.
MATERIALS AND METHODS
Plant preparation and chemical analysis
Nicotiana glauca (wild tree tobacco) was collected
along the Big Sandy River near Wikeup, Arizona, in
early June 1983 (83–1) and 1985 (85–2). It was in flower
and averaged about 12 ft in height. Green plant mate-
rial including leaves, stems, flowers, and woody parts
was collected and chopped coarsely in a tree branch
chopper and spread on tarps to facilitate drying. The
dry coarse-chopped material was further ground in a
hammer mill to pass through a 20-mesh screen (20
holes per square inch) and stored in plastic bags at
room temperature until use. Total alkaloid and percent-
age anabasine were measured by modifications of previ-
ously published methods (Keeler et al., ’81; Gardner
and Panter, ’94). Plant collection 83–1 contained 0.23%
anabasine and 85–2 contained 0.175% by dry weight.
Anabasine-rich extracts contained 2.4% anabasine. Al-
kaloid levels have been measured periodically since
1983 and just before this experiment began and have
remained consistent with no degradation.
Goat study
Twenty-seven crossbred Spanish-type female range
goats (caprine) were synchronized in estrus using intra-
 CLEFT PALATE INDUCTION BY N. GLAUCA IN GOATS AND SHEEP 205
TABLE 1. Cleft palate induced in goats gavaged with Nicotiana glauca or
anabasine-rich extract during different stages of gestation
Group No., wt* Treatment Gest days Clin. signs Results
A 4 (41.4) N. glauca 32–41 ⫹ to ⫹⫹⫹⫹ 9/9 CP
B 3 (41.7) Extract 32–41 ⫹ to ⫹⫹⫹⫹ 6/6 CP
1/6 MCC
C 4 (39) N. glauca 35–41 ⫹ to ⫹⫹⫹⫹ 6/8 CP
D 4 (41.7) N. glauca 36–40 ⫹ to ⫹⫹⫹⫹ 8/8 normal
E** 4 (42.5) N. glauca 37–39 ⫹ to ⫹⫹⫹⫹ 5/5 normal
F† 4 (42.3) N. glauca 38 ⫹ to ⫹⫹ 9/9 normal
G 4 (41.4) Control — — 7/7 normal
CP, cleft palate; MCC, multiple congenital contractures; ⫹, slight clinical signs, including
nervousness and slightly depressed; ⫹⫹, slight to moderate clinical signs, including
intermittent fine tremors of the legs, frequent urination, appetite still good; ⫹⫹⫹, moderate
clinical signs including regular tremors of the muscles, muscular weakness, frequent
urination and defecation, sternal recumbency; ⫹⫹⫹⫹, severe clinical signs, including
muscular weakness, ataxia, sternal recumbency, inability to stand for prolonged periods, off
feed, weakness in the neck muscles.
*Number of goats per treatment and average weight (in kg).
**Five of five normal kids from three goats, and one goat resorbed her fetus(es).
†Goats were dosed three times in one day at 7:00 a.m., Noon, and 3:30 p.m. for an average
single day dose of 14 mg anabasine/kg/goat.

TABLE 2. Average daily dose (mg/kg) of anabasine by day of gestation*

Anabasine dose (mg/kg) by day of gestation in goats
Treatment Days
group treated** 32 33 34 35 36 37 38 39 40 41
A 32–41 5 7.2 10.2 11.3 12.5 13.7 15.2 15.2 15 14.9
B 32–41 (ext) 8 9 10 11.5 13.5 15.5 17.5 19 18.5 17
C 35–41 — — — 7.8 10.3 12.9 14 13.8 12.8 14.5
D 36–40 — — — — 6.4 11.5 14.3 14.3 14.3 —
E 37–39 — — — — — 11.2 11.9 11.6 — —
F 38 — — — — — — 14 — — —
G Control — — — — — — — — — —
*Nicotiana glauca plant material contained 0.175–0.23% anabasine by dry weight. Anabasine-
rich extract (ext) contained 2.4% anabasine.
**Group B goats were administered anabasine-rich extract via methylcellulose capsules two
times daily.

vaginal sponges (Intervet International, Boxmeer, Hol-
land) containing 45 mg of a progesterone derivative
(fluorogestone acetate). The sponges were removed in
10–14 days, and the does were bred to Spanish-type
bucks 36–48 hr later. Pregnancy was confirmed by ultra-
sound 30–32 days after breeding (day of breeding ⫽ 0).
Seven treatments (groups A–G, Table 1), including
controls during five specific stages of gestation, were
selected around the normal palate closure period of
gestation day 38 (Evans and Sack, ’73). The gestational
periods for gavage included 32–41, 35–41, 36–40, 37–
39, and day 38 only (Table 1). These periods were
selected on the basis of previous feeding trials (Panter
and Keeler, ’92; Panter et al., ’90). The goats were
weighed before treatment and gavaged with the ground
plant material as a thick slurry mixed in water twice a
day AM and PM (Table 2). The doses were separated by
about 6–7 hr from AM to PM and 16–17 hr PM to AM.
Group B goats were administered anabasine-rich ex-
tracts from N. glauca via methyl cellulose capsules
from 32–41 days gestation 2 times daily on the same
schedule as those given ground plant by gavage (Table
2). Dosages were started low (5–8 mg anabasine per kg)
and adjusted daily up to an effective dose and then
readjusted to maintain moderate clinical signs of toxic-
ity during the feeding period (Table 2). Using ultra-
sound, we previously determined that fetal effects
(reduced fetal activity) from Nicotiana glauca were
evident 18 hr after dosing. The twice-a-day dosing
minimized maternal effects, yet optimized fetal effects
(K.E. Panter, unpublished data). At birth, newborn
goats were weighed, sex was determined, and each kid
was examined carefully for cleft palate, facial symme-
try, and skeletal defects.
Sheep study
Twenty-eight western crossbred ewes (ovine) were
synchronized in estrus similar to the goat study, except
that the intravaginal sponges contained 40 mg of the
progesterone derivative fluorogestone acetate. The
sponges were removed in 10–14 days and ewes bred to
Suffolk rams. Pregnancy was confirmed before treat-
ment using ultrasound. Three to four ewes were as-
signed to each of nine treatments (A–I, Table 3) and
206 PANTER ET AL.
TABLE 3. Effects of gavage of Nicotiana glauca to pregnant sheep at different
stages of gestation
Treatment
group No., wt* Treatment Gest. days Clin. signs** Results
A† 3 (58.6) Extract 34–41 ⫹ to ⫹⫹⫹⫹ 4/4 normal
B 3 (72.3) Extract 35–40 ⫹ to ⫹⫹⫹⫹ 4/4 normal
C 3 (68) Extract 35–41 ⫹ to ⫹⫹⫹⫹ 3/3 normal
D 3 (67.3) Plant 35–41 ⫹ to ⫹⫹⫹⫹ 5/5 normal
E 4 (63) Extract 36–41 ⫹ to ⫹⫹⫹ 5/5 normal
F‡ 3 (65.3) Plant 35–51 ⫹ to ⫹⫹⫹⫹ 3/4 normal
1/4 MCC
G§ 3 (68) Plant 37–50 ⫹ to ⫹⫹⫹ 4/5 normal
H§§ 3 (63.3) Plant 34–55 ⫹ to ⫹⫹⫹⫹ 1/5 CP
5/5 MCC
I 3 (66.7) Control — — 6/6 normal
*Number of ewes per treatment and average weight in kg.
**Clinical signs same as Table 1.
†One ewe resorbed her fetus, determined by ultrasound at the end of treatment.
‡One lamb had slight to moderate contractures in the carpal joints (buck knees), front legs
appeared normal after 3 weeks.
§One lamb had an asymmetrical head and shortened kinked tail.
§§Five of five lambs had moderate to severe contractures in the carpal joints (buck knees,
MCC) and one in five lambs had bilateral cleft palate (CP).

TABLE 4. Average daily dose (mg/kg) of anabasine by day of gestation*

Anabasine dose (mg/kg) by day of gestation in sheep
Days
Group** treated† 34 35 36 37 38 39 40 41 42 43–55
A 34–41 (ext) 8.7 6.3 9.5 10 9.1 14.2 14.9 4.9 — —
B 35–40 (ext) — 11.7 8.7 8.5 9.4 8.8 9.5 — — —
C 35–41 (ext) — 11.7 11.2 10.9 10.4 10.4 10.7 7.5 — —
D 35–41 — 7.8 9.7 9.4 11.9 14.3 9.7 8.8 — —
E 36–41 (ext) — — 12.1 11.1 11.4 11.5 11.7 12.4 — —
F 35–51 — 9.5 9.9 9.1 9.2 8.1 8.6 8.9 7.6 6.7 ⫾ 0.7
G 37–50 — — — 5.5 10.8 7.7 8.3 8 8.1 8.3 ⫾ 0.5
H 34–55 13.4 8.5 10.5 12.9 8.1 11.2 9.3 11.2 10.8 9.3 ⫾ 0.6
I Control — — — — — — — — — —
*Nicotiana glauca plant material contained 0.175–0.23% anabasine by dry weight. Anaba-
sine extract (ext) contained 2.4% anabasine.
**Group F averaged 6.7 ⫾ 0.7 mg/kg/day/goat over gestation days 43–51; group G averaged
8.3 ⫾ 0.5 mg/kg/day/goat over gestation days 43–50: group H averaged 9.3 ⫾ 0.6 mg/kg/day/
goat over gestation days 43–55.
†Group A, B, C, and E ewes were administered anabasine-rich extract via methylcellulose
capsules two times daily.

seven different stages of gestation selected around the
palate closure period of gestation day 38. Gestational
periods for gavage (34–41, 35–40, 35–41, 35–51, 34–55,
36–41, 37–50) were selected to establish frequency of
induced cleft palate in sheep and to define more pre-
cisely the period of induction. The gavage regimen was
based on previous trials in sheep and goats (Keeler et
al., ’84; Panter et al., ’88; Panter and Keeler, ’92). Either
ground plant material or anabasine-rich extracts there-
from were used. The animals were dosed by gavage or
capsule with ground plant or extracts, respectively
(Table 4). Ewes were dosed twice daily AM and PM, and
each dose was separated by about 6–7 hr AM to PM and
16–17 hr PM to AM. Each dosage was adjusted based on
the severity of the clinical signs. Dosing was adjusted to
induce moderate to severe maternal clinical signs of
toxicity and maintain optimum fetal effects throughout
the dosing period. At birth, lambs were weighed, sex
was determined, and each lamb was examined carefully
for cleft palate and skeletal abnormalities.
RESULTS
Goat study
Pregnant goats fed either Nicotiana glauca plant or
anabasine-rich extracts therefrom showed mild to se-
vere clinical signs which included temporary and inter-
mittent muscular weakness, ataxia, fine muscle trem-
ors, and episodes of sternal recumbency (Tables 1, 2).
Cleft palate occurred in 15 of 15 (100%) of the
newborn goats from seven does gavaged with N. glauca
or anabasine extracts during gestation days 32–41
(Table 1; Fig. 1). One (9%) of six of the newborn goats
from group B also had moderate to severe contractures
 CLEFT PALATE INDUCTION BY N. GLAUCA IN GOATS AND SHEEP
Fig. 1. Cleft palate goats showing slight variations in cleft widths.
Note the normal control goat on the right. These cleft palates were
induced when their mothers were fed N. glauca during gestation days
35–41.
Fig. 2. Occasional moderate to severe contractures associated with
cleft palates. One of 6 fetal goats had these types of malformations
from pregnant goats fed N. glauca during gestation days 32–41. Most
of the mild to moderate contractures resolve within 4–6 weeks
postpartum. If the feeding period is extended to day 60, these
contracture malformations are more severe and include the spine,
neck, and legs.
in the front legs (Fig. 2); these resolved partially
spontaneously within 2–6 weeks. Pregnant goats fed
during gestation days 35–41 had 6/8 (75%) newborn
goats with cleft palate (Table 1). The cleft palates were
primarily bilateral with minor variations in the width
of the cleft (Fig. 1), but all had complete detachment of
the vomer. One kid had a unilateral cleft palate in
which one side of the palate was attached to the palatal
midline. Occasionally, other craniofacial defects such as
maxillary hypoplasia and midfacial retrusion (Fig. 3)
occur, although not observed in this study. Most new-
born goats with cleft palate were able to nurse and
207
Fig. 3. Two cleft palate goats (twins) showing differences in facial
growth and development. The goat on the right shows the occasional
changes of midfacial development, including maxillary hypoplasia and
midfacial retrusion. Cephalometric measurements of the debrided
skulls are found in Weinzweig et al. (’99a,b).
survived to 6 months of age, at which time they were
sacrificed.
By contrast, pregnant does gavaged with N. glauca-
during gestation days 36–40, 37–39, and day 38 all had
normal kids with no observable malformations. One
doe treated during gestation days 37–39 had resorbed
fetus(es).
Sheep study
Pregnant ewes fed either Nicotiana glauca or anaba-
sine-rich extracts showed mild to severe clinical signs of
poisoning, including muscular weakness and tremors
similar to those described for goats (Tables 3, 4). All
lambs were normal in those groups that were fed
during gestation days 34–41, 35–40, 35–41, and 36–41,
except for fetal death and resorption, which occurred in
one ewe fed extract during gestation days 34–41 (Table
3). Among ewes fed plant material during gestation
days 35–51, one of four lambs had mild to moderate
contractures in the carpal joints, but no cleft palate.
The contractures resolved within 4–6 weeks postpar-
tum. Among ewes fed plant during gestation days
37–50, one of five lambs had an asymmetrical head and
shortened kinked tail, but no other gross anomalies.
Subsequent growth and development of this lamb ap-
peared normal. Among ewes fed plant during gestation
days 34–55, five of five lambs had contracture-type
skeletal defects in the carpal joints (Fig. 4) that were
moderate to severe; 1/5 had a bilateral cleft palate.
Therefore, only 1 (2.9%) of 35 lambs had cleft palate
from ewes fed Nicotiana glauca plant or extract during
what was considered a cleft palate susceptible stage of
pregnancy. Six (17%) of 35 had contracture-type defects
involving the front legs, most of which resolved sponta-
neously within 4–6 weeks postpartum. These contrac-
tures only occurred in the ewes gavaged during ex-
tended periods of gestation (days 34–55).
Nicotiana glauca plant material contained the terato-
genic alkaloid anabasine at 0.175–0.23%, dry weight,
and plant extract contained 2.4% anabasine. To achieve
a similar toxic maternal clinical response, anabasine
208 PANTER ET AL.
Fig. 4. Contracture defects (buck knees) in a lamb from an ewe fed N.
glauca during gestation days 34–55.
doses fed per body weight were on average higher in
goats (12.8 mg/kg/day) than in sheep (9.5 mg/kg/day).
DISCUSSION
This study was conducted to document the difference
in incidence of cleft palate between goats and sheep and
to define more precisely the gestational period for
inducing cleft palate. Previous research has deter-
mined that the fetal goat develops cleft palate after
maternal exposure to the piperidine alkaloid-contain-
ing plants Nicotiana glauca, Conium maculatum, and
Lupinus formosus (Panter et al., ’90; Panter and Keeler,
’92; Panter et al., ’94). Cleft palate has been induced
previously in sheep, although the experimental design
did not target specifically the period of cleft palate
induction (Keeler et al., ’84). However, compared with
cattle and pigs, the incidence in sheep was very low.
Interestingly, the grazing habits, diet selections, physi-
ology, fetal growth, and development of goats and sheep
are very similar, but their biological sensitivity to
various toxicants, or fetal sensitivity to this teratogen,
appears to be quite different. Whether this difference is
due to metabolism, maternal absorption/elimination,
degree of fetal susceptibility or something else is not
known.
While the sheep had previously been established as a
model to study methods of in utero scarless repair of
cleft palates and cleft lips, these clefts were created
surgically (iatrogenic) (Hedrick et al., ’96). Although the
surgical sheep model has been used successfully for
developing early techniques, certain clinical limitations
have been found when a surgical model versus an
intrinsic model is used. The limitation of the surgical
model was expressed by Hedrick et al. (’96) as follows:
‘‘Ideally, a large animal model that intrinsically forms
clefts would provide the best model with which to
evaluate the efficacy of fetal cleft repair . . . no such
practical intrinsic model exists . . . the fetal lamb is not
known to form clefts intrinsically in response to terato-
gens . . . clefts in this model must be created surgically.’’
While the fetal lamb forms intrinsically cleft palates, as
demonstrated in this study (1/35), the incidence is so
low and the occurrence so unpredictable that using the
sheep model is impractical. The goat model is more
efficient and resembles closely cleft palate deformities
in humans (Weinzweig et al., ’99a,b).
We believe that the primary mechanism responsible
for cleft palate induction in this study was an alkaloid-
induced reduction in fetal movement during the period
of palate closure (Panter et al., ’90; Panter and Keeler,
’92). Ultrasound imaging of fetuses in does gavaged
with Nicotiana glauca during the normal programmed
palate closure period of 35–41 days showed a tight
flexure of the head and neck, with negligible space
between the chin and sternum. These observations
were in contrast to the intermittent head and neck
extension movements in untreated fetal goats that
normally begin about days 35–38 of gestation (Panter
et al., ’91). Therefore, inhibited movement of the fetal
tongue with prolonged maintenance between palatal
shelves, combined with the hyperflexed position of the
head and neck, is believed to be the mechanical mecha-
nism of alkaloid-induced cleft palate formation in goats
(Panter and Keeler, ’92). This proposed mechanism is
similar to the theories offered for the pathogenesis of
cleft palate in the Pierre Robin syndrome (PRS), a
condition in humans attributed to malpositioning, which
prevents the tongue from descending from the nasal
cavity, preventing fusion of the palate at the midline
(Rintala et al., ’84).
In addition to the malpositioning induced by terato-
genic alkaloids, we suspect that the length of time of
continuous exposure of the fetus to the teratogen and
the sustained effects over the specific stages of gesta-
tion are important in the induction of cleft palate and
multiple congenital contractures. This is evident in this
experiment in which the high incidence was induced
during gestation days 32–41 and 35–41, but no clefts
were induced during days 36–40, 37–39, or 38 (Table 1).
The major difference in the incidence of induced cleft
palate between goats and sheep may be the length of
time of reduced fetal movement over the palate closure
period. Fetal goats are inhibited continuously between
doses, whereas fetal sheep have brief intermittent
periods of recovery (fetal movement) between doses
(Panter et al., ’88, ’90; Panter and Keeler, ’92).
While the incidence of cleft palate induction with N.
glaucais high during gestation days 32–41, other extra-
cranial anomalies (multiple congenital contractures)
have also been described after longer exposure to
teratogenic alkaloids (30–60 days). In an earlier study,
three clefted goats studied carefully 6 months after
birth had gross changes in midfacial development,
 CLEFT PALATE INDUCTION BY N. GLAUCA IN GOATS AND SHEEP
specifically maxillary hypoplasia and midfacial retru-
sion (Weinzweig et al., ’99a) (Fig. 3).
Ultrasound studies on pregnant sheep gavaged with
teratogenic plants to compare fetal movement with that
of the goat are still incomplete. More in-depth experi-
ments are needed to determine how long fetal move-
ment must be depressed to induce cleft palate and
whether short intermittent periods of fetal recovery
appear to allow palates to close (Panter et al., ’90;
Panter and Keeler, ’92). The fetal movement induction
concept is supported by studies in pregnant goats fed
fresh Conium maculatum plants and mature ground
seed in which fetal activity was monitored by ultra-
sound. While fetal movement was reduced in both
groups, as expected, it only lasted for 5 hr after dosing
of fresh plant material and normal fetal movement had
returned at 5–9 hr after treatment. Pregnant goats
treated with seed had a sustained inhibition of fetal
movement persisting for up to 18 hr, which was continu-
ous from dose to dose. All neonatal goats from does
gavaged with seed had cleft palate and severe multiple
congenital contractures, while none exposed to the
fresh plant had cleft palates and only temporary minor
to moderate contractures were noted in the front legs
(Panter et al., ’90). Conium maculatum seed and fresh
vegetative material contain different concentrations
and compositions of the coniine-type piperidine alka-
loids (Keeler and Balls, ’78).
While the mechanical mechanism appears to be a
major factor of cleft palate formation in goats, the
molecular mechanism remains unknown. We hypoth-
esize, however, that it may be due to a fetal pharmaco-
logic neuromuscular blockade. This hypothesis is sup-
ported by preliminary experiments in which two
compounds with known pharmacologic activity (succi-
nylcholine and curare extract) were infused via osmotic
minipumps into the amniotic sac of fetal goats during
susceptible periods of gestation (Panter, unpublished
data). Cleft palate and severe multiple congenital con-
tracture-type skeletal defects were induced. This evi-
dence is preliminary and further research is needed to
verify the biochemical mechanisms of action.
ACKNOWLEDGMENTS
The authors thank Terrie Wierenga for technical
assistance and Al Maciulis, Rex Probst, Danny Hansen,
and Seth Lundquist for plant preparation and animal
care and handling.
LITERATURE CITED
Adzick NS, Longaker MT. 1991. Animal models for the study of fetal
tissue repair. Am Surg Res 51:216–228.
Binns W, James LF. 1961. A congenital deformity in calves, similar to
‘‘crooked calf disease,’’ has been experimentally produced by feeding
heifers lupine and lead. Proc West Sec Am Soc Anim Prod 12(LXVI):
1–3.
Crowe MW. 1969. Skeletal anomalies in pigs associated with tobacco.
Mod Vet Pract 69:54–55.
209
Davis AM, Stout DM. 1986. Anagyrine in western lupine. J Range
Manage 39:29–30
Evans HE, Sack WO. 1973. Prenatal development of domestic and
laboratory mammals: growth curves, external features and selected
references. Anat Histol Embryol 2:11–42.
Gardner DR, Panter KE. 1994. Ammodendrine and related piperidine
alkaloid levels in the blood plasma of cattle, sheep and goats fed
Lupinus formosus. J Nat Toxins 3:107–116.
Hedrick MH, Rice HE, Vander Wall KJ, Adzick NA, Harrison MR,
Siebert J, Hoffman WY, Longaker MT. 1996. Delayed in utero repair
of surgically created fetal cleft lip and palate. Plast Reconstr Surg
97:900–907.
Keeler RF. 1976. Lupin alkaloids from teratogenic and non teratogenic
lupins. III. Identification of anagyrine as the probable teratogen by
feeding trials. J Toxicol Environ Health 1:887–889.
Keeler RF. 1979. Congenital defects in calves from maternal ingestion
of Nicotiana glauca of high anabasine content. Clin Toxicol 15:417–
426.
Keeler RF, Balls LD. 1978. Teratogenic effects in cattle of Conium
maculatum and Conium alkaloids and analogs. Clin Toxicol 12:
49–64.
Keeler RF, Crowe MW. 1984. Teratogenicity and toxicity of wild tree
tobacco, Nicotiana glauca in sheep. Cornell Vet 74:50–59.
Keeler RF, Panter KE. 1989. Piperidine alkaloid composition and
relation to crooked calf disease-inducing potential of Lupinus formo-
sus. Teratology 40:423–432.
Keeler RF, Balls LD, Panter K. 1981. Teratogenic effects of Nicotiana
glaucaand concentration of anabasine, the suspect teratogen in
plant parts. Cornell Vet 71:47–53.
Keeler RF, Crowe MW, Lambert EA. 1984. Teratogenicity in swine of
the tobacco alkaloid anabasine isolated from Nicotiana glauca.
Teratology 30:61–69.
Longaker MT, Adzick NS. 1991. The biology of fetal wound healing: a
review. Plast Reconstr Surg 87:788–798.
Morrow DA. 1986. Current therapy in theriogenology. Vol 2.Philadel-
phia: WB Saunders. p 1088.
Palotay JL. 1959. ‘‘Crooked calves.’’ West Vet 6:16–20.
Panter KE, Keeler RF. 1992. Induction of cleft palate in goats by
Nicotiana glaucaduring a narrow gestational period and the rela-
tion to reduction in fetal movement. J Nat Toxins 1:25–32.
Panter KE, Keeler RF, Buck WB. 1985a. Congenital skeletal malforma-
tions induced by maternal ingestion of Conium maculatum (poison-
hemlock) in newborn pigs. Am J Vet Res 46:2064–2066.
Panter KE, Keeler RF, Buck WB. 1985b. Induction of cleft palate in
newborn pigs by maternal ingestion of poison-hemlock (Conium
maculatum). Am J Vet Res 46:1368–1371.
Panter KE, Bunch TD, Keeler RF, Sisson DV. 1988. Radio ultrasound
observations of the fetotoxic effects in sheep from ingestion of
Conium maculatum (poison-hemlock). Clin Toxicol 26:175–187.
Panter KE, Bunch TD, Keeler RF, Sisson DV, Callan RJ. 1990.
Multiple congenital contractures (MCC) and cleft palate induced in
goats by ingestion of piperidine alkaloid-containing plants: reduc-
tion in fetal movement as the probable cause. Clin Toxicol 28:69–83.
Panter KE, Wierenga TL, Bunch TD. 1991. Ultrasonographic studies
on the fetotoxic effects of poisonous plants on livestock. In: Keeler
RF, Tu AT, editors. Handbook of natural toxins. Vol 6. New York:
Marcel Dekker. p 589–610.
Panter KE, Gardner DR, Molyneux RJ. 1994. Comparison of toxic and
teratogenic effects of Lupinus formosus, L. arbustus and L. caudatus
in goats. J Nat Toxins 3:83–93.
Panter KE, Gardner DR, Gay CC, James LF, Mills R, Gay JM, Baldwin
TJ. 1997. Observations of Lupinus sulphureus-induced ‘‘crooked calf
disease.’’ J Range Manage 50:587–592.
Panter KE, Gardner DR, Molyneux RJ. 1998a. Teratogenic and
fetotoxic effects of two piperidine alkaloid-containing lupines (L.
formosus and L. arbustus) in cows. J Nat Toxins 7:131–140.
Panter KE, Gardner DR, Shea RE, Molyneux RJ, James LF. 1998b.
Toxic and teratogenic piperidine alkaloids from Lupinus, Conium
and Nicotiana species. In: Garland T, Barr AC, editors. Toxic plants
and other natural toxicants. Wallingford, UK: CAB International.
p 345–350.
210 PANTER ET AL.
Panter KE, James LF, Gardner DR. 1999. Lupines, Poison-hemlock
and Nicotiana spp.: toxicity and teratogenicity in livestock. J Nat
Toxins 8:117–134.
Rintala A, Ranta R, Stegars T. 1984. On the pathogenesis of cleft palate in
the Pierre Robin Syndrome. Scand J Plast Reconstr Surg 18:237–240.
Shupe JL, Binns W, James LF, Keeler RF. 1967a. Lupine, a cause of
crooked calf disease. J Am Vet Med Assoc 151:198–203.
Shupe JL, James LF, Binns W. 1967b. Observations on crooked calf
disease. J Am Vet Med Assoc 151:191–197.
Shupe JL, James LF, Binns W, Keeler RF. 1968. Cleft palate in cattle.
Cleft Palate J. 1:346–354.
Wagnon KA. 1960. Lupine poisoning as a possible factor in congenital
deformities in cattle. J Range Manage 13:89–91.
Weinzweig J, Panter KE, Pantaloni M, Spangenberger A, Harper JS,
Edstrom LE, Gardner DR, Wierenga T. 1999a. The fetal cleft palate.
I. Characterization of a congenital model. Plastic Reconstr Surg
103:419–428.
Weinzweig J, Panter KE, Pantaloni M, Spangenberger A, Harper JS,
Lui F, James LF, Edstrom LE. 1999b. The fetal cleft palate. II.
Scarless healing following in utero repair of a congenital model.
Plast Reconstr Surg 104:1356–1364.