Effect of Alligator pepper (Zingiberaceae Aframomum melegueta) on first trimester pregnancy in Sprague Dawley rats

Abstract

Alligator pepper (Aframomum melegueta) is a spice that is widely used in many cultures for entertainment, religious rites, food flavor and as a part of many traditional doctors medications. Pregnant women are among those who ingest Alligator pepper in these activities. This experiment was carried out to determine the health risk or benefit of Alligator pepper to pregnant women if any. Fifteen male rats and fifteen female rats of proven fertility from a pilot study were randomly paired in fifteen cages in a well ventilated room. After three days of mating, the males were withdrawn from the females, which were allowed to stay in their separate maternity cages for 18-25 days. The females in the control group were fed with normal rat chow and clean drinking water ad libitum for the duration of the experiment. Each of the rats in the experimental group was served 20 g of rat chow mixed with 50mg of Alligator pepper for one day only and thereafter fed with normal rat chow and clean drinking water ad libitum for 18-25 days. The rats in the control group had a mean of 7 litters each, while the rats in the experimental group did not litter at all. It was concluded that ingestion of large quantities of Alligator pepper poses a health risk to women in their first trimester of pregnancy.

Full text

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Nigerian Journal of Physiological Sciences 24 (2): 161 -164 ©Physiological Society of Nigeria, 2009
Available online/abstracted at http://www.bioline.org.br/np; www.ajol.info/journals.njps; www.cas.org
EFFECT OF ALLIGATOR PEPPER (ZINGIBERACEAE AFRAMOMUM
MELEGUETA) ON FIRST TRIMESTER PREGNANCY IN SPRAGUE DAWLEY
RATS
U. INEGBENEBOR, M. I. EBOMOYI
1
, K. A. ONYIA
1
, K. AMADI, A. E.
AIGBIREMOLEN
2
Department of Physiology, College of Medicine, Ambrose Alli University, Ekpoma, Delta State, Nigeria E-mail:
druteinegbenebor@yahoo.com
Department of Physiology
1
, School of Basic Medical Sciences, College of Medicine, Benin City, Edo State,
Nigeria.
Department of Pharmacology
2
, College of Medicine, Ambrose Alli University, Ekpoma, Edo State, Nigeria
.
Summary: Alligator pepper (Aframomum melegueta) is a spice that is widely used in many cultures for
entertainment, religious rites, food flavor and as a part of many traditional doctors medications. Pregnant women
are among those who ingest Alligator pepper in these activities. This experiment was carried out to determine the
health risk or benefit of Alligator pepper to pregnant women if any. Fifteen male rats and fifteen female rats of
proven fertility from a pilot study were randomly paired in fifteen cages in a well ventillated room. After three
days of mating, the males were withdrawn from the females, which were allowed to stay in their separate
maternity cages for 18-25 days. The females in the control group were fed with normal rat chow and clean
drinking water ad libitum for the duration of the experiment. Each of the rats in the experimental group was
served 20 g of rat chow mixed with 50mg of Alligator pepper for one day only and thereafter fed with normal rat
chow and clean drinking water ad libitum for 18-25 days. The rats in the control group had a mean of 7 litters
each, while the rats in the experimental group did not litter at all. It was concluded that ingestion of large
quantities of Alligator pepper poses a health risk to women in their first trimester of pregnancy.
Key words : Effect, Alligator pepper, Herbal remedy, Nutrition, Pregnancy
Introduction
Alligator pepper is widely used by many
cultures in Nigeria for various purposes. It is served
along with Kola nuts to guests for entertainment,
members of the Iyayi (Faith) Society of Nigeria as
communion and used for religious rites by diviners
for invoking spirits. It is a common ingredient in
pepper soup, a spicy delight in most parts of West
Africa. Concoctions made of Alligator pepper are
often used by traditional doctors as medications for
various ailments. (Personal observation) Pregnant
women are not excluded from eating this widely
used substance. The constituents of an essential oil,
extractable by hydro-distillation from the seeds of
Aframomum melegueta include two sesquiterpene
hydrocarbons, humelene and caryophyllene, their
oxides and a few non-terpenoids. (Ajaiyieoba and
Ekundayo, 1999).
There is a need to know the danger, pregnant
women are exposed to, when this substance is used
as an ingredient in food or served during religious
rites and naming ceremonies. Hence, this
investigation aims to find out if granulated alligator
pepper poses any health risk to pregnant women
especially in their first trimester. It also aims to
determine other effects of alligator pepper in
pregnancy if any.
Materials and methods
Pilot study: selection of experimental rats
Twenty (20), six months old female and 20,
six months old male Sprague Dawley rats, of the
same strain; each weighing 125-150g and inbred in
Ambrose Alli University Animal House, were kept
in standard cages in a well ventilated room for
acclimatization, for a period of two weeks. During
the acclimatization period, normal rat chow and
clean drinking water were given to the rats ad
libitum.

162
After the acclimatization period, a pilot study
was carried out thus; Twenty female rats were
randomly put in 20 cages,( A -T ) Twenty male rats
of the same breed were randomly allocated to the
female rats and left to cohabit for three days, so that
mating could take place. Thereafter, the males were
withdrawn and were put in separate cages labeled
(MA, MB, ... MT) based on the female rat each
male rat mated with. The female rats were left in
their maternity cages for 18-25 days.
Experimental Study: To determine the effect of
Alligator pepper on pregnancy
Three weeks after the weaning of the
offsprings, the fertile female rats A, B, C, D, E, F,
G, J, L, M, N, P, Q, R and T from the above pilot
study, were selected, weighed and randomly
allocated into fifteen cages (1-15). Their weights
then ranged between 145g and 200g.
Experimental Animals
Fifteen female and fifteen male Sprague
dawley rats of proven fertility were used for this
study. The rats were now eight months old and were
kept in cages in a well ventilated laboratory. The
rats were fed with normal rat chow and clean water
ad libitum.
Experimental drug
Alligator pepper, obtained in Ekpoma market,
Edo state, Nigeria and ground into powder with a
clean grinding machine.
Method
The fifteen selected female rats of proven
fertility from the pilot study were randomly
allocated into fifteen cages (1-15). Each female rat
was kept in a separate cage so that there were fifteen
cages for the fifteen rats. Female rats (1 – 8) served
as the Control rats while female rats (9-15) served
as the Experimental rats. The fifteen selected male
rats of proven fertility from the pilot study were
randomly allocated to the fifteen cages containing
fifteen female rats so that each cage contained one
female rat and one male rat. They were left in their
various cages for three days in order to allow
mating to occur. After three days, the males were
withdrawn from the females. The male rats were
put in sepa\rate cages labeled (M1-M15) based on
the female rat each male rat mated with.
The female rats were left in their initial cages
so that each female rat occupied a separate
maternity cage.
Administration of Granulated Alligator pepper
Female rats 1-8 (Control group)
Female rats 1-8, which represented the control
group, were fed with normal rat chow and water ad
libitum. Alligator pepper was not administered to
any of these rats throughout the duration of the
experiment. The rats were observed in separate
maternity cages for 18 to 25 days.
Female rats 9-15 (Experimental group)
Female rats were starved of rat chow for three
hours after separation from the males on the fourth
day following first day of exposure to their male
partners. Thereafter, they were each served 20g of
rat chow mixed with 50 mg of Alligator pepper.
They were not served extra rat chow on the fourth
day. There were no left over rat chow by the
morning of the fifth day. From the fifth day to the
end of the experiment, they had normal rat chow
and clean water ad libitum. The rats were observed
in separate maternity cages for 18 to 25 days.
Statistical analysis
This was done using cross tabulation and
Daniel Soper’s free software for calculating One
Way Analysis of Variance (ANOVA) at 5% level of
significance (Soper, 2009)
U. INEGBENEBOR et al

163
Table 2: Effect of granulated alligator pepper on
pregnancy outcome in Sprague Dawley rats
Parameters Control
group
(n =8)
Experimental
Group (n=7)
Dose of granulated
alligator pepper
Nil 50mg
Dose of alligator
pepper (mg/Kg)
Nil 160g
Mean initial weight of
rats
165g 160g
Mean weight of rats
after 3 weeks
302.5g 212.6
Mean duration of
pregnancy
24 days Nil
Mean weight gain in
pregnancy
137.5
±17.66g
52.6*
±19.46g
Mean no of liters 7 Nil *
* P < 0.0001; n = no of rats in the group
Results
The results of the experiment are as shown in
Tables 2. The female rats in the control group (Rats
1 -8) had a mean of 7 litters each, while the female
rats in the experimental group ( Rats 9-15) did not
litter at all. This was found to be statistically
significant. (P<0.0001). There was also a significant
difference (p < 0.0001) between the weight gain in
the control and the experimental rats during
pregnancy. Weight gain in the experimental rats
declined after two weeks of pregnancy and
coincided with presence of copious blood stained
vaginal discharge.
Discussion
Results obtained from the investigation;
depicted by the absence of litters by the female rats
in the experimental group, their significant (p
<0.0001) weight loss compared to the controls and
the presence of copious blood stained vaginal
discharge, contradict the usual fluid retention and
weight gain during pregnancy.(Denton,1982) This
could imply that the ingestion of alligator pepper by
rats in the present experiment does not keep or
nurture pregnancy as evidenced by the fact that
none of the affected experimental rats littered even
one offspring.
Table 1: Selection of rats for experimental study
S/N Female Rat Male
Partner
Duration of
Pregnancy
No of Litters
delivered
SELECTION
A MA 24 DAYS 9 A, MA SELECTED
B MB 24 DAYS 9 B, MB SELECTED
C MC 27 DAYS 5 C MC SELECTED
D MD 27 DAYS 6 D MD SELECTED
E ME 23 DAYS 8 E, ME SELECTED
F MF 25 DAYS 3 F, MF SELECTED
G MG 23 DAYS 3 G, MG SELECTED
H MH 23 DAYS 1
I MI NIL NIL
J MJ 23 DAYS 6 J, MJ SELECTED
K MK NIL NIL
L ML 26 DAYS 4 L, ML SELECTED
M MM 23 DAYS 9 M, MM SELECTED
N MN 25 DAYS 7 N, MN SELECTED
O MO 23 DAYS 1
P MP 24 DAYS 6 P, MP SELECTED
Q MQ 23 DAYS 8 Q, MQ SELECTED
R MR 24 DAYS 5 R, MR SELECTED
S MS NIL NIL
T MT 25 DAYS 5 T, MT SELECTED
Effect of Alligator Pepper on Pregnancy

164
The dose of granulated Alligator pepper was
decided on because the rats tolerated the lower
doses of extract of Alligator pepper used in a
previous experiment on the effect of aqueous extract
of Alligator pepper on Gestational weight gain’.
(Inegbenebor et al, 2009). Previous reports have
shown that ingestion of 350mg of wholeseed
Alligator pepper (5-7mg/Kg body weight) caused
diplopia and blurred vision in healthy Igbo men
(Igwe et al, 1999). However, apart from the inital
loss of appetite, none of the 7 rats, which ingested
50mg of Alligator pepper ( 286-345mg/Kg body
weight) had serious side effects apart from the
discontinuaton of pregnancy.
Conclusion
When ingested in high doses, alligator pepper
causes discontinuation of first trimester pregnancies
in rats. (p < 0.0001) It will be unethical to carry out
this experiment in human subjects. However,
women in the reproductive age group might need to
avoid eating Alligator pepper if they are desirous of
childbirth or are in their first trimester of pregnancy.
Acknowledgement
We are grateful to Efuaose Okoduwa, Ehojie
Osamudiamhen, Monday Agbonifo, and Godspower
Oikhena, who assisted with the day to day activities
of this study.
References
Ajaiyieoba, O.E and Ekundayo, O.(1999): Essential
oil constituents of Aframomum melegueta
(Roscoe) K. Schum. seeds (alligator pepper)
from Nigeria. Flavour and Fragrance J. 14(2):
109-111.
Denton, D.A.(1982): The Hunger for Salt. Berlio
Springer-Verlag. 4:6-12.
Igwe S. A.; Emeruwa, I. C.; Modie, J. A.(1999):
Ocular toxicity of Aframomum melegueta
(alligator pepper) on healthy Igbos of Nigeria.
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Inegbenebor, U., Ebomoyi, M..I., Onyia, K.A,
Amadi, K. Aigbiremolen, A.E (2009). Effect
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14 December, 2009.
Received: September 1, 2009
Accepted: December 21, 2009
U.
INEGBENEBOR
et al