




































_____________________________________________________________________________________________________ 
 
*Corresponding author: E-mail: ndehforwah@gmail.com,  worcacccegroup@gmail.com; 
 
 
 

Asian Journal of Immunology 
 
4(1): 125-134, 2021; Article no.AJI.77165 
 

 
 

 

 

Fresh Raffia Palm Wine, Fermented Soya Bean Milk 
Supplementation, Enzymatic Antioxidant and Lipid 

Peroxidation Biomarkers in Lactating Female  
Wistar Rats 

 
Forwah Jacques Ndeh1*, Ebot Walter Ojong2, Uwem Okon Akpan1  

and Immaculate Ihuoma Ekeagba3  

 
1
Chemical Pathology Unit, Department of Medical Laboratory Science, Faculty of Allied Medical 

Sciences, University of Calabar, Nigeria. 
2
Department of Medical Laboratory Sciences, Faculty of Health Sciences, University of Buea,  

South West Region, Cameroon. 
3
WORCACCCE Group Integrated Healthcare Services, Technology ,Training and Innovative 

Research Foundation (WGIHSTAIRF), P.O.Box 45, Bamenda, North West Region, Cameroon. 
 

Authors’ contributions  
 

This work was carried out in collaboration among all authors. All authors read and approved the final 
manuscript. 

 
Article Information 

 
Editor(s): 

(1) Prof. Cynthia Aracely Alvizo Báez, Autonomous University of Nuevo Leon, Mexico. 
Reviewers: 

(1) Asmarani Kusumawati, Gadjah Mada University, Indonesia. 
(2) Isaac John Umaru, Federal University Wukari, Nigeria. 

(3) Julio Cesar Mendes Soares, Federal University of Santa Maria, Brazil. 
Complete Peer review History, details of the editor(s), Reviewers and additional Reviewers are available here: 

https://www.sdiarticle5.com/review-history/77165 

 
 
 

Received 04 October 2021  
Accepted 08 December 2021 
Published 29 December 2021 

 
 

ABSTRACT 
 

Aim: Lactation is the most crucial period characterized by intensive caloric requirements to boast 
up female’s nutritive system and breastfeeding life. Current study evaluated the effects of 
fermented soya bean milk (FSBM) and fresh palm wine (FPW) supplements on endogenous 
antioxidant enzymes activities and lipid peroxidation biomarker in thirty five (35) lactating Wistars 
rats.    
Methods: Animals were obtained from the Experimental Research Animal House of University of 
Calabar, Nigeria. At parturition, the animals were randomly divided into five groups containing five 

Original Research Article 



 
 
 
 

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126 

 

(5) rats (n=5). Oral treatment done as follow: Group I: Control group, was given normal feed of 
protein content 20mg/kg and distilled water (1 ml/kg), Group II: Metoclopramide (5 mg/kg), Group 
III: 10ml /kg of FPW. Group IV: Three (3) sub-groups of group four (4) received 10%, 20% and 
40% of FSBM respectively, Group V: Co-administration of 40% FSBM plus FPW (10ml /kg) 
supplements .Treatment was done every six hour daily for a duration of ten (10) days.  
Results: Serum malondialdehyde concentration, a biomarker of Lipid peroxidation was significantly 
increased (P < 0.05) in metoclopramide treated group than control group, FSBM and FPW 
supplemented groups. There was a significant decrease (P < 0.05) in serum activities of four 
endogenous antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) in 
metoclopramide treated group when compared to control group, FSBM and FPW supplemented 
groups. 
Conclusion: while fermented soya bean milk and fresh palm wine supplementation increased 
lactation and endogenous enzymatic antioxidant activities without lipid peroxidation, 
Metoclopramide at 5 mg/kg concentration increased lactation with lipid peroxidation. 
 

 
Keywords: Soya bean; palm wine; lactation; lipid peroxidation; endogenous antioxidant enzymes. 
 

1. INTRODUCTION  
 
According to the International Society of 
Antioxidant in Nutrition and Health (ISANH) and 
Medical Dictionaries, the term antioxidant was 
initially utilized to refer specifically to chemical 
products that prevent the consumption of oxygen 
[1- 3]. Endogenous antioxidants are agents 
which scavenge free radicals and prevent 
oxidative stress damage caused by excess free 
radicals in the human body.  They are also 
known as antiradicals [4]. Endogenous 
enzymatic antioxidants can be defined as 
enzymes  or molecules existing within the cellular 
fluid compartment of the body that  are capable 
of preventing, inhibiting or delaying harmful 
oxidative reactions caused by high levels of free 
radicals or oxidants, thus playing an important 
role in forming antioxidant defensive mechanism 
and protective system of the body [5]. In recent 
times, antioxidants in the body in required, 
acceptable and appropriate levels have been 
reputed to help the body in repairing, equipping 
and preventing the formation of and scavenging 
of free radicals and other potentially toxic 
oxidizing species in vivo [6-8]. The fundamental 
classes of antioxidants that form the total 
antioxidant status of an individual include the 
primary or chain breaking antioxidant enzymatic 
systems, secondary and tertiary antioxidants. 
The primary or chain breaking antioxidant 
systems include superoxide dismutase, catalase, 
glutathione peroxidase and reductases. The 
secondary or preventive antioxidants include 
glutathione, vitamin C or ascorbate, vitamin E, 
vitamin A, uric acid, etc .The tertiary or repairing 
antioxidants include small molecules and plasma 
proteins such as albumin, transferrin, 
ceruloplasmin and antioxidant micronutrients 

such as iron, copper, selenium and manganese 
[9]. A number of these compounds are of 
exogenous nature and are obtained from certain 
food substances namely: antioxidants like α-
tocopherol, β-carotene, and ascorbic acid, and 
some micro-nutrient elements such as zinc and 
selenium [10, 11]. Lactogenesis and lactation are 
essential for optimal feeding of infants during 
their first one year of dependable life and this has 
a direct impact on growth, development, 
immunity and health aspect in the neonatal 
period [12]. Mammalian lactating cells are 
therefore equipped with some sort of complex 
defiance mechanisms for radical detoxification 
[13].Oxidative stress (OS) is considered a 
metabolic disturbance that affects organ systems 
and its presence will affect not only the health 
status of the animals but also the quality and 
quantity of the final products, such as milk [14]. 
There is evidence that oxidative damage 
increases during lactation in some domesticated 
and laboratory animals [15, 16]. The soya bean 
is native to the Korean Peninsula and the 
Manchurian Area and has been one of the major 
sources of protein in Korean food [17]. Numerous 
foods are made by from soybeans, including 
doenjang (soya bean paste), ganjang (soya bean 
source), cheonggukjang (fast-fermented bean 
paste), bean curd, soybean milk, and bean-curd 
dregs [18]. Soybean consumption is effective for 
the prevention of osteoporosis, arteriosclerosis, 
strokes and dementia, and can reduce the risk of 
cancer and obesity [19, 20]. Palm wine is a 
whitish sap collected in vessels attached to the 
base of the palm tree from where some leaves 
have been removed. Fresh raffia palm wine 
(Raphia hooker) from these sources is sweet and 
contains little alcohol but, with fermentation, the 
alcohol content increases over time. Unbottled 



 
 
 
 

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palm wine has a slightly lower alcohol content 
(around 3%) than bottled palm wine (around 4%) 
[21,22]. Biochemical constituents of palm wine 
include  carbohydrates (sugars), protein, amino 
acid, lipid, lactic acid, alcohol, vitamins such as  
vitamin B 6 , B1, B2, vitamin  C and some 
antioxidants, mineral and trace elements like iron 
and selenium. The mean sugar content of fresh 
palm wine ranges between 0.10 in maltose and 
8.74 mg/100ml in sucrose [23- 27].  The present 
study was aimed at evaluating the effects of 
fermented soya bean and fresh palm wine 
supplement on lipid peroxidation biomarker and 
activities of four endogenous antioxidant 
enzymes (superoxide dismutase, catalase and 
glutathione peroxidase) in lactating Wistar rats in 
vivo. 
 

2. METHODS 
 
This experimental study was carried out from 
January 2018 to December 2018 at the 
Department of Physiology, Faculty of Basic 
Medical Sciences, College of Medical Science, 
University of Calabar, and the laboratory analysis 
of samples was carried out at the Chemical 
Pathology, and Haemalogy and Blood 
Transfusion Units, Department of Medical 
Laboratory Science, University of Calabar 
Teaching Hospital (UCTH) Calabar, Cross River 
State, Nigeria.   
 

2.1 Animal Grouping and Drug 
Administration 

 
Adult female apparently healthy Wistar rats 
weighing 180-200g were obtained from 
Biochemistry Department Experimental 
Research Animal House of University of Calabar, 
Calabar,Nigeria .Oral treatment was 
administered as follow:-  Group I: (Control 
group) was given normal vital feed pellets and 
distilled water, orally (1 ml/kg). Group II: 
Metoclopramide (5 mg/kg), Group III:  An oral 
treatment (through orogastric tube) of 10 ml/kg 
body weight per day of fresh raffia palm wine for 
10 days. The dosage was calculated as 10 
(ml)/1000 (g) × the body weight of the animal. 
Fresh undiluted palm wine from the sap of oil 
palm trees was collected at every 3 days interval 
from a local palm wine tapper. The palm wine on 
collection was administered to the animals and 
then conserved in a refrigerator for 3 days 
following which it was often replaced with a fresh 
collection.  
 

Group IV: The three (3) sub groups of group four 
(4) received 10%, 20% and 40% soya bean, 
respectively. Group V: was co-administered 20% 
soya bean supplement and fresh palm wine 
(10ml/kg). Administration was carried out orally 
for a period of ten (10) days.  
 

2.2 Sample Collection 
 
After ten days, the rats were anaesthetized by 
chloroform inhalation in a closed chamber and 
blood samples were obtained via cardiac 
puncture into specimen bottles and allowed to 
clot, and separated by centrifugation at 4000 
rev/per minutes for 10 minutes using Centrifuge 
Hettich (Universal 32, Made in Germany) at an 
average room temperature of 23˚C.  The 
supernatant obtained was used for biochemical 
assays. Sample haemolysis was avoided.  
 

2.3 Biochemical Assay of 
Malondialdehyde in Serum 

 
Malondialdehyde (MDA) is one of many low 
molecular weight end-products of lipid 
hydroperoxide decomposition and is most often 
measured as a biomarker and index of lipid 
peroxidation. MDA activity was estimated using  
the NWLSS™ Malondialdehyde  commercial 
assay kits  purchased  from Northwest Life 
Sciences Specialized Product NWK-MDA01, 
Vancouver WA, Specificity: Malondialdehyde, 
Sensitivity: 0.08 µ) based on the method adapted 
by Janero, [28]. The test principle is based on the 
reaction of MDA with thiobarbituric acid (TBA); 
forming an MDA-TBA2 adduct that absorbs 
strongly at 532 nm. 
 

2.4 Biochemical Assay of Serum 
Superoxide Dismutase Activities in 
the Serum 

 
Superoxidase Dismutase (SOD) activity was 
assayed using the North West Life Science 
Specialties (NWLSS™) SOD assays kit (product 
NWK-SOD02, Specificity: Cu/Zn, Mn and Fe 
superoxide dismutase, sensitivity: 5 U/mL). The 
method is based on the principle of superoxide 
inhibition of auto-oxidation rate of hematoxylin as 
originally described by, Martin, [29], with 
modifications to increase robustness and 
reliability.  About 920 μL of assay buffer was 
added to each cuvette. This was followed by 
addition of 40 μL of assay buffer (for blank) and 
40 μL of sample. The mixture was incubated for 
two (2) minutes. After which 40 μL hematoxylin 



 
 
 
 

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reagent was added and mixed quickly to start the 
auto-oxidation reaction. An absorbance was 
measured at 560 nm for every 10 seconds for 5 
minutes. 
                                                     

2.5 Biochemical Assay of Catalase 
Activities in the Serum 

 
Catalase (CAT) activity was estimated  using the 
North West Life Science Specialties (NWLSS™) 
NWLSS

TM
 Catalase assay kits (Product NWK-

CATO1, Specificity: 6.0 U Catalase/mL) based 
on the method of  Beers, (1952) [30], with 
modifications to increase robustness and 
convenience. The test is based on monitoring the 
consumption of H2O2 substrate. The ultraviolet 
absorption by hydrogen peroxide which is 
considered as a function of time and was used to 
follow the catalase-peroxide reaction. The 
absorption spectrum of hydrogen peroxide was 
measured spectrophotometrically at the 
wavelength of 240nm. The optical density as a 
measure of peroxide concentration, increases 
linearly with peroxide concentration and directly 
proportional to catalase in the sample in 
accordance with the Beer-Lambert law. The 
reaction products, oxygen and water, do not 
absorb light in this spectral region nor does 
catalase. 
 

2.6 Biochemical Assay of Glutathione 
Peroxidase Activity (GPx) in Serum 

 

Glutathione peroxidase (GPx) activity was 
assessed using the North West Life Science 
Specialties (NWLSS™) cGPx (GPX1) ELISA kit 
(Product NWK-GPX02, Specificity: Glutathione 
peroxidase, Sensitivity: 12.5 pg/ml) based on the 
method described by Avissar, [31]. The assay is 
based on a sandwich Enzyme-Linked 
Immunosorbent Assay, where sample GPx 
concentration was determined by comparing the 
450 nm absorbance sample wells to the 
absorbance of known standards. 
 

2.7 Statistical Analysis  
 

The analysis of data was computed using SPSS 
software version 20 (SPSS Inc., Chicago, USA) 
and Microsoft Excel (2007). The results of our 
study were analyzed with the aid of cross 
tabulations to explore proportional associations 
between variables and Chi square test was used 
to explore proportional association between 
groups. All data were expressed as mean ± 
standard error of the mean (mean ± SEM). 

Statistical significance was carried out using one 
way analysis of variance (ANOVA) followed by 
Tukey’s post-hoc test. Values of P < 0.05 were 
considered significant. 
 

3. RESULTS 
 
The results of the current study are shown in the 
following tables below. 
 
Table 1 shows the effects of ten (10) days 
administration of fermented soya bean milk and 
fresh palm wine supplementation on lipid 
peroxidation biomarker (serum malondialdehyde 
(MDA) concentration in female lactating         
wistar rats. These results show that  the mean 
values of MDA ( in  µmol/L) after 10 days   from 
Group I to Group V .The MTCL treated group 
had a higher level of MDA (P < 0.05) when 
compared to the control group, while Group V 
had the lowest level of MDA indicating the 
different degree of lipid peroxidation and 
lactation. 
 
As shown in Table 1, the MTCL treated group 
had a highest level of MDA (P < 0.05) compared 
to the control indicating a high degree of lipid 
peroxidation and lactation, while group V had the 
lowest level of MDA among the test groups, 
indicating a lowest degree of lipid peroxidation 
and lactation. Group III and IV had moderate 
levels of MDA. 
 
Table 2 shows the effects of ten (10) days 
administration of fermented soya bean milk and 
fresh palm wine supplements on the serum 
activities of four endogenous antioxidant 
enzymes in female lactating wistar rats namely:- 
 

3.1 Superoxide Dismutase (SOD) 
 
These results show that  the mean values of 
SOD (IU/L) after 10 days   from Group I to 
Group V. The MTCL treated group had the 
lowest level of SOD while group V had the 
highest level of SOD indicating the degree of 
elevation of endogenous antioxidants enzymes 
and less oxidative stress. 
 
Catalase (CAT): These results show that the 
mean values of catalase -CAT (IU/L) after 10 
days   from Group I to Group V . The MTCL 
treated group had the lowest level of catalase 
while Group V had the highest level of catalase 
indicating the degree of elevation of endogenous 
antioxidant enzymes and less oxidative stress.  



 
 
 
 

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Table 1.  Lipid peroxidation biomarker (Serum MDA levels ) in female lactating Wistar rats treated with fermented Soya bean milk and fresh palm 
wine supplements, metoclopramide, distilled water and normal feed 

 

Groups Animal handling   and treatment  of groups  

I II III IV V  

Control Treated 
with  

Treated 
with  

Treated  
with  

Treated 
with 

Treated  
with  

Treated 
with  

P-value 

Substances Administered Distilled 
water  

METCL  FPW FSBM FSBM FSBM FSBM  

Quantity  1ml/kg 5mg/kg 10ml/kg 10% 20% 40% 40%+FPW  
Mean±SEM  MDA after 10 days 
(µmol/L) 

1.33± 0.11 1.79± 0.15 1.53 ± 0.09 1.43 ± 0.11      1.61 ± 0.07   1.44 ± 0.05         1.42 ± 0.07          P< 
0.05* 

METCL = Metoclopramide, MDA = malondialdehyde, FPW =fresh palm wine, FSBM= fermented soya bean milk 
mean ± SEM= mean plus or minus standard error of the mean. 

* There was a significant difference in MDA levels in FSBM, and FPW treated groups (P < 0.05) compared to the control group, FSBM and FPW supplemented groups 
 

Table 2. Serum activities of four endogenous antioxidant enzymes (SOD , CAT and GPX ) in female lactating wistar rats treated with soya bean 
milk and fresh palm wine supplements, metoclopramide, distilled water and normal feed 

 

 Group  Animal handling  and treatment of groups  

I II III IV V P-value 

Control  Treated with  Treated with  Treated  with  Treated with Treated  with  Treated with  

Substances Administered Distilled water  METCL   FPW FSBM FSBM  FSBM FSBM 
 Quantity   1mg/kg 5mg/kg 10ml/kg 10% 20% 40% 40% +FPW 
Mean±SEM   2.25± 0.09 1.69±0.07  2.18 ± 0.10  1.76± 0.10 1.85±0.05 2.34±0.13 2.98 ± 0.12 P<0.05

α 

 activity (IU/L) 
Mean ± SEM Catalase activity  
(IU/L)after 10 days  

55.00 ± 0.71 48.20±1.55  66.80±1.59 65.00 ±1.14  67.20± 1.28 70.40± 1.17  78.20±1.02  P<0.05
β 

Mean ± SEM activity 
GPX(IU/L) after 10 days  

49.40±0.93), 45.20± 1.56  43.60±0.93 42.60 ±1.50  43.60± 1.50  49.00± 0.94 50.80 ± 0.86  P<0.05
γ 

SOD= superoxide dismutase, CAT= catalase, GPX =glutathione peroxidase, METCL = Metoclopramide, FPW =fresh palm wine, FSBM= Fermented soya bean milk , mean ± 
SEM= mean plus or minus standard error of the mean 

 



 
 
 
 

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Glutathione peroxidase (GPX): These results 
show that  the mean values of glutathione 
peroxidase (GPX)  (IU/L) after 10 days   from 
Group I to Group V. The MTCL treated group 
had the lowest level of Glutathione peroxidase 
(GPX) while group V had the highest level of 
glutathione peroxidase (GPX) indicating the 
degree of elevation antioxidants and less 
oxidative stress. 
 
α
There was a significant increase in SOD 

activities  in FSBM and FPW treated group (P < 
0.05) when compared to the control group and 
MTCL treated group.  In Table 2 above it is seen 
that the MTCL treated group had a statistically 
significant decrease (P < 0.05) activity of SOD 
when compared to the control group and the 
treated group with FSBM 10%, 20%, 40% and 
FPW while Group V had the highest activity of 
SOD (P < 0.05) indicating the different degree of 
elevation in the activities of endogenous 
antioxidant enzymes and lactation with less lipid 
peroxidation.  
 
β
 There was a significant increase in CAT 

activities  in FSBM and FPW   treated group (P < 
0.05)  when  compared to the control group  and 
MTCL treated groups. In Table 2 as shown 
above, the MTCL treated group had  a significant 
decrease in (P < 0.05) the activity of catalase 
when compared to the control, FSBM 10% , 
FSBM 20% and FPW treated groups, while  
Group V had the highest activity of catalase  
indicating different degrees in the  elevation of 
endogenous  antioxidant enzymes  activities  and 
lactation with less lipid peroxidation.  
 
γ
 There was a statistically significant increase in 

GPX levels in FSBM, and FPW treated group (P 
< 0.05)  when  compared to the control group  
and MTCL treated groups. In Table 2 as shown, 
the MTCL treated group had significant decrease 
(P < 0.05) in the activity of glutathione 
peroxidase (GPX) when compared to the control, 
FSBM 10%, 20%, 40% and FPW treated groups 
respectively, while group V had a significantly 
higher (P  <0.05) activity of glutathione 
peroxidase (GPX) indicating different degrees of 
elevation in endogenous antioxidant enzymes 
and lactation with less lipid peroxidation.  
 

4. DISCUSSION 
 

The  mean ± SEM  of serum MDA level after 10 
days of  the  current study was displayed  in 
Table 1.The  results showed a statistically 
significant increase (P < 0.05) in serum MDA 
levels. In  Group II subjects,  which was  treated 

with Metoclopramide as compared to  Group I  
subjects, the  control group  which was treated 
only with distilled water and normal feed of 
protein content 20mg/kg  Metoclopramide is a 
medication used to treat stomach disorders such 
as reflux, nausea, and vomiting (anti–emetic) 
[31], but in lactating subjects it can cause  
lactation induction  because of its galactagogue 
properties  and lactating  augmentative  action. 
These properties could be attributed to the 
mechanism of action of metoclopramide on its  
receptors  located in the gastro-intestinal, gastro-
esophageal mucosal, membranes of  
hepatocytes, breasts  and central nervous 
system including others target sites, where poly 
unsaturated fatty acids are  broken down with  
the resultant  increase of lipid peroxidation which 
can subsequently assessed by  the biomarker 
called malondialdehyde (MDA) formed  and 
released as a by-product [32]. The common side 
effects of metoclopramide are sleepiness, 
headache, or restlessness, depression and 
tardive dyskinesia which are signs of increase 
oxidative stress and increased lipid peroxidation 
[33]. The Subgroup of Group IV which was 
treated with 20% soya bean showed an 
increased in the level of serum MDA compared 
to the other treated Groups. This could also be 
due to a possible pro-activity of the supplements 
on oxidants resulting in increased lipid 
peroxidation of cells. It could have also been due 
to the inability of the supplement at this 
concentration that  effectively break down  the 
lipid peroxidation chain reaction since at higher 
concentrations it helps to reduce oxidative stress 
because of its high antioxidant content, daidzein , 
genistein  and aglycone  composition.These  are 
inline with [34,35]. However, the increase was 
not statistically significant compared to 
metoclopramide treated and the normal control 
groups (P > 0.05). Superoxide dismutase (SOD) 
activities as shown in Table 2 was decreases in 
the metoclopramide treated group compared to 
the control group although the results were not 
statistically significant. (P > 0.05).  This result 
could be due to the depletion of the endogenous 
antioxidant as a result of its utilization in 
combating the existing oxidative stress as 
suggested by the level of increased MDA in 
metoclopramide treated group. The result also 
could have been due to the inability of 
metoclopramide to stimulate the release of 
endogenous antioxidants as well as its tendency 
to cause increase in reactive oxygen species 
(ROS) within the system.  There was however, a 
remarkable increase in SOD activities , in the 
fresh palm wine  treated group as shown in Table 



 
 
 
 

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2 which could have been be due to its antioxidant 
activity in fresh  palm wine .These results agreed 
with [36]. The increase was however, rivaled by 
that of the soya bean 40% Group, which showed 
the highest increase in serum SOD activities. 
This result is suggestive of SB 40%  Group as 
having an antioxidant activity more than the 
standard anti-oxidant or possibly a more lasting 
effect than that of fresh palm wine and agreed 
with  [37]. This anti-oxidant activity of the 
supplement on serum SOD activities could be 
attributed to the presence of isoflavones [38]. Co-
administration of SB and fresh palm wine 
increased the activities of SOD more than when 
the supplement was given singlely in the 10% 
and 20% treated groups as shown in Table 2. 
This suggests that the co-administration of the 
supplement and fresh palm wine could be more 
potent than the supplement alone at lower doses 
of 10% and 20%. In Table 2 there was a 
statistically significant decrease in the level of 
catalase observed in the metoclopramide, and 
the supplement treated groups at 10% and 20%. 
This result suggests the inability of 
metoclopramide and the supplement at 10% and 
20% to specifically augment the release of 
endogenous catalase enzyme when compared to 
the activity of the supplement at 40%.This is 
expected because catalase and selenium-
containing glutathione peroxidase have been 
demonstrated in human milk but their levels in 
colostrum is higher than that in corresponding 
mature milk and this agreed with [39]. However, 
this highest levels in colostrum usually decline 
during early lactation, despite the fact that total 
lipids increase as reported by Yuksel (2015) [40]. 
As shown on Table 2, there was a statistically 
significant decrease in the activities of GPx in the 
supplement treated groups at 10% and 20% and 
also that of fresh palm wine. This result suggests 
that the supplement at these concentrations also 
do not seem to have antioxidant activity, 
specifically on GPx release or synthesis [41]. 
 

5. CONCLUSION 
 

Metoclopramide at 5 mg/kg concentration is 
capable of increasing lipid peroxidation according 
to this study and the FSBM supplement at 40% 
and fresh palm wine mixture showed an increase 
in endogenous enzymatic antioxidant activities 
with more lactation and less lipid peroxidation 
when compared to the control groups. 
 

WHAT IS KNOWN ABOUT THIS TOPIC 
 

In many African cultures and tradition there is 
long practice that fermented soya beam milk and 

fresh raffia palm wine and is capable of 
augmenting lactation and clinically, 
metoclopramide is known to be a good 
galactogogue and side effects .What is not 
known till the time of this is the effect of the 
fermented soya bean milk and fresh raffia palm 
wine on the oxidative stress and the endogenous 
antioxidant enzymes and lipid peroxidation. 
 

WHAT THIS STUDY ADDS 
 
Findings from this study have  help highlighted 
the role  of  the combine effects of  fresh raffia  
palm wine and fermented soya bean milk 
supplementation during lactation in viviparous 
mammals. In addition the long standing 
traditional dilemma and the superstitious belief 
about the usage of fresh palm wine fermented 
soya bean milk in humans have been elucidated 
by the findings of the current study . 
 

AVAILABILITY OF DATA AND 
MATERIALS 
 
Data sets generated and analyzed in this study 
are available from the corresponding author on 
request. 
  

DISCLAIMER 
 

The products used for this research are 
commonly and predominantly use products in our 
area of research and country. There is absolutely 
no conflict of interest between the authors and 
producers of the products because we do not 
intend to use these products as an avenue for 
any litigation but for the advancement of 
knowledge. Also, the research was not funded by 
the producing company rather it was funded by 
personal efforts of the authors. 
 

ETHICAL APPROVAL 
 

Ethical approval was obtained from the Ethical 
Committee of University of Calabar on Animal 
Handling, consistent with standard animal 
welfare guidelines. 
 

COMPETING INTERESTS 
 

Authors have declared that no competing 
interests exist. 
 

REFERENCES 
 
1. 18th ISANH Middle East Antioxidants 

World Congress, Beirut Antioxidants 2017. 
[(Accessed on 21 June 2019)];  



 
 
 
 

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Available: https://www. isanh-me.com 
2. Definition of antioxidants by NCI's 

Dictionary of Cancer Terms, at the 
National Institutes of Health, online 2020,  
Available:https://www.cancer.gov/publicati
ons/dictionaries/cancer-
terms/def/antioxidant accessed 2 Nov. 
2020. 

3. “Antioxidant.” Merriam-Webster.com 
Dictionary, Merriam-Webster.  
Available: https://www.merriam-
webster.com/dictionary/antioxidant. 
Accessed 2 Nov. 2020, 

4. Khiena Brainina, Natalia Stozhko, and 
Marina Vidrevich, Antioxidants: 
Terminology, Methods, and Future 
Considerations, Antioxidants (Basel). 2019 
Aug;8(8):297. PMCID: PMC6720181, 
PMID: 31404992, 
DOI:10.3390/antiox8080297, 

5. Arun Rasheed, Rinshana Fathima Abdul 
Azeez, A review on natural antioxidants, 
traditional and complementary medicine, 
Cengiz Mordeniz, Intech Open, (December 
13th 2019).  
DOI: 10.5772/intechopen.82636.  

6. De La Fuente M. Effects of antioxidants on 
immune system ageing. Europe Journal of 
Clinical  Nutrition. 2002;56(3):S5–S8. 

7. Ergul Belge Kurutas. The importance of 
antioxidants which play the role in cellular 
response against oxidative/nitrosative 
stress: current state, PMCID: 
PMC4960740, PMID: 27456681, 2016 Jul 
25. DOI: 10.1186/s12937-016-0186-5.  

8. Valko M, Rhodes CJ, Moncol J, Izakovic 
M, Mazur M. Chem biol interact. Free 
radicals, metals and antioxidants in 
oxidative stress-induced cancer. Epub. 
2006;160(1):1-40.  
DOI: 10.1016/j.cbi.2005.12.009.  

9. Manal Azat Aziz, Abdulkareem Shehab 
Diab, Abeer Abdulrazak Mohammed. 
Antioxidant categories and mode of action 
IntechOpen; November 6th 2019. 
DOI: 10.5772/intechopen.83544. 

10. Alejandro Chehue Romero, Elena G. 
Olvera Hernández, Telma Flores Cerón, 
Angelina Álvarez Chávez. The exogenous 
antioxidants, oxidative stress and chronic 
degenerative diseases - a role for 
antioxidants, José A. Morales-González, 
Intech Open, May 22nd 2013,  
DOI: 10.5772/52490/IntechOpen.  

11. Miguel Roehrs, Juliana Valentini, Clóvis 
Paniz, Angela Moro, Mariele Charão, 
Rachel Bulcão, Frnando Freitas, Natália 

Brucker, Marta Duarte, Mirna Leal, Geni 
Burg, Tilman Grune, Solange Cristina 
Garcia. The relationships between 
exogenous and endogenous antioxidants 
with the lipid profile and oxidative damage 
in hemodialysis patients, BMC Nephrology. 
2011;12:Article number: 59.  

12. Margaret C. Neville, et al., Lactation and 
Neonatal Nutrition: Defining and Refining 
the Critical Questions, J Mammary Gland 
Biol Neoplasia. 2012 Jun;17(2):167–188., 
PMCID: PMC3428522, PMID: 22752723, 
DOI: 10.1007/s10911-012-9261-5,  

13. Tinu Mary Samuel, Qianling Zhou, 
Francesca Giuffrida, Daniel Munblit, 
Valérie Verhasselt, and Sagar K. Thakkar, 
Nutritional and Non-nutritional Composition 
of Human Milk Is Modulated by Maternal, 
Infant, and Methodological Factors, Front 
Nutr. 2020;7:576133. PMCID: 
PMC7557356, PMID: 33117843 DOI: 
10.3389/fnut.2020.576133.  

14. Castillo, C., Hernández, J. and Valverde, I. 
Plasma Malonaldehyde (MDA) and Total 
Antioxidant Status (TAS) during Lactation 
in Dairy Cows. Research in Veterinary 
Science. 2006;80:133-139.  

15. Antoine Stier, Sophie Reichert, Sylvie 
Massemin, Pierre Bize & François 
Criscuolo ,Constraint and cost of oxidative 
stress on reproduction: correlative 
evidence in laboratory mice and review of 
the literature, Frontiers in Zoology. 
2012;9:Article number: 37.  

16. Upreti K, Chaki SP, Misro MM. Evaluation 
of peroxidative stress and enzymatic 
antioxidant activity in liver and kidney 
during pregnancy and lactation in rats. 
Health Population Perspective Issues. 
2002;25:177-185. 

17. Kim GY, Moon HK, Lee SW, Moon JN, 
Yoon WJ. Effect of mixed soybeans 
materials on quality characteristics of 
traditional soybean paste (Doenjang) 
during aging. Korean Journal of Food and 
Cookery Science. 2010;26:314-322. 

18. Malgwil IS, Tanko Y, Kawu MU, Eze ED, 
Salami HA, Emmanuel NS, Mohammed A. 
Lipid Peroxidation and Antioxidant 
Enzymes Evaluation in Lactating Female 
Albino Rats Following Supplementation 
with Fermented Soya Bean and Vitamin C 
Advances in Enzyme Research. June 
2017;5(2). 

19. Burneo Palacios ZL. Determination of total 
phenolic compounds and antioxidant 
activity of total extracts of twelve native 

https://www.isanh-me.com/
https://www.cancer.gov/publications/dictionaries/cancer-terms/def/antioxidant
https://www.cancer.gov/publications/dictionaries/cancer-terms/def/antioxidant
https://www.cancer.gov/publications/dictionaries/cancer-terms/def/antioxidant
https://www.merriam-webster.com/dictionary/antioxidant%20.Accessed%202%20Nov.%202020
https://www.merriam-webster.com/dictionary/antioxidant%20.Accessed%202%20Nov.%202020
https://www.merriam-webster.com/dictionary/antioxidant%20.Accessed%202%20Nov.%202020
https://www.ncbi.nlm.nih.gov/pubmed/?term=Brainina%20K%5BAuthor%5D&cauthor=true&cauthor_uid=31404992
https://www.ncbi.nlm.nih.gov/pubmed/?term=Stozhko%20N%5BAuthor%5D&cauthor=true&cauthor_uid=31404992
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6720181/
https://www.ncbi.nlm.nih.gov/pubmed/31404992
https://dx.doi.org/10.3390%2Fantiox8080297
https://www.ncbi.nlm.nih.gov/pubmed/?term=Kurutas%20EB%5BAuthor%5D&cauthor=true&cauthor_uid=27456681
https://www.ncbi.nlm.nih.gov/pubmed/27456681
https://dx.doi.org/10.1186%2Fs12937-016-0186-5
https://pubmed.ncbi.nlm.nih.gov/16430879/
https://pubmed.ncbi.nlm.nih.gov/16430879/
https://pubmed.ncbi.nlm.nih.gov/16430879/
https://www.ncbi.nlm.nih.gov/pubmed/?term=Neville%20MC%5BAuthor%5D&cauthor=true&cauthor_uid=22752723
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3428522/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3428522/
https://www.ncbi.nlm.nih.gov/pubmed/22752723
https://dx.doi.org/10.1007%2Fs10911-012-9261-5
https://www.ncbi.nlm.nih.gov/pubmed/?term=Samuel%20TM%5BAuthor%5D&cauthor=true&cauthor_uid=33117843
https://www.ncbi.nlm.nih.gov/pubmed/?term=Zhou%20Q%5BAuthor%5D&cauthor=true&cauthor_uid=33117843
https://www.ncbi.nlm.nih.gov/pubmed/?term=Giuffrida%20F%5BAuthor%5D&cauthor=true&cauthor_uid=33117843
https://www.ncbi.nlm.nih.gov/pubmed/?term=Munblit%20D%5BAuthor%5D&cauthor=true&cauthor_uid=33117843
https://www.ncbi.nlm.nih.gov/pubmed/?term=Verhasselt%20V%5BAuthor%5D&cauthor=true&cauthor_uid=33117843
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7557356/
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7557356/
https://www.ncbi.nlm.nih.gov/pubmed/33117843
https://dx.doi.org/10.3389%2Ffnut.2020.576133
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133 

 

plant species in southern Ecuador (Thesis) 
Loja, Ecuador: Universidad Técnica 
Particular de Loja; 2009.   

20. Felipe PT, Juleana VBJ, Zulma TR. 
Pharmacological overview of 
galactogogues. Veterinary Medicine 
International. 2014;66:505-511. 

21. Gire JT, Dimah A. Cultural transformation 
and the changing pattern of alcohol 
consumption among the Tiv of Central 
Nigeria. African Journal of Drug and 
Alcohol Studies. 2001;1(2):125–137. 

22. Bennett LA et al. Alcoholic beverage 
consumption in India, Mexico, and Nigeria. 
Alcohol Health and Research World. 
1998;22(4):243–252. 

23. Stanley PC, Odejide AO. Socio-
demographic and forensic characteristics 
of alcohol abusers in Jos,Nigeria. Nigerian 
Journal of Medicine. 2002;11(3):113–117. 

24. Obot IS et al. Drug and alcohol 
consumption by out-of-school Nigerian 
adolescents. African Journal of Drug and 
Alcohol Studies, 2001;1(2):98–109  

25. Haard NF, et al. Fermented cereals: a 
global perspective. Rome, Food and 
Agriculture Organization of the United 
Nations; 1999.  
Available:http://www.fao.org/docrep/x2184
e/x2184e07.htm, accessed 27 March 
2004). 

26. Stringini M, et al. Yeast diversity during 
tapping and fermentation of palm wine 
from Cameroon. Food Microbiol. 
2009;26(4):415-20. 

27. Isaac UE, Akpuaka FC, Ndukwe GU. The 
effect of intake of palm wine on body 
weight and testicular microarchitecture in 
adult wistar rats. J Exp Clin Anat [serial 
online]; 2017 [cited 2020 Nov 3];16:12-7.  

28. Janero DR. Malondialdehyde and 
thiobarbituric acid-reactivity as diagnostic 
indices of lipid peroxidation and 
peroxidative tissue injury. Free Radical 
Biology and Medicine. 1990;9:515-540.   

29. Martin JP, Daily M, Sugarrnan E. Negetive 
and positive assay of superoxide 
dismutase based on hematoxylin 
autoxidation. Archives of Biochemistry and 
Biophysics. 1987;255:329-336.  

30. Beers RF, Sizer IW. A Spectrophotometric 
method for measuring the breakdown of 
hydrogen peroxide by catalase. Journal of 
Biological Chemistry. 1952;195:133-140. 

31. Avissar N, Eisenmann C, Breen JG. 
Human placenta makes extracellular 
glutathione peroxidase secretes it into 

maternal circulation. American Journal of 
Physiology. 1994;267:68-76. 

32. Cynthia Mannion, Deborah Mansell, 
Gideon Koren. Drugs during pregnancy 
and lactation; new solutions to serious 
challenges. Obstetrics and Gynecology 
International. 2012;2012:Article ID 56270.  

33. Mannion C, Mansell D. Breastfeeding self-
efficacy and the use of prescription 
medication: a pilot study. Obstet Gynecol 
Int. 2012;2012:562704. 

34. Kuan-I Chen, Mei-Hui Erh, Nan-Wei Su, 
Wen-Hsiung Liu, Cheng-Chun Chou, 
Kuan-Chen Cheng. Soyfoods and soybean 
products: from traditional use to modern 
applications, Appl Microbiol Biotechnol. 
2012;96:9–22. 

35. Van Dyck S., in Oxidation in foods and 
beverages and antioxidant applications: 
Understanding Mechanisms of Oxidation 
and Antioxidant Activity. 2010;57-75. 
DOI:10.1533/9780857090447.1.57, 
PUBLISHER(S), Elsevier BV 

36. Ogunro PS,  Ologunagba PO. The effect of 
palm wine on lipid peroxidation and 
antioxidant status of rural dwellers in South 
West Nigeria. Niger Postgrad Med J. 2011 
Sep;18(3):186-90.PMID: 21909148 Actions 

37. Shuhong Li, Dan Zhu, Kejuan Li, Yingnan 
Yang, Zhongfang Lei, Zhenya Zhang, 
"Soybean Curd Residue: Composition, 
Utilization, and Related Limiting Factors", 
International Scholarly Research Notices, 
2013;2013:8:Article ID 423590 

38. Yuksel S, Yigit AA, Cinar M, Atmaca N, 
Onaran Y. Oxidant and antioxidant status 
of human breast milk during lactation 
period, Dairy Sci. & Technol. 2015;95:295–
302. 
DOI 10.1007/s13594-015-0211-z 

39. Kuan-Chen Cheng, Jiun-Yi Wu, Jiun-Tsai 
Lin, Wen-Hsiung Liu. Enhancements of 
isoflavone aglycones, total phenolic 
content, and antioxidant activity of black 
soybean by solid-state fermentation with 
Rhizopus spp. Eur Food Res Technol. 
2013;236:1107–1113.  

40. Eze Christopher Osita, Berebon Dinebari 
Philip, Gugu Thadeus Harrison, Nworu 
Chukwuemeka Sylvester, Esimone 
Charles Okechukwu. Effects of 
Lactobacillus spp. isolated from the sap of 
palm tree Elaeis guineensis (palm wine) on 
cellular and innate immunity, African 
Journal of Microbiology Research, 14 
January. 2019;13(2):33-39, Article 
Number: 401717059818,  

https://www.hindawi.com/journals/ogi/
https://www.hindawi.com/journals/ogi/
https://www.hindawi.com/journals/ogi/contents/year/2012/
https://doi.org/10.1155/2012/562704
https://doi.org/10.1155/2012/562704
https://doi.org/10.1155/2012/562704
https://www.sciencedirect.com/book/9781845696481
https://www.sciencedirect.com/book/9781845696481
https://www.sciencedirect.com/book/9781845696481
https://www.sciencedirect.com/book/9781845696481
http://doi.org/10.1533/9780857090447.1.57
https://pubmed.ncbi.nlm.nih.gov/?term=Ogunro+PS&cauthor_id=21909148
https://pubmed.ncbi.nlm.nih.gov/?term=Ologunagba+PO&cauthor_id=21909148


 
 
 
 

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41. Kazuhiro Suzuki, Hidekazu Koike, Hiroshi 
Matsui, Yoshihiro Ono, Masaru Hasumi, 
Haruki Nakazato Hironobu Okugi, 
Yoshitaka Sekine, Kazuya Oki, Kazuto Ito 
Takumi Yamamot ,Genistein, a soy 

isoflavone, induces glutathione peroxidase 
in the human prostate cancer cell lines 
LNCaP and PC‐ 3. International Journal of 
Cancers. 20 June 2002;99(6):846-852.  

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