The effect of
oral vitamin E supplementation on the health status of liver and kidney of
growing pigs during oxidative stress induced by hypothermia was studied. 27
Landrace pigs, with an average weight of 14 ±1.02 were used. Pigs were
randomly assigned to three treatments (T1, T2, and T3) with 9 pigs per treatment.
Each treatment had 3 replicates with 3 pigs per replicate and the study
lasted for six (6) weeks. All treatments received similar diets (Pfizer
Growers Marsh TM) except that T3 was administered vitamin E orally from the
4th week at the rate of 470mg/kg body weight daily. 24 hours after the last
administration, T3 and T2 were immersed into a water body temperature of
120C up to their neck while same was done for T1 in temperature of 360C
water. Immediately after immersion, body temperatures were taken and blood
samples were collected and preserved. Results showed that Alanine Amino Transferase (ALT) had higher significant differences
(P< 0.05) between T3 and T2. The T3 and T1 groups had similarities (43.66
and 44.32) in Aspartic Amino Transferase (AST)
while T2 had significant differences (P<0.05) in figures. Alkaline
phosphatase had significant differences (P < 0.05) in all treatments.
Blood urea nitrogen (BUN), T2 had significant differences (p< 0.05) and
T3 and T1 had similar values (p < 0.05).Creatinine had higher value in T2 (P <
0.01) with T1 and T3 with similar values (p < 0.01).
Accepted:29/09/2023
Published: 02/10/2023
*Corresponding
Author
Diri Moses
E-mail: dirimoses@ yahoo.com
Keywords: Hypothermia,
Kidney; Liver; Oxidative Stress; and Vitamin E
INTRODUCTION
It is an established
fact that oxidative stress could lead to death of cells in liver, kidney and
other visceral organs in animals if intensive measure is not taken. To this
point therefore, it is pertinent to work out modalities on how to effectively
care for pigs during a case of oxidative stress that may be induced by
hypothermia. Johnson et al., (2020) demonstrated
that presence of hypothermia induced oxidative stress to weaner
rabbits. Also, the media is inundated with literatures confirming that vitamin
E (dl- Alpha tocopheryl Acetate) has antioxidant
properties and as such can be used to combat the effect of lipid peroxidation
in animals when supplemented in diets (Fulyaet al., 2012). This is achieved by its
donation of hydrogen atom from the hydroxyl group and the hydrogen atom reacts
with hydroxyl radicals thereby detoxifying them to water and rendering reactive
oxygen species inactive to cells and tissues of animals ( Aslan
and Meral, 2007; Johnson et al. 2023).
During cold weather,
like the hammatan and rainy seasons which mimics
winter, the tissues and visceral organs of pigs may be compromised as a result
of the production of reactive oxygen species in the cells caused by cold
conditions and this may lead to apoptosis (programmed cell death) or even death
of pigs if proper measure is not taken to cushion the effect of cold which may
be achieved by detoxification of free radicals produced by cold conditions. Aslan and Marel, (2007) showed
that vitamin E was able to up-regulate the activities of liver enzymes in pigs.
High ALT and AST
level in blood serum of animals is an indication of abnormality in liver functionality.
Thus, this has proved that ALT and AST are the major liver enzymes, confirmed
to be a good biomarker of oxidative stress induced by extremely cold season.
Also, blood urea nitrogen and creatinine
concentrations are higher in the serum when the kidney has compromised its
functions (Fulya et al., 2012; Lalita
et al.,2016 ), thus, making them good kidney
biomarkers of oxidative stress.
The objective of this
study is to determine the effect of oral vitamin E supplementation on the
health statuses of liver and kidney of growing pigs during oxidative stress induced
by hypothermia.
MATERIAL
AND METHOD
The study was
conducted at the piggery section of The Rivers State University, Teaching and
Research Farm, Port Harcourt. Pigs were purchased in a local market within Port
Harcourt City.Twenty seven (27) landrace
pigs with an average weight of 14 ± 1.02 kg were purchased and used for this
study. Prior to the arrival of the animals, the floor of the pens were
thoroughly washed with water and hypochlorite and allowed to dry for one week
before introducing the animals into their pens. They were randomly placed to
their individual pens and were given a week of adjustment to enable them acclimatizing
with the new environment. All animals were similarly managed during the
adjustment period. Thus, during the period the animals were similarly fed ad-libitum
prior to the commencement of the experiment.Proper hygiene was observed throughout the study period. The Pigs were
placed on three dietary treatments having nine (9) pigs per treatment. Each
treatment had 3 replicate, with 3 pigs in each of the replicate and the
experiment lasted for 6 weeks.All
pigs were fed Pfizer Grower MarshTM and
vitamin E (dl- Alpha tocopheryl acetate) was
administered at the rate of 470mg/kg body weight in diets of T2 and
T3 from the last two (2) weeks of the study whereas, T1 (
control diet) was not supplemented with Vitamin E. Twenty-four (24) hours after
the last administration of Vitamin E, pigs in the control (T1) group
were dipped into a water of body temperature of 360C with their head
up for 5 minutes while hypothermia was introduced to T2 and T3by
dipping them into a water body of 120C up to their neck for 5
minutes without any form of tranquilizer or sedative, after which their body
temperatures were taken rectally and blood samples were collected.At the last day of the sixth week of
the study, blood samples were taken from all treatment groups using 2ml syringe
via their external jugular vein and placed in a well labeled non-ethylene diaminetetracetic acid (non-EDTA)
treated vial tubes between 7 and 8 am and immediately placed inside a deep
freezer for later analysis.
Liver Enzymes (ALT, AST
and ALP) were analyzed. ALT and AST were analyzed by the method of Reitman and
Frankel (1957) while ALP was determined by the method of Aaron (1930). Kidney Biomarkers: Test for blood
urea nitrogen and creatinine was tested for; to
determine the functionality of the kidney. Urea concentration was determined by
the method of Machado and Horizonte (1958) while creatinine
was determined by the method of Max (1886)
Statistical
Analysis
Data obtained from
the study was subjected to analysis of variance (ANOVA) using the general
linear model of SAS (SAS Inst., Cary, NC, 1988). Treatment means were compared
using turkey test. Completely Randomized Design was used as the experimental
design with linear model: Yij = u + Xi + Eij
Where Eij is error involved in monitoring animal
receiving ith treatment in the study.
Xi is effect of ith level of treatment
u is the population
mean
Yij is the individual observation
of any animal receiving treatment.
RESULTS
and DISCUSSION
Pigs in all the
treatment groups were fed normal and seen to be in good condition all through
the study period as they were monitored regularly. After introduction of cold,
mean body temperature for T1 group was 36.8 ± 0.5, T2 group
had 33 ± 1.2 and T3 group was 33.2 ± 0.8 respectively. This showed
that slight cold was experienced for T2 and T3treatment
groups respectively. Data in Table 1 show that hypothermia and vitamin E
actually had effect on the liver biomarkers. For ALT and AST, there were
significant differences (P < 0.05)
in T3 compared to T2. However, T3 and T1
have similarities. For ALP, there were significant differences (P < 0.05) across board in all
treatment groups.
ALT is one of the
chief enzymes found in the liver of animals. ALT aids in breaking down food
into energy. A damaged liver will release more ALT into the blood stream and
this is why its test is conducted in most cases to determine the functionality
of the liver, thus making it a biomarker of oxidative stress. T2
group had a higher significant difference (p
< 0.05) compared to other treatment groups. This is a clear indication
that cold affected ALT and oxidative stress was induced by the presence of
cold. This is in conformity with the findings of Johnson, et al (2019). Also there were similarities between T1
and T3 groups, this could have been that vitamin E supplemented to T3
group was able to reverse the effect of oxidative stress caused by cold.
This agrees with findings of some researchers (Aslan
and Meral, 2007; Johnson et al., 2019; Johnson et al.,
2023) which posited that oral supplementation of vitamin E reversed the effect
of oxidative stress induced by cold and crude oil respectively by reducing ALT
levels in blood serum.
AST is one of the
major enzymes domiciled in the tissues of liver organ of animals. Reactive
oxygen species produced by some toxic factors caused damage to the liver
thereby releasing large volume of AST in the blood stream (Ekhato,
et al., 2014). This is an indication
that AST just like ALT is a good biomarker of oxidative stress as such induced
by hypothermia. From the result of this study, oxidative stress was induced in
T2 group and oral vitamin E supplementation was able to normalize
the effect of hypothermia as evidenced in the T3 group and T1 group
having similarities in value. This consolidates findings that confirms that
vitamin E up-regulated the activities of antioxidants in blood serum of animals
subjected to oxidative stress (Mahaswari, et al., 2015; Johnson, et al., 2023)
ALP is an enzyme
found in almost all part of the body, especially in the liver. ALP test is
usually done when there is a suspected case of liver, bone or kidney
abnormality. The higher the volume in the blood serum, the
tendency of a major damage in the liver. Judging the result of this
study, T2 group had a higher value of ALP which is an indication
that presence of hypothermia actually had a negative effect on the liver. Also
in T3 group, vitamin E was able to reduce the effect of hypothermia
as evidenced in the lower value when compared to T2 group. However,
vitamin E didn’t actually bring down the value to normalcy as evidenced in the
result when T3 group was compared to T1 group. This may be that the introduction of
hypothermia may have not only affected the liver but have had little or no
effect on the bones of the pigs, since hypothermia was introduced by immersion.
Table1.Serum
Levels of Liver enzymes with or without hypothermia.
Item
T1(control)
T2(Hypo)
T3(vit E + Hypo)
SEM
P-Value
ALT [iu/l]
102.55a
112.20b
104.12a
0.8
0.026*
AST [iu/l]
44.32b
51.20a
43.66b
0.6
0.001**
ALP [iu/0
43.68c
47.86a
45.50b
0.5
0.012**
a,b,c, means within the
same row with different superscripts are significantly (P<0.05)different; SEM = standard error of mean. Iu/l means international units per liter, and Hypo means
hypothermia
In Table 2 BUN had a
significant increase (p < 0.05)in T2 pigs compared to T1
and T3 that had similarities as BUN was significantly reduced (p< 0.05). Creatinine
was significantly higher (p < 0.01)
in T2 pigs compared with T1 and T3 which had
similarities in their concentrations (p
< 0.01).
. BUN occurs in
larger volume in the serum as evidenced in T2 group compared to
other treatment groups. This may be that the kidney failed to eliminate some
nitrogenous wastes via urine. In other words, the kidney compromised its
function and this is a clear indication that cold induced oxidative stress
which led to the kidney compromising. This agrees with the research work which
shows that rabbits exposed to hypothermic conditions actually had a higher
volume of BUN in their serum which kept the rabbits in an oxidative stress
conditions (Johnson and Diri, 2020). Also, the
similar values obtain in T3 and T1 groups showed that the
presence of vitamin E was able to reverse the effect of oxidative stress
induced by cold. This corresponded with the research which showed that vitamin
E reduced the level of creatinine in the serum of
rabbits on oxidative stress induced by hypothermia (Johnson and Diri, 2020). However, this did not agree with the study which
showed that there were no significant differences on rabbits administered
vitamin E during oxidative stress induced by partial bladder outlet obstruction
when compared to their control group (Lin, et
al., 2011)
Creatinine compound are by
products of creatinephosphatate. It is produced in
the body during metabolism of creatine and usually
excreted by the urine after it has been filtered by the kidney. Increased level
of creatinine in the serum is an indication that the
kidney malfunctioned as seen in T2 group compared to other treatment
groups. In other words, T2 had a higher significant differences (p< 0.01) compared to T3
and T1 groups. Vitamin E had effect on creatinine
volume as it was able to reduce creatinine level as T3
group had similarities (p < 0.01)
with T1 group. This result disagreed with this study of Lin, et al (2011) but agrees with the studies
of Mahaswari, et
al., 2015; Johnnson and Diri
2020; and Johnson, et al., 2023.
Table 2.Serum
levels of kidney biomarkers with or without hypothermia
Items
T1(control)
T2(Hypo)
T3(vit E + Hypo)
SEM
P-Value
BUN (mmol/l)
4.42b
6.01a
4.45b
0.1
0.046*
Creatinine (mmol/l)
78.42b
89.22a
76.25b
1.2
0.001**
Means with different
superscripts within the same row are significantly different (P < 0.05); (P < 0.01) SEM = error of mean, and mmol/l
means milimole/liter
CONCLUSION
Oral vitamin E supplementation at 470mg/kg
body weight can help reduce the effect of oxidative stress in pigs during
hypothermic conditions such as the raining, winter and hammatan
seasons.
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Cite this Article:
Diri M; Fakae, LB; Ogbamgba, VM; Johnson, NC (2023). Vitamin E, Liver and
Kidney Functions of Growing Pig During Hypothermia. Greener Journal of Agricultural Sciences, 13(3): 213-216.
