Potential effects of different levels of egg derived choline

on reproductive health of male albino rats

Presented By : Ruhma Fazal

Roll No: 2221014

Department Of Nutritional Sciences,

GCUF
SUPERVISORY
COMMITTEE

SUPERVISOR 1
PROF. DR. MAHR-UN-NISA
SUPERVISOR 2
Dr. Huma Ambreen
SUPERVISOR 3
Dr. Nazir Ahmed
 HEADER
REWIEW OF
INTRODUCTION
LITERATURE

NEED OF PROJECT METHODOLOGY

HYPOTHESIS OBJECTIVES

REFERENCE
 INTRODUCTION

 Infertility are often complicated and difficult to identify, health and

lifestyle factors affect the ability of both men and women to reproduce.
 In men, abnormalities infertility is referred as the abnormality in sperm
count, morphology and motility all have an impact on successful
fertilization. (Alejandra M. Weidman et al., 2018)
 Choline is one of the major nutrient that contribute in health of male
healthy sperm by donating the methyl group. (Steven H. Zeisel MD,
2015)
 NEED OF PROJECT

• Nearly 50 million couples worldwide experience infertility. According to

the 2009–2010 Canadian Community Health Survey, estimates of the
prevalence of infertility range from 10% to 15.5% of couple.
• Male factor infertility accounts for 20–30% of total cases, with an
additional 20–30% being due to a combination of male and female factor
infertility.
 OBJECTIVE

• The main objective of study is to investigate the effect of egg

extracted choline on male reproductive hormones and sperm health
such as sperm morphology, motility and sperm count
 HYPOTHESIS

Choline rich phospholipid may improve the male

reproductive health by improving their motility,
sperm count and morphology
 REVIEW OF LITERATURE

• Choline deficiency in male reduce the sperm motility due to changes the
phospholipid co centration and the level of male hormones.
• Genetic variations, particularly in the choline metabolizing gene CHDH,
such as the rs12676 variant associated with reduced sperm motility,
contribute to infertility risk.
• Single nucleotide polymorphisms in folate and choline metabolizing
genes increase the demand for dietary choline, impacting
phosphatidylcholine biosynthesis. (Weidman et al ., 2018)
 METHODOLOGY

Ethical approval:
 Ethical approval will be taken from ethical committee of the university.
Research area:
 The research will be conducted in the department of nutritional science, government
college university in Faisalabad.
 Animal will be kept and treaded according to the principles of laboratory animal care.
 They will be under relative humid condition temperature 25˚C±5 degree under 12
hours of light and 12 hours of dark cycle. (NRC,2010)
 STUDY DESIGN

• Total duration of the study will be 33 days in which 5 days will be

adjustment period.
• Thirty adult albino rats will be divided into 5 groups (n=6)by completely
randomized design(CRD).
• The name of different levels will be given as C1, C2, C3 ,C4 and C5.
• Basal diet will be given to group C1 without egg choline.
Extraction Of Choline From Egg Yolk
 Remaini
20g The
ng
fresh Supern methanol
material Place Pl will
Add 30 extract
egg atant will be Transfe centrifu be
ml of will be Centrif
yolk will be dissolve r to ge obtaine
methan place in uge it
will be filtered in 35ml centrifu bottle d in the
ol and vacuum
of for 25
put in or hot air ge in cold form of
centrifu Repeat Hexane min
centrifu dryer bottle bath for precipit
ge it 3 times and 80ml
ge over
of
15 min ation
bottle night
Acetone
 Study protocol

Group Condition Treatment

C1 Healthy rats Control diet

C2 Healthy rats Choline(egg) as per RDA

from synthetic source
C3 Healthy rats Choline(egg) as per RDA
(1.1g/kg)
C4 Healthy rats Choline(egg) 5%↑then
RDA
C5 Health rats Choline(egg) 10%↑then
RDA
 Feed and Nutrients Digestibility Analysis
Feed and Nutrients Digestibility Analysis

 Feed and water intake will be measured daily, weight changes will be recorded weekly
while digestibility will be performed at the end of the trial by proximate analysis
 Feed and Nutrient Digestibility analysis
 Nutrient digestibility(%)=Nutrient intake- Nutrient in feces x 100 / Nutrient intake
(Chen et al., 2016)
 Feces samples will be collected in such a way that collect samples accurately and feces
will be stored as -20 (Manzar et al., 2021)
 Physical Parameters

Feed and water intake

• Feed and water intake will be accessed on daily basis
Weight gain
• Weight of rats will be weighed by using precious balance

(Pieczynska et al., 2022)

The different assays will be performed to
evaluate the effect of treatment like
 Statistical analysis
The data from all the experimental
sources will be subjected to statistical
analysis using Statistics 8.1
(sterling et al., 2006)
 References

 Steven H. Zeisel MD, P. (2015). The fetal origins of memory: The role of dietary choline in optim
 Wiedeman AM, Barr SI, Green TJ, et al. Dietary Choline Intake: Current State of Knowledge
Across the Life Cycle.
 Dietary Reference Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, Folate, Vitamin B12,
Pantothenic Acid, Biotin, and Choline. The National Academies Collection: Reports funded by
National Institutes of Health. Washington (DC),
 Ducker GS, Rabinowitz JD. One-Carbon Metabolism in Health and 1Disease.
 Friso S, Udali S, De Santis D, et al. One-carbon metabolism and epigenetics. Mol Aspects Med
201727. Johnson AR, Lao S, Wang T, et al. Choline dehydrogenase polymorphism rs12676 is a
functional variation and is associated with changes in human sperm cell function. PLoS One.
2012;7:e36047.
 References
 Canada; Go. Fertility 2013 [updated Feb 4 2013.
 Mascarenhas MN, Flaxman SR, Boerma T, et al. National, regional, and global trends in infertility
prevalence since 1990: a systematic analysis of 277 health surveys.
 Bushnik T, Cook JL, Yuzpe AA, et al. Estimating the prevalence of infertility in Canada. Hum
Reprod
 Agarwal A, Mulgund A, Hamada A, et al. A unique view on male infertility around the globe.
Reprod Biol Endocrinol 2015;13:37. 10.1186/s12958-015-0032-1
 Buhling KJ, Grajecki D. The effect of micronutrient supplements on female fertility. Curr Opin
Obstet Gynecol 10. Gaskins AJ, Chavarro JE. Diet and fertility: a review. Am J Obstet Gynecol
2018;218:379-89.
 References

 Chen, Yu-ming, Yan Liu, Rui-fen Zhou, Xiao-ling Chen, Cheng Wang, Xu-ying Tan, Li-jun Wang,
Rui-dan Zheng, Hong-wei Zhang, and Wen-hua Ling. 2016. “Associations of Gut-Flora-Dependent
Metabolite Trimethylamine-N-Oxide, Betaine and Choline with Non-Alcoholic Fatty Liver Disease
in Adults.” Scientific Reports 6(1):1–9
 National Research Council. (2010). Guide for the care and use of laboratory animals .