COMBINED EFFECT OF ETHANOL EXTRACT OF Mucuna urens (HORSE-EYE BEANS SEED) AND ETHANOL EXTRACT OF Zingiber officinale (GINGER RHIZOME) ON SPERM MOTILITY IN MALE WISTAR ALBINO RATS

COMBINED EFFECT OF ETHANOL EXTRACT OF Mucuna urens (HORSE-EYE BEANS SEED) AND ETHANOL EXTRACT OF Zingiber officinale (GINGER RHIZOME) ON SPERM MOTILITY IN MALE WISTAR ALBINO RATS

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LIST OF TABLES

Tables                                                     Title                                                           Pages

Table 2.1: Taxonomical Classification of Mucuna urens        -           -           -           7

Table 2.2: Mineral Composition of Mucuna urens     -           -           -           -           9

Table 2.3: Taxonomical Classification of Zingiber officinale  -           -           11

Table 2.4: Chemical Constituent of Zingiber officinale          -           -           -           13

Table2.5: Normal values of Semen Parameters           -           -           -           -           19


LIST OF FIGURES

Figure                                          Title                                                                       Pages  

Figure 1:          Mucuna urens seed     -           -           -           -           -           -           8

Figure 2:          Ginger Rhizome (Zingiber officinale) -           -           -           12

Figure 3:          Chemical Structure of Gingerol and Shogaol             -           -           14

Figure 4:          Comparative Effect of MU and ZO on Total Cell Concentration     30       

Figure 5:          Comparative Effect of MU and ZO on Total Motile Sperm  -           32

Figure 6           Comparative Effect of MU and ZO on Velocity of Active Path       34

Figure 7           Comparative Effect of MU and ZO on Percent Motile Sperm -        36

Figure 8           Comparative Effect of MU and ZO on Sperm Progressivity   -         38

Figure 9           Comparative Effect of MU and ZO on Total Cell Detected   -         40       


ABSTRACT

Semen analysis of albino rats exposed to ethanol extract of Mucuna urens seed and ethanol extract of Zingiber officinale  were investigated in fifty male adult rats weighing 120g-200g.the rats were divided into ten groups (n=5). Group I was control, Group II, Group III, Group IV, Group V, Group VI, Group VII, Group VIII, Group IX and Group X were the experimental groups. 2000mls and 1000mls of ethanol were for the extraction of Mucuna urens and Zingiber officinalerespectively. Filtered and concentrated in water bath at 450 C. 1000mg of the extracts were dissolved in 10mls distil water. The extracts were administered orally for twenty-one days. Group I was administered 5mls water, group II was administered 500mg/kg Mucuna urens, group III was administered 1000mg/kg Mucuna urens, group IV was administered 1500mg/kg Mucuna urens, group V was administered 86.6mg/kg Zingiber officinale, group VI was administered 173.21mg/kg Zingiber officinale, group VII was administered 259.81mg/kg Zingiber officinale, group VIII was administered 500mg/kg Mucuna urens+86.6mg/kg Zingiber officinale, group IX was administered 1000mg/kg Mucuna urens +173.21mg/kg Zingiber officinale and group X was administered 1500mg/kg+259.81mg/kg Zingiber officinale. After sacrifice freshly collected semen samples was analyzed using computer assisted sperm analyzer. Total concentrated cell and percent motile sperm were increased in all experimental groups, progressivity decreased in Mucuna urens only treated groups and increased in other experimental groups. Severity was dose dependent. Data were analyzed using one-way ANOVA (p<0.05). Ethanol seed extract of Mucuna urens impaired sperm motility and Zingiber officinale extract improves sperm motility.


CHAPTER ONE

INTRODUCTION

1.1       Background of the Study

The understanding of male reproductive function and the importance of male factor in infertility has advanced significantly in the recent time. The reproductive function of male is divided into three major subdivisions: spermatogenesis, performance of the male sexual act and regulation of male reproductive functions by the various hormones. Association with these function are the effects of male sex hormones on the accessory sexual organs, cellular metabolism, growth and other body functions (Guyton and Hall, 2011). Infertility is a disease of reproductive system defined by failure to achieve the clinical pregnancy after 12 months or more of regular unprotected sexual intercourse (Zegers-Hochschild et al., 2009). Infertility is one of the major health challenges in life approximately 30% of infertility is due to male factor (Isidori et al., 2006).

Factors affecting male infertility are classified into pre-testicular, testicular and post-testicular (Balen,2008). This may cause the testicles producing a decreased number of sperm or disturbance along the path preventing cells from maturing into sperm production or reaching the woman fallopian tube where fertilization occur. Germ cells are gradually transformed into spermatozoa. This occurs in the seminferous tubules of the testes. It is divided into three stages (Cuninghan and Kelvin, 2007). Proliferative phase: Spermatogonia-Spermatocytes, Meiotic phase: spermatocytes - spermatids, Differentiation phase: (Spermiogenesis) spermatids-spermatozoa (Hess, 1999).Several conditions interfere with spermatogenesis and reduce sperm quality and production,

Semen is an organic fluid that may contains spermatozoa. It is secreted by the gonads (sexual glands). Seminal fluid contains spermatozoa, proteolytic enzymes and fructose which promote the survival of spermatatozoa and provide a medium through which they move or swim (Ali et al., 2015). Male infertility can be assess through semen analysis and reproductive hormonal profile (Jungwirth, et al., 2012). Male infertility is commonly due to deficiencies in the semen and semen quality (Cooper et al.,2010).

Males with sperm parameters below the WHO normal values are considered to have male factor infertility. Most significant of these are low sperm concentration (oligospermia), poor sperm motility (asthenospermia), and abnormal sperm morphology (tetratospermia). Factors less associated with infertility include semen volume and other serminal markers of epididymal, prostatic and serminal vesicle function (Harris et al.,2011). Positive association exit between abnormal semen parameters and sperm count (Sebra et al., 2014).

Abnormality in sperm count, motility and morphology arises from disarray in control mechanism, including pre-testicular, testicular and post testicular factors (Wamoto et al.,2007). Semen analysis remains the single most useful and fundamental investigation to detect men with a genuine problem of male infertility (Butt et al.,2013). It is a simple test that assesses the formation and maturity of sperm as well as how the sperm interacts in the serminal fluid. It also provides insight not only on sperm production (count), but the sperm quality (motility and morphology) as well (Fisch,2008). As many as 2% men exhibits suboptimal sperm parameters, which may be one or a combination of low sperm concentration, poor sperm motility, or abnormal morphology (Kumar et al.,2015).

Mucuna urens also known as Ox-eye beans is a common soup thickener consumed mostly in South Eastern states in Nigeria, it is a plant belonging to the family fabaceae, commonly found in home and gardens in the south eastern part of Nigeria, West Africa, where the Efiks, Ibibio and Igbos uses the seed as a major soup condiment for thickening (Achivewhu, 1984). It is incorporated into the normal feed for farm animals in the North due to its protein content (Umoren et al., 2007). Mucuna urens is called “Ibaba” by the Efik/Ibibios and “Ukpor” by the Igbos and usually sold in the local markets during the harvest season ie January (Eilitta et al., 2003). It may be found throughout the year. Mucuna urens requires support for growth hence cultivated near trees. This enables production of many seeds per plant (Sridhar et al.,2007). Other names for Mucuna urensinclude Horse eye bean, Ox-eye bean and devil bean. Mucuna urens had been found to contain some endogenous toxic factors. High concentration of tannis, phytic acid, cynogenic glucoside, oxalate and gossypol has been reportedin Mucuna urens (Laurena et al., 1994).

Toxic compounds including L-DOPA (3,4-dihydroxy-L-phenylalanine), nicotine, physostigmine and serotonine are found in Mucuna urens, which also act as anti-nutritive value of Mucuna urens (Umoren et al.,2007). Mucuna urens seed extract causes sperm degeneration in testicular tubules,collapse of villi in the prostate gland and seminal vesicles of male guinea-pigs (Udoh and Ekpeyong, 2001).

Ginger rhizome (Zingiber officinale Roscoe; Family: Zingibercease) is used worldwide as a spice. It has both anti-oxidative and androgenic activities in animal models (Sekiwa et al.,2000, Kantch Oung et al., 2002). The local names of ginger includes; ‘Afa-ije’ in Yoruba and ‘Jinja’ in Igbo/Efik/Ibibios. Ginger rhizome contains active ingredients such as Zingerone, Gingerdiol, Zingibren,Gingerol and shogaols which have antioxidants activity (Zulican et al.,2002). Ginger is a great source of manganese, essential to man’s sex drive and sperm count by helping the body produce testosterone. Ginger oil has protective effect on DNA damage induced by H2O2 and might be used as antioxidant (Grzarana et al., 2005). Antioxidant protects DNA and other important molecules from oxidation and damage which can improve sperm quality and consequently increase fertility rate in men (Rajeer et al., 2006).

Ginger is a native of South Eastern Asia but over the centuries has been spread to various part of the world including Africa. In Nigeria, the highest producer of ginger is Kaduna state, others includes Gombe, Bauchi, Benue, Nassarawa among others. Ginger is available in various forms; Fresh ginger rhizome, powder ginger and dry ginger rhizome. A mature ginger rhizome is fibrous and has a striated texture. The outer skin of the rhizome is brownish in colour while the inner flesh depending on the variety may be red, yellow or white. For effective cultivation of ginger the following conditions are required; mulched fertile soil, loam is the most preferred soil type, ridges and a minimum annual rainfall of about 1500mm. An average temperature of about 300C, viable ginger rhizomes with buds and good drainage prevents water logging/flooding.

1.2       Statement of the Problem

Despite the degenerative effect of Mucuna urens seed on spermatogenesis and seminal parameters, it is of great benefit to humanity, agriculturally it serves as a cover crops, to food technologist it is used as a thickening agent in soup; medically sap from the cut liana is rubbed on sprains, rheumatic areas, contusions, sore muscles and used for children’s fever. It also has anti-anaemic effect, antibacterial effect and inhibitory activity on some enzyme. Ginger is been used daily as food spices and in herbal medicine which increases semen volume, sperm count and testosterone level. This research project shall provide further insights into the causes and treatment of male infertility through semen analysis caused by the effect of Mucuna urens and Zingiber officinale.

1.3       Significance of the Study

The result of this study will increase knowledge to the existing literature about the effect of ethanol seed extract of Mucuna urens and ginger on semen analysis.

1.4       Aim and Objective of the Study

This research was aimed at investigating the serious danger of consumption of Mucuna urens seed and the counter effect of ginger rhizome on semen analysis.   Its objectives include;

i.        To investigate changes in semen analysis in male albino rats administered with ethanol seed extract of Mucuna urens.

ii.      To investigate changes in semen analysis in male albino rats administered with ethanol extract of Ginger rhizome.

iii.    To investigate the counter effect of ethanol extract of ginger rhizome on semen abnormalities induced by the administration of ethanol seed extract of Mucuna urens in male albino rats.

1.5       Scope of the Study

The scope of this study includes;

i.        Ethanol extraction of Mucuna urens seed and Zingiber officinale Roscoe.

ii.      Concentration of ethanol extract of Mucuna urens seed and ethanol extract of Zingiber officinale in a water bath.

iii.    Determination of Lethal Dose (LD50) for Mucuna urens seed extract and Zingiber officinale using mice.

iv.    Determination of body weight of experimental animals.

v.      Administration of ethanol seed extract of Mucuna urens and ethanol extract Zingiber officinale to experimental animals.

vi.    Monitoring of body weight and size of experimental animals with respect to dosage of the extracts.

vii.  Collection of semen and determination of semen analysis.



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