ABSTRACT
Extracts of Hibiscus Sabdariffa (HS) and Azadirachta Indica (AI) are widely used for medicinal purposes and have been shown to affect postnatal growth of the offspring through mechanisms not yet defined. Breast milk creamatocrit is the length of the cream column separated from milk by centrifugation and expressed as a percentage of the length of the total milk column. It is a simple accurate inexpensive technique and useful measure of the lipid concentration and the energy content of breast milk. No data has shown the effect of these two extracts on breast milk creamatocrit which is a vital factor for the early postnatal growth of offspring. The study was designed to investigate the effects of the two extract on breast milk creamatocrit and early postnatal growth of offspring. 40 pregnant rats weighing 150-200g were used for this study. They were divided into control, HS and AI groups. HS and AI groups were subdivided into two subgroups of low and high dose. Extract administration commenced on day 1 of pregnancy and ended on postnatal day 21. On the day of delivery litter sizes, length and weights were recorded. Litter weight and length were subsequently recorded at postnatal days 7, 14 and 21. Breast milk was collected from 4 dams in each group into a plain sample container at postnatal day 7, 14 and 21 for determination of breast milk creamatocrit. Milk sample volume collected ranged between 0.4ml-0.5ml per dam. The study revealed that the aqueous extract of Hibiscus Sabdariffa and Azadirachta Indica increased consumption of food and fluid during pregnancy and lactation, increased maternal weight gain during pregnancy and lactation and also caused a progressive increase in breast milk creamatocrit during lactation days 7, 14, 21. The progressive increase in breast milk creamatocrit values peaked greatly in the third week of lactation. The aqueous extract of Hibiscus Sabdariffa and Azadirachta Indica caused a progressive increase in BMI of the extract treated group that became significantly greater than that of the control at weaning. From the results of the present study, it can be concluded that consumption of extracts of HS and AI during pregnancy and lactation increases fluid, food intake and weight gain while accelerating the postnatal growth of the offspring through increased breast milk creamatocrit.
TABLE OF CONTENTS
Title page . . . . . . . . . . i
Attestation . . . . . . . . . . ii
Certification . . . . . . . . . . iii
Dedication . . . . . . . . . . iv
Acknowledgment . . . . . . . . . v
Abstract . . . . . . . . . . viii
Table of contents . . . . . . . . . x
List of Table . . . . . . . . . xviii
List of Figures . . . . . . . . . xx
CHAPTER ONE
1.0 Introduction . . . . . . . . . 1
1.1 Statement of Problem . . . . . . . 4
1.2 Aim and Objectives . . . . . . 5
1.2.1 Aim . . . . . . . . . . 5
1.2.2 Objectives . . . . . . . . . 5
1.3 Operational Definition of Terms . . . . . 6
CHAPTER TWO
2.0 Literature Review . . . . . . . 8
2.1 Hibiscus sabdariffa . . . . . . . 8
2.1.1 Botanical Description . . . . . . . 8
2.1.2 Geographical Distribution and Common Names . . . 9
2.1.3 Composition of Hibiscus sabdariffa . . . . . 10
2.1.3.1 Nutritive Value . . . . . . . . 10
2.1.3.2 Phytochemical Constituents . . . . . . 11
2.1.4 Pharmacological Properties of Hibiscus sabdariffa . . 12
2.1.4.1 Antioxidant Properties . . . . . . . 12
2.1.4.2 Nephroprotective Properties . . . . . . 13
2.1.4.3 Delayed Puberty Activity . . . . . . 14
2.1.5 Uses of Hibiscus sabdariffa . . . . . . 15
2.1.5.1 Medicinal Use . . . . . . . . 15
2.1.5.2 Food Use . . . . . . . . 16
2.1.5.3 Non Medicinal Use . . . . . . . 17
2.2 Azadirachta indica . . . . . . . 17
2.2.1 Botanical Description . . . . . . . 17
2.2.2 Geographical Distribution and Common Names . . . 19
2.2.3 Composition of Azadirachta indica . . . . . 19
2.2.3.1 Phytochemical Constituents . . . . . . 19
2.2.4 Pharmacological Properties of Azadirachta indica . . . 20
2.2.4.1 Anti-Bacterial Properties . . . . . . 20
2.2.4.2 Antioxidant Properties . . . . . . . 21
2.2.5 Uses of Azadirachta indica . . . . . . 21
2.2.5.1 Medicinal Use . . . . . . . . 21
2.2.5.2 Non Medicinal Use . . . . . . . 22
2.3 Breast Milk . . . . . . . . . 22
2.3.1 Development of Breast (Mammary Gland) In Rats . . . 22
2.3.2 Onset of Lactation in Rats . . . . . . 25
2.3.3 Stages of Lactation . . . . . . . 26
2.3.4 Composition of Breast Milk . . . . . . 27
2.3.5 Factors Affecting Breast Milk Composition . . . 29
2.4 Postnatal Development in Rats . . . . . . 30
2.4.1 Factors Affecting Early Postnatal Development . . . 32
CHAPTER THREE
3.0 Material and Methods . . . . . . . 38
3.1 Plant Collection and Identification . . . . . 38
3.2 Preparation of Plant Extract . . . . . . 38
3.3. Phytochemical Analysis . . . . . . . 39
3.4 Experimental Animals . . . . . . . 39
3.5 Induction of Pregnancy in Rats . . . . . . 40
3.6 Experimental Design . . . . . . . 40
3.7 Extract Administration . . . . . . . 41
3.8 Maternal Body Weight Measurement . . . . . 41
3.9 Measurement of Food and Fluid Intake . . . . 42
3.10 Measurement of Litter Size, Length and Weight . . . 42
3.11 Breast Milk Extraction . . . . . . . 42
3.12 Breast Milk Creamatocrit Measurement . . . . 43
3.13 Statistical Analysis . . . . . . . 44
CHAPTER FOUR
4.0 Results . . . . . . . . 45
4.1 Extract Yield . . . . . . . . 45
4.2 Phytochemical Screening . . . . . . 46
4.3.1 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa on Food Intake During Pregnancy . . . . . 49
4.3.2 Effect of Consumption of Aqueous Extract of Azadirachta indica on Food Intake During Pregnancy . . . . . 51
4.3.3 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa
on Fluid Intake During Pregnancy . . . . . 53
4.3.4 Effect of Consumption of Aqueous Extract of Azadirachta indica on Fluid Intake During Pregnancy . . . . . . . 55
4.3.5 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa on Maternal Weight During Pregnancy . . . . . 57
4.3.6 Effect of Consumption of Aqueous Extract of Azadirachta indica on Maternal Weight During Pregnancy . . . . . 59
4.3.7 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa
on Food Intake During Lactation . . . . . 61
4.3.8 Effect of Consumption of Aqueous Extract of Azadirachta indica
on Food Intake During Lactation . . . . . 63
4.3.9 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa
on Fluid Intake During Lactation . . . . . 65
4.3.10 Effect of Consumption of Aqueous Extract of Azadirachta indica
on Fluid Intake During Lactation . . . . . 67
4.3.11 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa on Maternal Weight During Lactation . . . . . 69
4.3.12 Effect of Consumption of Aqueous Extract of Azadirachta indica
on Maternal Weight During Lactation . . . . 71
4.3.13 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa on Creamatocrit Value of the Breast Milk (Cream/Lipid)
In Percentage(%) . . . . . . . . 73
4.3.14 Effect of consumption of aqueous extract of Azadirachta indica
on creamatocrit value of the breast milk (cream/lipid)
in percentage (%) . . . . . . . . 75
4.3.15 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa During Lactation on Body Weight of Offspring . . . . 77
4.3.16 Effect of Consumption of Aqueous Extract of Azadirachta indica During Lactation on Body Weight of Offspring . . 79
4.3.17 Effect of Consumption of Aqueous Extract of Hibiscus sabdariffa
During Lactation on BMI of Offspring . . . . 81
4.3.18 Effect of Consumption of Aqueous Extract of Azadirachta indica During Lactation on BMI of Offspring . . . . 83
CHAPTER FIVE
5.0 Discussion and Conclusion . . . . . . 85
5.1 Maternal Study . . . . . . . . 85
5.1.1 Food and Fluid Intake in Dams That Consumed Hibiscus
sabdariffa During Pregnancy and Lactation . . . . 85
5.1.2 Food and Fluid Intake in Dams That Consumed Azadirachta
indica During Pregnancy and Lactation . . . . 87
5.1.3 Weight of Dams That Consumed Hibiscus sabdariffa During
Pregnancy and Lactation . . . . . . 89
5.1.4 Weight of Dams That Consumed Azadirachta indica During
Pregnancy and Lactation . . . . . . 90
5.1.5 Breast Milk Creamatocrit of Dams That Consumed Hibiscus
Sabdariffa and Azadirachta indica During Lactation . . 91
5.2 Offspring Study . . . . . . . . 92
5.2.1 Postnatal Growth of Offspring Whose Mother Consumed
HS and AI During Lactation . . . . . 92
5.3 Conclusion . . . . . . . . 93
5.4 Recommendation . . . . . . . 94
Summary of Findings . . . . . . 95
References . . . . . . . . 96
CHAPTER ONE
1.0 INTRODUCTION
Medicinal plants over the years has been part and parcel of human society to combat diseases (Kausik et al., 2002). These plants has been a good source of food because they provide essential nutritional values, medicinal properties and notable physiological effect to life (Dalziel, 1973) .
Hibiscus sabdariffa and Azadinachta indica are used as medicinal and food ingredients in many parts of the world including Nigeria. Both plants are highly sourced as food vegetable particularly because of their health promoting and diseases- preventing properties strongly suspected to be due to the presence of many phytochemicals in them (Ijeomah et al, 2012). These phytochemicals present like alkaloid, saponin, glycoside, tannin, phenol, flavonoid, steroid , reducing sugar, nimbidin, sodium nimbidate, nimbin, gedunin, delphinidin 3- sambubioside and protocatechuic acid in the two plants have been found to be protective and preventive against many degenerative diseases and pathological process such as in ageing (Burns et al., 2001). Hibiscus sabdariffa (family: malvaceae) commonly known as zobo in Nigeria, is an annual herbaceous shrub, cultivated for its flowers, leaves and seeds. It is found in the tropics, subtropics and other parts of the world (Dalziel, 1973) but it is utilized beyond these areas of cultivation globally. In folk medicine this medicinal herb is used for the treatment of hypertension (Wang et al., 2000; Odigie et al., 2003). The plant is reported to have hepatoprotective, antihypertensive, antihyperlipidemic, anticancer and antioxidant properties (Mahadevan and Shivali, 2009).
Azadirachta indica which is commonly referred to as Dogonyaro or neem plant is an evergreen robust tree belonging to the family meliaceae. It is mostly found in tropics and sub- tropical areas of the world African and Asia (Ejoba, 2012) .The tree occurs in medium to large size and has dark grey bark and a dense rounded row of pinnate leaves (Ogbuewu, 2008). All parts of the neem tree (leaves, flowers, seeds, fruits, roots and bark) are widely used in traditional medicine. Azadirachta indica is used for the treatment of many health related problems and also known to extent anticancer, antioxidant, wound – healing and antimicrobial properties (Hybercyclopedia, 2014). According to (Ogbuewu, 2008; Mossini et al., 2004), all parts of this plant are useful and has been used to treat diseases ranging from tooth decay, ulcer, swollen liver, malaria and dysentery.
Breast milk is the normative standard for infant feeding and nutrition because it has been associated with a reduction in the incidence of gastrointestinal tract infection, respiratory tract infection and otitis media (AAP, 2012; WHO, 2013). It is important to note that breast milk is widely accepted to be the optimal source of nutrition for the newborn infants because it contains appropriate amount of protein, lipid, carbohydrate, micronutrients, trace elements, vitamins, minerals, digestive enzyme and hormones (Chantry et al., 2011) required for early postnatal growth of offspring, development and immune protection (Ostrom, 1990). However it is now universally recognized that there is no commercial formula that can equal breast milk. The relationship of breast milk feeding with a significant reduction in the incidence of necrotizing enterocolitis (NEC) has been suggested in preterm infants (Sullivan, 2010).
Additionally there is extensive evidence that offspring who had been breastfed with breast milk show lower risk of same non- communicable diseases later in life (AAP, 2012; WHO, 2013). Indeed breast milk given to infant at the early postnatal stage has been associated with lower risk of obesity, Lower levels of arterial blood pressure, lower total and LDL-Blood cholesterol level in adulthood and lower risk of developing type – 2 diabetes (AAP, 2012; WHO, 2013). Breast milk is not only associated with improved parameters of growth in offspring but also with a better neuronal-behavioral development (Mortensen et al., 2002; AAP, 2012). Breast milk creamatocrit is the cream column separated from the milk by centrifugation and expressed as a percentages of the length of the total milk column. It is a simple method of estimating the calorific value of breast milk ( Lucas, 1978; Wang et al., 1999).
Infant growth during the early postnatal period is obviously the result of the development of complex functional, structural and metabolic mechanisms, which are however not yet entirely clarified (Patel and Srinivasan, 2010). However this period is characterized by a great plasticity with critical windows in which any inadequate insult or intervention may cause both positively and adversely postnatal growth and development (Ostadelova and Babicky, 2012). This study is to determine the effect of consumption of aqueous extract of Hibiscus sabdariffa and Azadirachta indica during pregnancy and lactation on breast milk creamatocrit and early postnatal growth of offspring, considering the fact that the extract is widely used socially.
1.1 STATEMENT OF PROBLEM
The sweetened aqueous extract of Hibiscus sabdariffa is consumed by both males and females including pregnant and lactating mothers not necessarily for medicinal purposes but as a substitute for carbonated drinks (Iyare and Adegoke, 2011) while Azadinachta indica is commonly and indiscriminately used in traditional medicine for the treatment of malaria in Nigeria (Ucheya et al., 2011). These plant extract are consumed without regards to the physiological state of the body (Iyare and Adegoke, 2011). There is paucity of information on the effect of these two plant extracts on breast milk creamatocrit which is a vital factor for the early postnatal growth of offspring.
1.2 AIM AND OBJECTIVES
1.2.1 AIM
This study was aimed at investigating the effect of consumption of aqueous extract of Hibiscus sabdariffa and Azadirachta indica during pregnancy and lactation on breast milk creamatocrit and early postnatal growth of offspring.
1.2.2 OBJECTIVES
The objectives of this study included the following:
- To determine the effect of the aqueous extract of Hibiscus sabdariffa and Azadirachta indica on food and fluid intake during pregnancy and lactation.
- To determine the effect of the aqueous extract of Hibiscus sabdariffa and Azadirachta indica on maternal weight during pregnancy and lactation.
- To determine the effect of the aqueous extract of Hibiscus sabdariffa and Azadirachta indica on breast milk creamatocrit at postnatal day 7, 14 and 21
- To determine the effect of the aqueous extract of Hibiscus sabdariffa and Azadirachta indica on body parameters such as, litter weight, litter length and BMI of the offspring at birth, postnatal day 7, 14 and 21.
1.3 OPERATIONAL DEFINITION OF TERMS
For the purpose of clarification and better understanding of this study, some concepts require definition, they include
PREGNANCY: The period from conception to birth after the egg has been fertilized by a sperm and then imported into the lining of the uterus which develops into embryo and later into a fetus.
LACTATION: The secretion of milk from the mammary gland after parturition and the period of time that a mother lactates to feed her young.
BREAST MILK: It is the milk produced by the breast (or mammary glands) of a female mammal (humans & animals) for their infant offspring. Milk is the primary source of nutrition for newborns before they are able to eat and digest other food.
BREASTMILK CREAMATOCRIT: It is the length of cream column separated from milk by centrifugation and expressed as a percentage of the length of the total milk column.
EARLY POSTNATAL GROWTH: The period of growing or development from birth when the offspring begins to develop complex functional, structural and metabolic mechanisms.
OFFSPRING: The progeny or descendant of a person or an animal.
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