Introduction

Human milk (Stanya) is an excellent source of nutrition for infants, as it contains essential nutrients in the correct balance and numerous immunological, biochemical, cellular component, probiotic, and stem cells. Human milk has been also considered as live tissue. The milk has a unique composition which is species specific, evolved over millions of years to suit the needs of infants. There is individual variations in human milk composition, which are attributed to the stage of lactation, the degree of breast fullness, infant feeding, the health of the breastfeeding dyad, and other factors.^1,2 Research has shown a close association between milk fat and cell contents that changes with the degree of breast fullness. It may be associated with the maternal diet and environment, and potentially with genetic factors.³

Studies have also reported that the lactation strategies along with milk synthesis and its composition vary among individuals within species_. In this way infants receive “personalized” milk from their mother.^4,5

WHO (2019) recommends exclusive breastfeeding for the first six months of life, after which, infants should receive nutritionally adequate and safe complementary food while breastfeeding continues for up to two years of age or beyond.⁶ Exclusive breastfeeding for the first six months of life is associated with a decreased prevalence of infections and persistent diseases.^{7, 8}

Ayurveda is an ancient holistic, traditional health care system of India. The most fundamental principle of Ayurveda is “Tridosha” or the three vital humours i.e. Vata, Pitta and Kapha. These are the three biological entities that regulate all the biological activities and are responsible for determination of Prakriti (psychosomatic constitution) of an individual. The phenotypic expression of genotype differs as per the predominance of Dosha resulting in variation in physical, physiological and psychological characteristics in an Individual.⁹ Ayurveda has categorized human population into seven categories as per Doshika dominance and their permutations. The seven types of Prakriti are namely Vatika, Paittika, Kahaja, Vata-Paittika, Pitta-kaphaja, Vata-kaphaja, and Samadosha or Samaprakriti(Ch. S. Vi. 8/).¹⁰ In every person these three Dosha differ in permutations and combinations resulting in metabolic and physiological variations. Since human milk (Stanya) is also a product of metabolism that’s why there are differences in values of constituents of human milk as per Doshika psychosomatic constitution also. The importance of pure or normal mother milk for health, unimpeded growth and development, longevity of body organs, disease free state in infants was well recognized by ancient Ayurveda scholars and comprehensive descriptions were made about the characteristics of pure and vitiated mother milk,^10,11,12 measures for purification of vitiated milk, effect of diet and lifestyle on quality of mother milk and its effect on infants.

Recent studies have reported the substantial inter individual variation within populations, which have been associated with maternal characteristics. The human milk synthesis and its composition depends on maternal factors such as diet, parity, and body fat exert greater influence on milk synthesis.^13,14,15

Keeping these views in mind, this study was carried out with the aim to find out the analysis of normal and vitiated human milk and its composition among lactating mothers of different Prakriti.

Material and Methods

Study Design and Participants

For this study a cross sectional analytical study design was followed. 200 lactating Indian mothers (15^th to 25^th days of delivery) were registered after, getting informed written consent. Participants were selected by purposive sampling method from OPD of Department of Kaumarbhritya/Balaroga.

Study place: Department of Kryia Sharir, Department of Kaumarbhritya, Sir Sunder Lal Hospital, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.

Study Period: year 2015 to 2017

Ethical Approval: The present study proposal was approved from ethical committee of the institute vide letter number dean/2014-15/EC/1322 dated on 02/09/2015.

Inclusion Criteria

• Age Group 20-35 years.
• Mothers, who were enrolled in OPD of PrasutiTantra for the follow up after delivery and no acute or chronic disease was noted by the gynecologist/obstetrician on the OPD slip.
• Mothers were screened also in terms of history like present or past illness, drug history, and routine question related to health¹⁶ (K.S.Kh.S. 5/6-8) in addition to mother’s discharge slip.

Exclusion Criteria

• Maternal age below 20 years and above 35 years.
• Mother has known gestational diabetes, history of pre-eclampsia.
• Mother with contraindicated breast feeding
• Mother taking any specific drug for any illness or suffering from any disease including moderate (9.97 to 7.00g/dl) to severe (6.94 to 4.00 g/dl) anemia.¹⁷

Assessment of Psychosomatic Constitution (Prakriti) of Mother

It was assessed through a reliable and validated Performa developed by Tripathi PK et al, 2016.¹⁸ This Performa was based on the characteristics of Vata, Pitta and Kapha Prakriti individuals as described in various Ayurvedic texts. Eka Doshaja Prakriti was assigned on the basis of highest percentage of Dosha (more than 40%) and difference of at least 7% between two Dosha.

Method of collection of Human milk samples

After getting informed consent, under all the recommended precautions, the mothers were asked to wash their hands and breast particularly nipple and areola part with soap water properly and were allowed to dry. Physical examination was done for the absence of cracked nipple, breast abscess etc. The milk sample was collected between 10.00 am to 12.00 noon. Mothers were asked to express their one breast completely (average quantity 50 ml) in a sterile container with the help of human milk pump (Philip Avent Manual Breast Pump).¹⁹ After complete emptying of one breast, milk was transferred to another sterile glass container for further instant evaluation. An average time for initiation of test was 1 minute to avoid false report.

Assessment and Analysis of Human milk

As per Ayurveda, Pure milk has certain characteristics viz. white in color, natural odor, sweet taste and cold in touch, dissolves uniformly in water, and after digestion provides nourishment, unimpeded growth and development, strength, longevity of body organs and disease free state i.e. healthy state of the child.^10,16

To assess the quality of mother milk Jala-pariksha as an objective parameter was adopted since it is simple method and can be performed in OPD or bed side in natural light with tap water. Use of tap water was taken by considering the effect of Desha. In this study, water test (Jal-pariksha) of mother milk was done for differentiating non-vitiated mother milk with vitiated mother milk before doing analysis by lactometer for other objective parameters.

Human milk examinations by water test (Jala Priksha)

Jala Pariksha^{10, 12, 16} was carried out to test the characteristics of mother milk.

Method of Water test (Jala Pariksha)

The vitiation / purity of milk was assessed by Jala Pariksha and categorized accordingly into Vata, Pitta, Kapha Dosha vitiated or pure milk. One drop of milk sample was poured over water surface in a beaker of 500 ml capacity, filled with tap water. Time was noted when the lowest milk fiber touched first time at the bottom of beaker and the time taken for complete dissolution in stagnant water of beaker was recorded.

Interpretation

• When a drop of human milk is put on the surface of water, it gets mixed uniformly without floating over the surface of water or settles down at the bottom of beaker, was considered pure mother milk.
• When a drop of milk is put on the surface of water, floats in a scattered manner and spreads like an umbrella is suggestive of Vata vitiated human milk.
• When a drop of milk submerged in water with yellow color (hue) streaks, was considered Pitta vitiated human milk.
• When a drop of human milk is put on the surface of water, it settles down rapidly (< 10 second) towards the bottom of beaker and then persists for a long period (> 180 second) before dissolution indicates the Kapha vitiated human milk.²⁰

Method for Human Milk Analysis

Analysis of milk was done through Eco milk Analyzer (EON Trading LLC, USA Company) in Department of Animal Husbandry and Dairy, Institute of Agriculture Sciences, BHU. The working principle of analyzer is based on ultra sound technology and does not require any reagent or chemicals for the test.

To analyze the mother milk, the sucker point of the analyzer was washed with the distilled water by allowing the water to be drawn through it. The milk was taken in three different test tubes in equal quantity; one tube was placed in pH electrode, second tube in temperature slot for their finding, while milk of 3^rd tube by keeping in given slot, was allowed to be sucked for the assessment of the various parameters viz.fat (%) ± 0.1%, solid non-fat (SNF %) ± 0.2%, milk density (g/cm3)± 0.0005 g/cm3, protein (%) ± 0.2%, lactose (%) ± 0.2%, freezing point (°C) ±0.015°C, conductivity (mS/cm) ±1 % (18°C)and pH ±0.02.

Statistical Tools

Statistical analysis was done by using Statistical Package for Social Sciences (SPSS) Software Version 16.1.Data tabulated and appropriate statistical test viz. mean, standard deviation, Independent‘t’ -test, One Way ANOVA and Post Hoc test were applied in order to draw meaningful inferences such as difference in composition of milk as per Prakriti of mother in case of both vitiated and non-vitiated milk.

Observations and Result

Incidence of Prakriti types in Mothers

In this study population, out of 200 lactating healthy mothers, 26.5% were of Pitta Prakriti, 32%of Vata Prakriti and 43.5% belonged of Kapha Prakriti.

Table 1: Mean height, weight and BMI of lactating mothers as per Prakriti.

In Vata, Pitta and Kapha Prakriti lactating mothers, on applying One Way ANOVA, the mean values of weight and BMI varied significantly as per Prakriti (p<0.0001) although all the mothers were in normal range of BMI. The significant groups observed through Post Hoc test were Vata v/s Pitta (p=0.0003) and Vata v/s Kapha (p<0.0001) (Table No.1)

Table 2: Composition wise comparison of normal and vitiated milk of Vata Prakriti mothers.

The mean values of Fat%,SNF%,Density(g/cm3),Protein %, and Freezing Point(°C) were more in vitiated milk of Vata Prakriti mothers as compared to pure milk, while the mean values of conductivity(mS/cm)and pH were observed higher in pure milk of Vata Prakriti mothers. On applying independent sample t –test to compare the composition of both types of milk, values of only fat and protein percentage varied significantly (p=0.02, p=0.005).(Table No.2)

Table 3: Composition wise comparison of normal and vitiated milk of Pitta Prakriti mothers.

The mean values of Fat%,SNF%, Density(g/cm3) and Protein% were found higher in vitiated milk of Pitta Prakriti mothers as compared to normal (pure) milk while values of Freezing Point(°C)., Lactose (%),Conductivity (mS/cm) and pH were found higher in pure milk of Pitta Prakriti mothers. However, on applying independent sample t –test, none of these mean values were found to vary significantly in pure as well as in vitiated milk. (Table No.3)

Table 4: Composition wise comparison of normal and vitiated milk of Kapha Prakriti mothers.

The mean values of Fat%, SNF%, Density (g/cm3), Protein %, Conductivity and Freezing Point were also observed higher in vitiated milk of Kapha Prakriti mothers as compared to pure milk while in case of lactose %, conductivity (mS/cm) and pH the mean values were observed higher in pure milk of Kapha Prakriti mothers. On applying independent sample t –test to compare the composition of both types of milk, values of fat percentage(p=<0.0024) and Protein% (p=<0.0001) were found to vary significantly(Table No.5).

Table 6: Variations in normal human milk compositions as per Prakriti.