Effect of Storage on Bio-Chemical Composition of Bee Bread Collected from Apis mellifera Colonies of Indora Region (District-Kangra) Himachal Pradesh

Master's Thesis, 2019
30 Pages, Grade: 7.8








3.1 Collection & Preservation of bee bread
3.2 Physical appearance of bee bread
3.3 Biochemical analysis
3.3.1 pH
3.3.2 Electrical Conductivity
3.3.3 Moisture Content
3.3.4 Ash Content
3.3.5 Protein Content
3.3.6 Carbohydrate content: Reducing Sugar (IS method)
3.3.7 Total Sugar





“Presentation, Inspiration & Motivation have always played a important role in in success of venture”

Foremost , I want to offer his endeavour to our GOD Almighty for the wisdom he bestowed upon me, the strength, peace of my mind and good health in order to finish this project work.

It is a great pleasure to acknowledgment my deepest thanks and gratitude to Mrs. Vijayta Sharma HOD of life sciences.

Dr. Rajesh Kumar and PhD scholar Indu Sharma of Arni University for suggesting me a topic for this project work and their kind supervision. It is a great honour and pleasure to work under their supervision.

I would like to express my sincere gratitude towards and appreciation to Prof. (Dr. Sarika Sharma, Mr. Sudesh Kumar Banyal, Mr. Sachin Sharma, Dr. Arup Giri, Miss Priyanka Sharma, Mrs. Neha Sharma ( Faculty, Department of Life Science) for their encouragement affection & everyday gesture, which make it possible for me to each this level. I am gratefully thanks to my non teaching staff of the department for their help during the project work.

I want to express my gratitude to Indu Kumari (Research Scholar) for their supportive suggestions. I am immensely obliged to my friends (Sonali Singh, Rimpi Devi, Shapali Sharma, Anu Dhiman, Mamta Devi, Shalini Sharma, Khushboo and Ajay Bhatia) for their elevating inspiration, encouraging, guidance, and kind supervision in the completion of my project work.

I would like to express my deep gratitude to my family members for their encouragement which helped me in completion of this project work. My beloved parents Mr. Nand Lal and Mrs. Soma Devi who are always by my side and my lovable brother Mr. Arun Bangga for their support and help during my project work.

Thank you...

List of Tables

Difference between the different parameters of fresh bee breed and stored bee breed

List of Figures

1. Collected Bee Bread from Bee hive

2. Physical Appearance of Bee Bread

3. Instrument used to Detect the electrical Conductivity

4. Graphical Representation of value of Biochemical Parameters

5. pH meter showing reading of the sample

6. EC meter showing the result of the sample

7. Measuring the weight of Ash Content

8. Worked at moisture content in Hot Air Oven

9. Protein Content of bee bread using Lowry’s Method

10. Working At analysis of Carbohydrate content


Honey bees are one of the most beneficial creatures in the animal kingdom. They derive pollen and nectar from a large number of plant species for their food. The present study was carried out by using bee bread sample collected from Arni University, Indora (Kangra), during 2017-2018. In this study, two types of bee bread samples i.e. stored bee bread and fresh bee bread was used. For the biochemical analysis parameters like pH, electrical conductivity (EC), Moisture content, Ash content, Protein content, Carbohydrate content were measured. It was concluded that the end of the study that electrical conductivity and moisture content increase in stored bee bread as compare to Fresh bee bread. But in the case of Ash content, protein And Carbohydrate content reported being higher in fresh bee bread as compared to stored bee bread. The reason behind this might be the effect of time on the storage of bee bread as well as poor storage condition which lead to the degradation of biochemical component of bee bread.


Honey bees are one of the most beneficial creatures in the animal kingdom. They derive pollen and nectar from a large number of plant species for their food. Energy required for all the activities is provided by the oxidation of food they consume. Insects mainly feed on the plants and also perform pollination. Honey bees, wasps, butterflies and aphids are main pollinators of the crop plants. Honey bees collects nectar from different flora which acts as energy source for them whereas, need of protein, carbohydrates, lipids, minerals and vitamins for brood rearing and development is fulfilled by pollen they collect from the flower.

Bee bread is fermented naturally preserved pollen. The process of bee bread formation starts with gathering of pollen, then a bee mixes it with flower nectar or honey and saliva and carries to the bee hive, where non flying bees fills the mixture into the honey comb cells for ¾ of the cell volume. Residual cell volume is filled with honey, thus protecting the pollen mass from oxygen. An anaerobic lactic fermentation process takes place and the bee bread is forming. Bee bread is differs from pollen by lower phi (3.9- 4.9).

In most parts of our country, natural resource of pollen and nectors become scarces during summers popularly known as “Death Period”. The ornamental plants do not bear flowers during this season while the available agriculture crops like cotton, rice etc are frequently sprayed with toxic insecticides. Quite often beekeepers harvest excess amount of honey before dearth period so colonies cannot sustain in shortage of food. Periodical dearth of pollen results into low nutrition reserves which adversely affect the colonies performance due to the reduction of the brood rearing. The longevity of adult bees is also affected which causes decline in colony strength. The lack of food reduces the energy of forager bees which they requires fetching out pollen, water and resins from distant places. Food shortage may failure of bee’s colonies.

Larvae of the honey bees mainly feed on bee bread which acts as main source of protein for them. Young adults also consumes higher amount of pollen as enabling their hypo pharyngeal gland to produce jelly for feeding larvae, after the transition to foraging they mainly feed on carbohydrates.

Nutritional value of bee bread is more than of pollen so it may be more preferable than pollen. Bee bread acts as an atherosclerotic flushing agent that it enhances the stamina of the individual by increasing the allowance of more oxygen reach to the body and brain cells.

Bee bread provides a rapid increase in hemoglobin level of the body so it is very useful in anemia and helps in flushing out the impurities from the body. Bee bread protects any deficiencies in vitamins, minerals and amino acid. Bee bread has antimicrobial, antioxidant hepatoprotective, immune modulating and anti-radiation activity apoptogenic properties. It stimulates protective forces of a human body normalizes metabolism and has a positive influence on the liver, nervous and endocrine system functions and enhances regeneration of tissues, physical and mental persistence of human body (Bogdanovian,2015).

Now a day’s Bee bread is growing in commercial interest due to its high nutritional properties. The exact composition of bee bread varies according to the flora honey bee forages.

Review of Literature

Bee bread is a collection of pollen gathered by the bees and mixed with the bees digestive enzymes, which was later carried back to the hive packed into pellets or stacks of pollen and preserved with a tiny bit of honey and wax. (Nagai et al., 2004).

A a study conducted at the department of Zoology, Punjab University, revealed that maximum constituents were extracted from bee pollen & honey with water as solvent, which is valuable for research and drug development in the field of pharmacy (Kour et al., 2013)

Egorova (1971) and pain & maugenet (1966) revealed after microbial examination of stored pollen, it has a specific bacterial flora associated with it. Pollen of different plants do not have the same physiological effects (Maurizio, 1954; Todd & Bretherick, 1942).

Todd &Bretherick (1942) reported that all the ash content of bee-collected pollen ranged from 0.91 to6.36% with a mean of 2.70% and the content of crude protein ranged from 7.02 to 29.87 %.The pH range of pollen is reduced from 4.8 to an average of 4.07 in bee bread, which facilitates the growth of various microorganisms ( Egorova,1971).

Information by Herbert and Shimanuki (1978) suggests that pollen or bee bread might be used as a standard in the formulation of our future basal diets.

Pollen stored in comb cells & through microbial mediated lactic acid fermentation gets chemically changed (Gilliam et al., 1989).

A study done by De Grandi-Hoffman et al. (2012) showed that bee bread made by European honey bees are more acidic than the bee bread made by the African honey bees & European honey bees but is lower than in Pollen.

Fuenmayor, (2009) and Risco et al, 2012 reported that bee bread has increased the availability of nutrients and bioactive components in comparison to pollen. It is reported that specialized devices and materials have been designed to extract bee bread from the hive ( Wilara, 2014).

A loss in the values of bioactive compounds are reported from the study of Zuluaga et al (2016)for bee bread as compared to Bee pollen due to the breaking of the outer layer of the pollen grains, which shield it to be degraded in environmental condition.

In Lithuania, a study performed by Cepsteryte (2008) analyzed 22 fatty acids in Bee bread sample include five omega 3, four omega 6,& three omega 9 polyunsaturated fatty acid.

Chevtchik (1950) and pain &Maugenet (1966) describes a series of microbes & chemical changes occur during fermentation of pollen. It is reported that bacterial action occurring which is aided by contents of pollen. Bee bread differs from pollen by lower pH (3.8 – 4.3), it contains less protein & fats but more carbohydrates and lactic acid.

Bee bread has better bioavailability because the walls of pollen which cannot be destructed by gastrointestinal liquid has been partly destructed by fermentation (Mizrahi and Lensky, 1997).

According to the study of Rubik and Buchmann (1984), the water content of nectar can vary greatly according to environmental condition. Bee prefers to collect concentrated nectars, which maximize their energy gain.

There is an increase in soluble proteins due to the fermentation process of pollen which converts into bee bread (Langer, 1931).

The study of reported by Cobet (2003) the energy content of a flowerʼsnector depend upon its volume and sugar concentration.

Vasquez & Olofsson, 2009 suggests that Lactic acid bacteria are already present in bee-collected pollen and are the source of vitamin and the digestive fluids from the bee are found to rich in with lactic acid bacteria.

It is reported in a study Seeley, 1995 that a single wild Honey bee colony will harvested 120kg of nectar and 20kg of pollen annually in a temperature region; this amount may be in managed colonies.

In addition to preservation (Anderson et al.2014) the fermentation process of the pollen also renders its nutrients more available (Mattila et al., 2012). Some protein is broken down into amino acids, starches are metabolized into simple sugars, and vitamins become more bio-available (Hoffman et al., 2013; Herbert and Shimanuki, 1978). In this sense, bee bread is even more health-giving than the more commonly available fresh bee pollen.

A bouda et al., 2012 reported that bee bread has antibacterial activities against both Gram-positive and Gram negative bacteria such as E.coli, Staphylococcus aureus, and Pseudomonas aeruginosa.

The lactic acid fermentation that occurred in bee bread was similar to that in production of yogurt where it enables the end product to be more digestible & enriched with new nutrients (Krell,1966).

Mutsaers et al., 2005 concludes in their work that bee the bread was easier to be absorbed by the body as compared to the original pollen although it contains less amount of protein. Some of the components present in the bee bread were essential amino acids, several vitamins, some pigments, carotenoids & anthocyanin, several types of enzymes, flavonoids as well as vast numbers of different minerals.

Habryka et al., 2016 reveals in their work that bee bread can be considered as valuable medicinal product to the consumers.

The taste of the bee bread comes from plant resins which the bees used to build their hives& honey pots and its varies at different times of the year depending on the flowers & tree visited. Bee bread contains enzymes which are essential biological catalysts during the digestive process. The enzymes found in bee bread include amylase, catalase, cozymase, cytochrome, dehydrogenase, diaphorase, diastase, lactic acid, pectase and phosphatase (Combey, 2017).

It was reported in a study that honey bees (Apis mellifera L.) use pollen as their nutritional source of protein, fatty acids, lipids, sterols, vitamins, minerals and certain carbohydrates (Dietz,1975).

According to Herbert &Shimanuki (1978) and pain &Maugenet (1966), there is no any increase in nutritional value due to the fermentation process although Herbert & Shmanuki reported earlier consumption & brood rearing with bee bread.

It was reported by Lukoschuset al., 1968 that there is a large change in titrable acidity is probably due to the addition of mandibular & hypopharyngeal gland secretions.

Gilliam (1979 a, 1979 b), reported that the loss of ascorbic acid during fermentation of the bee bread, undoubtedly influenced by the large changes in microbial flora.

Nectar provides carbohydrates and pollen supplies the remaining dietary requirements such as protein, lipids, vitamins, and minerals to bee (Brodschneiderand crailsheim, 2010).

Pain &Maugenet (1966) found three microbial genera in bee bread: Pseudomonas, Lactobacillus and Saccharomyces. These suggest that microorganisms are probably involved in the storage of pollen.

Beutler and opfinger (1949) revealed that bee lived longer on pollen removed from comb than from pollen traps.

Material and Methods

The present study was carried out by using bee bread sample which was collected from Arni University, Indora (Kangra), during 2017-2018. The methodology used during the study was as under:

3.1 Collection & Preservation of bee bread:

Bee bread was collected from the bee hives manually. For this purpose, complete frame of bee was taken out and bees were removed from the frame with help of a brush. The fame was then brought to lab for the extraction of the bee bread. Bee bread was removed with the help of spatula & collected upon butter paper. Then it was stored in airtight containers. Containers were kept in refrigerator till further use.

Abbildung in dieser Leseprobe nicht enthalten

Figure1: Collected Bee Bread from Bee hive

3.2 Physical appearance of bee bread:

Bee bread was visually analyzed for physical appearance color and odor. Color of the bee bread was observed before refrigeration and after dry ing.

Abbildung in dieser Leseprobe nicht enthalten

Figure 2: Physical Appearance of Bee Bread

3.3 Biochemical analysis

The biochemical analysis of stored bee bread was carried out in the laboratory, Dept. of Bio sciences. Following parameters were analyzed.

1. pH
2. Electrical conductivity
3. Moisture content
4. Ash content
5. Protein content
6. Carbohydrate content


Excerpt out of 30 pages


Effect of Storage on Bio-Chemical Composition of Bee Bread Collected from Apis mellifera Colonies of Indora Region (District-Kangra) Himachal Pradesh
MSc Zoology
Catalog Number
ISBN (eBook)
effect, himachal, district-kangra, region, indora, colonies, apis, collected, bread, composition, bio-chemical, storage, pradesh
Quote paper
Rajesh Kumar (Author)Anjali (Author), 2019, Effect of Storage on Bio-Chemical Composition of Bee Bread Collected from Apis mellifera Colonies of Indora Region (District-Kangra) Himachal Pradesh, Munich, GRIN Verlag, https://www.grin.com/document/494530


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