**1. Introduction**

Melissopalynology is a branch of plant sciences that studies pollen found in honey. Precision in interpreting pollen data recovered from the honey has always been a primary goal of those who study pollen and honey. We are using pollen count to determine the nectar source of a honey sample and recognize the types and percentage of recovered pollen in the honey. This study is the fact that honey bees utilize certain natural raw materials which are identifiable in honey. These natural raw materials include pollen and nectar [1]. The growth and development of honey bees depend on nectar as the source of carbohydrates and pollen as the source of proteins [2, 3]. Palynological analyses of honey and pollen loads are used to know about honey bee foraging ecology, habitat and vegetation [4–11]. Pollen of honey samples provides reliable information on floral resources of honeys along with the relative preferences of bees among the diverse assemblages of plant species flowering synchronously [12, 13].

The significant melissopalynological works have been reported from different sectors of this state, dealing with pollen analysis of honey [14–16]. Similar melissopalynological research work has been worked out in Karnataka [17–19],

Af-44, and Af-64), 2 from Munchingiputtu (Af-14 and Af-54), 2 from G.Madugula (Af-3 and Af-48), 3 from Peddabayalu (Af-11, Af-53, and Af-65), and one each from Araku Valley (Af-37), Dumbriguda (Af-9), and Hukumpeta (Af-51). For palynological assessment, the honey samples were chemically processed using the acetolysis method, i.e., 1 ml of honey sample was dissolved in 10 ml of distilled water and centrifuged. The supernatant liquid was drawn out. The resultant sediment was treated with 5 ml of glacial acetic acid and centrifuged. After decanting acetic acid, the sediment was treated with acetolysis mixture (5 ml). It was prepared by nine parts of acetic anhydride and one part of con. sulfuric acid and then heated under water bath until the liquid turned chestnut-brown color. After cooling it was again centrifuged and the supernatant liquid was decanted. The sediment then treated with glacial acetic acid, later centrifuged, and the supernatant liquid was decanted off. And the sediment was washed with distilled water, and 50% aqueous glycerin (5 ml) was added and centrifuged for 10 min. The supernatant liquid was decanted off, and the tubes were inverted upside down on a filter paper for a few

*Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam…*

The pollen sediment was taken on a pellet of glycerin jelly and transferred to the center of the slide. After being warmed slightly, the melted jelly with pollen sediment was covered by cover slip. Cover glass was later sealed with paraffin wax and labeled with their respective codes. Three slides were prepared for each sample and

Identification of pollen recovered from honey samples were carried out through the consultation of reference pollen slides available in Paleobotany and Palynology lab, UCS, Saifabad, OU. Quantitative pollen analysis was based on the method recommended by the International Commission for Bee Botany [6]. Pollen contents were taken at random, covering the maximum mounted area to avoid repletion. Once identified and counted, the pollen grains were placed into one of following pollen frequency classes-predominant pollen types (>45%), secondary pollen types (16–45%), important minor pollen types (3–15%), and minor pollen types (<3%). Honey samples containing more than 45% of a single type of pollen were considered as unifloral honey. The pollen types were placed into arboreal and non-arboreal taxa for making honey pollen spectra (**Figure 2**). A detailed list which included sample number, locality, nature and type of honey, collection season, and frequency of

*Pollen spectra showing frequency of non-arboreal and arboreal taxa from different honeys of Paderu forest*

minutes [34].

**Figure 2.**

*division.*

**51**

studied critically for their pollen contents.

*DOI: http://dx.doi.org/10.5772/intechopen.88908*

pollen types recovered is given in **Table 1**.

**Figure 1.** *Study area of Paderu forest division in Visakhapatnam district, Andhra Pradesh.*

Bihar [20], Madhya Pradesh [21], Maharashtra [22], Uttarakhand [23–26], Uttar Pradesh [27, 28], and West Bengal [29–32], but the information is still rather sketchy. Qualitative and quantitative melissopalynological analyses in the east coast regions of India demonstrate that these regions are rich in bee plants with potential for producing adequate unifloral honeys, have an extended honey flow period, and thus can be utilized commercially for a moderate- to large-scale apiculture enterprises [33].

The study area includes the Paderu forest division of Visakhapatnam district, Andhra Pradesh. This division is the higher altitude zone in the hilly tracts of Eastern Ghats of Andhra Pradesh. It has the second highest tribal population in Andhra Pradesh. Paderu forest division (**Figure 1**) lies in between latitudes of 17°-51″ and 18°-35″ north and longitude of 82°-17″ and 83°-1″ east with a total geographical area of 3,24,965 ha, out of which the forest area under the control of the division is 104811.91 ha. The division comprises of a series of hills with an average annual rainfall of 2800 mm and a rich diversity of plant wealth.

## **2. Materials and methods**

The materials for the present study are 17 honey samples (50 ml each) that were procured from 8 different blocks of Paderu forest division during 2011–2013, i.e., 3 samples from Ananthagiri (Af-13, Af-34, and Af-57), 4 from Paderu (Af-10, Af-40, *Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam… DOI: http://dx.doi.org/10.5772/intechopen.88908*

Af-44, and Af-64), 2 from Munchingiputtu (Af-14 and Af-54), 2 from G.Madugula (Af-3 and Af-48), 3 from Peddabayalu (Af-11, Af-53, and Af-65), and one each from Araku Valley (Af-37), Dumbriguda (Af-9), and Hukumpeta (Af-51). For palynological assessment, the honey samples were chemically processed using the acetolysis method, i.e., 1 ml of honey sample was dissolved in 10 ml of distilled water and centrifuged. The supernatant liquid was drawn out. The resultant sediment was treated with 5 ml of glacial acetic acid and centrifuged. After decanting acetic acid, the sediment was treated with acetolysis mixture (5 ml). It was prepared by nine parts of acetic anhydride and one part of con. sulfuric acid and then heated under water bath until the liquid turned chestnut-brown color. After cooling it was again centrifuged and the supernatant liquid was decanted. The sediment then treated with glacial acetic acid, later centrifuged, and the supernatant liquid was decanted off. And the sediment was washed with distilled water, and 50% aqueous glycerin (5 ml) was added and centrifuged for 10 min. The supernatant liquid was decanted off, and the tubes were inverted upside down on a filter paper for a few minutes [34].

The pollen sediment was taken on a pellet of glycerin jelly and transferred to the center of the slide. After being warmed slightly, the melted jelly with pollen sediment was covered by cover slip. Cover glass was later sealed with paraffin wax and labeled with their respective codes. Three slides were prepared for each sample and studied critically for their pollen contents.

Identification of pollen recovered from honey samples were carried out through the consultation of reference pollen slides available in Paleobotany and Palynology lab, UCS, Saifabad, OU. Quantitative pollen analysis was based on the method recommended by the International Commission for Bee Botany [6]. Pollen contents were taken at random, covering the maximum mounted area to avoid repletion. Once identified and counted, the pollen grains were placed into one of following pollen frequency classes-predominant pollen types (>45%), secondary pollen types (16–45%), important minor pollen types (3–15%), and minor pollen types (<3%). Honey samples containing more than 45% of a single type of pollen were considered as unifloral honey. The pollen types were placed into arboreal and non-arboreal taxa for making honey pollen spectra (**Figure 2**). A detailed list which included sample number, locality, nature and type of honey, collection season, and frequency of pollen types recovered is given in **Table 1**.

Bihar [20], Madhya Pradesh [21], Maharashtra [22], Uttarakhand [23–26], Uttar Pradesh [27, 28], and West Bengal [29–32], but the information is still rather sketchy. Qualitative and quantitative melissopalynological analyses in the east coast regions of India demonstrate that these regions are rich in bee plants with potential for producing adequate unifloral honeys, have an extended honey flow period, and thus can be utilized commercially for a moderate- to large-scale apiculture enter-

*Study area of Paderu forest division in Visakhapatnam district, Andhra Pradesh.*

*Modern Beekeeping - Bases for Sustainable Production*

The study area includes the Paderu forest division of Visakhapatnam district, Andhra Pradesh. This division is the higher altitude zone in the hilly tracts of Eastern Ghats of Andhra Pradesh. It has the second highest tribal population in Andhra Pradesh. Paderu forest division (**Figure 1**) lies in between latitudes of 17°-51″ and 18°-35″ north and longitude of 82°-17″ and 83°-1″ east with a total geographical area of 3,24,965 ha, out of which the forest area under the control of the division is 104811.91 ha. The division comprises of a series of hills with an average annual rainfall of 2800 mm and a rich diversity of plant wealth.

The materials for the present study are 17 honey samples (50 ml each) that were procured from 8 different blocks of Paderu forest division during 2011–2013, i.e., 3 samples from Ananthagiri (Af-13, Af-34, and Af-57), 4 from Paderu (Af-10, Af-40,

prises [33].

**50**

**Figure 1.**

**2. Materials and methods**


**S. no Sample**

**53**

**Locality**

 **Nature of**

**Type of**

**Season**

**Predominant**

**Secondary**

 **pollen**

**Important minor pollen types**

**Minor pollen types**

**Pollen present**

**(0.5–<1%)**

**(1–3%)**

**(3–15%)**

*Ocimum sanctum* *Schleichera oleosa* *Eucalyptus globulus* (4.94%),

*Polygonum barbatum*

*Blumea oxyodonta*

*Casuarina equisetifolia* (3.18%)

(3.53%),

*Solanum nigrum* (1.76%), *Celosia*

*argentea* *Lagerstroemia*

*parviflora* *Commelina*

*suffruticosa* *Xanthium strumarium*

(1.48%)

5

6

 V-P-MUN-L-AF-14

Munchingiputtu

 Multifloral

 Squeezed

 Autumn —

 —

*Eucalyptus globulus* *Tridax procumbens* (8.77%),

*Centipeda minima*

*Ageratum conyzoides* (7.41%),

*Terminalia*

*Psidium guajava* *Crotalaria juncea* *Syzygium cumini* (6.95%),

*Commelina*

*Sapindus emarginatus* (4.84%),

*Erythrina variegata* *Leucas aspera* (3.78%)

(3.02%),

(1.66%), *Lantana*

*camara* (1.47%)

 *suffruticosa*

(6.95%),

*stricta* *Lannea* 

*coromandelica*

(1.96%),

 *arjuna*

(6.80%),

(6.80%),

(6.95%),

(8.32%),

(13.46%),

*Datura stramonium*

—

(2.26%), *Cassia*

*occidentalis* (2.42%),

*Vernonia cinerea* (2.11%), *Citrullus*

*lanatus* (2.11%),

*Albizia lebbeck* (1.96%), *Conyza*

V-P-AG-Ch-AF-13

Ananthagiri

 Unifloral

 Squeezed

 Autumn

*Schleichera*

—

*Mimosa pudica* *Eucalyptus globulus* (8.65%),

*Phyllanthus*

*Dendrophthoe*

*Leucas aspera* (3.69%)

 *falcata*

(3.07%),

*Careya arborea*

(1.69%), *Vicoa indica*

(1.50%), *Dillenia*

*pentagyna* (1.34%)

 *emblica*

(6.92%),

*guajava*

(1.73%),

(0.46%)

(13.07%),

*Manilkara zapota*

*Clerodendrum*

*Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam…*

*inerme*

(2.88%), *Psidium*

*oleosa*

(55.00%)

(1.41%),

(1.06%),

(1.41%),

*DOI: http://dx.doi.org/10.5772/intechopen.88908*

(3.53%),

(6.00%),

*Vernonia cinerea* (2.12%), *Cajanus*

*cajan*

(2.12%),

(5.30%),

**honey**

**honey**

**collected**

**pollen type**

**types (16–45%)**

**(>45%)**

**code**


*Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam… DOI: http://dx.doi.org/10.5772/intechopen.88908*

**S. no Sample**

**52**

**Locality**

 **Nature of**

**Type of**

**Season**

**Predominant**

**Secondary**

 **pollen**

**Important minor pollen types**

**Minor pollen types**

**Pollen present**

**(0.5–<1%)**

**(1–3%)**

**(3–15%)**

**honey**

**honey**

**collected**

**pollen type**

**types (16–45%)**

**(>45%)**

**code**

1

2

3

4

 V-P-PB-PB-AF-11

Peddabayalu

 Multifloral

 Squeezed

 Autumn —

 —

*Coriandrum*

*Mimosa pudica* *Syzygium cumini*

*Ageratum conyzoides*

(15.19%),

(12.36%),

(11.30%),

*murex*

(2.47%),

 *sativum*

(15.90%),

*Vicoa indica* *Amaranthus*

(2.47%), *Pedalium*

 *spinosus*

(2.47%),

—

 V-P-P-Sa-AF-10

Paderu

 Unifloral

 Squeezed

 Spring

*Ageratum conyzoides*

—

(68.25%)

V-P-DG-Vb-AF-9

Dumbriguda

 Multifloral

 Squeezed

 Spring

—

*Mimosa pudica*

*Crotalaria juncea* *Tridax procumbens* (8.57%),

*Psidium guajava*

*Andrographis*

*Terminalia*

*Schleichera oleosa*

*Commelina*

*Ageratum conyzoides* (5.20%)

*Schleichera oleosa*

*Parkinsonia*

*Citrullus lanatus* (3.17%)

 *aculeata*

(6.03%),

(7.93%),

*Peltophorum*

*Cyperus*

*pterocarpum* *Crotalaria juncea* (2.22%), *Ocimum*

*basilicum* *Tridax procumbens*

(1.90%), *Mimosa*

*pudica* *Terminalia*

(1.58%)

 *arjuna*

(1.58%),

(2.53%),

(0.95%),

*Pedalium*

*murex*

(0.69%)

(2.22%),

*rotundus*

(0.95%),

*Acacia chundra*

 *suffruticosa*

(5.57%),

 *arjuna*

(7.94%),

(2.23%)

(7.06%),

 *echioides* (8.17%),

(8.55%),

(13.01%),

*Dendrophthoe*

(2.60%), *Acacia*

*chundra* *Pedalium murex*

(2.48%),

 *falcata*

—

(28.62%)

 V-P-GM-Bp-AF-3

G.Madugula

 Multifloral

 Squeezed

 Spring

—

*Carum copticum*

Urticaceae type (15.02%),

*Eucalyptus globulus*

*Syzygium cumini*

*Psidium guajava* *procumbens* (6.05%), *Blumea*

*oxyodonta* (4.03%), *Brassica nigra* *Vernonia cinerea* (3.65%)

(3.81%),

(4.70%), *Bombax ceiba*

(13.22%),

*Spilanthes calva*

—

(2.69%), *Hakea*

*laurina*

(1.79%),

(12.78%),

(9.40%),*Tridax*

*Saccharum*

*officinarum*

(1.12%)

*Modern Beekeeping - Bases for Sustainable Production*

(21.74%)


**S. no Sample**

**55**

**Locality**

 **Nature of**

**Type of**

**Season**

**Predominant**

**Secondary**

 **pollen**

**Important minor pollen types**

**Minor pollen types**

**Pollen present**

**(0.5–<1%)**

**(1–3%)**

**(3–15%)**

**honey**

**honey**

**collected**

**pollen type**

**types (16–45%)**

**(>45%)**

**code**

14 V-P-MUN-Sk-AF-54

15 16 V-P-P-Rb-AF-64

Paderu

 Multifloral

 Squeezed

 Autumn

—

*Tridax*

*Eucalyptus globulus* *Leucaena leucocephala* Urticaceae type (5.47%), *Cocos*

*nucifera*

*spinosus* (3.45%),

*falcata* (3.21%)

(4.32%),

*Amaranthus*

*Dendrophthoe*

(12.96%),

—

 —

*Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam…*

(6.34%),

*procumbens*

(27.95%),

*Schleichera oleosa*

(19.88%),

*Erythrina*

*variegata*

(16.42%)

17 V-P-PB-L-AF-65

**Table 1.** *Pollen content in honey samples of Paderu forest division in* 

*Visakhapatnam*

 *district.*

Peddabayalu

 Multifloral

 Squeezed

 Autumn

—

*Mimosa pudica*

*Hakea laurina*

(12.97%)

—

 —

(41.0%),

*Schleichera oleosa*

(26.61%),

*Dendrophthoe*

*falcata*

(19.42%)

V-P-AG-Mt-AF-57

Ananthagiri

 Multifloral

 Squeezed

 Autumn

—

*Syzygium cumini*

*Spilanthes calva*

*Mimosa pudica* *Centipeda minima*

*Cyathocline*

*Leucaena leucocephala*

*Tridax procumbens* (5.71%),

*Vernonia cinerea* (5.39%)

 *purpurea*

(13.40%),

—

 —

*DOI: http://dx.doi.org/10.5772/intechopen.88908*

(13.24%),

(12.43%),

(12.76%),

(10.10%),

(26.97%)

Munchingiputtu

 Multifloral

 Squeezed

 Autumn

—

*Syzygium cumini*

Urticaceae type (11.82%),

*Vernonia cinerea* *procumbens* (5.84%)

(7.74%),*Tridax*

—

 —

(37.63%),

*Centipeda minima*

(18.70%), *Mimosa*

*pudica*

(18.27%)


*Melissopalynological Analysis of Honeys from Paderu Forest Division of Visakhapatnam… DOI: http://dx.doi.org/10.5772/intechopen.88908*

> **Table 1.**

 *Pollen content in honey samples of Paderu forest division in Visakhapatnam district.*

**S. no Sample**

**54**

**Locality**

 **Nature of**

**Type of**

**Season**

**Predominant**

**Secondary**

 **pollen**

**Important minor pollen types**

**Minor pollen types**

**Pollen present**

**(0.5–<1%)**

**(1–3%)**

**(3–15%)**

**honey**

**honey**

**collected**

**pollen type**

**types (16–45%)**

**(>45%)**

**code**

7

8

9

10 V-P-P-Vm-AF-44

11 V-P-GM-Ak-AF-48 12 V-P-HP-R-AF-51 13 V-P-PB-L-AF-53

Peddabayalu

 Multifloral

 Squeezed

 Spring

—

*Syzygium cumini*

*Mimosa pudica* *Vernonia cinerea* Urticaceae type (6.80%)

(14.59%),

*Caesalpinia*

(1.36%), *Bombax*

*ceiba*

(2.33%),

*Dendrophthoe*

(1.36%)

 *falcata*

 *bonduc*

*Sida acuta* (0.97%), *S.*

*cordata*

(0.82%)

(11.08%),

(41.82%),

*Centipeda minima*

(18.87%)

Hukumpeta

 Multifloral

 Squeezed

 Spring

—

*Eucalyptus*

*Hygrophila auriculata*

(14.83%),

—

 —

> *globulus*

*Mimosa pudica*

(17.41%).

(35.48%),

*Cocos nucifera*

*flabellifer*

*sativus* (9.71%).

(10.96%), *Cucumis*

(11.61%), *Borassus*

G.Madugula

 Multifloral

 Squeezed

 Spring

—

*Mimosa pudica*

*Pedalium murex* *suaveolens* (3.56%),

(5.24%), *Hyptis*

*Tribulus terrestris*

—

*Saccharum*

(2.72%)

(32.91%), *Conyza*

*stricta* *Tridax*

*procumbens*

(17.81%).

(28.30%),

*officinarum* (3.14%), *Ageratum conyzoides*

(3.14%)

(3.18%), *Sida acuta*

Paderu

 Unifloral

 Squeezed

 Spring

*Mimosa pudica*

—

*Hyptis suaveolens*

*Conyza stricta*

(13.59%)

(14.56%),

*Tridax procumbens*

—

(1.26%),

*Cardiospermum*

*halicacabum*

(1.02%)

(69.57%)

 V-P-P-Sp-AF-40

Paderu

 Unifloral

 Squeezed

 Spring

*Ageratum conyzoides*

—

*Dendrophthoe*

 *falcata* (4.98%)

(92.83%)

V-P-AR-Me-AF-37

Araku Valley

 Unifloral

 Squeezed

 Spring

*Psidium guajava*

*Mimosa pudica*

*Terminalia*

*Delonix regia* (5.85%)

 *arjuna*

(10.53%),

*Acacia chundra*

—

(2.34%), *Gardenia*

*lucida* (1.66%) *Tridax procumbens*

—

*Modern Beekeeping - Bases for Sustainable Production*

(2.19%)

(24.59%)

(55.03%)

V-P-AG-Gb-AF-34

Ananthagiri

 Unifloral

 Squeezed

 Spring

*Ageratum conyzoides*

—

*Phyllanthus*

*Madhuca indica* (5.86%)

 *emblica*

(12.06%),

*Saccharum*

—

*officinarum*

(1.05%)

(81.03%)
