Journal of Phytopathology and Pest Management 6(1): 24-31, 2019
pISSN:2356-8577 eISSN: 2356-6507
Journal homepage: http://ppmj.net/
Corresponding author:
Mohamed O. M. Omar,
E-mail: mohamed.omar@aun.edu.eg
24
Copyright © 2019
Diversity of pollen species in commercial honey
samples collected from different governorates of
Egypt
Salah H. Rateb, Heba R. Abd-El Karim, Mohamed O.M. Omar*, Mostafa H. Hussein
Plant Protection Department, Faculty of Agriculture, Assiut University, 71526 Assiut, Egypt
Abstract
Keywords: pollen identification, qualitative pollen analysis, bee honey.
25
1. Introduction
For many years, varietal identification of
bee honeys has been a research subject in
many scientific centers (Bogdanov,
1999). Using pollen analysis to assess
quality and botanical origin of honey was
discussed by Lukacs (1997). Quality
control methods have been found to be
able to classify honeys from different
geographical regions. Bee honeys were
considered monofloral honey whenever
the dominant pollen type was found to be
over 45% of total pollen grains in the
tested honey (Amaral et al., 2003). Also,
both pollen identification and count have
been used to determine the authentication
of honey according to the floral type
(Serrano et al., 2004). Results of Wen et
al. (1995) indicated that, about 30% of
samples were adulterated with sugar
syrup or other products. Due to its
simplicity, pollen analysis was used
extensively to identify different types of
honey samples from different botanical
origins in different countries (Devillers et
al., 2004; Marini et al.,
2004; Cordella et
al., 2002; 2003). Pollen spectra of honey
were influenced by time of the year and
location (Ponnuchamy et al., 2014). In
Egypt, the qualitative pollen analysis of
honeys was done by many authors (El-
Metwally, 2015; Rateb, 2005; Nour et al.,
1991; Nour, 1988). Pollen analysis of
sixty Egyptian honey samples was done
by Nour (1988); he found that the main
pollen sources of Egyptian honeys were
clover (
Trifolium alexandrinum
L.),
eucalyptus (
Eucalyptus
spp.),
Citrus
sp.
date palm (
Phoenix dactylifera
L.), maize
(
Zea mays
L.), sunflower
(
Helianthus
annus
L.) and faba bean
(
Vicia fabae
L.).
Cotton pollens were estimated at less than
1% of the total pollen found in market
cotton honey samples. Rateb (2005)
identified fifteen pollen types in honey
samples from Assiut region, most of them
(80%) from necteriferous plants. The
main polleneferous plants of pollen
spectrum in the studied honeys were
Zea
mays, Phoenix dactylifera
and
Casuarina
equistifolia
L. Also, the same author
reported that pollen density of honey
collected by Egyptian honey bee race
was 9.6, 13.6, 5.1 and 16.1 times more
than those collected by Italian,
Caucaseca, Carniolan hybrid and
Carniolan honey bee races, respectively.
Furthermore, pollen density varied and
depended on the collection locality.
Morever, Egyptian bee honey samples
were examined by El-Metwally (2015)
and fourteen types of pollen were
classified. Clover pollen was found in
high percentage (30.2%) while date palm
and umbliferus pollen were recorded in
considerable percentages as 13.20 and
9.39%, respectively. The present work
aims to evaluate and classify some
commercial honeys (trademark) produced
in different localities in Egypt according
to their botanical origin.
2. Materials and methods
Laboratory works of the present
investigation were carried out at Plant
Protection Department, Faculty of
Agriculture, Assiut University, Egypt
during 2017 season.
2.1 Collection of honey samples
A total of 16 commercial honey samples
with trademark were collected from
markets in different regions of Egypt
during 2017 season (Table 1) for
evaluation and classification.
2.2 Pollen content determination
The method recommended by the
International Commission of Bee Botany
(ICBB) for pollen analysis was followed
26
to determine pollen grains in honey
samples (Louveaux et al., 1978). Ten
grams from each honey sample were
mixed with 10 ml hot distilled water
(40°C), then the solution was centrifuged
for 20 min at 4500 rpm. The sediment
was smeared in two slides, mounted in
Fucsin-glycerin gel and examined
microscopically for pollen analysis as
described by Nair (1960). Pollen species
were identified with the help of reference
slides prepared from local flora during
the present study, in addition to relevant
literature (Rateb, 2005; Hussein, 1983).
Table 1: Commercial honey samples with trademark
were collected from markets in different regions of
Egypt for evaluation during 2017 season.
Code No.
Trademark
Locality of production
1
Isis
Different localities
2
Hero
Different localities
3
Shad Alaeyun
Different localities
4
Soud (1)
El-Manofia
5
Soud (2)
El-Manofia
6
Alryhan
El-Minia
7
Altemsah
Tanta, Cairo, Alex
8
Jana
Cairo
9
Aimtinan
Different localities
10
Khayer Zman (1)
Different localities
11
Khayer Zman (2)
Different localities
12
Dabur
El-Gharbia
13
Alhut
Alexandria
14
Alsafa
Different localities
15
Alassad
Cairo Alexandria
16
Alwaha
El-Minia
2.3 Pollen diversity determination
Total number of pollen grains in 10
microscopic fields was counted
separately as an indicator of pollen grain
density. Abundance percentages of pollen
species in each sample were calculated.
After pollen analysis, examined honey
samples can be defined as monofloral or
polyfloral honeys. If the honey sample
contains more than 45% of the total
pollen count from one plant species;
honey was considered as predominant
class, secondary (16-45%), important (3-
16%) and minor pollen types (less than
3%) as described by Maurizio (1975),
Bryant and Jones (2001) and Amaral et
al. (2003). However, classification of
citrus honey was a special case. A
minimum of 10% of citrus pollen grains
in honey sample was enough to consider
the honey as monofloral citrus honey as
described by Serra and Ventura (1995)
and Terrab et al. (2003). The diversity
index (α) was calculated according to the
following formula (Williams, 1947):
S =

 

Where S = number of species. N =
number of individuals and α = diversity
index.
3. Results and Discussion
Sixteen commercial honeys (trademark
honeys) were collected from different
Egyptian regions. After pollen analysis
of honey samples, twenty-seven pollen
species were identified. Classified pollen
species are belonging to 18 families
(Table 2). As shown in table (3), ten
pollen species were represented by
percentages ranged from 1 to 36%.
Seventeen pollen species which everyone
has less than 1% were represented 6.6%
from the total pollen count. Clover (
T.
alexandrinum
) pollen was the
predominant one (36.2%) among the rest
of other pollen types. Eucalyptus (
E.
globulus
), date palm (
P. dactylifera
),
alfalfa (
M. sativa
) and faba bean (
V.
fabae
) pollen grains had the following
percentages, 16.9, 12.6, 9.0 and 7.7%,
respectively. Date palm is known as
polleneferous sources only for honey bee
colonies. However, other field crops
which cultivated in large areas in
27
different regions of Egypt were the
sources of both nectar and pollen. Thus,
we considered that the commercial honey
produced in Egypt were mainly collected
from clover, alfalfa, faba bean,
eucalyptus and partially from medicinal
plants specially Umbelliferae, citrus and
sunflower fields.
Table 2: Classification of pollen grain species which isolated from commercial honey
samples.
No.
Common name
Scientific name
1
Acacia
Acacia arabica L.
2
Alfalfa
Medicagoi sativa L.
3
Apple
Malus domestica Borkh.
4
Anise
Pimpinella anisum L.
5
Aster
Aster tataricus L.
6
Basil
Ocimum sp.
7
Cabbage
Brassica oleracea var. capitate L.
8
Eucalyptus
Eucalyptus globulus Labill.
9
Capparis
Capparis spinosa L.
10
Casuarina
Casuarina equistifolia L.
11
Citrus
Citrus spp. L.
12
Coriander
Coriandrum sativum L.
13
Cotton
Gossypium spp.
14
Dahlia
Dahlia hybrid
15
Date palm
Phoenix dactylifera L.
16
Egyptian clover
Trifolium alexandrinum L.
17
Faba beans
Vicia faba L.
18
Gazania
Gazania joseph Gaertner.
19
Helichrysum
Helichrysum stoechas (L.) Moench
20
Iberis
Iberis gibraltarica L.
21
Lettuce
Lactuca sativa L.
22
Maize
Zea mays L.
23
Onion
Allium cepa L.
24
Sesame
Sesamum indicum L.
25
Sonchus
Sonchus oleraceus L.
26
Sunflower
Helianthus annuus L.
27
Zinnia
Zinnia elegans L.
To determine the nectar source of honey,
pollen grain percentages in every sample
were calculated and placed into one of
the following four pollen frequency
classes: predominant source (more than
45%); secondary source (16-45%);
important minor source (3-16%) and
minor source (less than 3%). As shown in
Table (4). The predominant pollen more
than 45% were recorded in 13 honey
samples. Eight predominant pollen
sources were identified in tested honey
samples. Faba bean pollen was recorded
in samples no. 8, 9 and 10 with 46, 67
and 84%, respectively. The sample no. 2,
4 and 13 contained clover pollen with 56,
57 and 97% respectively. Alfalfa pollen
had predominant percentages in three
samples no. 7, 12 and 15 with 54, 48 and
50%, respectively. Date palm, coriander,
eucalyptus and sunflower pollens were
recorded in samples no. 6, 11, 14 and 16,
respectively, in predominant percentage
> 45%. The sample no. 5 was the only
one contained citrus pollen with 19% and
it considered as citrus honey following
Serra and Ventura (1995) and Terrab et
al
.
(2003) as a special case; whereas, the
pollen frequencies of 1645% was noted
for 9 taxa. Important minor sources of
28
pollen which had percentages from 3 to
16% were noted for 16 taxa. The lowest
pollen frequencies <3% were noted for
12 taxa. The present data indicated that
faba bean plants were considered a good
source of nectar and pollen for honey bee
colonies in the early spring period in
many governorates in Egypt. Also, the
forage crops clover and alfalfa cultivated
in large areas are the important major
source of honey bees forage in summer
season.
Table 3: Representation % of different pollen species in supermarket honeys collected
from different regions of Egypt during 2017 season.
Common name
Total pollen count
Representation (%)
Clover
452
36.2
Eucalyptus
211
16.9
Date palm
158
12.6
Alfalfa
113
9.0
Faba bean
96
7.7
Capparis
52
4.2
Coriander
43
3.4
Sunflower
15
1.2
Citrus
15
1.2
Cabbage
13
1.0
Other pollen types (17 species)
82
6.6
Total of count (27 species)
1250
100
Table 4: Pollen spectrum of sixteen honey samples (Trademark) collected from different Egyptian
regions during 2017 season.
Code
No.
Tradem-ark
Number of
pollen Species
Predominant (>45%)
Secondary (16-45%)
Important (3-16%)
Minor (<3%)
1
Isis
9
Egyptian clover (32)
Gazania (12)
Iberis (2)
Capparis (26)
Helichrysum (12)
Cabbage (4)
Alfalfa (5)
Sunflower (4)
Basil (4)
2
Hero
6
Egyptian clover (56)
Alfalfa (13)
Cabbage (9)
Basil (9)
Capparis (6)
Helichrysum (6)
3
Shhad Alaeyun
3
Alfalfa (42)
Egyptian clover (32)
Sunflower (25)
4
Soud 1
9
Egyptian clover (57)
Cabbage (22)
Alfalfa (7)
Zinnia (2)
Iberis (6)
Sunflower (2)
Basil (2)
Dahlia (2)
Aster (0.78)
5
Soud 2
8
Citrus (19)
Date palm (37)
Eucalyptus (13)
Basil (3)
Lettuce (11)
Coriander (2)
Alfalfa (10)
Cabbage (6)
6
Alryhan
3
Date palm (65)
Eucalyptus (29)
Acacia (6)
7
Altemsah
4
Alfalfa (54)
Maize (15)
Cotton (15)
Sesame (15)
8
Jana
6
Faba bean (46)
Eucalyptus (23)
Sonchus (8)
Casuarina (3)
Apple (18)
Acacia (5)
9
Aimtinan
3
Faba bean (67)
Eucalyptus (25)
Alfalfa (7)
10
Khayer zman 1
4
Faba bean (84)
Cabbage (8)
Acacia (4)
Alfalfa (4)
11
Khayer zman 2
2
Coriander (87)
Alfalfa (13)
12
Dabur
6
Alfalfa (48)
Date palm (41)
Onion (4)
Eucalyptus (2)
Citrus (2)
Anise (2)
13
Alhut
4
Egyptian clover (97)
Capparis (1)
Sunflower (1)
Alfalfa (0.28)
14
Alsafa
2
Eucalyptus (72)
Date palm (28)
15
Al'assad
4
Alfalfa (50)
Maize (21)
Sesame (14)
Capparis (14)
16
Alwaha
3
Sunflower (71)
Egyptian clover (14)
Capparis (14)
29
According to the present results alfalfa
plants were the only pollen source
which was identified in 12 samples from
sixteen tested honey samples (75%)
followed by clover plants and
eucalyptus which everyone was found
in six samples (37.5%). Only two honey
samples (no. 1 and 3) were classified as
polyfloral honey. However, fourteen
samples were classified as monofloral
honey. Generally, it could be
summarized from the present results
that commercial Egyptian honeys could
be considered as monofloral honeys.
The sixteen honey samples were
divided into two groups (spring and
summer honeys) after botanical
classification of predominant pollen in
every sample. The spring honeys
included samples no. 5, 6, 8, 9, 10, 11,
12 and 14, while the summer honey
included samples no. 1, 2, 3, 4, 7, 13,
15 and 16. The diversity index (α) of
pollen species in spring and summer
honey samples were represented in
Tables (5 and 6).
Table 5: The diversity index (α) of pollen species in commercial spring honeys in Egypt
collected in 2017season.
Code No. of
sample
Predominant
pollen
No. of
pollen
species
Total count of
pollen grains
(no/10 fields)
Diversity
index (α)
5
Citrus
8
63
2.43
6
Date Palm
3
31
0.82
8
Faba bean
6
40
1.96
9
Faba bean
3
55
0.68
10
Faba bean
4
40
1.02
11
Coriander
2
46
0.43
12
Alfalfa
6
123
1.32
14
Eucalyptus
2
232
0.30
Total
14
640
2.53
Table 6: The diversity index (α) of pollen species in commercial summer honeys in Egypt
collected in 2017 season.
Code No. of
sample
Predominant
pollen
No. of
pollen
species
Total count of
pollen grains
(no/10 fields)
Diversity
index (α)
1
Polyfloral
9
57
3.01
2
Clover
6
32
2.18
3
Polyfloral
3
11
1.36
4
Clover
9
72
2.72
7
Alfalfa
4
13
1.97
13
Clover
4
349
0.63
15
Alfalfa
4
14
1.87
16
Sunflower
3
7
1.99
Total
16
555
3.08
The results indicated that the values of
the diversity (α) of spring honeys in
pollen grains were relatively low in
comparison with summer honey. The
diversity index (α) of spring honeys
ranged between 0.30 and 2.43. Eight
honey samples contained pollen species
from spring flowering plants and they
were monofloral honeys according to the
pollen classification. The results of
summer honey represented in Table (6)
showed that the values of α diversity
30
index ranged between 0.63 and 3.01.
Two samples, no. 1 and 3, recorded as
polyfloral honeys. The pollen analysis of
summer honeys indicated that the
diversity index (α) of pollen grain in
tested honey samples were relatively
higher than spring honeys. The present
results provides ageneral picture on a
nectar sources of Egyptian hones. From
pollen analysis, all tested honey samples
had a natural sources. Clover,
Eucalyptus, Alfalfa were the main
sources for honeybee colonies to produce
honey. Pollen diversity in honey samples
was higer in summer than in spring
honeys.
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