Grana

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Comparison of the melissopalynological,

antimicrobial and organoleptic composition of
honey samples from Apis mellifera L. and Apis
florea Gaertn. in Sudan

Seif Eldin A. Mohammed, Abdalmaged Osman Dafalla, Mogbel El-Niweiri &

Gamal E.B. El Ghazali

To cite this article: Seif Eldin A. Mohammed, Abdalmaged Osman Dafalla, Mogbel El-Niweiri
& Gamal E.B. El Ghazali (2023) Comparison of the melissopalynological, antimicrobial and
organoleptic composition of honey samples from Apis mellifera L. and Apis florea Gaertn. in
Sudan, Grana, 62:5-6, 342-351, DOI: 10.1080/00173134.2024.2311865

To link to this article: https://doi.org/10.1080/00173134.2024.2311865

Published online: 23 Feb 2024.

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Grana, 2023
Vol. 62, Nos. 5–6, 342–351, https://doi.org/10.1080/00173134.2024.2311865

Comparison of the melissopalynological, antimicrobial and

organoleptic composition of honey samples from Apis mellifera L. and
Apis florea Gaertn. in Sudan

SEIF ELDIN A. MOHAMMED 1, ABDALMAGED OSMAN DAFALLA1,

MOGBEL El-NIWEIRI1,2 & GAMAL E.B. EL GHAZALI 3
1
Department of Bee Research, Environment and Natural Resources & Desertification Research Institute, Khartoum, Sudan,
2
Unit of Bee Research and Honey Production, Faculty of Science, King Khalid University, Abha, Saudi Arabia, 3Medicinal
and Aromatic Plants Research Institute, National Centre for Research, Khartoum, Sudan

Abstract
Honey from Apis mellifera and Apis florea was collected from western and eastern regions of Sudan and investigated for
quantitative/qualitative pollen analysis, organoleptic characterisation, and antibacterial analysis. Nearly 29 different pollen
taxa belonging to 19 families were identified. Based on the frequencies determined: Leguminosae (≥ 10) was the
dominant family, Poaceae and Amaranthaceae were sporadic (9–3) whereas Cyperaceae was rare (< 3). Herbs
represented 52%, trees 38%, and shrubs 10%. Pollen from both cultivated and wild vegetation was identified. The taxa
identified are angiospermous and no gymnospermous species were encountered. Four anemophilous pollen types were
observed. Pollen of particular interests, such as Borassus aethiopum and Hyphaena thebaica, was also found. Calotropis
procera and Leptadenia spp. were also observed in the current investigation; these pollen taxa are of infrequent occurrence
in honey especially in Sudan and the nectar of these plants are claimed to contain poisonous substances. Pollen grains
of the Combretaceae with a unique heterocolpate sculpture were found only, in Al Leeri representing a clear
uniflorality of honeys in the area. The organoleptic analysis revealed statistically significant differences in pH (p ≤ 0.001),
ash (p ≤ 0.03), sucrose (p ≤ 0.04), and electrical conductivity (p ≤ 0.05). Respectively, honey from A. florea had higher
sodium (17.647, 6.753 mg/kg) and higher phosphorus (2.900, 2.080 mg/kg) than honey from A. mellifera. Whereas honey
from A. mellifera, had higher manganese (0.218–0.151 mg/kg) than honey from A. florea, both honeys exhibited similar
antibacterial potency against Staphylococcus aureus and Salmonella typhi.

Keywords: pollinia, heterocolpate, anemophilous, poisonous nectar, antemophily, meta-reticulate

Estevinho et al. (2008) reported that one of the
factors that affects the economic value of honey is
its designation of both botanical and geographical
origin. The most frequently used tool for determin-
ing botanical origin is the palynological content
and the pollen profile can be used to determine the
geographical region or origin (Louveaux et al.
1978). However, Hermosin et al. (2003) mentioned
that it is, sometimes, difficult to ascertain the exact
botanical taxa depending only on palynology. Pollen
analyses of honey (melissopalynology) and bee
loads are widely exploited to understand honeybee
foraging ecology and habitat, changes in honeybee
food sources, and the geographical region of the
hive location (Terrab et al. 2004). Melissopalynology
can predicts changes in honeybee nectar and pollen
sources and thus ascertain changes in habitat compo-
sition. Turnover of minor plant sources can reduce

Correspondence: Seif Eldin A. Mohammed, Department of Bee Research, Environment and Natural Resources & Desertification Research Institute, P. O. Box
6096 – Khartoum, Sudan. E-mail: seif_san@yahoo.com

(Received 20 October 2022; accepted 9 January 2024)

© 2024 Collegium Palynologicum Scandinavicum


the quality and quantity of the harvest and dramati-
cally affect the hive dynamics and honeybee popu-
lations. Any loss on honeybee populations limits the
pollination role of honeybees, reduce honey yields,
and potentially cause food shortages (Jones 2012).
As honeybee populations decline worldwide, melis-
sopalynology can help monitor changes in nectar
and pollen sources and may help understand the
reasons for these changes.
Honey is a natural super-saturated mixture of
organoleptic and bioactive compounds. It consists
mainly of an energy source (invert sugar and
sucrose) and minor components like vitamins, pro-
teins, enzymes, amino acids, organic and non-
organic acids, flavonoid, phenolic acids, traces of
lipids and metals.
The antimicrobial activity of honey has both been
scientifically documented and practically
implemented in medicine since ancient times. It has
proved antimicrobial activity towards nearly 60
aerobes and anaerobes bacterial strains, including
Gram-positive and Gram-negative bacteria (Taor-
mina et al. 2001).
The mineral composition of honey depends on the
metal constituents of flowers foraged by honeybees.
Mineral concentrations in different honey types
vary from 1.0 to 1.1% in Apis florea honey and 0.16
to 0.26% in A. mellifera honey (Al-Ghamdi et al.
2019). Its contents in honey can possibly provide
useful information for its environmental and botani-
cal identity (Celli & Maccagnani 2003). Therefore,
many researchers have successfully shown corre-
lations between the elemental profiles of honey and
its origin by applying various statistical procedures
on palynological, organoleptic, and sensory data
(Necemer et al. 2009).
The majority of research has been oriented
towards honey gathered by the European honeybee
Apis mellifera and, until now, relatively little attention
has been devoted to honey from A. florea. Apis melli-
fera L. and A. florea Fabr. in Sudan which have been
coexisting symbiotically for many decades. However,
most of the commercialised honey and wax in the
country comes from the European honeybee
A. mellifera, and the coexisting A. florea plays vital
roles in pollination of important cultivated crops. A
related previous study pointed out that both honey
types vary in their organoleptic characteristics due
to different nesting habitats, different ways of proces-
sing the honey, and foraging from different nectar
sources (Al-Ghamdi et al. 2019). Thus, this investi-
gation, aimed to focus on a detailed quantitative/
qualitative pollen analysis, some organoleptic proper-
ties, and establishing possible linkages between the
Honey samples from Sudan 343
antimicrobial potency of both honeys and the
foraged plant taxa.
Material and methods
Collection of honey samples
Ten samples were collected from different districts of
Sudan, with four samples originating from Apis melli-
fera honeycombs gathered from western Sudan
(Wadi Salih, Umm Dafoug, Al Leeri, and Umm
Dukhn) and one sample from eastern Sudan
(Gadarif). Similarly, five samples originating from
A. florea honeycombs were gathered from Al-Faw
(eastern Sudan). All samples originated from bees
reared in traditional hives or natural nests.
Sample preparation
Any plant debris, pollen grains, and broods were
removed away from the combs. Each honeycomb
was separately pressed between hands to extract the
honey and filtered via screen mesh (5 mm pore
size). Then the honey was left in a container over-
night to settle. Samples were saved in plastic contain-
ers and coded (WAS, UMDA, ELRI, UMDU, and
FA6–FA10).
Quantification of pollen
The preparation of slides and pollen counts followed
the standard method of Louveaux et al. (1978). The
pollen types encountered in the samples were
assigned to one of three frequency classes: dominant
pollen with ≥ 10 pollen types, sporadic with 9–3
pollen types and rare pollen with ˂ 3 pollen types
per slide.
Pollen identification
Three slides per sample were examined under a com-
pound light microscope (Zeiss 3136002552 Axio lab
A1; Carl Zeiss MicoImaging GmbH, Gottingen,
Germany). Taxa were identified using 100× phase
contrast (ph) objective, 10× eyepiece and phase con-
trast condenser (NA = 1.4). The pollen grains in each
honey sample were identified based on identification
keys, light microscopy (LM) and scanning electron
microscopy (SEM) micrographs and comparison
with the reference material provided by the previous
study on the pollen flora of the Sudan (El Ghazali
1993) and relevant studies (Bonnefille & Riollet
1980). All taxa were included in this investigation
344 S. E. A. Mohammed et al.
and no attempt was made to exclude pollen types
interpreted as wind pollinated (anemophilous).
Knowing the floristic composition of the various
areas under study, it was possible to distinguish the
pollen taxa in each pollen sample up to the species.
However, in some cases, identification was only poss-
ible to the generic or family levels. Unidentified
pollen grains were found in all samples but in low
numbers. Members of the families Poaceae, Cypera-
ceae and Combretaceae appear to possess similar
pollen grains and were clumped into a single pollen
type each. Pollen grains belonging to Amaranthaceae
including Chenopodiaceae were differentiated into
two categories: Amaranthus-type and Alternanthera-
type, based on the presence of meta-reticulate sculp-
turing (APG 2016; El Ghazali 2021). In the Astera-
ceae, only tricolporate pollen grains were
encountered, and were designated as Pulicaria-type
in view of the dominance of members of the Pulicaria
in the sampling areas. The accepted nomenclatures
of the plants identified in the honey samples were ver-
ified using ‘The Plant List’ published at (http://www.
theplantlist.org/, version 1:1 (2003)).
Organoleptic analysis of honey
Invert sugar and sucrose were processed according to
the FAO (1995) methods. The carbohydrate content
was determined according to Antia et al. (2006) by
the following formula:
Carbohydrate = 100%–(%Moisture + %Crude Fat
+%Crude Protein + %Ash).
Moisture, ash, pH, acidity, and fat were determined
according to the standard procedures of the Associ-
ation of the Official Analytical Chemists (AOAC
1990). Electrical conductivity was measured as
described by Mohammed and Babiker (2009). The
protein content was estimated with the micro-Kjeld-
hal procedure to determine the total nitrogen
content, and the protein content was calculated by
multiplying the nitrogen by a factor of 6.25.
Minerals were determined according to Hernan-
dez et al. (2005). Briefly, about 5.0 g of honey (ten
samples) was put in a pre-weighed porcelain crucible
and warmed first in a stove and then in a furnace. The
obtained ash was weighed, dissolved in 3 ml of con-
centrated nitric acid and diluted with de-ionised
water in a 25 ml conical flask. This solution was
employed to estimate the elements using atomic
absorption spectrophotometer (Perkin-Elmer 2380,
UK). The metal ions studied (potassium, sodium,
calcium, magnesium, copper, iron, manganese, and
zinc) were estimated by comparing the atomic spec-
troscopic signal for each honey sample with that for
a standard solution of the similar ion. Sulphur was
determined following the titrimetric method and
phosphorus was estimated according to the flame
photometry using the molybdovanadate method.
Antibacterial test
The cup-plate agar diffusion method as described by
Kavanagh (1972) was used with some minor modifi-
cations to assess the antibacterial activity of the honey
samples against the Gram-positive Staphylococcus
aureus (ATCC 25923) and the Gram-negative
Salmonella typhi (ATCC 25922). First 1 ml of the stan-
dardised bacterial stock suspension (108 –109 CFU/ml)
was thoroughly mixed with 100 ml of molten sterile
nutrient agar which was maintained at 45 °C. Then
20 ml aliquots of the inoculated nutrient agar were dis-
tributed into sterile Petri-dish plates. Next four cups
(10 mm in diameter) were cut using a sterile cork
borer (No. 4) and agar discs were removed. Each cup
was filled with 0.1 ml of the sample and allowed to
diffuse at room temperature for 2 h. The plates were
then incubated in an upright position at 37 °C for
18 h. Two replicates were carried out for each sample
against each of the test organisms. After incubation,
the diameters of the resultant growth inhibition zones
(in millimetres) were measured and averaged.
Results
Pollen frequencies and identification
The map depicts distribution of the locations of the
collected honey samples (Figure 1). Twenty-nine
different pollen types belonging to 19 families were
identified from the samples examined. Seventeen
types were identified to the plan species, six types
were identified to the generic taxa, three types were
identified to the family level, two types were recog-
nised in the Amaranthaceae, and one pollen type
was recognised in the Asteraceae (Table I). The
number of pollen taxa per sample varied from six in
Al Leeri to ten in Al-Faw, herbs (52%) and trees
(38%) shared dominance in the pollen taxa of these
samples, while pollen grains belonging to shrubs rep-
resented only (10%) of the total floristic composition
of the taxa identified (Table II). The dominant plant
family identified in these samples was the Legumino-
sae with five pollen types; four of these types were
identified to the plant species, whereas the Acacias
were grouped into a single type. Pollen grains of

Figure 1. Map of the honey collection sites.
both cultivated and wild vegetation were identified in
the samples under consideration. Cultivated plants
were represented by Hibiscus species (mainly
H. sabdarifa) and Sesamum alatum.
All taxa identified were angiospermous (23 dicoty-
ledons and two monocotyledons), and no gymnos-
permous or fungus spores were encountered in the
samples. Four non-melliferous (wind pollinated)
pollen taxa were identified. None of these types
were dominant, but Poaceae and Amaranthaceae
types were sporadic in five samples. Whereas Cyper-
aceae was rare, found only in honey from Gadarif.
Members of the family Leguminosae were identified
Honey samples from Sudan 345
as having the highest number of pollen types (five
taxa) whereas the remaining families were rep-
resented by one to two pollen taxa each. Samples
from Al Leeri were distinctly dominated by
members of the family Combretaceae, whereas the
Epilobium hirsutum pollen grains were of sporadic
occurrence (9–3), and the pollen grains of the rest
of the taxa were rare > 3 (Table I).
Organoleptic composition of honey
Honey collected and processed by the European hon-
eybee Apis mellifera and the dwarf honeybee A. florea
346 S. E. A. Mohammed et al.

Table I. Pollen types identified in the ten localities with their frequencies.

Taxa identified frequency

Sample location Sample code Dominant (≥ 10) Sporadic (9‒3) Rare (< 3)

Wadi Salih WAS Khaya senegalensis Alternanthera-type Abutilon panosum

Amaranthus-type
Justicia spp.
Monsonia senegalensis
Poaceae
Polygala spp.
Prosopis africana
Pulicaria-type
Strychnos spinosa
Syzygium guineese
Tribulus terristris
Umm Dafoug UMDA Acacia spp. Khaya senegalensis Combretaceae.
Dalbergia melanoxylon
Epilobium hirsutum
Hibiscus spp.
Polygala spp.
Pulicaria-type
Syzygium guineese
Al Leeri ELRI Combretaceae. Epilobium hirsutum Acacia spp.
Hibiscus spp.
Hyphaena thebaica
Umm Dukhn UMDU Acacia spp. Amaranthus-type. Combretaceae
Dichrostachys cinera
Ficus spp.
Hyphaena thebaica
Peristropha bicalyculata
Poaceae
Polygala spp.
Polygonium aviculare L.
Gadarif GARI Sesamum alatum Acacia spp. Cyperaceae
Dichrostachys cinera
Hibiscus spp.
Leptadinia spp.
Mimosa pigra
Pulicaria-type
Al Faw FA6–FA10 Epilobium hirsutum Combretaceae Borassus aethiopum
Justicia spp. Poaceae Calotropis procera
Hibiscus spp.
Hyphaena thebaica
Pulicaria-type
Syzygium guineese

was studied for their organoleptic properties ‘proxi-
mate composition and mineral contents’ (Tables
III, IV). Generally, the results were within the inter-
national honey standards (Codex 2001). There were
significant differences between the honey processed
by A. mellifera and the dwarf honeybee A. florea.
The values showed the statistical differences were
the pH (p ≤ 0.001), ash (p ≤ 0.03), sucrose (p ≤
0.04), and the electrical conductivity (p ≤ 0.05).
However, both honey types showed similar results
in moisture, protein, fat, carbohydrate, acidity, and
invert sugar (Table III).
Up to 31 different minerals essential for human
dietary and health care have been identified in honey
from various plants and geographical sources (Taor-
mina et al. 2001). However, the quantity and diversity
of minerals found in honeys are mainly dependent, on
their availability in the soil and absorbability by the
plants (Gonzalez-Miret et al. 2005). A total of ten
major and trace element were analysed. Their abun-
dances in descending order were as follows: potass-
ium, magnesium, sodium, calcium, sulphur,
phosphorus, iron, copper, manganese, and zinc
(Table IV). The comparison between the honeys
 Honey samples from Sudan 347

Table II. A list of the taxa identified from the ten honey samples and their habit.

No. Family Taxa identified Habit

1. Amaranthaceae Alternanthera-type Herbs

2. Amaranthaceae Amaranthus-type Herbs
3. Acanthaceae Juss. Justicia L. Herbs
4. Acanthaceae Peristropha bicalyculata (Retz.) Nees. Herbs
5. Apocynaceae Calotropis procera (Ait.) Dryand. Shrubs
6. Apocynaceae Leptadinia R.Br. Shrubs
7. Arecaceae Borassus aethiopum Mart Trees
8. Arecaceae Hyphaena thebaica (L.) Mart. Trees
9. Asteraceae Pulicaria-type Herbs
10. Combretaceae Combretaceae R.Br. Trees
11. Cyperaceae Cyperaceae Herbs
12. Geranaceae Monsonia senegalensis Guill. et Perr. Herbs
13. Leguminosae Acacia Mill. Trees
14. Leguminosae Dalbergia melanoxylon Guill. et Perr. Trees
15. Leguminosae Dichrostachys cinera (L.) Wight. et Arn. Trees
16. Leguminosae Mimosa pigra L. shrubs
17. Leguminosae Prosopis africana Guill. et Perr. Trees
18. Loganaceae Strychnos spinosa Lam. Trees
19. Malvaceae Abutilon panosum (G. Forst.) Schltdl. Herbs
20. Malvaceae Hibiscus L. Herbs
21. Meliaceae Khaya senegalensis (Desv.) A. Juss. Trees
22. Moraceae Ficus L. Trees
23. Myrtaceae Syzygium guineese (Willd.) DC. Trees
24 Onagraceae Epilobium hirsutum L. Herbs
25. Pedaliaceae Sesamum alatum Thonn. Herbs
26. Poaceae Barnhart Herbs
27. Polygalaceae Polygala L. Herbs
28. Polygonaceae Polygonum aviculare L. Herbs
29. Zygophyllaceae Tribulus terristris L. Herbs

produced by Apis melliera and A. florea honeybees
showed significant statistical differences in sodium
(p ≤ 0.000), phosphorus (p ≤ 0.012) and manganese
(p ≤ 0.031). Respectively, honey from the dwarf hon-
eybees showed higher sodium (17.647, 6.753 mg/kg)
and higher phosphorus (2.900, 2.080 mg/kg) than
honey from the European honeybees, whereas honey
from the later honeybees, showed higher manganese
(0.218–0.151 mg/kg) than honey from the former
respectively (Table IV).

Table III. Comparison of physical and chemical properties between Apis mellifera and Apis florea honey samples collected from different
areas of Sudan.

Apis mellifera Apis florea

honey (n = 5) honey (n = 5)
No. Factor Mean ± standard deviation Mean ± standard deviation p-Value

1. Moisture (%) 18.19 ± 3.49 20.21 ± 0.74 0.09 NS

2. Ash (%) 0.33 ± 0.22 0.17 ± 0.03 0.03 *
3. Protein (%) 0.93 ± 0.02 0.93 ± 0.01 0.72 NS
4. Fats (%) 1.16 ± 0.13 1.21 ± 0.12 0.38 NS
5. Carbohydrates (%) 79.37 ± 3.53 77.40 ± 0.86 0.10 NS
6. pH 4.21 ± 0.09 4.00 ± 0.05 0.00 **
7. Acidity (%) 0.46 ± 0.23 0.42 ± 0.08 0.61 NS
8. Invert sugars (%) 60.26 ± 4.48 61.54 ± 4.34 0.52 NS
9. Succors (%) 3.41 ± 1.25 5.20 ± 2.29 0.04 *
10. Electrical conductivity (mS/cm) 0.34 ± 0.33 0.56 ± 0.04 0.05 *

Note: NS = No significant difference, * = significant difference, ** = highly significant difference.

348 S. E. A. Mohammed et al.

Table IV. Comparison of mineral constituents (mg/kg) between Apis mellifera and Apis florea honey samples collected from different areas of
Sudan.

Apis mellifera honey (n = 5) Apis florea honey (n = 5)

No. Factor Mean ± standard deviation Mean ± standard deviation p-Value

1. Calcium 8.340 ± 4.398 8.108 ± 3.388 0.896 NS

2. Copper 0.224 ± 0.105 0.252 ± 0.134 0.617 NS
3. Iron 1.072 ± 0.502 0.904 ± 0.241 0.354 NS
4. Potassium 418.571 ± 339.972 350.558 ± 78.262 0.545 NS
5. Magnesium 52.809 ± 34.547 32.770 ± 2 5.293 0.156 NS
6. Manganese 0.218 ± 0 .070 0.151 ± 0.057 0.031 *
7. Sodium 6.753 ± 4.225 17.647 ± 5.236 0.000 **
8. Zinc 0.158 ± 0.046 0.191 ± 0.075 0.252 NS
9. Phosphorus 2.080 ± 0.308 2.900 ± 0.871 0.012 *
10. Sulphur 5.180 ± 3.547 3.060 ± 1.814 0.110 NS

Note: NS = No significant difference, * = significant difference, ** = highly significant difference.

Antibacterial activity Poaceae, the scarcity of distinctive morphological

Through agar diffusion, all honeys produced by both
honeybee species prohibited both the growth of the
Gram-positive (Staphylococcus aureus) and the
Gram-negative (Salmonella typhi) bacteria (Table
V). The largest inhibition zone was observed for Sta-
phylococcus aureus; the mean values were 30.0 and
30.1 mm for Apis mellifera and A. florea honeys,
respectively. Both honeys provided relatively
smaller inhibition zones against Salmonella typhi;
the mean values were 27.8 and 25.5 mm respectively;
for A. mellifera and A. florea honeys. However, there
was no statistical significance between both types of
honeys against both Gram-positive and Gram-nega-
tive bacteria investigated.
Discussion
Pollen taxa
The palynological analysis of honey samples from
western and eastern Sudan revealed the presence of
29 pollen morphs belonging to 19 plant families.
Almost half of the pollen types found (13/29 =
45%) were only identified to the plant species.
Those taxa identified only to the generic or family
levels, conceal a diverse pollen type in the samples
examined; e.g. in the samples from western Sudan
(four honey samples), Poaceae was represented in
the flora of the area by 206 species, Cyperaceae by
47 species, Amaranthaceae by 14 species and Com-
bretaceae by 11 species (Wickens 1976).
Pollen identification to the plant species is made
difficult by many factors; e.g. the lack of distinctive
morphological characters for the resolution and
identification up to the species level in many plants
(Rahl 2008; Salmaki et al. 2008), and in the
characters leads to the use of quantitative character-
istics instead of qualitative ones (Faegri & Iversen
1989). These quantitative characters vary according
to the chemical treatments used in the preparation
of the samples (Faegri & Iversen 1964) and were
regarded by Borsch (1998) as not suitable for diag-
nostic characters.
Examination with LM is regarded as the main
routine technique used in pollen analysis (Jones
2012). In many cases, LM, however, does not allow
easy distinction of the pollen morphs of the con-
sidered taxa due to the many similarities shared at
the magnification power limitation (Angelini et al.
2014).
Four anemophilous pollen morphs were identified
in the present study. These were Poaceae, Amar-
anthaceae (two types) and Cyperaceae. These ane-
mophilous taxa do not produce nectar (Friedman &
Barrett 2008) but produce large quantities of pollen
(Ghazoul 2004). In spite of the abundant amount
of pollen grains, they are not dominant in the
samples examined, in agreement with previous
studies (Bryant & Jones 2001). Pollen intrudes into
the hive via various means: (i) regurgitation of the
honeybees of the collected nectar and deposition of
it into the honeycomb cells; (ii) a honeybee while
grooming herself, can cause pollen accidentally to
fall into the open honeycomb cells; (iii) airborne
pollen from taxa not visited by honeybees can enter
the hive on air currents and fall into the open honey-
comb cells; (iv) pollen can fall into the honeycomb as
it is being removed by the beekeeper.
Borassus aethiopum and Hyphaena thebaica are
dioecious trees belonging to Arecaceae (Palmae).
These palms exhibit both anemophily and ante-
mophily (Salomon-Torres et al. 2021). They have

Table V. Comparison of antibacterial activity (inhibition zone mm) between Apis mellifera and A. florea honey samples collected from
different areas of Sudan.
No. Factor Bacterial Gram
1. Salmonella typhi _ 27.80 ± 3.64
2. Staphylococcus aureus + 30.00 ± 4.66
Note: NS = No significant difference.
characteristics related to anemophily such as the
reduction in attractive flower parts and the pro-
duction of large numbers of pollen grains (Bertin
1989), and characters linked to entemophily such
as fragrance, production of nectar, and the flowers
being relatively open and easily accessible by insects
(Silberbauer-Gottsberger 1990). In addition, palms
are attractive to insects through various incentives
such as food rewards of pollen grains/nectar and
shelter and oviposition sites (Barfod et al. 2011).
These peculiar pollination characteristics of B. aethio-
pum and H. thebaica highlighted that they were of fre-
quent occurrence in honeys harvested from their
natural habitats (Dukku 2013; Orijemie 2017).
Calotropis procera and Leptadenia spp. family Apoc-
ynaceae, subfamily Asclepiadaceae; (APG 2016),
identified in this investigation, are of particular inter-
est because of their specialised floral morphology
and their pollen grains are packed into pollinia. The
latter are mainly carried by the mouthparts or legs
of honeybees, and although they were encountered
in various honey samples (Maclvor et al. 2017),
they are of infrequent occurrence especially in
Africa. In C. procera and Leptadenia spp., honeybees
are attracted by the nectar secreted in the stigmatic
chamber of their flowers (Eisikowitch 1985) and
the pollinia are accidentally collected. The nectar of
these plants is claimed to constitute poisonous com-
pounds (Al Sulaibi et al. 2021).
Pollen grains of the Combretaceae possess a
unique heterocolpate and tricolporate pollen grains
(El Ghazali et al. 1998). Heterocolpate pollen
grains found only, in Al Leeri represented a clear
dominance (unifloral) with respect to the low
number of taxa identified in the honey sample.
Persano Oddo (1995) outlined that palynological
examinations (quantitative and qualitative) are not
sufficient to establish the uniflorality of the honey
sample; a reliable diagnosis may be achieved by paly-
nological examination in addition to organoleptic
analysis and physiochemical properties (Escuredo
et al. 2019). Therefore, although the main objective
of this investigation focused on palynology, a few
Honey samples from Sudan 349
Apis florea
Apis mellifera honey (n = 5) honey (n = 5)
Mean ± standard deviation Mean ± standard deviation p-Value
25.50 ± 5.72 0.29 NS
30.10 ± 3.34 0.95 NS
diversions were made to include the organoleptic
analysis of the samples. Moreover, heterocolpate
pollen grains encountered in these honey samples
were assigned as Combretaceae according to the
dominant plants taxa belonging to this family in the
studied area. Heterocolpate pollen grains exist in a
wide array of families including at least some
genera of the families Acanthaceae, Boraginaceae,
Lythraceae, Melastomaceae, etc. (El Ghazali & Krzy-
winski 1989). Hence, the identification of the source
of vegetation of these heterocolpate pollen grains
may not be reliably identified, and it is far from
being ascribed as a honey of unifloral origin.
Organoleptics
The results obtained by the organoleptic analysis of
the samples were within the specification limits of
honey standards. The variation between Apis florea
and A. mellifera honeys in the organoleptic and
sensory characteristic have been reported by several
researchers (de Almeida-Muradian et al. 2013;
Krishnasree & Ukkuru 2015). Thus, the variations
between both honeys in pH, ash, sucrose, and electri-
cal conductivity corroborate the previous findings
(Al-Ghamdi et al. 2019).
Antibacterial activity
In view of the results that showed no statistical sig-
nificance between Apis mellifera and A. florea
honeys against Staphylococcus aureus and Salmonella
typhi; it is important to note that all pollen spectra
observed in the dwarf honeybee honey, were rep-
resented in A. mellifera honey. Two pollen types
(Calotropis procera and Borassus aethiopum) were
observed particularly in A. florea honey. Although
in this investigation, the antibacterial compounds in
the honey samples were not determined, the differ-
ences observed in the antibacterial activity of both
honey types could be attributed in part to the differ-
ent plant taxa foraged by the bees which differ in the
composition of the active compounds. A literature
350 S. E. A. Mohammed et al.
survey of antibacterial capacities in honeys from
different plant taxa indicated clearly that the phyto-
chemical composition of honey was responsible for
the degree of bacteriostatic and bactericidal capacity
(Lusby et al. 2005).
Disclosure statement
No potential conflict of interest was reported by the author(s).
ORCID
Seif Eldin A. Mohammed http://orcid.org/0000-0001-
5632-1194
Gamal E.b. El Ghazali http://orcid.org/0000-0003-
2040-7054
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