Yahya Al Naggar1,2, Anja Vogt2, Garry Codling2, El Saied Naiem1,
Mohamed Mona1, Amal Seif1 and John P. Giesy2
1 Department
2 Toxicology
of Zoology, Faculty of Science, Tanta University 31527, Tanta, Egypt.
Centre, University of Saskatchewan, 44 Campus Drive, Saskatoon, SK, S7N 5B3,
Canada.
 Provide an overview about OPs
residues in honey, pollen and bees
in Egypt.
 Characterization of 14 current use OPs
in Bee Matrices
 Estimate potential hazard of OPs to
bee hives.
 Are there OPs residues in samples of
honey, pollen and bees collected from
Egypt during spring and summer 2013?
 Are
there
concentrations
differences
between
spring
in
and
summer?
 Is there a potential hazard from direct
and/or dietary exposure to these OPs
to bee colonies?
Toxicology Centre
Oxon
Diazinon
Malathion
Chlorpyrifos
Methyl
Profenofos
Ethoprop
Fenamiphos
Dicrotophos
Dimethoate
Coumaphos
Title or place of presentation
Dichlorvos
Phorate
Fenthion
Date of presentation
Toxicology Centre
 35% of all arable crops need honey bees
to pollinate and 90% of wild plants need
bees too [1].
 Egypt has about 1.3 million hives[2] :
 7,700 are mud hives
 270,000 beekeepers
 Stats on Bee keeping in Egypt are
limited but it is considered to be one of the
most influential in the Middle East and
Africa [3].
Title or place of presentation
Date of presentation
 Colony Collapse Disorder (CCD):
 Normally 15% of hives fail over
winter, today its ~30%
 Egypt has reported incidence of
CCD along River Nile [4].
 To
date,
no
clear
single
explains colony loss in bees [5]
factor
 Parasites and Pathogens.
 Malnutrition and Loss of habitat.
 Genetic factors and Beekeeping practice.
 Pesticides
 Combination of all factors.
Toxicology Centre
 There are no clear guidelines for use in Egypt
 OPs have become the major compound group used in pest control, over
80% of all insecticide used in Egypt are OPs [6]
World market share of insecticides, by class (2008)
Title or place of presentation
Date of presentation
 Study areas
Map of study sites (S1-15) in the Nile Delta governorates of Egypt.
[7]
Quick
Easy
Cheap
Effective
Rugged
Safe
Weigh 3g of matrix in a 50 ml
falcon tube
Fortified with 100
µL of Dimethoate
d6 (PCS)
Extraction
Add 27 mL of extraction solution
(44% deionized water, 55% acetonitrile,
and 1% glacial acetic acid).
Salting-out
Add 6 g MgSO4 + 1.5 g NaAc,
centrifuge (4000 rpm, 10 min)
Clean-up
C18 SPE cartridges + PSA
Quantification
LC-MS/MS
Shaking (1 min)
Assessment of Hazard
The hazard characterization scheme applied was based on methods
proposed by the USEPAs Office of Chemical Safety and Pollution
Prevention for assessing risks of foliar sprayed pesticides.[8]
 Uncertainty was assessed by calculating :
Best case
In the best case scenario
concentrations of OPs less
than the limit of
detection (LOD) were set
to zero (0.0)
Worst
case
In the worst case
scenario concentrations
of OPs less than the LOD
were set to LOD
Total daily intake (TDI) of pesticides received by bees via food was
calculated (Equation 1):
TDI = OP concentration detected in honey and pollen x Proposed
total food consumption rate of adult workers (292 mg d-1)

(1)
HQs for individual OPs were calculated (Equation 2):
HQs = TDI (honey + pollen )/ Acute oral LD 50
(2)
 HQs based on lethality of bees exposed to OPs directly were estimated
(Equation 3):
HQs = OPs detected in bee body burden /Acute oral LD 50
(3)
Organophosphorous pesticides (OPs) (ng/g, wm) detected in honey, pollen and
honey bees
Spring
No. of
positive
samples
Mean
Conc.
Dimethoate
4/19
3.36
Dichlorvos
1/19
2.8
OP
Honey
Summer
Mean
Conc.
Diazinon
1/20
0.25
Dicrotophos
Profenofos
1/20
2/20
0.34
0.28
Chlorpyrifos
1/20
3.27
OP
Malathion
Profenofos
6/14
5/14
0.61
1.45
Diazinon
Malathion
5/17
6/17
0.16
2.91
Chlorpyrifos
1/14
23.63
Dimethoate
1/17
0.43
Profenofos
16/17
11.56
Chlorpyrifos
11/17
26.44
Ch. Methyl
Fenthion
Diazinon
2/17
217
5/18
17.5
5.74
0.19
Fenamiphos
2/18
0.36
Profenofos
2/18
6.85
Chlorpyrifos
1/18
31.04
Pollen
Bees
No.
of positive
samples
Diazinon
4/16
0.42
Chlorpyrifos
1/16
32.72
Toxicology Centre
Proportion of positive samples (%)
Diazinon
100
Chlorpyrifos
90
Malathion
80
Profenofos
Spring honey
Summer honey
Spring pollen
70
60
50
40
0
10
20
30
Frequencies of detections (%) (n=104)
Summer pollen
Spring bees
Summer bees
30
20
10
0
Percentages (%) of positive samples of OPs detected in honey, pollen and honey bees
Title or place of presentation
Date of presentation
Tier-1 HQs for lethality of bees exposed to OPs in honey and pollen consumed by
bees during spring
OP
Ref.LD50
(ng. bee-1)
Total Daily Intake (TDI) (ng. bee-1 day-1)
Honey
Honey
Pollen
Pollen
(best case) (worst case)
(best case)
(worst case)
0.00
0.04
0.00
0.02
HQs (honey & pollen)
best
case
0.000
worst
case
0.000
Diazinon
168.0
Dicrotophos
137.6
0.00
2.10
0.00
1.26
0.000
0.024
Ethoprop
5560.0
0.00
0.09
0.00
0.17
0.000
0.000
Malathion
335.2
0.00
0.13
0.18
0.18
0.001
0.001
Dimethoate
129.6
0.98
0.98
0.00
3.56
0.008
0.035
Coumaphos
14390.0
0.00
0.13
0.00
0.13
0.000
0.000
Phorate
196.0
0.00
0.01
0.00
0.58
0.000
0.003
Dichlorvos
218.4
0.82
0.82
0.00
0.16
0.004
0.004
Fenamiphos
1870.0
0.00
0.04
0.00
0.11
0.000
0.000
Profenofos
95.0
0.00
0.04
0.42
0.42
0.004
0.005
Chlorpyrifos
67.8
0.00
0.04
6.90
6.90
0.102
0.102
Ch. methyl
110.0
0.00
0.82
0.00
0.91
0.000
0.016
Fenthion
251.2
0.00
0.43
0.00
1.17
0.000
0.006
0.1
10
0.2
5
Sum
Margin of Exposure (MOE)
Tier-1 HQs for lethality of bees exposed to OPs in honey and pollen consumed by
bees during summer.
pesticide
Ref.LD50
(ng. bee-1)
Total Daily Intake (TDI) (ng. bee-1 day-1)
Honey
(worst case)
0.07
Pollen
(best case)
0.05
Pollen
(worst case)
0.05
HQs
(honey & pollen)
best
worst
case
case
0.00
0.00
Diazinon
168.0
Honey
(best case)
0.07
Dicrotophos
137.6
0.10
0.10
0.00
1.26
0.00
0.01
Ethoprop
Malathion
Dimethoate
Coumaphos
Phorate
Dichlorvos
5560.0
335.2
129.6
14390.0
196.0
218.4
0.00
0.00
0.00
0.00
0.00
0.00
0.09
0.13
0.99
0.13
0.01
6.29
0.00
0.85
0.13
0.00
0.00
0.00
0.17
0.85
0.13
0.13
0.58
0.16
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.00
0.01
0.00
0.00
0.03
Fenamiphos
1870.0
0.00
0.04
0.00
0.11
0.00
0.00
Profenofos
95.0
0.08
0.08
3.38
3.38
0.04
0.04
Chlorpyrifos
67.8
0.95
0.95
7.72
7.72
0.13
0.14
Ch. methyl
Fenthion
110.0
251.2
0.00
0.00
0.82
0.43
5.12
1.68
5.12
1.68
0.05
0.01
0.05
0.01
0.2
5
0.3
3.3
Sum
Margin of Exposure (MOE)
Tier-1 HQs for lethality of bees from direct exposure to OPs during spring and
summer.
Conc. (ng g-1, wm)
Ref.LD50
(ng. bee-1)
spring
summer
168
0.42
Dicrotophos
137.6
Ethoprop
HQs
0.19
spring
best
worst
case
case
0.000
0.000
summer
best
worst
case
case
0.000
0.000
ND
ND
0.000
0.002
0.000
0.002
5560
ND
ND
0.000
0.000
0.000
0.000
Malathion
335.2
ND
1.11
0.000
0.000
0.000
0.000
Dimethoate
129.6
ND
ND
0.000
0.007
0.000
0.007
Coumaphos
14390
ND
ND
0.000
0.000
0.000
0.000
196
ND
ND
0.000
0.000
0.000
0.000
Dichlorvos
218.4
ND
ND
0.000
0.001
0.000
0.001
Fenamiphos
1870
ND
0.36
0.000
0.000
0.000
0.000
95
ND
6.85
0.000
0.000
0.006
0.006
67.76
32.72
31.04
0.039
0.039
0.037
0.037
110
ND
ND
0.000
0.003
0.000
0.003
251.2
ND
ND
0.000
0.001
0.000
0.001
0.04
25
0.1
10
0.04
25
0.1
10
OP
Diazinon
Phorate
Profenofos
Chlorpyrifos
Ch. methyl
Fenthion
Sum
Margin of Exposure (MOE)
0.6
Best case
Worst case
Summer
Spring
0.5
0.4
0.3
0.2
0.1
0
Spring
Summer
Comparison of HQs for lethality of bees from direct exposure or and
dietary exposure to OPs .
 Samples collected in summer were more contaminated with OPs.
 Pollen was most contaminated with OPs.
 Profenofos, chlorpyrifos, malathion and diazinon were the most
frequently detected OPs.
 Coumaphos, the most common OP used directly on hives was not
detected.
 The OPs investigated
pose a minimal threat to bees in Egypt at
measured concentrations, higher-tier assessments (Tier II and Tier III)
were not indicated by the results of this study.
 Literature derived concentrations of OP insecticides daizinon, malathion,
profenofos and chlorpyrifos in honey were compiled and were tested as a
mixture at two different concentrations, the median and the 95th percentile
for best and worst case to assess their effects on:
1.
Learning behavior and memory of bee foragers after 24 h
exposure.
2.
Survival, AChE, detoxification and immunity genes transcripts
of honey bee workers after 5 days exposure.
[1] Klein, A.M., Vaissie`re, B.E., Cane, J.H., Steffan-Dewenter, I., Cunningham, S.A., Kremen, C.,
Tscharntke, T., 2007. Importance of pollinators in changing landscapes for world crops. Proc. R. Soc. B
274,303–313
[2] The first international Forum for the Egyptian Beekeepers, 2009. (For the Best Future for Beekeeping
Industry). http://pcela.rs/1st_forum_Egyptian.htm
[3] http://www.beekeeping.com/articles/us/arab_countries.htm
[4] Fairbrother, A., Purdy, J., Anderson, T., Fellk, R., 2014. Risks of Neonicotinoid Insecticides to
Honeybees. Environ. Toxicol. Chem. 33(4), 719-731.
[5] Hassan, A.R., 2009. “Proceedings of the 4th COLOSS Conference”.
http://www.unep.org/dewa/Portals/67/pdf/Global_bee_colony_disorder_and_threats_insect_pollinators.pdf.
[6] Mansour, S.A., 2004. Pesticides exposure- Egyptian scene. Toxicol. 198, 91–115
[7] Al Naggar, Y., Vogt, A., Codling, G., Naeim, E., Mona, M., Seif, A., Giesy, J (In press).
Organophosphorus insecticides in honey, pollen and bees (Apis mellifera L.) and their potential hazard to
bee colonies in Egypt. Ecotoxicology and Environmental Safety.
[8] USEPA, 2012. White Paper in Support of the Proposed Risk Assessment Process for Bees. Office of
Chemical Safety and Pollution Prevention, Office of Pesticide Programs, Environmental Fate and Effects
Division, United States Environmental Protection Agency, Washington, DC.
Contact
PhD researcher,
Toxicology center, Saskatchewan University
44 campus drive, Saskatoon, S7N 5B3
Phone: 3067156328
Assistant Lecturer
Zoology Department, Faculty of Science
Tanta University 31527
Egypt.
E-mail:Yehia.elnagar@science.tanta.edu.eg
:Yaa007@mail.usask.ca
https://www.researchgate.net/profile/Yahya_Al_Naggar
https://usask.academia.edu/yahyaAlNaggar