Transcript What is Bee Venom?

MANAGEMENT OF MULTIPLE SCLEROSIS: APITHERAPY
IN THE NAME OF GOD
Is bee venom having antibacterial activity?
Ahmed G. Hegazi and Samira Shawky
ahmed@ahmedhegazi com
[email protected]
www.ahmedhegazi.com
National Research Center, Dokki,
Giza, Egypt
The bilogy of honey bee(apis mellifera)
Kingdom: Animalia
Phylum: arthropoda
Class: insecta
Family: apidae
Genus: apis
Species: apis mellifera
Apis mellifera
(honey bee)
Historic Record
• Humans eventually began providing
cavities for honey bees to nest in.
• Earliest records are Egyptian
Egyptian beekeeping
Harvesting honey combs (right) and packing honey (left)
ca. 1450 BC, West Bank, Luxor
The earliest
record of man’s
interest in honey
comes from
cave paintings
like this one in
Spain circa 4000
BC.
7
Prehistoric relations
• Prehistoric records show a honey
bee/human connection going back
6,000 BP
• Humans were essentially another
predator of the honey bee
Smoking the hive
Motopo Hills, Zimbabwe, ca. 10,000 yrs ago
Mesolithic cave painting
Cueva de la Arana, Bicorp, Spain
BEE VENOM
Insect Sting
= Injection of up to 100 g protein
(or 0.10 mg or 0.0001 gram)
What is Bee Venom?
• Bee venom is a complex composition of
enzymes, proteins and amino acids.
• It is a colorless clear liquid, with a sweet
taste and a little bitter.
• It is soluble in water, insoluble in alcohol
and ammonium sulphate.
• If is comes in contact with air, it forms,
opaque or grayish-white crystals.
Bee venom
pH 5-5.5
A:liquid
Colorless
sharp-bitter tasting
B:Dried
Yellowish brown
LD50:2.8mg/kg (IV) (In mice)
Cold resistance
Heat resistance (when dry)
Bee venom composition
PEPTIDES
melittin (family)
melittin F
apamin
mast-cell degranulation peptide
401 (MCD)
secarpin
tertiapin
adolapin
protease inhibitor
procamine A, B
minimine
cardiopep
ENZYMES
phospholipase A2
hyaluronidase
acid phosphomonoesterase
glucosidase
lysophospholipase
ACTIVE
AMINES
histamine
dopamine
norepinephrine
leukotriens
NON-PEPTIDE
COMPONENTS
carbohydrates like:
Glucose
Fructose
LIPIDS
6 phospholipids
AMINO-ACIDS
r-aminobutyric acid
B-aminoisobutyric acid
Dried bee venom composition
MOL. Wt.
% (Dry Venom)
Hyaluronidase
38,000
1.5-2.0
Phospholipase A2
19,000
10-12
Glucosidase
170,000
0.6
Acid
Phosphomonoesterase
55,000
1.0
Lysophospholipase
22,000
1.0
ENZYMES
COMPONENT
MOL. Wt.
% (Dry
Venom)
PEPTIDES
Melittin
2,840
40-50
Apamin
2,036
2-3
MCD-Peptide 401
2,588
2-3
Adolapin
11,500
1.0
Protease inhibitor
9,000
< 0.8
Secarpin
0.5
Tertiapin
0.1
Melittin F
0.01
Procamine A, B
1.4
Minimine
Cardiopep
6,000
2-3
< 0.7
ACTIVE AMINES
Histamine
Dopamine
0.13-1.0
Norepinephrine
0.1-0.7
NON-PEPTIDE
COMPONENTS
Carbohydrates:
Glucose &
Fructose
< 2.0
B.V. SUBSTANCES AND THEIR EFFECTS:
•radioprotective activity;
•mastocytolitic;
•histamine release;
•blood pressure depressants
Phospho
•antigenic properties;
lipase A
it is the major BV allergen ;
(enzyme)
•antagonistic effect on staphylococic
alfa-toxin and tetanus toxin;
•antitumoural effect
•acts on biological membranes
B.V. SUBSTANCES AND THEIR EFFECTS:
•selectively attacks tissue
hyaluronic acid polymers;
•increase the capillary
permeability (Neumann and
Hyaluronidase Habermann);
•immune response and tissuespread properties;
•antigenic;
•anaphylactogene
Apamin (a
•antigenic and;
polypeptide with 18 •anti-inflammatory properties
amino acids)
B.V. SUBSTANCES AND THEIR EFFECTS:
Mast Cell
Degranulating
peptide
Adolapin
In many animal studies, in
comparison studies with
hydrocortisone, this peptide was
100 times more potent as an antiinflammatory agent in suppressing
the development of adjuvantinduced arthritis. (Simics p 13) &
quot.
•analgesic (Shkenderov, 1982);
•anti-inflammatory (Shkenderov,
1982)
B.V. SUBSTANCES AND THEIR EFFECTS(continued)
•antibacterial;
•antifungal;
•anti-lyme disease (in vitro experiment)
Melittin
(a polypeptide
also consisting of
26 amino acids
which represents
40-60% of the
bee venom)
•antitumoural;
•central nervous system inhibitory;
•block nerve muscle and ganglial synapses;
•contraction of the striated and smooth muscles;
•histamine releasing;
•mastocytololysic;
•radio protecting (against X-irradiation; study on mice, Shipman and Cole,
1967);
•vascular permeability increasing;
•haemolysis;
•lowers blood pressure;
•anti-inflammatory;
•mellitin (which represents 40-60 % from the B.V. substances) has no antigenic
properties (Orlov); otherwise, according to Artemov, the bee enemies would
have gotten a specific immunity;
•stimulate the pituitary - adrenal axis to release both cathecolamines and cortisol
(Brooks et al.);
•increase plasma cortisol levels
•acts on biological membranes
Presently, it is one of the most potent anti-inflammatory agents known, and it
can be useful in treating arthritis and rheumatism.
B.V. SUBSTANCES AND THEIR EFFECTS(continued)
Cardiopep
•increase both the force of
contraction (beta-adrenergic) and
the heart rate with little or no effect
on coronary circulation (Brooks et
al.);
•anti-arrhythmic properties (Brooks et
al.);
•stimulate the pituitary - adrenal axis
to release both cathecolamines and
cortisol (Brooks et al.)
Bee Venom
• Uses
–As Pure Bee Venom for use in
desensitization
–As treating a variety of
neurological disorders.
Biomedical Applications:
Folk remedies and anecdotal
reports suggest that honey bee
venom has beneficial effects for
patients with:
Multiple Sclerosis
Arthritis
Infectious Diseases
RESEARCH FOCUS:
Does Bee Venom (BV) have
useful antimicrobial activity?
• Antibacterial activity of snake, scorpion and
bee venoms: a comparison with purified
venom phospholipase A2 enzymes
• Perumal et al., 2007
• indicate that have significant antibacterial
effects against gram (+) and gram (−) bacteria,
which result of the primary antibacterial
components of the PLA2 enzymes.
Atomic Force Microscopy Study of the Effect of
Antimicrobial Peptides on the Cell Envelope of
Escherichia coli
M. Meincken1, D. L. Holroyd2 and M. Rautenbach2, 2005
• bee venom melittin on Escherichia coli as the
target cell were studied by atomic force
microscopy (AFM).
• peptides are lytic to E. coli, peptide causes
distinct morphological changes in the outer
membrane and in some cases the inner
membrane,
• probably as a consequence of different
mechanisms of action..
Peptides
Volume 33, Issue 1, January 2012, Pages 18–26
Toxicity study of antimicrobial peptides from wild bee
venom and their analogs toward mammalian normal
and cancer cells
Jiřina Slaninová, Veronika Mlsov. Hilda Kroupová, Lukáš AlánTereza
Tůmová, Lenka Monincová, Lenka Borovičková, Vladimír Fučík, Václav
Čeřovský
isolated and characterized remarkable
antimicrobial peptides (AMPs) from the
venom reservoirs of wild bees.
display high antimicrobial activity against
Gram-positive and -negative bacteria,
antifungal activity
Modulation of the Activity of Secretory
Phospholipase A2 by Antimicrobial
Peptides
Hongxia Zhao and Paavo K. J. Kinnunen, 2003
• the concerted action of antimicrobial
peptides and sPLA2 could improve
the efficiency of the innate response
to infections.
Antimicrobial Activity of Honey Bee
Venom against Select Infectious Fish
Pathogens
North American Journal of Aquaculture Volume 75, Issue 3, 2013
• Sang Mi Hana, Kwang Gill Leea, Kwan Kyu
Parkb & Sok Cheon Pakc
BV inhibits the growth and survival of
bacterial strains and that BV may be a
useful complementary antimicrobial agent
against fish pathogenic bacteria.
• A strong antibacterial activity of bee
venom against both Gram negative and
Gram positive bacteria has been
reported [Perumal et al., 2007].
• Nakatuji et al. [2009] also reported that
bee venom could control the growth of
Staphylococcus aureus.
• Bee venom exhibited antibacterial
activities against skin bacteria such
as
• Propionibacterium acnes,
• Staphylococcus epidermidis and
• Streptococcuspyogenes pyogenes
[Han et al., 2010].
Antibacterial activities of bee venom, propolis, and
royal jelly produced by three honey bee, Apis mellifera
L., hybrids reared in the same environmental conditions
Annals of Agric Sci., Moshtohor, Vol. 45, No. 2: 895-902 pp., 2007.
Khaled M. Attalla , Ayman A. Owayss and Karem M. Mohann
There Gram (+) bacteria; Staphylococcus aureus, Bacillus subtilis
and Listeria monocytogenes and two Gram (-); Escherichia coli and
Salmonella enteritidis were used
• Bee venom seemed to be the most active followed by propolis
then royal jelly.
• Gram (+) bacteria was more sensitive to these products than
Gram (-) ones.
• The use of these, natural, cheap and safe bee products as
alternative food preservatives and in some pharmaceutical
application is promising,
Sung-Won Park et al., Altern Integ Med 2013, 2:10
Antimicrobial activities of honey bee venom against
pathogens isolated from clinical bovine mastitis in
Korea
• In this study, the concentration of three major
active components had a little frustration among
three venom products,
• It was clearly demonstrated that the honey bee
venom inhibited the growth of seventeen gram
positive bacteria strains and two gram negative
strains isolated from bovine mastitis in Korea.
However, the pore forming on cell membrane
wasn’t observed by using bee venoms and
sensitive gram positive bacteria.
African Journal of Microbiology Research Vol.
5(18), pp. 2765-2772, 16 September, 2011
N. S. Surendra, G. N. Jayaram and M. S. Reddy
• Antimicrobial activity of crude venom extracts in
honeybees (Apis cerana, Apis dorsata, Apis florea)
tested against selected pathogens
The selected bacteria and fungal species were Pseudomonas
aeruginosa, Klebsiella pneumoniae, Escherichia coli, Xanthomonas
subtilis, Proteus vulgaris, Salmonella typhimurium and Candida
albicans
• The results showed that, ABV has significant
antimicrobial effects and could be a potential
alternative antibiotic.
Antimicrobial Effect of Melittin Isolated from Syrian
Honeybee (Apismellifera) Venom and its Wound
Healing Potential
Int. J. Pharm. Sci. Rev. Res., 21(1), Jul – Aug 2013; n° 54, 318-324
Omran Alia, Massouh Laila , Al-DaoudeAntonious
• In this study, bee venom (BV) was collected using electric
shock method.
• Melittin, was isolated and identified using RP-HPLC C18
column and MALDI-TOF-MS analysis.
• The obtained melittin exhibited a potent antibacterial activity
particularly against Gram-positive bacteria as its MIC was
12.5µg/ml for Listeria monocytogenes compared with
200µg/ml for Yersinia kristensenii (a Gram-negative
bacterium) indicating that melittin has significant
antibacterial effects.
World Applied Sciences Journal 30 (3): 266270, 2014
Evaluation of the Antibacterial Activity
of Bee Venom from Different Sources
Ahmed Hegazi, Amr M. Abdou, Sherein. Abd ElMoez and Fyrouz Abd Allah
• evaluate the antibacterial activity of bee venom from different
sources against selected Gram-positive and Gram-negative
bacterial strains of medical importance.
• Three different samples of bee venom (BV) collected from the
honeybee Apis mellifera as well as whole bee extract were
assessed for their potential use as antibacterial agents against
five pathogenic bacterial strains
•
•
•
•
•
•
•
pathogenic bacterial strains including
Staphylococcus aureus,
Streptococcus pyogenes,
Klebsiella pneumoniae,
Escherichia coli
Pseudomonas aeruginosa.
Both bee venom and whole bee extract exhibited
antibacterial activity against all five bacterial strains
with different levels according to the type.
• The results of the current study indicate that BV inhibits
the growth and survival of bacterial strains and that BV
can be used as complementary antimicrobial agent
against pathogenic bacteria
Table (1): Influence of bee venom on growth
inhibition of different bacteria
Treatment
Bacterial
Normal growth
Apitox
Vacsera
Ethanolic sac
extract
Whole Bee
extract
Tetracycline
(50ug)
Staphylococcus Streptococcus
aureus
pyogenes
1.701±
0.015*
0.167 ±
0.003
0.267 ±
0.001
0.351 ±
0.001
0.401 ±
0.009
0.369 ±
0.004
0.901 ±
0.15
0.106 ±
0.001
0.116 ±
0.001
0.501±
0.001
0.306 ±
0.001
0.349 ±
0.001
Klebsiella
Pneumoniae
1.559 ±
0.005
0.155 ±
0.003
0.305 ±
0.007
0.552±
0.002
0.415 ±
0.005
0.095 ±
0.001
Pseudomonas
aeruginosa
Escherichia
Coli
1.400 ± 1.450 ±
0.001 0.005
0.125 ± 0.114 ±
0.007
0.029
0.325 ± 0.664 ±
0.004
0.009
0.317±
0.910 ±
0.006
0.012
0.457±
0.681 ±
0.005
0.004
0.057±
1.049 ±
0.002
0.003
Table (2): The minimal inhibitory concentration
(MIC) of bee venom of different bacteria
Treatment
Staphylococcus Streptococcus
aureus
pyogenes
Klebsiella
Pneumoniae
Pseudomonas
aeruginosa
Escherichia
Coli
Bacterial Normal
growth
----
----
----
----
----
Apitox
1600 *
2800
3600
2600
1000
1000
1000
2400
1000
1800
1200
1000
2400
1700
1800
2400
2600
3600
2400
2100
2800
2100
3800
2400
4400
Vacsera
Ethanolic sac
extract
Whole Bee
extract
Tetracycline
(50ug)
Int.J.Curr.Microbiol.App.Sci (2015) 4(4): 141-1
Antibacterial Activity of Bee Venom Collected from Apis
mellifera Carniolan Pure and Hybrid Races by Two
Collection Methods
Ahmed G. Hegazi, EL-Feel M. A. , Eman H.
Abdel-Rahman and Abed Al-Fattah M. A
The objective of this investigation was to
evaluate the antibacterial activity of bee
venom against selected Gram-positive and
Gram-negative bacterial strains of medical
importance.
Ahmed G. Hegazi, EL-Feel M. A. , Eman H. AbdelRahman and Abed Al-Fattah M. A
• Antibacterial activity of bee venom evaluated
against five pathogenic bacterial strains,
including
• Staphylococcus aureus,
• Streptococcus pyogenes,
• Klebsiella pneumoniae,
• Escherichia coli
• Pseudomonas aeruginosa.
Table 1 Bee venom collected weight gm. / colony
from pure and hybrid Carniolan honey bee race.
Treatment
1st week
Sample
2nd week
Sample
3rd week
Sample
4th week
Sample
Over all mean
Apis mellifera
Carniolan race
Hybrid race
Fiber
Latex
Fiber
Latex
52
89
82
74
32
41
84
100
72
111
132
145
29
42
111
105
102±11.91
106±14.66
46±10.03 75±17.47
Table (2): Influence of bee venom collected by two different
methods (fiber and latex) on growth inhibition of different
bacteria
Treatment
Staphylococcus
aureus
Streptococcus
pyogenes
Klebsiella
Pneumoniae
Pseudomonas
aeruginosa
Escherichia
coli
Normal growth
Fiber
1.550 ±
0.005
Latex
1.550 ±
0.005
Fiber
1.400 ±
0.001
Latex
1.400 ±
0.001
Fiber
0.900 ±
0.150
Latex
0.900 ±
0.15
Fiber
1.700 ±
0.150
Latex
1.70 ±
0.150
Fiber
1.550 ±
0.005
Latex
1.55 ±
0.005
Tetracycline
(50ug)
0.095 ±
0.001
0.095
±0.001
0.057 ±
0.002
0.057 ±
0.002
0.349 ±
0.001
0.349 ±
0.001
0.069 ±
0.004
0.069 ±
0.004
0.049 ±
0.003
0.049 ±
0.003
1st week Sample
0.104±
0.005
0.155 ±
0.005
0.500±
0.008
0.125 ±
0.002
0.178±
0.003
0.101 ±
0.001
0.197±
0.0025
0.167±
0.001
0.173±
0.004
0.114 ±
0.009
2nd week Sample
0.155 ±
0.005
0.305 ±
0.005
0.257±
0.002
0.325 ±
0.002
0.106 ±
0.001
0.116 ±
0.001
0.187±
0.001
0.267±
0.001
0.114 ±
0.009
0.134 ±
0.009
3rd week Sample
0.352±
0.002
0.552±
0.002
0.3117±
0.002
0.3117±
0.002
0.201±
0.001
0.109 ±
0.001
0.201±
0.009
0.401±
0.009
0.510 ±
0.012
0.510 ±
0.012
4th week Sample
0.404±
0.005
0.115 ±
0.005
0.615±
0.008
0.257±
0.002
0.173±
0.003
0.106 ±
0.001
0.143±
0.002
0.267±
0.001
0.273±
0.004
0.104 ±
0.009
Table (3): Influence of bee venom collected by two
different methods (fiber and latex) on minimal inhibitory
concentration
Treatment Staphylococcus
aureus
Tetracycline
(50ug)
1st week
Sample
2nd week
Sample
3rd week
Sample
4th week
Sample
Streptococcus
pyogenes
Klebsiella
Pneumoniae
Pseudomonas
aeruginosa
Escherichia
coli
Fiber Latex Fiber Latex Fiber Latex Fiber Latex Fibe Late
r
x
1.000 1.00 1.60 1.60 1.80 1.80 1.200 1.200 4.40 4.40
1.600 2.80
1.80
2.40
2.20
2.20 1.600 1.900 2.10 2.20
2.800 2.80
2.40
2.60
2.00
2.40 2.600 1.700 2.80 2.40
3.600 3.20
3.60
3.60
2.40
2.50 1.800 1.800 3.80 3.80
2.600 2.80
2.30
2.40
2.70
2.60 2.600 1.900 2.40 2.60
Ahmed G. Hegazi, EL-Feel M. A. , Eman H. AbdelRahman and Abed Al-Fattah M. A
• The results revealed that the amount of bee
venom collected from the pure Carniolan race
was 46±10.03 mg / colony of venom vz 102
±11. 91 mg / colony of venom in hybrid.
• Both bee venom of pure and hybrid bees
exhibited antibacterial activity against all five
bacterial strains and differs according to the
type.
Ahmed G. Hegazi, EL-Feel M. A. , Eman H. AbdelRahman and Abed Al-Fattah M. A
• Bee venom exhibited antibacterial activity
against all five bacterial strains.
• The minimum inhibitory concentration of BV
was determined.
• These results indicate that BV inhibits the
growth and survival of bacterial strains and
that BV may be a useful complementary
antimicrobial agent against pathogenic bacteria
even if bee venom collected by different
methods
Synergy with Antibiotics
Normal Bacterial Growth
Optical Density
BV only
Penicillin only
With Penicillin and BV
Time
Is bee venom having antibacterial activity?
Is bee venom having antibacterial activity?
• Prof. Dr. Ahmed Hegazi
Professor of Microbiology and Immunology
National Research Center, Dokki, Giza, Egypt
President of Egyptian Environmental Society for uses and production of bee products
Secretary of Egyptian Society of Apitherapy
Secretary General of African Federation of Apiculture Associations
Member of Apitherapy Commission , APIMONDIA
E mail: [email protected]
and [email protected]
www.ahmedhegazi.com
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