The intestinal microbiota of tephritid fruit flies as a potential tool … · 2010. 11. 3. · The...
Transcript of The intestinal microbiota of tephritid fruit flies as a potential tool … · 2010. 11. 3. · The...
The intestinal microbiota of tephritid fruit flies as a potential tool to improve
rearing and the SIT
Dept. of Entomology, Dept. of Microbiology and Plant Diseases, The Robert H. Smith Faculty of Agriculture, Food and Environment,
The Hebrew University of Jerusalem, Rehovot, Israel
Michael Ben-Yosef, Eyal Ben-Ami, Sagi Gavriel, Edouard Jurkevitchand Boaz Yuval
Outline
1. Tephritid - bacteria associations2. Bacteria and adult fruit fly fitness:
The olive fly as an example3. Bacteria and the Sterile Insect Technique:
The Medfly as a case study4. Final conclusions
Superfamily Tephritoidae Family Tephritidae
General saprophagy Plant saprophagy Phytophagy
Evolution of Tephritid – bacteria associations
Plant saprophagy:Self-rotting vegetation
Phytophagy:Fly mediated rot in living
plant tissuesDiaz-Fleischer et al. 2000
Olive fly-bacteria association
Petri 1910Capuzzo et al. 2005 Petri 1910
Adult flies feed opportunistically but mainly on honeydew
Effect of bacteria on olive fly fitness
Examining the effect of bacteria and diet on female fecundity
Mated wild females:
Sucrose Sucrose + non-essential amino-acids
Full diet (sucrose + Yeast
hydrolysate)
Ben-Yosef et al., Proc. R. Soc. B, 2010
+ / - Antibiotics
Effect of bacteria on olive fly fitness
Ben-Yosef et al., Proc. R. Soc. B, 2010
05101520
mean
no. o
f egg
s / fe
male P = 0.028
Honeydew
• Bacterial contribution to fecundity is diet dependent.
• Bacteria are needed only when females feed on an unbalanced amino-acid diet
intestinal bacteria contribute amino-acids or protein to the fly
Allows flies to subsist on low quality food such as honeydew
Other fruit flies may similarly depend on their intestinal bacteria
(Dacus, Ceratitis, Rhagoletis, Anastrepha)
Bacteria and adult fruit fly fitness: conclusions
• Egg disinfection • Diet Acidification • Use of antibiotics and preservatives
Disruption of symbiotic relationships during mass rearing
• Irradiation of pupae
The irradiation affects the gut microbiota
Reduced mating competence of sterile Medfly males
Irradiation effect on medfly gut microbiotaV8 (Day 1) Sterile V8 (Day 1) Sterile V8 (Day 5) Wild (Day 1)
1. Gut bacterial community profiling by Amplified rDNA restriction analysis (ARDRA)
Enterobacter sp.
Klebsiella sp.
EnterobacteriaceaePseudomonads
Enterobacter sp.
Klebsiella sp.
Enterobacter sp.
Klebsiella sp.
18.67%
4%
Ben-Ami et al. ISME Journal 2009
Irradiation effect on medfly gut microbiotaV8 (Day 1) Sterile V8 (Day 1) Sterile V8 (Day 5) Wild (Day 1)
EnterobacteriaceaePseudomonads
Enterobacter sp.
Klebsiella sp.4 %Klebsiella
sp.23%
Enterobacter sp.
Ben-Ami et al. ISME Journal 2009
1. Gut bacterial community profiling by Amplified rDNA restriction analysis (ARDRA)
2. Monitoring the establishment of Klebsiella oxytoca in the gut
Effect of “probiotic” diet on medfly gut microbiota
StrpRKlebsiella oxytoca
ChromoAgarMedium Strp Medium
Klebsiella treatmentSucrose only
Klebsiella + Enterobacter
Sterile V8 (Day 1)
0.00E+002.00E+074.00E+076.00E+078.00E+071.00E+081.20E+081.40E+081.60E+081.80E+082.00E+08
TO T3 T5 T7Days
Num
ber o
f col
onie
s
0 3 5 7
*
* *20
16
12
8
4
0
No. o
f colo
nies (
x107 )
Days
Klebsiella treatmentSucrose only
Pseudomonas spp.
0
4
8
12
16
0 3 5 7Days
No. o
f colo
nies (
x10 6
)
*
Ben-Ami et al. ISME Journal 2009
2. Monitoring the establishment of Klebsiella oxytoca in the gut
Effect of “probiotic” diet on medfly gut microbiota
StrpRKlebsiella oxytoca
ChromoAgarMedium Strp Medium
16SrRNA Sequencing:Klebsiella oxytoca
All StrpR colonies turned blue on ChromAgar
Sterile V8 (Day 1)
0.00E+00
2.00E+07
4.00E+07
6.00E+07
8.00E+07
1.00E+08
1.20E+08
1.40E+08
1.60E+08
1.80E+08
T0 T3 T5 T7Days
Num
ber o
f col
onie
s
0 3 5 7Days
16
12
8
4
0
No. o
f colo
nies (
x107 )
DaysBen-Ami et al. ISME Journal 2009
Effect of “probiotic” diet on male competitiveness
3. Examining copulatory success of sterile males fed with Klebsiella oxytoca
Live bacteria
Sterile V8 (Day 1)
Autoclaved bacteria
Sucrose only
Sucrose only
“Sade” strain“Sade” strain
X5 days feeding on:
Ben-Ami et al. ISME Journal 2009
020406080
100
Livebacteria
SadeStrain
(control)
Autoclavedbacteria
Sucroseonly
Laten
cy to
mate
(min)
*0.00.10.20.30.40.50.60.70.80.91.0
Failin
g0 20 40 60 80 100 120 140 160 180
Time to mate
• The intestinal microbiota contributes significantly to fruit fly fitness.
• Mass reared V8 flies differ from wild flies in their gut bacterial community structure.
• The sterilizing irradiation affects the bacterial community within the gut.
• Klebsiella oxytoca added to the post-irradiation diet significantly improve sterile male performance.
Final conclusions
Colonization
Production
Sterilization
Holding & emergence
Release
Supplementing pre-release diets with
bacteria
Supplementing adult diets with bacteria
Applying bacteria in larval rearing procedures
• Minimizing larval requirements of nutrients•Reducing microbial contamination
• Enhancing sterile male performance•Extending life through nutrient provisioning, minimizing infections
• Helping newly established colonies to adapt to artificial rearing conditions
Implementing bacteria in mass rearing of fruit flies
• Shlomit Shloush and Batya Kamenski• Bio-Fly facility: Inbar Shouster-Dagan• BARD and IAEA/FAO for financial support
Many thanks to