Stephen Tawanda Musasa

Chinhoyi University of Technology

Zimbabwe
Master of philosophy degree in Agricultural Sciences
(Postharvest Science)
Principal Supervisor: Prof A.B Mashingaidze

Co-supervisor: Dr R. Musundire
Fruit fly identification and invasion pathway
in sweet orange orchards, Rusitu Valley -
Zimbabwe

Presented at the 2017 Research Seminar for Post-

graduate students
School of Agricultural Sciences and Technology
08 February 2017
Background to the study

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 What is sweet orange (Citrus sinensis) ?
• Sweet orange is a member of the citrus family
(Rutaceae).
• Pre-historic era;- sweet oranges were cultivated in
several locations including areas now occupied by
the modern China, India, Bhutan, Burma, and
Malaysia (Webber, 1943).
• Globally - leading producers include Brazil,
European Union and China.
• In Africa - leading producers include Egypt, South
Africa, Morocco and Zimbabwe.
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 Sweet orange production in Zimbabwe
• Zimbabwe’s geographic position and climate
makes it ideal to produce early maturing varieties
of oranges - target markets earlier than
neighbouring competing countries
• Oranges are mainly produced in areas within or
surrounding Limpopo, Save, Mazowe, and Rusitu
valleys.
• Rusitu Valley:- more than 30% orange fruit post-
harvest losses were reported (Musasa et al., 2015).
• Losses resulted from degradation of aesthetic and
market value of oranges due to fruit flies. 5
 How does fruit flies attack orange fruits?

• Damage is done by female adult fruit fly

• Punctures orange and deposit eggs
• Rotting is initiated by microbes
• Eggs hatch and developed larvae feeds on fruit
• Fruit drop
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 Distribution and record of Bactrocera spp
(EPPO, 2016)
• B. dorsalis–
believed to
be of Asian
origin.
 First
recorded in
Africa on the
coastal area
of Kenya
and Tanzania
Key  In about 10
No record years -
First recorded 2003
First recorded 2004
establish in
First recorded 2005
more than 33
First recorded 2007 African
First recorded 2008 nations
First recorded 2010
First recorded 2013 7
 Why are we worried about sweet orange fruit
production in Zimbabwe?

• Important for local consumption and export.

• Exports declined to US$23,5 million in 2015 from
US$40 million registered in 2013 (Foastat, 2015).
• Major raw material in several fruit based products.
• Livelihoods in several rural communities - centred
on fruit production especially sweet oranges.
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 Aim

• To verify the fruit fly taxonomy and invasion

source in Rusitu Valley, Zimbabwe

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Materials and methods

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Study Site

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• Study was carried out during the 2014 fruiting seasons
of sweet oranges.
• Twenty study locations were selected for fruit fly
trapping.
• Fruit flies were trapped using bait trap of methyl-
eugenol and Malathion solution.
• The trapped fruit flies were identified using
morphological traits
• Molecular techniques used to confirm and infer
invasion pathway.
- mtDNA COI region was amplified, cloned and sequenced
from 11 randomly chosen individuals.
-Additional sequences were retrieved from GenBank to
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perform a Haplotype Network analysis
Results and discussion

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 A, B, C, and D were identified as B. dorsalis
A B C D

Photos show the morphological traits of individuals

present in Zimbabwe
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 Wing analysis

• A mean wing length of 4.3 mm (± 0.2) was observed

for Zimbabwean populations of B. dorsalis
• Lower than 6 mm and 7.3 mm observed,
respectively for B. dorsalis of Asian and American
origin (Drew et al., 2008; Weems et al., 2012).
• Supporting the idea of morphological differences
across B. dorsalis complex as recently observed by
several authors (Frey et al., 2013; Hendrichs et al.,
2015; Schutze et al., 2012; Schutze et al., 2015a,b).

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 Molecular confirmation of the species identification

• 10 sequences clustered into two large groups,

namely;
– six sequences were identical to 101 B. dorsalis
sequences available in GenBank, four were identical to
129 B. dorsalis sequences available in GenBank
• One sequence showed three nucleotide differences
from the latter group.
• Therefore, the collected flies were clearly B.
dorsalis
• Source of invasion is likely from neighbouring east
African countries and not from an independent
invasion. 16
Zimbabwe samples cluster into the two clades - likely
representing two original main outbreaks with an east
African origin, probably from Kenya and Tanzania
(Khamis et al., 2009). 17
Nigerian B. dorsalis samples
cluster with B. kandiensis
samples and with B. dorsalis
samples from Sri Lanka,
India and Myanmar

Nigerian population - an
independent invasion from
Sri Lanka, India or
Myanmar.

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Conclusions

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• The positive morphological and molecular
identification of Zimbabwe flies as B. dorsalis helps to
join one more piece into the puzzle of the African
invasion by this species.
• With the exception of Lesotho, Malawi and Somalia, B.
dorsalis has now been reported in every African country
where a suitable habitat has been predicted for this
species .
• It should be noted that, it is unclear whether in Lesotho,
Malawi and Somalia there was any attempt to look for
B. dorsalis.

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• The invasion of Zimbabwe’s Rusitu Valley was not
an independent invasion.
• The source of the invasion in Zimbabwe is likely
from neighbouring east African countries.
• Zimbabwe’s invasion pathway is linked to the two
reported clear east African outbreaks whose source
was revealed to be Asian countries east of Sri Lanka,
India, and Myanmar.
• It can be noted that, ten years after the first report in
Kenya of B. dorsalis, the complete or near complete
invasion of Africa has been achieved, since in
northern Africa the distribution is clearly limited by
the Sahara desert.
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References

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• Drew, R.A.I., Raghu, S., and Halcoop, P. 2008. Bridging the
morphological and biological species concepts: studies on
the Bactrocera dorsalis (Hendel) complex (Diptera: Tephritidae:
Dacinae) in South-east Asia. Biological Journal of the Linnean
Society, 93 (4), 217-226
• European and Mediterranean Plant Protection Organization
(EPPO), 2016 Bactrocera invadens (Diptera: Tephritidae) A new
fruit fly species.
www.eppo.int/QUARANTINE/Alert_List/insects/BCTRIN.htm
• FAOSTAT. 2015. Value of Agricultural Production; citrus
production, Zimbabwe. Food and Agriculture Organisation.
[Online] http://faostat3.fao.org
• Frey, J.E., Guillen, L., Frey, B., Samietz, J., Rull, J., and Aluja, M.
2013. Developing diagnostic SNP panels for the identification of
true fruit flie (Diptera: Tephritidae) within the limits of CO1-based
species delimitation. BMC Evolution Biology – speciation and
evolution genetics 24
• Hendrichs, J., Vera, M.T., De Meyer, M., and Clarke A.R. 2015. Resolving
cryptic species complexes of major tephritid pests. In: De Meyer M, Clarke
AR, Vera MT, Hendrichs J (Eds) Resolution of Cryptic Species Complexes
of Tephritid Pests to Enhance SIT Application and Facilitate International
Trade. ZooKeys, 540, 5–39.
• Khamis, F.M., Karam, N., Ekesi S., M. De Meyer, M., Bonomi, A.,
Gomulski, L. M., Scolari, F., Gabrieli, P., Siciliano, P., Masiga, D., Kenya,
E. U., Gasperi, G., Malacrida, A. R., And Guglielmino, C. R. 2009.
Uncovering the tracks of a recent and rapid invasion: the case of the fruit fly
pest Bactrocera invadens (Diptera: Tephritidae) in Africa. Molecular
Ecology, 18, 4798–4810.
• Musasa S.T, Musundire R, Mashingaidze A.B, and Makuza S.M
(2015) A preliminary study of the orange (Citrus sinensis) fruit
value-chain in Chimanimani Rural District, Zimbabwe. African
Journal of Agricultural Research 10 (35) pp 3507-3516
• Schutze, M.K., Krosh M.N., Armstrong, K.F., Chapman, T.A.,
Englezou, A., Chomic, A., Cameroon, S.L., Hailstone, D., and
Clarke A.R. 2012. Population structure of Bactrocera dorsalis s.s.,
B. papayae and B. philippinensis (Diptera: Tephritidae) in
southeast Asia: evidence for a single species hypothesis using
mitochondrial DNA and wing-shape data. BMC Evolutionary
Biology,12,130 25
• Schutze, M.K., Aketarawong, N., Amornsak, W., Armstrong, K.F.,
Augustinos, A.A., Barr, N., Bo, W., Bourtzis, K., Boykin, L.M., Cáceres, C.,
Cameron, S.L., Chapman, T.A., Chinvinijkul, S., Chomič, A., De Meyer, M.,
Drosopoulou, E., Englezou, A., Ekesi, E., Gariou-Papalexiou, A., Geib,
S.M., Hailstones, D., Hasanuzzaman, M., Haymer, D., Hee, A.K.W.,
Hendrichs, J., Jessup, A., Ji, Q., Khamis, F.M., Krosch, M., Leblanc, L.,
Mahmood, K., Malacrida, A.R., Mavragani-Tsipidou, P., Mwatawala, M.,
Nishida, R., Ono, H., Reyes, J., Rubinoff, D., San Jose, M., Shelly, T.E.,
Srikachar, S., Tan, K.H., Thanaphum, S., Haq, I., Vijaysegaran, S., Wee,
S.L., Yesmin, F., Zacharopoulou, A., and Clarke A.R. 2015a.
Synonymization of key pest species within the Bactrocera dorsalis species
complex (Diptera: Tephritidae): taxonomic changes based on a review of
20 years of integrative morphological, molecular, cytogenetic, behavioural
and chemoecological data. Systematic Entomology,40(2), 456-471
• Schutze, M.K., Mahmood, K., Pavasonic, A., Bo W., Krosh, M.N., and
Cameroon, S.L. 2015b. One and the same: integrative taxonomic evidence
that Bactrocera invadens (Diptera: Tephritidae) is the same species as the
Oriental fruit fly Bactrocera dorsalis. Systematic Entomology, 40, 472–486
• Webber H.J (1943). History and Development of Citrus Industry. [Online]
http://websites.lib.ucr.edu/agnic/webber/Vol1/Chapter1.htm
• Weems, H.V., Heppner, J.B., Nation, J.L., and Fasulo, T.R. 2012. Oriental
Fruit Fly (Bactrocera dorsalis (Hendel) (Insecta: Diptera: Tephritidae)
Featured Creatures - Entomology and Nematology, University of Florida.
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I THANK YOU ALL

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