FYP Poster

Anukhcha Wannaket A/L Peetak (SEE110004); Supervisor: Prof. Dr. Hiroyuki Takaoka;

Co-supervisor: Dr. Low Van Lun Division of Ecology and Biodiversity, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.

Email: [email protected]

Simulium tuberosum species-group is generally found in Palearctic,

Nearctic, and Oriental Regions, and at least 27 morphologically similar

nominal species exist in the Oriental Region (Adler & Crosskey, 2014).

Certain taxa of the family Simuliidae are morphologically inseparable, but

biologically distinct. In this study, the phylogenetic relationships of Simulium

tani species-complex from Vietnam, Taiwan, Malaysia, and Thailand were

inferred from mitochondrial cytochrome c oxidase subunit I (COI) and II

(COII) DNA sequences.

This study examines the phylogenetic relationships of Simulium tani and Simulium sp. of S. tani species-complex of Vietnam. Simulium sp. is

morphologically different from S. tani and similar to S. suzukii from Taiwan and Japan by the pupal gill arrangement. Ten specimens of S. tani collected from

Stream 1, Lam Dong, Vietnam; 12 specimens of Simulium sp. from Stream 9, Lam Dong, Vietnam, 10 specimens of S. suzukii from Taiwan, were used for

DNA analysis using mitochondria encoded cytochrome c oxidase subunit I (COI) and II (COII). Isolates of S. tani species-complex from Malaysia and

Thailand available from the NCBI GenBank database were included for analysis. A maximum parsimony phylogenetic tree for both COI and COII genes

revealed that both S. tani and Simulium sp. from Vietnam are placed in the same cluster, which is distinct in lineage from those of S. tani species-complex

from Taiwan, Malaysia and Thailand.

Figures 8 and 9. Results of maximum parsimony trees indicate distinct

species of Simulium tuberosum species-group from various geographical

areas. Bootstrap values with only more than 50% are shown in the tree.

Abstract

Introduction Results

*Specimens collection from:

• Streams 1 and 9, Lac Duong District, Lam Dong Province, Vietnam,

• Daping River, Beipu Township, Taiwan, and

• Malaysia and Thailand (Sequences available from NCBI GenBank)

DNA extraction, amplification, purification, and sequencing used here,

follow those of Low et al. (2014).

Materials and Methods

To clarify the species status of two taxa (S. tani species-complex &

Simulium sp. of S. tuberosum species-group) collected from Vietnam, and

to infer their phylogenetic relationships with isolates of S. tani species-

complex from Thailand and Malaysia and with S. suzukii from Taiwan.

Objective

I would like to express my deepest appreciation to my supervisor, Prof. Dr.

Hiroyuki Takaoka and my co-supervisor, Dr. Low Van Lun who have guided

me in all aspects of this study. I also thanks to Ms. Vinnie Siow Wei Yin and

Ms. Zubaidah Ya’cob for guidance and support.

Acknowledgement

• It is indicated that two taxa from Vietnam are not separable genetically,

although they are morphologically distinguishable from each other.

• The result obtained is unexpected and represents a striking example

rarely occurring in the family Simuliidae. This may have been caused by

low genetic differentiation between the taxon of S. tani species-complex

and Simulium sp, or the weak sensitivity of the genes used.

• The phylogenetic trees show the relationship of the two taxa from

Vietnam is closer to isolates of S. tani species-complex from Thailand

and Malaysia then to S. suzukii from Taiwan, as expected from the

geographical point of view.

• Further studies on other isolates of S. tani species-complex from various

localities in Vietnam are needed to clarify the species status, and to infer

their phylogenetic relationships with other isolates of S. tani and other

related species of the S. tuberosum species-group.

Discussions

All specimens from Vietnam, Taiwan, Malaysia, and Thailand

populations are placed in different clusters of both phylogenetic trees.

Both S. tani and Simulium sp. from Vietnam are genetically not

separable from each other, though they are found to be distinct from

S. tani from Thailand and Malaysia, and also from S. suzukii from

Taiwan.

Conclusion

• Adler, P. H. & Crosskey, R. W. (2014). World blackflies (Diptera: Simuliidae): a

comprehensive revision of the taxonomic and geographical inventory [2014].

http://www.clemson.edu/cafls/biomia/pdfs/blackflyinventory.pdf

• Low, V. L., Adler, P. H., Takaoka, H., Ya’cob, Z., Lim, P. E., Tan, T. K., Lim, Y. A. L.,

Chen, C. D., Norma-Rashid, Y., & Sofian-Azirun, M. (2014). Mitochondrial DNA

markers reveal high genetic diversity but low genetic differentiation in the black fly

Simulium tani Takaoka & Davies along an elevational gradient in Malaysia. PloS ONE,

9(6), e100512.

References

Figure 1: Specimens Collection*

Figure 2: Specimens Figure 3: DNA

Extraction

Figure 4: Polymerase

Chain Reaction

Figure 5: Electrophoresis Figure 6: Band Visualization

Figure 7: Data Analysis

Figure 8: Maximum Parsimony Phylogenetic Tree COI

Figure 9: Maximum Parsimony Phylogenetic Tree COII

DNA Analysis of Simulium tani

Species-Complex (Diptera: Simuliidae) from Vietnam