Novel molecular evidence of population structure in Anopheles (Kerteszia) bellator from Brazilian Atlantic Forest

Anopheles bellator is a primary malaria vector in the Atlantic Forest. Partial sequences of timeless and Clock genes were used to assess the genetic differentiation of five Brazilian populations, which showed strong population structure (e.g. high F _ST values and fixed differences) in all pairwise comparisons between Bahia sample and the others from Paraná, São Paulo and Rio de Janeiro states. Also, the resulting phylogenetic trees clearly grouped the sequences from Bahia in a different cluster with high bootstrap values. Among southern and southeastern populations low levels of genetic differentiation were found suggesting a general stability of the genetic structure.

Key words:
Anopheles (Kerteszia) Bellator; bromeliad-malaria; Brazilian Atlantic Forest

Authorship

Kamila Voges

Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, BrasilUniversidade Federal de Santa CatarinaBrazilFlorianópolis, SC, BrazilUniversidade Federal de Santa Catarina, Centro de Ciências Biológicas, Departamento de Biologia Celular, Embriologia e Genética, Florianópolis, SC, Brasil

Marcela Possato Correa da Rosa

Betina Westphal-Ferreira

Universidade Federal do Paraná, Setor de Ciências Biológicas, Departamento de Zoologia, Curitiba, PR, BrasilUniversidade Federal do ParanáBrazilCuritiba, PR, BrazilUniversidade Federal do Paraná, Setor de Ciências Biológicas, Departamento de Zoologia, Curitiba, PR, Brasil

Mario Antonio Navarro-Silva

Carime Lessa Mansur Pontes

Universidade Federal de Santa Catarina, Centro de Ciências Biológicas, Departamento de Microbiologia, Imunologia e Parasitologia, Florianópolis, SC, BrasilUniversidade Federal de Santa CatarinaBrazilFlorianópolis, SC, BrazilUniversidade Federal de Santa Catarina, Centro de Ciências Biológicas, Departamento de Microbiologia, Imunologia e Parasitologia, Florianópolis, SC, Brasil

André Nóbrega Pitaluga

Fundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Biologia Molecular de Parasitas e Vetores, Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz-FiocruzBrazilRio de Janeiro, RJ, BrazilFundação Oswaldo Cruz-Fiocruz, Instituto Oswaldo Cruz, Laboratório de Biologia Molecular de Parasitas e Vetores, Rio de Janeiro, RJ, Brasil

Imperial College London, Department of Life Sciences, London, United KingdomImperial College LondonUnited KingdomLondon, United KingdomImperial College London, Department of Life Sciences, London, United Kingdom

Instituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, RJ, BrasilInstituto Nacional de Ciência e Tecnologia em Entomologia MolecularBrasilRio de Janeiro, RJ, BrasilInstituto Nacional de Ciência e Tecnologia em Entomologia Molecular, Rio de Janeiro, RJ, Brasil

Carlos José de Carvalho-Pinto

Luísa DP Rona ⁺+ Corresponding author: luisa.rona@imperial.ac.uk; luisa.rona@ufsc.br

http://orcid.org/0000-0002-3400-3950

+ Corresponding author: luisa.rona@imperial.ac.uk; luisa.rona@ufsc.br

LDPR, KV, MPCR and CLMP participated in data generation and analysis; LDPR, BWF, MANS, CJCP and ANP carried mosquito collections and morphological identification; LDPR drafted the manuscript; ANP and CJCP helped in the paper drafting by critically reading the original manuscript; LDPR was the principal investigator, participated in its design and coordination. All authors read and approved the final manuscript.

SCIMAGO INSTITUTIONS RANKINGS

Figures | Tables

Figures (2)
Tables (1)

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Fig. 1:
Anopheles bellator collection sites in the Brazilian Atlantic Rainforest. (A): map of Brazil. (B): magnification of the black box from (A) displaying the sites where An. bellator samples were collected. The x and y axes drawn in the map (B) display the longitude and latitude, respectively.

Thumbnail

Fig. 2:
maximum likelihood trees of Anopheles bellator timeless and Clock sequences (TIM3e+G+I and HKY+G models, respectively). These phylogenies show that two structured groups were identified in the An. bellator populations from Brazil: An. bellator B occurs in the North-East Atlantic Rainforest (Camacan) and An. bellator A is found in the other more Southern Brazilian regions. Numbers on the nodes represent the percentage bootstrap values based on 1000 replications. Lowercase letters in the haplotype names specify the two alleles (A or B) obtained from each individual.

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TABLE
Genetic differentiation between all Anopheles bellator populations

Fig. 1: Anopheles bellator collection sites in the Brazilian Atlantic Rainforest. (A): map of Brazil. (B): magnification of the black box from (A) displaying the sites where An. bellator samples were collected. The x and y axes drawn in the map (B) display the longitude and latitude, respectively.

Fig. 2: maximum likelihood trees of Anopheles bellator timeless and Clock sequences (TIM3e+G+I and HKY+G models, respectively). These phylogenies show that two structured groups were identified in the An. bellator populations from Brazil: An. bellator B occurs in the North-East Atlantic Rainforest (Camacan) and An. bellator A is found in the other more Southern Brazilian regions. Numbers on the nodes represent the percentage bootstrap values based on 1000 replications. Lowercase letters in the haplotype names specify the two alleles (A or B) obtained from each individual.

TABLE Genetic differentiation between all Anopheles bellator populations

Populations	Gene	Km	F _ST	Dxy	Da	Ss	Sf
Abraão (RJ) x Sítio Forte (RJ)	timeless	10.89	-0.1039	0.0177	-0.0018	11	00
Abraão (RJ) x Sítio Forte (RJ)	Clock		-0.0157	0.0322	-0.0005	08	00
Ilha do Mel (PR) x Abraão (RJ)	timeless	496.5	0.0411	0.0215	0.0009	09	00
Ilha do Mel (PR) x Abraão (RJ)	Clock		0.1134	0.0404	0.0046	07	00
Ilha do Mel (PR) x Sítio Forte (RJ)	timeless	488.6	0.0695	0.0224	0.0016	11	00
Ilha do Mel (PR) x Sítio Forte (RJ)	Clock		0.0904	0.0377	0.0034	08	00
Ilha do Mel (PR) x Cananéia (SP)	timeless	70.1	0.1147	0.0233	0.0027	10	00
Ilha do Mel (PR) x Cananéia (SP)	Clock		-0.0508	0.0376	-0.0019	10	00
Cananéia (SP) x Sítio Forte (RJ)	timeless	425.7	0.1158^*	0.0221	0.0026	08	00
Cananéia (SP) x Sítio Forte (RJ)	Clock		0.0001	0.0364	0.0000	09	00
Cananéia (SP) x Abraão (RJ)	timeless	434.1	0.1311^*	0.0222	0.0029	06	00
Cananéia (SP) x Abraão (RJ)	Clock		0.0086	0.0383	0.0003	08	00
Camacan (BA) x Ilha do Mel (PR)	timeless	1449	0.6471^*	0.0621	0.0402	00	07
Camacan (BA) x Ilha do Mel (PR)	Clock		0.6482^*	0.1168	0.0757	02	05
Camacan (BA) x Cananéia (SP)	timeless	1379	0.6694^*	0.0622	0.0417	00	09
Camacan (BA) x Cananéia (SP)	Clock		0.6287^*	0.1163	0.0731	02	05
Camacan (BA) x Abraão (RJ)	timeless	987.1	0.6722^*	0.0628	0.0422	01	10
Camacan (BA) x Abraão (RJ)	Clock		0.6236^*	0.1050	0.0655	02	03
Camacan (BA) x Sítio Forte (RJ)	timeless	990.1	0.6758^*	0.0641	0.0434	00	09
Camacan (BA) x Sítio Forte (RJ)	Clock		0.6562^*	0.1105	0.0725	02	05

FST: pair-wise estimates of population differentiation. The significance of FST values was evaluated by 1,000 random permutations (* p < 0.05). Dxy: average number of nucleotide substitutions per site between populations; Da: number of net nucleotide substitutions per site between populations.29Ss: number of shared polymorphisms between the two populations. Sf: number of fixed differences between the two populations. Km: the approximated geographic distances between localities in km.

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Novel molecular evidence of population structure in Anopheles (Kerteszia) bellator from Brazilian Atlantic Forest

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[1] + Corresponding author: luisa.rona@imperial.ac.uk; luisa.rona@ufsc.br