Hawaria et al.

Malaria Journal (2023) 22:373 Malaria Journal

https://doi.org/10.1186/s12936-023-04813-x

RESEARCH Open Access

First report of Anopheles stephensi

from southern Ethiopia
Dawit Hawaria1*, Solomon Kibret2, Daibin Zhong3, Ming‑Chieh Lee3, Kidane Lelisa4, Belayneh Bekele5,
Muntasha Birhanu6, Mathe Mengesha7, Hiwot Solomon8, Delenesaw Yewhalaw9 and Guiyun Yan3

Abstract
Background Anopheles stephensi is an emerging exotic invasive urban malaria vector in East Africa. The World
Health Organization recently announced an initiative to take concerted actions to limit this vector’s expansion
by strengthening surveillance and control in invaded and potentially receptive territories in Africa. This study sought
to determine the invasion of An. stephensi in southern Ethiopia.
Methods A targeted entomological survey, both larvae and adult, was conducted in Hawassa City, southern Ethiopia
between November 2022 and February 2023. Anopheles larvae were reared to adults for species identification. CDC
light traps and BG Pro traps were used indoors and outdoors overnight at selected houses to collect adult mosquitoes
in the study area. Prokopack aspirator was employed to sample indoor resting mosquitoes in the morning. Adults
of An. stephensi was identified using morphological keys and then confirmed by PCR.
Results Larvae of An. stephensi were found in 28 (16.6%) of the 169 potential mosquito breeding sites surveyed.
Out of 548 adult female Anopheles mosquitoes reared from larvae, 234 (42.7%) were identified as An. stephensi
morphologically. A total of 449 female anophelines were caught, of which 53 (12.0%) were An. stephensi. Other
anopheline species collected in the study area included Anopheles gambiae sensu lato (s.l.), Anopheles pharoensis,
Anopheles coustani, and Anopheles demeilloni.
Conclusion This study confirmed the presence of An. stephensi in southern Ethiopia. The presence of both larval
and adult stages of this mosquito attests that this species established sympatric colonization with native vector
species such as An. gambiae (s.l.) in southern Ethiopia. The findings warrant further investigation on the ecology,
behaviour, population genetics, and role of An. stephensi in malaria transmission in Ethiopia.
Keywords Anopheles stephensi, Malaria, Mosquito breeding, Mosquito habitat, Ethiopia

*Correspondence:
Dawit Hawaria
[email protected]
1
School of Environmental Health, Hawassa University, Hawassa, Ethiopia
2
West Valley Mosquito and Vector Control District, Ontario, CA, USA
3
Program in Public Health, University of California at Irvine, Irvine, CA
92697, USA
4
Department of Biology, Dilla University, Dilla, Ethiopia
5
Disease Prevention Department, Sidama Regional Health Bureau,
Hawassa, Ethiopia
6
Hawassa City Administration Health Department, Hawassa, Ethiopia
7
Sidama Regional Public Health Institute, Hawassa, Ethiopia
8
Ministry of Health, Addis Ababa, Ethiopia
9
Tropical and Infectious Diseases Research Centre (TIDRC), Jimma
University, Jimma, Ethiopia

© The Author(s) 2023. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which
permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the
original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or
other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line
to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory
regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this
licence, visit http://​creat​iveco​mmons.​org/​licen​ses/​by/4.​0/. The Creative Commons Public Domain Dedication waiver (http://​creat​iveco​
mmons.​org/​publi​cdoma​in/​zero/1.​0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
Hawaria et al. Malaria Journal (2023) 22:373 Page 2 of 8

Background elevation of 1708 m (5604 ft) above sea level. The city’s
Anopheles stephensi is one of several important malaria population was 502,980 in 2022 [14]. Being situated at
vectors of urban environments in Southeast Asia, the the shore of Lake Hawassa, the city has seen rapid growth
Middle East, and the Arabian Peninsula [1]. Since its in recent years. The city constantly sees new construction
initial discovery in Djibouti in 2012 [2], this vector sites following its swift industrial boom in recent years.
species has been found in eastern Ethiopia [3, 4], Sudan The average annual temperature and precipitation are
[5], Somalia [1], and Kenya [6]. about 21 °C and 961 mm, respectively. A shorter rainy
Its adaptation to the urban environment [7] coupled season occurs between March and May followed by a
with the rapid spread of An. stephensi in Eastern Africa longer wet season between July and October. Common
poses serious challenges for controlling and eliminating livestock in the area include cattle, chickens, goats, sheep,
malaria in Africa’s rapidly urbanizing nations. Malaria donkeys, and horses. Malaria is seasonal in the area
outbreaks have been documented following its recent and the main malaria transmission period is between
colonization of the Horn of Africa region [8] and the September and December [14].
increase in malaria incidence in Djibouti was attributed
to An. stephensi [9]. According to geospatial modelling Malaria profile review
studies, many cities in sub-Saharan Africa have favorable Laboratory-confirmed malaria data was obtained from
environmental conditions for An. stephensi to proliferate, the Hawassa City Health Department. The malaria
putting an additional 126 million Africans at risk of morbidity data, which was sorted by age, sex, parasite
contracting malaria if the spread of An. stephensi is species, and residence for the previous nine years was
not curbed [10]. Another modelling study predicted collected and analysed, and the annual transmission
that if tailor-made An. stephensi interventions are not pattern was established.
implemented, and the number of malaria cases in
Ethiopia might double [11]. The geographical range of Mosquito larval survey and rearing
An. stephensi may spread into rural areas, although it is Any water collections were surveyed for mosquito larvae
mostly an urban vector [12]. It is, therefore, crucial to and pupae between November 2022 and February 2023.
describe the rural–urban transitions as a continuum To increase the likelihood of discovering An. stephensi,
rather than as distinct contexts given the ongoing urban target sampling was conducted. Mosquito larvae and
centre expansion in rural and urban areas. pupae were dipped from likely larval breeding habitats,
Determining the geographic range of An. stephensi including man-made water containers, freshwater
helps design tailored vector interventions. Given the pools, lake margins, discarded tires, plastic containers,
importance of An. stephensi to transmit urban malaria and concrete water collection tankers at construction
and its potential effects on public health in Africa, the sites. Dipping was done under larval search strategies
World Health Organization (WHO) recently launched an recommended by the WHO [15]. The larvae and pupae
initiative to take coordinated actions to limit the vector’s were brought in jars to the environmental laboratory at
spread by improving An. stephensi surveillance and Hawassa University, where they were placed in trays
control in Africa [1]. While the spread of An. stephensi and raised to adult in preparation for morphological
has recently been documented in several localities in identification. Each enamel tray containing larvae was
eastern, northeastern, and central Ethiopia [3, 4], its labeled for the habitat types from where the larvae were
occurrence and distribution in the area of southern obtained to identify the species after adult emergence.
Ethiopia is still unknown. Balkew et al. [13] surveyed The larvae were allowed to develop in the water drawn
larval habitat in southern Ethiopia in 2019 and could not from the field to maintain the same aquatic environment.
find An. stephensi in the surveys. Unfortunately, the study The pupae were sorted and transferred with pipettes
relayed a one-time survey that lasted only for a few days. from the enamel trays to beakers with modest volumes
The present study aimed to explore the invasion of An. of water, then kept inside cages. A dissecting microscope
stephensi in Hawassa, southern Ethiopia. was used to identify emerging adults to the species using
identification key [16]. All identified specimens were
Methods preserved individually in Eppendorf tubes for further
Study sites analysis.
Entomological survey was conducted from November
2022 to February 2023 in Hawassa City in southern Adult mosquito survey
Ethiopia (Fig. 1). Hawassa City is the largest city in the Three different types of traps were used to collect adult
middle of the Great Ethiopian Rift Valley and is the mosquitoes: CDC Light Traps (Model: John W. Hock
capital of Sidama Regional State. It is located at an CDC Light Trap 512, USA); Bioagents (BG-pro) Traps
Hawaria et al. Malaria Journal (2023) 22:373 Page 3 of 8

Fig. 1 Map of the study site and Anopheles stephensi larval habitat distribution

with lure; and Prokopack aspirator (John W. Hock 1418, further sorted into species using a morphological key
USA). Collection with CDC Light Trap and BG-pro [16].
were made overnight from 18:00 to 06:00 h. A trap was
suspended 1.5 m above the ground near a sleeping area Molecular identification of An. stephensi
where the people are protected by LLINs. Mosquito A subset of the morphologically identified An. stephensi
sampling took place both indoors and outdoors. specimens were molecularly analyzed to confirm the
Prokopack aspirator was employed to sample indoor species. DNA was extracted from a single leg using the
resting mosquitoes in the morning from 6:30 h to 8:00 h. Chelex method [17] with modification. Two methods
Animal shelters were also surveyed. were used to identify the species: (i) PCR endpoint
All collected adult mosquitoes were brought to the assay using the internal transcribed spacer 2 (ITS2)
lab for species identification. Mosquitoes were killed locus; and (ii) sequencing portions of cytochrome
by placing them in a refrigerator. The specimens were c oxidase subunit 1 (cox1) and cytochrome B gene
then sorted into culicines and Anopheles. Culicines were (cytb) loci. ITS2 endpoint assay was performed as
counted, recorded, and discarded. All anophelines were previously described using the primers 5.8SB (5′-ATG
Hawaria et al. Malaria Journal (2023) 22:373 Page 4 of 8

CTT AAA TTT AGG GGG TAG TC-3′) and 28SC NCBI’s GenBank. A threshold limit of 98% sequence
(5′-GTC TCG CGA CTG CAA CTG-3′) and the similarity for cox1 was used to classify sequences into
following modifications: final reagent concentrations species [20].
and components were 0.5 μM for each primer;
1× DreamTaq Green Master Mix (Thermo Fisher Results
Scientific); and water for a total reaction volume of Malaria morbidity
17 μl. PCR reaction conditions were set as denaturation Over the last nine years, 128,946 malaria cases were
at 95 °C for 3 min, 35 cycles of 94 °C for 30 s, annealing reported in the study area. Plasmodium falciparum and
at 55 °C for 30 s, extension at 72 °C for 30 s, and a final Plasmodium vivax were prevalent in the area and nearly
step at 72 °C for 6 min. Anopheles stephensi specimens had identical proportions with 66,570 (51.6%) and 62,376
were identified by visualization of the 522-bp band with (48.4%), respectively. According to the data, malaria
gel electrophoresis; non-An. stephensi specimens did transmission dropped for three years in a row, from 2014
not amplify and no band was present [3]. Portions of the to 2016. However, from 2017 to 2019, there was a slight
cox1 and cytb loci were also amplified for sequencing increment. Then, after a drop in the year 2020 number of
using previously detailed methods [18]. PCR products malaria cases rose steeply in 2022 (Fig. 2).
were purified and sequenced using Sanger technology
by Genewiz Inc (South Plainfield, NJ). Sequences were Larval habitat distribution, positivity, and Anopheles
cleaned and analysed using CodonCode (CodonCode species composition
Corporation, Centerville, MA, USA). Next, cox1 and Anopheles stephensi larvae were detected in 28 (16.6%)
cytb sequences from An. stephensi were submitted of the 169 mosquito breeding sites surveyed. Breeding
as queries to the National Center for Biotechnology habitats where An. stephensi discovered were water
Information’s (NCBI) Basic Local Alignment Search tanks, including plastic and metal, concrete water
Tool (BLAST) [19] against the nucleotide collection in cisterns at construction sites, and car wash facilities

25000

20000
Malaria case number

15000

10000

5000

0
2014 2015 2016 2017 2018 2019 2020 2021 2022
Year

Pf Pv
Fig. 2 Annual malaria cases in Hawassa City, southern Ethiopia, 2023
Hawaria et al. Malaria Journal (2023) 22:373 Page 5 of 8

Table 1 Anopheles mosquito larval habitat positivity and species composition by habitat types, Hawassa, southern Ethiopia, 2023
Habitat types Surveyed habitats Positive for any Positive for Positive for Anopheles spp.
n (%) mosquito larvae Anopheles An. stephensi
n (%) larvae n (%)
n (%)

Water tanks 57 (31.5) 24 (42.1) 20 (35.1) 20 (35.1) An. stephensi

Concrete water cisterns 44 (26.2) 20 (45.5) 11 (25.0) 6 (13.6) An. stephensi and An. gambiae (s.l.)
at construction sites
Concrete water collection box 10 (6.0) 3 (30.0) 2 (20.0) 2 (20.0) An. stephensi
for carwash
Shoreline/lake edge 9 (5.4) 8 (88.9) 7 (77.7) No An. gambiae (s.l.), An. pharoensis,
and An. coustani
Drainage ditch 8 (4.8) 4 (50.0) 2 (25.0) No An. gambiae (s.l.)
Discarded buckets 7 (4.2) 3 (42.8) 2 (28.6) No An. gambiae (s.l.)
Manmade pools 5(3.0) No No No
Swamps/marshes 4 (2.4) 4 (100.0) 4 (100.0) No An. gambiae (s.l.), An. pharoensis,
and An. coustani
Discarded tires 4 (2.4) 2 (50.0) No No
Tire tracks/road paddles 4 (2.4) 2 (50.0) 2 (50.0) No An. gambiae (s.l.)
Excavated ground for road 3 (1.8) 3 (100.0) 3 (100.0) No An. gambiae (s.l.)
construction
Others 14 (8.3) 7 (50.0) 1 (7.1) No
no not observed, spp. species

(Table 1). During the survey, 2012 Anopheles larvae were confirmed as An. stephensi. Only two of the 50
were collected and reared, and 548 (27.7%) adult females (4.0%) morphologically identified An. stephensi were
Anopheles emerged. Morphologically, 234 (42.7%) of 558 not confirmed with the PCR endpoint assay. Sanger
were identified as An. stephensi. Other Anopheles species sequencing was performed for 30 confirmed and 2
identified include Anopheles gambiae sensu lato (s.l.) unconfirmed specimens. Of the 30 An. stephensi, 11
(n = 167; 30.5%), Anopheles pharoensis (n = 98; 17.9%), were found to be cox1 haplotype 2 (GenBank accession
and Anopheles coustani (n = 49; 8.9%). Figure 1 depicts OQ865406) and cytb haplotype 2 (GenBank accession
the distribution of the breeding habitats of An. stephensi. OQ863377), while 19 were cox1 haplotype 3 (GenBank
accession OQ865407) and cytb haplotype 1 (GenBank
Adult collection’s Anopheles species composition accession OQ863376). The two specimens not confirmed
The adult survey captured 449 female Anopheles, 53 by ITS2 PCR were identified as Anopheles arabiensis by
(12.0%) of them were An. stephensi. Anopheles stephensi cox1 sequencing.
was captured by the BG Pro Trap and the ProkoPack
aspirator. From 53 An. stephensi, 33 (62.3%) were Discussion
collected indoors and 20 (37.7%) were outdoors; 38 The recent colonization of An. stephensi in Eastern
(71.6%) of them were blood-fed. No An. stephensi was Africa has called for additional molecular surveillance to
captured by the CDC light trap. Anopheles gambiae s.l., delineate the expansion of this new species in Africa [3,
An. pharoensis, An. coustani, and Anopheles demeilloni 4]. To the best of our knowledge, this is the first report
were other Anopheles species collected during the survey. to confirm the presence of An. stephensi in southern
Anopheles gambiae s.l. was the predominant species Ethiopia. Recent increases in malaria transmission in this
(53.7%) followed by An. pharoensis (23.3%) and An. region coincided with the expansion of this species in
coustani (11.5%) (Fig. 3). Ethiopia.
Such rapid spread of An. stephensi to many areas
Molecular identification of An. stephensi of Ethiopia raises the question of whether it is truly
Of the 50 morphologically identified An. stephensi new arrival or has been present for a while, but
specimens analysed, ITS2 PCR results were obtained gone unnoticed. Scholars have forwarded several
from all of them. The morphological identification to hypotheses, one of which is that An. stephensi was
the ITS2 PCR endpoint assay results were compared. recently brought to Ethiopia from the neighbouring
With the PCR endpoint assay, 48 (96%) specimens country Djibouti, which Ethiopia uses as a primary
Hawaria et al. Malaria Journal (2023) 22:373 Page 6 of 8

300

An. gambiae (s.l.)

250 An. stephensi
An. pharoensis
Number of Anopheles spp.

200 An. coustani

An. demeilloni

150

100

50

0
BG CDC ProkoPack
Trapping Methods
Fig. 3 Number of Anopheles species caught in different trapping methods, Hawassa City, southern Ethiopia, 2023

commodity importation route [10]. According to to map An. stephensi’s geographic distribution in the
the recent data, the Ethiopian isolate is most closely country.
related to an isolate from Pakistan [4]. The second Water tanks at construction sites, concrete water
hypothesis involved An. stephensi being present in cisterns, and concrete water collection boxes for
Ethiopia for an extended period yet has gone unnoticed carwashes were habitats found to harbour An. stephensi
[20]. Given the existing gaps in molecular vector in the study area. All these habitats were man-made and
surveillance, it is necessary to carefully re-examine produced as a result of urbanization. Previous studies in
the trend of entomological surveillance practices several countries reported that artificial containers are
that are entirely based on morphological keys for a common larval habitat for An. stephensi [3, 4, 21, 22].
species identification. The morphological similarity Hawassa is one of Ethiopia’s major cities, which has been
of An. stephensi to other Anopheles species, such as rapidly expanding in recent years. Massive development
An. arabiensis as documented in this study, plus the projects are currently undergoing, opening the door
infrequent utilization and cost of molecular methods for the proliferation of new An. stephensi breeding
for identification, such as PCR and sequencing, could environments. Building strong collaborations across
explain why the species have been overlooked [4, many sectors, including health, trade and industry,
12]. This study was carried out in southern Ethiopia, housing agencies, education, and municipalities, is
which is far from the location where An. stephensi critical in achieving successful vector control responses
colonization was initially recorded in the country in such metropolitan contexts. As the World Health
[4], suggesting that An. stephensi colonization is still Organization recommends, an integrated vector control
ongoing. Recently, Balkew et al. [12] conducted a larval strategy should be promoted in the area [1].
survey in southern Ethiopia in 2019 and did not find Discovering An. stephensi in Hawassa has important
An. stephensi in the surveys. Unfortunately, the study public health implications. A survey done in 2019 did not
was a one-time survey that lasted only for a few days. find An. stephensi in the area [12], which was reflected in
In the present study, more intensive surveys were done. the WHO’s malaria threats map as an absence [23]. The
This finding underscores the critical need to expand present study however delineated its presence in the area.
molecular vector surveillance in all malaria-prone areas Coincidentally, the city has witnessed a spike in malaria
Hawaria et al. Malaria Journal (2023) 22:373 Page 7 of 8

cases in recent years [13]. Both Plasmodium vivax and Consent for publication
Not applicable.
P. falciparum are actively transmitted in the study area.
Due to the well-documented vectorial competence of Competing interests
An. stephensi [7], the presence of this species might have The authors declare that they have no competing interests.
contributed to the increase in malaria transmission in
the study area. Previous studies have also indicated that Received: 15 June 2023 Accepted: 2 December 2023
malaria incidence increased following the establishment
of An. stephensi in eastern Ethiopia and Djibouti [8, 9].
However, the present study did not look into the vectorial
capacity of An. stephensi in malaria transmission. Future
studies need to determine the human blood index and References
1. WHO. Initiative to stop the spread of Anopheles stephensi in Africa.
Plasmodium sporozoite rate of this species in Ethiopia Geneva: World Health Organization; 2022. https://​www.​who.​int/​publi​
and the region. The relative contribution of An. stephensi catio​ns/i/​item/​WHO-​UCN-​GMP-​2022.​06.
to the city’s rising malaria incidence also need to be 2. Faulde MK, Rueda LM, Khaireh B. First record of the Asian malaria vector
Anopheles stephensi and its possible role in the resurgence of malaria in
addressed as it coexists alongside native vector species Djibouti, Horn of Africa. Acta Trop. 2014;139:39–43.
such as An. arabiensis, An. coustani, and An. pharoensis. 3. Balkew M, Mumba P, Dengela D, Yohannes G, Getachew D, Yared S,
The survey result suggests the need for further study Chibsa S, Murphy M, George K, Lopez KJD. Geographical distribution of
Anopheles stephensi in eastern Ethiopia. Parasit Vectors. 2020;13:35.
to elucidate this invasive species’ ecology, behavior, 4. Carter TE, Yared S, Gebresilassie A, Bonnell V, Damodaran L, Lopez K, et al.
response to insecticides, and population genetics. A First detection of Anopheles stephensi Liston, 1901 (Diptera: Culicidae)
better understanding of the extent of its distribution and in Ethiopia using molecular and morphological approaches. Acta Trop.
2018;188:180–6.
role in malaria transmission helps develop tailor-made 5. Ahmed A, Pignatelli P, Elaagip A, Hamid MM, Alrahman OF, Weetman
vector intervention strategies in urban settings. D. Invasive malaria vector Anopheles stephensi mosquitoes in Sudan,
2016–2018. Emerg Infect Dis. 2021;27:2952–4.
6. Ochomo E, Milanoi S, Abong’o B, Onyango B, Muchoki M, Omoke D, et al.
Conclusion Molecular surveillance leads to the first detection of Anopheles stephensi
Anopheles stephensi coexists with native vector species in Kenya. Research Square (preprint). 2023.
7. Tadesse FG, Ashine T, Teka H, Esayas E, Messenger LA, Chali W, et al.
in southern Ethiopia. Further research is needed to Anopheles stephensi mosquitoes as vectors of Plasmodium vivax and
determine its relative role in malaria transmission in falciparum, Horn of Africa, 2019. Emerg Infect Dis. 2021;27:603–7.
the region. Heath authorities need to revise the existing 8. Vogel G. Invasive mosquito adds to Africa’s malaria toll. Science.
2022;378:582–3.
vector control strategies to target Anopheles stephensi 9. de Santi VP, Khaireh BA, Chiniard T, Pradines B, Taudon N, Larréché S, et al.
alongside the native malaria vector species. Role of Anopheles stephensi mosquitoes in malaria outbreak, Djibouti,
2019. Emerg Infect Dis. 2021;27:1697–700.
Acknowledgements 10. Sinka ME, Pironon S, Massey NC, Longbottom J, Hemingway J, Moyes CL,
The authors of this manuscript would like to thank the field teams for their et al. A new malaria vector in Africa: predicting the expansion range of
efforts toward collecting and organizing mosquito collections. The authors Anopheles stephensi and identifying the urban populations at risk. Proc
would like to thank Hawassa University, Sidama Regional State Health Office, Natl Acad Sci USA. 2020;117:24900–8.
Hawassa City Administration Health Department, and Sidama Region Public 11. Hamlet A, Dengela D, Tongren JE, Tadesse FG, Bousema T, Sinka M,
Health Institute for facilitating the study. et al. The potential impact of Anopheles stephensi establishment on the
transmission of Plasmodium falciparum in Ethiopia and prospective
Author contributions control measures. BMC Med. 2022;20:135.
DH, SK, and GY contributed to the conception and design of the study. DH, KL, 12. Mnzava A, Monroe AC, Okumu F. Anopheles stephensi in Africa requires a
BB, MB, and MM organized and led the collection of specimens. DH analysed more integrated response. Malar J. 2022;21:156.
the data and drafted the manuscript. MCL is involved in data analysis. DZ 13. Balkew M, Mumba P, Yohannes G, Abiy E, Getachew D, Yared S, et al. An
conducted laboratory analysis. HS contributed to the writing of the paper. update on the distribution, bionomics, and insecticide susceptibility of
SK, DY, and GY critically reviewed the manuscript for intellectual content. All Anopheles stephensi in Ethiopia, 2018–2020. Malar J. 2021;20:263.
authors read and approved the final manuscript. 14. HCAHD. Annual malaria morbidity report. Hawassa, Ethiopia; 2022.
15. WHO. Manual on practical entomology: part II method and techniques.
Funding Geneva: World Health Organization; 1975.
This study was funded by the National Institutes of Health (Grants U19 16. Coetzee M. Key to the females of Afrotropical Anopheles mosquitoes
AI129326 and D43 TW001505). (Diptera: Culicidae). Malar J. 2020;19:70.
17. Onyango SA, Ochwedo KO, Machani MG, Olumeh JO, Debrah I, Omondi
Availability of data and materials CJ, et al. Molecular characterization and genotype distribution of
The datasets used and analysed during the current study are available from thioester-containing protein 1 gene in Anopheles gambiae mosquitoes in
the corresponding author upon reasonable request. western Kenya. Malar J. 2022;21:235.
18. Carter TE, Yared S, Hansel S, Lopez K, Janies D. Sequence-based
identification of Anopheles species in eastern Ethiopia. Malar J.
Declarations 2019;18:135.
19. Altschul SF, Gish W, Miller W, Myers EWLD. Basic local alignment search
Ethics approval and consent to participate tool. J Mol Biol. 1990;215:403–10.
Ethical clearance was obtained from the Institutional Review Board (IRB) of 20. Lobo NF, St. Laurent B, Sikaala CH, Hamainza B, Chanda J, Chinula D,
College of Medicine and Health Science, Hawassa University. The study did not et al. Unexpected diversity of Anopheles species in Eastern Zambia:
involve any endangered or protected species.
Hawaria et al. Malaria Journal (2023) 22:373 Page 8 of 8

implications for evaluating vector behavior and interventions using

molecular tools. Sc Rep. 2015;5:17952.
21. Gayan Dharmasiri AG, Perera AY, Harishchandra J, Herath H, Aravindan
K, Jayasooriya HTR, et al. First record of Anopheles stephensi in Sri Lanka:
a potential challenge for prevention of malaria reintroduction. Malar J.
2017;16:326.
22. Manouchehri AV, Javadian E, Eshighy N, Motabar M. Ecology of Anopheles
stephensi Liston in southern Iran. Trop Geogr Med. 1976;28:228–32.
23. Global Malaria Programme. WHO initiative to stop the spread of
Anopheles stephensi in Africa. Glob Malar Program. Geneva: World Health
Organization; 2023. https://​www.​who.​int/​publi​catio​ns/i/​item/​WHO-​UCN-​
GMP-​2022.​06.

Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.

Ready to submit your research ? Choose BMC and benefit from:

• fast, convenient online submission

• thorough peer review by experienced researchers in your field
• rapid publication on acceptance
• support for research data, including large and complex data types
• gold Open Access which fosters wider collaboration and increased citations
• maximum visibility for your research: over 100M website views per year

At BMC, research is always in progress.

Learn more biomedcentral.com/submissions