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Background  The major malaria vector in Sri Lanka is reported to be  Anopheles culicifacies  with  Anopheles subpictus, Anopheles annularis , and  Anopheles varuna  considered as potential vectors. The occurrence of  Anopheles stephensi , which is the key vector of urban malaria in India and the Middle East, had never been reported from Sri Lanka.    Methods  A series of entomological investigations were carried out by the Anti Malaria Campaign, Ministry of Health, Sri Lanka during December 2016 to April 2017 in two localities of the Mannar District in the Northern Province of the country. Adult mosquito collections were done through indoor and outdoor resting collections, animal and human biting collections and emergence traps. Potential mosquito breeding sites were investigated through larval surveys. The larvae and adults of  An. stephensi  were initially identified using morphological keys, and subsequently confirmed by sequencing the barcode region of the cytochrome c oxidase I ( COI ) gene.    Results  This is the first report of the presence of  An. stephensi  in the island of Mannar in the Northern Province of Sri Lanka.  Anopheles stephensi  (36.65%) was the most abundant anopheline species in the larval habitats in Mannar. It was found breeding together with  An. culicifacies  (20.7%),  An. subpictus  (13.5%) and  An. varuna  (28.13%).  Anopheles stephensi  was found to be abundantly breeding in built wells used for domestic purposes. Adult females of  An. stephensi  were observed in emergence trap collections (93.9%), human landing catches all night (79.2%), pyrethrum spray sheet collections (38.6%), outdoor collections (8.3%), donkey-baited trap collections (14.3), and cattle-baited net trap collections (0.7%).    Conclusions  Sri Lanka was certified as malaria-free by the WHO in September 2016, however, this new finding may pose a serious challenge to the efforts of the Ministry of Health to prevent the re-introduction of malaria transmission in the country, considering the role that  An. stephensi  could play in urban and high vulnerability areas of Sri Lanka.

First record of Anopheles stephensi in Sri Lanka: a potential challenge for prevention of malaria reintroduction