Mosquito identification and haemosporidian parasites detection in the enclosure of the African penguins (Spheniscus demersus) at the SANBI zoological garden

The National Zoological Gardens (NZG) is a facility of the South African National Biodiversity Institute (SANBI) and the largest zoo in southern Africa. Among the 9000 captive animals kept by the NZG, is the endangered African penguin (Spheniscus demersus). There have been several post-mortem reports on deaths of penguins in the NZG due to haemosporidian infections, however, the haemosporidian lineages involved and possible insect vector are unknown. Haemosporidians are apicomplexan parasites that infect vertebrates through blood-sucking dipteran insects. Therefore, the current study aimed to identify mosquitoes that are potential vectors found within the African penguin enclosure as well as to detect the haemosporidian parasites from these insects using nested-PCR and real-time PCR (qPCR) analyses. Mosquito samples were collected using an overnight UV-light trap setup for 3 months. From the 65 pooled samples representing 325 mosquitoes, morphological and molecular analysis showed that Culex pipiens (52.31%) was the dominant species followed by Cx.theileri (30.77%) and Cx. quinquefasciatus (16.92%). Nested-PCR detected parasite DNA of Leucocytozoon sp. and Plasmodium sp. The Cx. pipiens had the highest minimum infection rate (MIR) of 5.88% by nested-PCR and 9.41% by qPCR whilst Cx. quinquefasciatus had MIR of 3.64% in both assays and no haemosporidian parasites were detected from Cx.theileri. One Cx. pipiens sample had a co-infection of both Plasmodium sp. and Leucocytozoon sp. detected by nested-PCR. These findings suggest that effective control measures for blood-sucking dipteran insects is required at the NZG and more studies should be conducted to determine the actual prevalence of these haemosporidian parasites among other bird species within NZG.