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.

国家动物园（NZG）是南非国家生物多样性研究所（SANBI）和南部非洲最大的动物园的设施。NZG饲养的9000只圈养动物中，濒临灭绝的非洲企鹅（Spheniscus demersus）。已有几份死后报告报道了由于血友病感染而导致的NZG企鹅死亡，但是尚不清楚涉及的血友病谱系和可能的昆虫媒介。血孢子虫是apiplexplexan寄生虫，可通过吸血的二倍体昆虫感染脊椎动物。因此，当前的研究旨在鉴定蚊子，它们是在非洲企鹅围栏中发现的潜在载体，并使用巢式PCR和实时PCR（qPCR）分析来检测这些昆虫的血孢子虫。使用过夜紫外线灯诱集装置收集蚊子样品3个月。在代表325个蚊子的65个样本中，形态和分子分析表明，淡色库蚊（52.31％）是优势种，其次是Cx。 t heileri（30.77％）和Cx。quinquefasciatus（16.92％）。巢式PCR检测到Leucocytozoon sp。的寄生虫DNA 。和疟原虫属。该CX。巢式PCR的pipiens最高最低感染率（MIR）为5.88％，而qx PCR的pipiens最高为9.41％。倦有MIR的在两个测定中3.64％和没有检测到寄生虫haemosporidian CX。 t heileri。一个Cx。pipiens样本同时感染了两个疟原虫。和白细胞抗原sp。通过巢式PCR检测。这些发现表明，NZG需要采取有效的吸血二翅类昆虫防治措施，还应开展更多研究以确定这些血吸虫寄生虫在NZG中的其他鸟类中的实际患病率。