Transcriptome sequencing and analysis of Plasmodium gallinaceum reveals polymorphisms and selection on the apical membrane antigen-1

BACKGROUND: Plasmodium erythrocyte invasion genes play a key role in malaria parasite transmission, host-specificity and immuno-evasion. However, the evolution of the genes responsible remains understudied. Investigating these genes in avian malaria parasites, where diversity is particularly high, offers new insights into the processes that confer malaria pathogenesis. These parasites can pose a significant threat to birds and since birds play crucial ecological roles they serve as important models for disease dynamics. Comprehensive knowledge of the genetic factors involved in avian malaria parasite invasion is lacking and has been hampered by difficulties in obtaining nuclear data from avian malaria parasites. Thus the first Illumina-based de novo transcriptome sequencing and analysis of the chicken parasite Plasmodium gallinaceum was performed to assess the evolution of essential Plasmodium genes. METHODS: White leghorn chickens were inoculated intravenously with erythrocytes containing P. gallinaceum. cDNA libraries were prepared from RNA extracts collected from infected chick blood and sequencing was run on the HiSeq2000 platform. Orthologues identified by transcriptome sequencing were characterized using phylogenetic, ab initio protein modelling and comparative and population-based methods. RESULTS: Analysis of the transcriptome identified several orthologues required for intra-erythrocytic survival and erythrocyte invasion, including the rhoptry neck protein 2 (RON2) and the apical membrane antigen-1 (AMA-1). Ama-1 of avian malaria parasites exhibits high levels of genetic diversity and evolves under positive diversifying selection, ostensibly due to protective host immune responses. CONCLUSION: Erythrocyte invasion by Plasmodium parasites require AMA-1 and RON2 interactions. AMA-1 and RON2 of P. gallinaceum are evolutionarily and structurally conserved, suggesting that these proteins may play essential roles for avian malaria parasites to invade host erythrocytes. In addition, host-driven selection presumably results in the high levels of genetic variation found in ama-1 of avian Plasmodium species. These findings have implications for investigating avian malaria epidemiology and population dynamics. Moreover, this work highlights the P. gallinaceum transcriptome as an important public resource for investigating the diversity and evolution of essential Plasmodium genes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/1475-2875-13-382) contains supplementary material, which is available to authorized users

Interactions of Water Quality and Integrated Groundwater Management: Examples from the United States and Europe

Groundwater is available in many parts of the world, but the quality of the water may limit its use. Contaminants can limit the use of groundwater through concerns associated with human health, aquatic health, economic costs, or even societal perception. Given this broad range of concerns, this chapter focuses on examples of how water quality issues influence integrated groundwater management. One example evaluates the importance of a naturally occurring contaminant Arsenic (As) for drinking water supply, one explores issues resulting from agricultural activities on the land surface and factors that influence related groundwater management, and the last examines unique issues that result from human-introduced viral pathogens for groundwater-derived drinking water vulnerability. The examples underscore how integrated groundwater management lies at the intersections of environmental characterization, engineering constraints, societal needs, and human perception of acceptable water quality. As such, water quality factors can be a key driver for societal decision making