Acanthamoebae, pathogenic free-living amoebae, can cause Granulomatous Amoebic Encephalitis (GAE) and keratitis, and for both types of infection, no adequate treatment options are available. As the metabolism of pathogens is an attractive treatment target, we set out to examine the energy metabolism of Acanthamoeba castellanii and studied the aerobic and anaerobic capacities of the trophozoites. Under anaerobic conditions, or in the presence of inhibitors of the electron-transport chain, A. castellanii trophozoites became rounded, moved sluggishly and stopped multiplying. This demonstrates that oxygen and the respiratory chain are essential for movement and replication. Furthermore, the simultaneous activities of both terminal oxidases, cytochrome c oxidase and the plant-like alternative oxidase, are essential for normal functioning and replication. The inhibition of normal function caused by the inactivity of the respiratory chain was reversible. Once respiration was made possible again, the rounded, rather inactive amoebae formed acanthopodia within 4 h and resumed moving, feeding and multiplying. Experiments with radiolabelled nutrients revealed a preference for lipids over glucose and amino acids as food. Subsequent experiments showed that adding lipids to a standard culture medium of trophozoites strongly increased the growth rate. Acanthamoeba castellanii trophozoites have a strictly aerobic energy metabolism and β-oxidation of fatty acids, the Krebs cycle, and an aerobic electron-transport chain coupled to the ATP synthase, producing most of the used ATP. The preference for lipids can be exploited, as we show that three known inhibitors of lipid oxidation strongly inhibited the growth of A. castellanii. In particular, thioridazine and perhexiline showed potent effects in low micromolar concentrations. Therefore, this study revealed a new drug target with possibly new options to treat Acanthamoeba infections.
The origin and significance of host specificity are intriguing questions in parasitology. In the case of single-host versus multiple-host parasites, this topic integrates with the concept of the specialist/generalist trade-off. We use the model of sucking lice Polyplax serrata and rodent hosts Apodemus, to address these concepts. Polyplax serrata was shown to form a complex genetic structure, with a strictly specific S lineage living on Apodemus flavicollis, and a less specific N lineage on A. flavicollis and Apodemus sylvaticus. Moreover, the S lineage formed two mitochondrial clades with geographically exclusive distributions and a narrow hybrid zone, providing an opportunity to test the hypothesis that hybrids suffer a decrease in fitness. We sampled 451 individual lice from two host species at 103 localities. We used prevalences and intensities as proxies of fitness, which the parasites realize on their host. The S lineage, strictly specific to Apodemus flavicollis, reached significantly higher prevalences and intensities on its host compared with the N lineage. Conversely, the N lineage occurred with high prevalence and intensity on A. sylvaticus but tended to use also A. flavicollis when the louse populations became too dense. We discuss possible mechanisms behind this difference (particularly interspecific competition as a typical phenomenon in the specialist/generalist systems). We conclude that a parasite's "choice", not accessibility of the host or interspecific competition, is the main factor affecting the louse prevalences. We suggest that historical differences in geographic distribution of both lice and mice may provide a possible explanation for the observed life strategy differences. In contrast to the convincing picture in S and N lineage prevalences, we did not detect an expected drop in fitness in hybrids. We consider instability of the hybrid zone, or decline in abundance of the respective hosts, as possible explanations for this result.
The neotropical fish genus Astyanax (Characidae) and its associated helminths migrated northward from South America following the Great American Biotic Interchange (GABI): ca. 150 Astyanax spp. are found throughout South and Central America, up to the Mexico-USA border. Most characids are distributed south of the Trans-Mexican Volcanic Belt (TMVB), which bisects the country and represents a major transition zone between the neotropical and nearctic realms. Here, we characterize parasites of the monogenean genus Gyrodactylus infecting Astyanax spp. in Mexico: Astyanax aeneus south of the TMBV, Astyanax mexicanus north of it. Based on morphological, phylogenetic (internal transcribed spacer (ITS) and cytochrome oxidase subunit II (cox 2)) and statistical analyses of morphometric data, we confirmed the validity of Gyrodactylus pakan and Gyrodactylus teken, and erected two new species, Gyrodactylus aphaa n. sp. and Gyrodactylus ricardoi n. sp. These four gyrodactylids are part of a complex of morphologically cryptic species, which are phylogenetically closely related to each other, and sister species to Gyrodactylus carolinae and Gyrodactylus heteracanthus, parasites of characins in Brazil. Four gyrodactylid lineages (G. pakan, G. ricardoi n. sp., G. teken, Gyrodactylus sp. A) are distributed north of the TMVB; G. pakan is also widely distributed south of the TMVB, together with G. aphaa n. sp. Based on the ITS phylogeny, Brazilian parasites form a sister clade to all Mexican gyrodactylids, whose derived clades are distributed in progressively more northerly latitudes in Mexico - the three most-derived species north of the TMVB. This would suggest that gyrodactylid species diverged gradually, presumably as their characid fish hosts colonized and adapted to new environments north of the TMVB.
Haemoproteus species (Haemosporida, Haemoproteidae) are cosmopolitan blood parasites that affect bird fitness and health. Recent discoveries based on the application of molecular markers showed that exo-erythrocytic or tissue stages of haemoproteids damage various internal organs including the brain. However, the patterns of exo-erythrocytic development remain unclear for most of the described species. This study aimed to understand the exo-erythrocytic development of Haemoproteus parasites in naturally infected Thrush nightingales Luscinia luscinia (Muscicapidae). Infections were confirmed in eight bird individuals by microscopic examination and PCR-based methods. Organs were examined using histology and in situ hybridization, which applied genus-specific and lineage-specific oligonucleotide probes targeting the 18S rRNA of the parasites. Exo-erythrocytic meronts of Haemoproteus attenuatus (lineage hROBIN1) were found and described for the first known time in this avian host. Most meronts were seen in the lungs, with a few also present in the liver, heart, and pectoral muscle. The available data suggest that this parasite produces only meronts, and not megalomeronts. However, numerous megalomeronts at different stages of development were observed in the gizzard and the heart of one individual. Based on the morphology, location in organs, and diagnostics using the lineage-specific probes, the megalomeronts were attributed to Haemoproteus majoris (lineage hWW2). Two cases of empty capsular-like walls of megalomeronts were seen in the gizzard, indicating that the megalomeronts had already ruptured and degenerated. The extensive microscopic examination did not reveal gametocytes of H. majoris, obviously indicating an abortive development. Abortive haemosporidian infections were often speculated to occur in wildlife but have not been documented in naturally infected birds. This study recognised patterns in the exo-erythrocytic development of H. attenuatus, and is to our knowledge the first documentation of abortive Haemoproteus infection in a naturally infected bird during exo-erythrocytic development.