Antimicrobial effect of auranofin against Acanthamoeba spp.

Acanthamoebae are opportunistic pathogens that cause serious infections, including Acanthamoeba keratitis, a sight-threatening disease affecting mainly contact lens wearers, and granulomatous amoebic encephalitis, an infection of the central nervous system that occurs mostly in immunocompromised individuals. Although these infections are rare, they are a challenge for healthcare providers. In the last decade, the search for and implementation of novel treatment approaches against these parasites and the infections they cause have intensified, but current options are still unsatisfactory. The aim of this study was to investigate the in vitro activity of the gold-based compound auranofin against Acanthamoeba spp. The study showed that auranofin has potent antimicrobial activity against Acanthamoeba spp., with an IC50 ranging from 2.9 to 3.48 µM, and thus may be useful in the prevention and control of Acanthamoeba infections.

Protein profiles and immunoreactivities of Acanthamoeba morphological groups and genotypes.

Acanthamoeba is a free-living protozoan found in a wide variety of habitats. A classification of Acanthamoeba into currently eighteen genotypes (T1-T18) has been established, however, data on differences between genotypes on the protein level are scarce. The aim of this study was to compare protein and immunoreactivity profiles of Acanthamoeba genotypes. Thirteen strains, both clinical and non-clinical, from genotypes T4, T5, T6, T7, T9, T11 and T12, representing three morphological groups, were investigated for their protein profiles and IgG, IgM and IgA immunoreactivities. It was shown that protein and immunoreactivity profiles of Acanthamoeba genotypes T4, T5, T6, T7, T9, T11 and T12 are clearly distinct from each other, but the banding patterns correlate to the morphological groups. Normal human sera revealed anti-Acanthamoeba antibodies against isolates of all investigated genotypes, interestingly, however only very weak IgM and virtually no IgA immunoreactivity with T7 and T9, both representing morphological group I. The strongest IgG, IgM and IgA immunoreactivities were observed for genotypes T4, T5 and T6. Differences of both, protein and immunological patterns, between cytopathic and non-cytopathic strains, particularly within genotype T4, were not at the level of banding patterns, but rather in expression levels.

Linguatula serrata tongue worm in human eye, Austria.

Linguatula serrata, the so-called tongue worm, is a worm-like, bloodsucking parasite belonging to the Pentastomida group. Infections with L. serrata tongue worms are rare in Europe. We describe a case of ocular linguatulosis in central Europe and provide molecular data on L. serrata tongue worms.

Acanthamoeba strains show reduced temperature tolerance after long-term axenic culture.

Acanthamoeba is a genus of free-living organisms that can be found in various habitats. We investigated the physiological characteristics of 15 Acanthamoeba isolates, representing five genotypes (T4, T5, T6, T7, and T11) of both clinical and nonclinical origins. Moreover, in order to evaluate possible alterations from long-term culture, old and fresh isolates were included, and results were compared to a previous study. We found that there is no significant difference in physiological characteristics between genotypes. However, Acanthamoeba strains that had been grown in axenic culture over long periods of time adapted to axenic growth. Overall growth rates under-agarose migration and particularly, temperature tolerance decrease after long-term axenic culture at room temperature. The only trait that remained rather constant was the cytopathic effect.

Acanthamoeba castellanii : growth on human cell layers reactivates attenuated properties after prolonged axenic culture.

The free-living, but potentially pathogenic, bacteriovorous amoebae of the genus Acanthamoeba can be easily grown axenically in a laboratory culture. This, however, often leads to considerable losses in virulence, and encystment capacity, and to changes in drug susceptibility. We evaluated potential options for a reactivation of a number of physiological properties, attenuated by prolonged axenic laboratory culture, including encystment potential, protease activity, heat resistance, growth rates and drug susceptibility against N-chlorotaurine (NCT). Toward this end, a strain that had been grown axenically for 10 years was repeatedly passaged on human HEp-2 cell monolayers or treated with 5'-azacytidine (AzaC), a methyltransferase inhibitor, and trichostatin A (TSA), a histone deacetylase inhibitor, in order to uplift epigenetic gene regulation. Culture on human cell monolayers resulted in significantly enhanced encystment potentials and protease activities, and higher susceptibility against NCT, whereas the resistance against heat shock was not altered. Treatment with AzaC/TSA resulted in increased encystment rates and protease activities, indicating the participation of epigenetic mechanisms. However, lowered resistances against heat shock indicate that possible stress responses to AzaC/TSA have to be taken into account. Repeated growth on human cell monolayers appears to be a potential method to reactivate attenuated characteristics in Acanthamoeba.

Molecular identification and classification of Cochlonema euryblastum, a zoopagalean parasite of Thecamoeba quadrilineata.

Free-living amoebae can serve a great variety of organisms, predominantly bacteria but to a certain extent also fungi, as a suitable host supplying them with nutrients and protecting them from adverse environmental conditions. In the current study 18S rDNA sequencing was performed to identify a fungal parasite in a Thecamoeba quadrilineata isolate. This parasite morphologically resembled Cochlonema euryblastum, a member of the order Zoopagales, which comprises parasitic species on fungi and invertebrates. Sequence analysis corroborated the morphological identification and the fungal parasite clearly can be assigned to the Zoopagales. Phylogenetic analysis revealed C. euryblastum clustering with two representatives of the mycoparasitic family Piptocephalidaceae. This zooparasitic-mycoparasitic clade represents a sister group of a clade including another member of the Piptocephalidaceae and two other zooparasitic families. Thus, the addition of C. euryblastum to the zoopagalean tree further confirms the finding that molecular data do not support the traditional classification of the Zoopageles.

Onchocerca jakutensis filariasis in humans.

We identified Onchocerca jakutensis as the causative agent of an unusual human filariasis in a patient with lupus erythematosus. To our knowledge, this is the first case of human infection with O. jakutensis and the first human case of zoonotic onchocercosis involving >1 worm.