Ciliates from the faeces of the free-ranging dromedary from Oman: Morphology and molecular phylogeny.

Ciliates Infundibulorium cameli from the faeces of the free-ranging dromedary from Oman were studied using a set of methods of the light and immunofluorescence microscopy and molecular phylogeny. With the use of molecular genetic methods, it was confirmed that the cysts found in the samples simultaneously with trophozoites actually belong to the species I. cameli. Tubulin cytoskeleton organization of trophozoites and cysts of this species were described for the first time. A striking morphological similarity between species I. cameli and Buxtonella sulcata was demonstrated, including the organization of ciliature. Different isolates of I. cameli and B. sulcata formed a common clade on the phylogenetic tree. The level of evolutionary divergence between the 18 S rRNA sequences of I. cameli, B. sulcata and species closest to them according to the results of molecular phylogenetic analysis was estimated. It was demonstrated that the divergence between I. cameli and B. sulcata is extremely low compared to members of other genera included in the analysis. Taxonomic position of I. cameli and B. sulcata was discussed in according to the data of comparative morphology and molecular phylogeny.

If host is refractory, insistent parasite goes berserk: Trypanosomatid Blastocrithidia raabei in the dock bug Coreus marginatus.

Here we characterized the development of the trypanosomatid Blastocrithidia raabei in the dock bug Coreus marginatus using light and electron microscopy. This parasite has been previously reported to occur in the host hemolymph, which is rather typical for dixenous trypanosomatids transmitted to a plant or vertebrate with insect's saliva. In addition, C. marginatus has an unusual organization of the intestine, which makes it refractory to microbial infections: two impassable segments isolate the anterior midgut portion responsible for digestion and absorption from the posterior one containing symbiotic bacteria. Our results refuted the possibility of hemolymph infection, but revealed that the refractory nature of the host provokes very aggressive behavior of the parasite and makes its life cycle more complex, reminiscent of that in some dixenous trypanosomatids. In the pre-barrier midgut portion, the epimastigotes of B. raabei attach to the epithelium and multiply similarly to regular insect trypanosomatids. However, when facing the impassable constricted region, the parasites rampage and either fiercely break through the isolating segments or attack the intestinal epithelium in front of the barrier. The cells of the latter group pass to the basal lamina and accumulate there, causing degradation of the epitheliocytes and thus helping the epimastigotes of the former group to advance posteriorly. In the symbiont-containing post-barrier midgut segment, the parasites either attach to bacterial cells and produce cyst-like amastigotes (CLAs) or infect enterocytes. In the rectum, all epimastigotes attach either to the cuticular lining or to each other and form CLAs. We argue that in addition to the specialized life cycle B. raabei possesses functional cell enhancements important either for the successful passage through the intestinal barriers (enlarged rostrum and well-developed Golgi complex) or as food reserves (vacuoles in the posterior end).

Obligate development of Blastocrithidia papi (Trypanosomatidae) in the Malpighian tubules of Pyrrhocoris apterus (Hemiptera) and coordination of host-parasite life cycles.

Blastocrithidia papi is a unique trypanosomatid in that its life cycle is synchronized with that of its host, and includes an obligate stage of development in Malpighian tubules (MTs). This occurs in firebugs, which exited the winter diapause. In the short period, preceding the mating of overwintered insects, the flagellates penetrate MTs of the host, multiply attached to the epithelial surface with their flagella, and start forming cyst-like amastigotes (CLAs) in large agglomerates. By the moment of oviposition, a large number of CLAs are already available in the rectum. They are discharged on the eggs' surface with feces, used for transmission of bugs' symbiotic bacteria, which are compulsorily engulfed by the newly hatched nymphs along with the CLAs. The obligate development of B. papi in MTs is definitely linked to the life cycle synchronization. The absence of peristalsis allow the trypanosomatids to accumulate and form dense CLA-forming subpopulations, whereas the lack of peritrophic structures facilitates the extensive discharge of CLAs directly into the hindgut lumen. The massive release of CLAs associated with oviposition is indispensable for maximization of the infection efficiency at the most favorable time point.

Life cycle of Blastocrithidia papi sp. n. (Kinetoplastea, Trypanosomatidae) in Pyrrhocoris apterus (Hemiptera, Pyrrhocoridae).

Blastocrithidia papi sp. n. is a cyst-forming trypanosomatid parasitizing firebugs (Pyrrhocoris apterus). It is a member of the Blastocrithidia clade and a very close relative of B. largi, to which it is almost identical through its SSU rRNA gene sequence. However, considering the SL RNA gene these two species represent quite distinct, not even related typing units. Morphological analysis of the new species revealed peculiar or even unique features, which may be useful for future taxonomic revision of the genus Blastocrithidia. These include a breach in the microtubular corset of rostrum at the site of contact with the flagellum, absence of desmosomes between flagellum and rostrum, large transparent vacuole near the flagellar pocket, and multiple vacuoles with fibrous content in the posterior portion of the cell. The study of the flagellates' behavior in the host intestine revealed that they may attach both to microvilli of enterocytes using swollen flagellar tip and to extracellular membranes layers using hemidesmosomes of flagellum. Laboratory experiments on B. papi transmission in P. apterus demonstrated that the parasite may be transmitted vertically (via contaminated surface of eggs) and horizontally (via contaminated substrate and/or necrophagy). We argue that the parasite exploits transmission mechanisms intended for obligate bacterial symbionts of P. apterus.