Dentition patterns and molecular diversity of Mastophorus muris (Gmelin, 1790) (Nematoda: Spiruroidea) support a host-associated subdivision.

Mastophorus muris (Gmelin, 1790) is a globally distributed parasitic nematode of broad range mammals. The taxonomy within the genus Mastophorus and the cryptic diversity among the genus are controversial among taxonomists. This study provides a detailed morphological description of M. muris from Mus musculus combined with a molecular phylogenetic approach. Moreover, descriptions and molecular data of M. muris from non-Mus rodents and wildcats complement our findings and together provide new insights into their taxonomy. The analysis of M. muris was based on light microscopy and scanning electron microscopy. The morphological description focused on the dentition pattern of the two trilobed pseudolabia. Additionally, we described the position of the vulva, arrangement of caudal pairs of papillae, spicules and measured specimens from both sexes and the eggs. For the molecular phylogenetic approach, we amplified the small subunit ribosomal RNA gene and the internal transcribed spacer, and the cytochrome c oxidase subunit 1. Mastophorus morphotypes based on dentition patterns and phylogenetic clustering indicate a subdivision of the genus in agreement with their host. We recognize two groups without a change to formal taxonomy: One group including those specimens infecting Mus musculus, and the second group including organisms infecting non-Mus rodents. Our genetic and morphological data shed light into the cryptic diversity within the genus Mastopohorus. We identified two host-associated groups of M. muris. The described morphotypes and genotypes of M. muris allow a consistent distinction between host-associated parasites.

An apicoplast-resident folate transporter is essential for sporogony of malaria parasites.

Malaria parasites are fast replicating unicellular organisms and require substantial amounts of folate for DNA synthesis. Despite the central role of this critical co-factor for parasite survival, only little is known about intraparasitic folate trafficking in Plasmodium. Here, we report on the expression, subcellular localisation and function of the parasite's folate transporter 2 (FT2) during life cycle progression in the murine malaria parasite Plasmodium berghei. Using live fluorescence microscopy of genetically engineered parasites, we demonstrate that FT2 localises to the apicoplast. In invasive P. berghei stages, a fraction of FT2 is also observed at the apical end. Upon genetic disruption of FT2, blood and liver infection, gametocyte production and mosquito colonisation remain unaltered. But in the Anopheles vector, FT2-deficient parasites develop inflated oocysts with unusual pulp formation consisting of numerous single-membrane vesicles, which ultimately fuse to form large cavities. Ultrastructural analysis suggests that this defect reflects aberrant sporoblast formation caused by abnormal vesicular traffic. Complete sporogony in FT2-deficient oocysts is very rare, and mutant sporozoites fail to establish hepatocyte infection, resulting in a complete block of parasite transmission. Our findings reveal a previously unrecognised organellar folate transporter that exerts critical roles for pathogen maturation in the arthropod vector.

A lipocalin mediates unidirectional heme biomineralization in malaria parasites.

During blood-stage development, malaria parasites are challenged with the detoxification of enormous amounts of heme released during the proteolytic catabolism of erythrocytic hemoglobin. They tackle this problem by sequestering heme into bioinert crystals known as hemozoin. The mechanisms underlying this biomineralization process remain enigmatic. Here, we demonstrate that both rodent and human malaria parasite species secrete and internalize a lipocalin-like protein, PV5, to control heme crystallization. Transcriptional deregulation of PV5 in the rodent parasite Plasmodium berghei results in inordinate elongation of hemozoin crystals, while conditional PV5 inactivation in the human malaria agent Plasmodium falciparum causes excessive multidirectional crystal branching. Although hemoglobin processing remains unaffected, PV5-deficient parasites generate less hemozoin. Electron diffraction analysis indicates that despite the distinct changes in crystal morphology, neither the crystalline order nor unit cell of hemozoin are affected by impaired PV5 function. Deregulation of PV5 expression renders P. berghei hypersensitive to the antimalarial drugs artesunate, chloroquine, and atovaquone, resulting in accelerated parasite clearance following drug treatment in vivo. Together, our findings demonstrate the Plasmodium-tailored role of a lipocalin family member in hemozoin formation and underscore the heme biomineralization pathway as an attractive target for therapeutic exploitation.

A case of Intersexuality in the Parthenogenetic Marmorkrebs (Decapoda: Astacida: Cambaridae).

We describe an intersex specimen of the Marmorkrebs, the only obligate parthenogenetic freshwater crayfish with an all-female population. The individual was a fully functional female which possessed male-like first pleopods. Nevertheless, it reproduced successfully and the offspring were normally developed parthenogenetic females, lacking any trace of male traits. The general rarity of aberrant sexual traits in freshwater crayfishes, in particular in Procambarus, is discussed. We suggest that a dysfunction of the sex determining system, which controls the anlagen of the androgenic glands during development, caused the partial male-like phenotype of this Marmorkrebs specimen. The application of this organism for investigations of sex determination and differentiation is recommended.

The parthenogenetic Marmorkrebs (marbled crayfish) produces genetically uniform offspring.

Genetically identical animals are very much in demand as laboratory objects because they allow conclusions about environmental and epigenetic effects on development, structures, and behavior. Furthermore, questions about the relative fitness of various genotypes can be addressed. However, genetically identical animals are relatively rare, in particular, organisms that combine a high reproduction rate and a complex organization. Based on its exclusively parthenogenetic reproduction mode, it has been suggested that the Marmorkrebs (Crustacea, Decapoda, Astacida), a recently discovered crayfish, is an excellent candidate for research addressing the aforementioned questions. However, until now, a study using molecular markers that clearly proves the genetic uniformity of the offspring has been lacking. Here, with this first molecular study, we show that this crayfish indeed produces genetically uniform clones. We tested this with 19 related individuals of various generations of a Marmorkrebs population by means of six different microsatellite markers. We found that all examined specimens were identical in their allelic composition. Furthermore, half of the analyzed loci were heterozygous. These results and the absence of meioses in previous histological studies of the ovaries lead us to conclude the Marmorkrebs propagates apomictically. Thus, a genetically uniform organism with complex morphology, development, and behavior is now available for various laboratory studies.