The stone crayfish, Austropotamobius torrentium, is a native European freshwater species sensitive to habitat alterations such as stream channelization and urban development, putting its populations at risk. This study aims to assess how habitat alteration and subsequent water quality changes affect the stone crayfish population in the Kustošak Stream (Croatia) through three selected sites under different levels of anthropogenic disturbance: (1) undisturbed; (2) recently modified, and (3) long-time modified sites. At each site, crayfish were captured, measured, and marked to estimate population size, structure, and crayfish condition. Additionally, we examined whether water quality (18 measured physicochemical parameters) affects relative crayfish abundance. We also used mitochondrial sequencing and microsatellite genotyping to assess species genetic diversity and population connectivity and to compare results among the sites. The results showed that habitat alteration caused an overall reduction in population abundance and changes in size structure; however, it had not yet resulted in detectable genetic differences. Partial least squares regression showed that crayfish abundance was affected by the physicochemical parameters of water, including, among others, oxygen, calcium ions, nitrates, pH, and water temperature. We discuss our findings in the context of the effects of anthropogenic disturbance on the viability and persistence of this EU priority species.

Clinostomum is a genus of parasitic trematodes found worldwide, infecting a wide range of hosts, including freshwater fishes, snails, birds and occasionally humans. In this study, clinostomid metacercariae were collected from Nile tilapia raised in fish farms in the Upper Tana River region, Kenya. The prevalence of infection was 17.2%, with metacercariae infecting the skin, gills and buccal cavity of the fish. Using light microscopy, scanning electron microscopy (SEM) and molecular methods targeting both nuclear ribosomal (ITS1, 5.8S, ITS2) and mitochondrial (COI) regions, the metacercariae were identified as C. cutaneum, C. phalacrocoracis, C. tilapiae and Euclinostomum heterostomum. The three species of Clinostomum have previously been reported to infect fish or piscivorous birds in Kenya, while this is the first report of E. heterostomum in this country. SEM analysis revealed new ultrastructural features of C. cutaneum, including an excretory pore surrounded by minute spiny papillae, an everted cirrus and dome-shaped papillae on the tegumental area around the genital pore. The cirrus lacked basal papillae, showing morphological variation between the adult and metacercarial stages. Our study, therefore, provides new insights into the phenotypic identification of flukes that may be pathogenic to fishes and humans and, therefore, of scientific and practical importance.

Dragonflies and damselflies are important indicator species for quality and health of (semi-)aquatic habitats. Hitherto, 78 species of Odonata have been reported for Austria. Ecological data, Red List assessments, and a dragonfly association index exist, but population- and species-level genetic data are largely lacking. In this study, we establish a comprehensive reference DNA barcode library for Austrian dragonflies and damselflies based on the standard barcoding marker COI. Because of the increasing significance of environmental DNA (eDNA) analyses, we also sequenced a segment of the mitochondrial 16S rRNA gene, a marker often used in eDNA metabarcoding approaches. In total, we provide 786 new COI barcode sequences and 867 new 16S sequences for future applications. Sequencing success was >90 percent for both markers. Identification success was similar for both markers and exceeded 90 percent. Difficulties were only encountered in the genera Anax Leach, 1815, Chalcolestes Kennedy, 1920, Coenagrion Kirby, 1890 and Somatochlora Selys, 1871, with low interspecific genetic distances and, consequently, BIN (barcode index number) sharing. In Anax, however, individual sequences clustered together in species-specific groups in the COI tree. Irrespective of these challenges, the results suggest that both markers perform well within most odonate families in terms of sequencing success and species identification and can be used for reliably delimiting Austrian species, monitoring, and eDNA approaches.

A ParasiteBlitz event offers a brief, intense opportunity to discover diverse parasite species and to reveal life cycles of heteroxenous parasite taxa. In this study, we describe Monomyxum ligophori n. sp., a hyperparasitic myxozoan (Monomyxidae) proliferating in two dactylogyrid monopisthocotylan flatworms ( Ligophorus saladensis, Ligophorus mugilinus ) infecting mugilid fishes ( Mugil cephalus, Mugil curema ) on the Atlantic coast of North America. Furthermore, we used DNA barcoding to infer the parasite’s complex life cycle, matching its hyperparasitic myxospore stages with actinospore stages infecting the polychaete Streblospio benedicti found in the same locality during the ParasiteBlitz and also reported previously from the same region. Thus we report the first life cycle of a myxozoan that most likely does not require a vertebrate host. Hyperparasitic myxozoans are rare with only five species reported worldwide to infect flatworms. This study provides more information on the previously discussed host specificity towards monopisthocotylan hosts of these monomyxid myxozoan hyperparasites. Notably, Monomyxum ligophori n. sp. was detected in two out of four gill-infecting parasitic flatworms (being absent in Ligophorus uruguayensis and Metamicrocotyla macracantha ) found infecting the same fish individuals during the ParasiteBlitz. Our molecular data and phylogenetic analysis support the previously suggested common origin of Monomyxum species infecting monopisthocotylan flatworms, and contribute to understanding the life cycle and host interactions of this unique hyperparasitic myxozoan lineage.

Clinostomum is a genus of parasitic trematodes found worldwide, infecting a wide range of hosts, including freshwater fishes, snails, birds and occasionally humans. In this study, clinostomid metacercariae were collected from Nile tilapia raised in fish farms in the Upper Tana River region, Kenya. The prevalence of infection was 17.2%, with metacercariae infecting the skin, gills and buccal cavity of the fish. Using light microscopy, scanning electron microscopy (SEM) and molecular methods targeting both the ribosomal (ITS1, 5.8S, ITS2) and mitochondrial (COI) regions, the metacercariae were identified as C. cutaneum, C. phalacrocoracis, C. tilapiae and Euclinostomum heterostomum. The three species of Clinostomum have previously been reported to infect fish and piscivorous birds while E. heterostomum is the first report for Kenya. SEM analysis revealed new ultrastructural features of C. cutaneum, including an excretory pore surrounded by minute spiny papillae, an everted cirrus and dome-shaped papillae on the tegumental area around the genital pore. The cirrus lacked basal papillae, showing morphological variation between the adult and metacercarial stages. Our study, therefore, provides new insights into the phenotypic identification of flukes that may be pathogenic to fishes and humans and, therefore, of scientific and practical importance.