Gurltia paralysans: a neglected parasite of domestic cats

Gurltia paralysans (order Strongylida; family Angiostrongylidae) is a metastrongyloid parasite that causes chronic meningomyelitis in domestic cats in South America. The geographic distribution of G. paralysans includes rural and peri-urban areas of Chile and Argentina. However, feline gurltiosis has recently been reported in other South American countries, including Uruguay, Colombia, and Brazil, and was also recently reported in Tenerife, Canary Islands (Spain). Feline gurltiosis is increasingly detected in domestic cats in southern Chile and its apparent geographic range is also increasing, together with an awareness of the disease among veterinarians. The life cycle of the parasite is unknown, but is probably indirect, involving gastropods as the intermediate host, as in other metastrongyloid nematode species. The clinical signs of G. paralysans infection include progressive pelvic limb ataxia, paraparesis, paraplegia, faecal or urinary incontinence, and/or tail paralysis. A definitive diagnosis of feline gurltiosis is still challenging and only possible with necropsy, when adult G. paralysans nematodes are detected within the spinal cord vasculature, together with macroscopic lesions, and characteristic morphological features. A semi-nested PCR method was recently developed for the in vivo diagnosis of this neglected parasite. Current treatment options include macrocyclic lactones and mylbemicn oxime, but the prognosis is poor in severe cases. In this article, we review G. paralysans infection in cats, focusing on the diagnosis shortcomings and the future directions of research into its biology and the associated neurological disease. Comprehensive updates on the epidemiology and clinical features, diagnosis, treatment, and prevention of feline gurltiosis are provided

Suppression of artifacts and barcode bias in high-throughput transcriptome analyses utilizing template switching

Template switching (TS) has been an inherent mechanism of reverse transcriptase, which has been exploited in several transcriptome analysis methods, such as CAGE, RNA-Seq and short RNA sequencing. TS is an attractive option, given the simplicity of the protocol, which does not require an adaptor mediated step and thus minimizes sample loss. As such, it has been used in several studies that deal with limited amounts of RNA, such as in single cell studies. Additionally, TS has also been used to introduce DNA barcodes or indexes into different samples, cells or molecules. This labeling allows one to pool several samples into one sequencing flow cell, increasing the data throughput of sequencing and takes advantage of the increasing throughput of current sequences. Here, we report TS artifacts that form owing to a process called strand invasion. Due to the way in which barcodes/indexes are introduced by TS, strand invasion becomes more problematic by introducing unsystematic biases. We describe a strategy that eliminates these artifacts in silico and propose an experimental solution that suppresses biases from TS

Gastrointestinal Parasites and Bacteria in Free-Living South American Sea Lions (Otaria flavescens) in Chilean Comau Fjord and New Host Record of a Diphyllobothrium scoticum-Like Cestode

Present study aimed to characterize gastrointestinal parasites and culturable bacteria from free-living South American sea lions (Otaria flavescens) inhabiting waters of Comau Fjord, Patagonia, Chile. Therefore, a total of 28 individual fecal samples were collected from sea lions within their natural marine habitat during several diving expeditions. Using classical parasitological techniques, study revealed infections with five different gastrointestinal parasite genera. In addition, bacterial cultures showed presence of at least 28 different bacterial genera. Referring to parasites, protozoan, and metazoan species were found with some of them bearing anthropozoonotic potential and/or pathogenic impact for these marine mammals. As such, four of identified parasite genera harbored zoonotic potential (i.e., Entamoeba, Balantidium, Diphyllobothrium, Anisakis) and one genus (Parafilaroides) represented a specific lungworm of marine pinnipeds. Proglottids from fecal samples showed high morphological homology to “Diphyllobothrium” scoticum (Rennie and Reid, 1912; Meggitt, 1924), which was found in Antarctic sea leopards (Hydrurga leptonyx; Phocidae), but contained eggs of smaller size. Molecular characterization revealed 97–100% identity to a new “Diphyllobothrium” species which was recently isolated from a Californian sea lion (Zalophus californianus; Otariidae) in San Francisco. As such, O. flavescens represents a new host record for this parasite species. Furthermore, potential zoonotic bacteria (i.e., Clostridium, Escherichia, Vibrio, Yersinia, Salmonella) were identified amongst others in O. flavescens indicating a reservoir role for these pinnipeds in marine ecosystem. Current data should be considered as a baseline study for future monitoring surveys on anthropozoonotic pathogens circulating in wild free-living sea lions and their possible impact on public health issues and marine wildlife

Antischistosomal Activity of Trioxaquines: In Vivo Efficacy and Mechanism of Action on Schistosoma mansoni

Schistosomiasis is among the most neglected tropical diseases, since its mode of spreading tends to limit the contamination to people who are in contact with contaminated waters in endemic countries. Here we report the in vitro and in vivo anti-schistosomal activities of trioxaquines. These hybrid molecules are highly active on the larval forms of the worms and exhibit different modes of action, not only the alkylation of heme. The synergy observed with praziquantel on infected mice is in favor of the development of these trioxaquines as potential anti-schistosomal agents

The Syk Kinase SmTK4 of Schistosoma mansoni Is Involved in the Regulation of Spermatogenesis and Oogenesis

The signal transduction protein SmTK4 from Schistosoma mansoni belongs to the family of Syk kinases. In vertebrates, Syk kinases are known to play specialized roles in signaling pathways in cells of the hematopoietic system. Although Syk kinases were identified in some invertebrates, their role in this group of animals has not yet been elucidated. Since SmTK4 is the first Syk kinase from a parasitic helminth, shown to be predominantly expressed in the testes and ovary of adult worms, we investigated its function. To unravel signaling cascades in which SmTK4 is involved, yeast two-/three-hybrid library screenings were performed with either the tandem SH2-domain, or with the linker region including the tyrosine kinase domain of SmTK4. Besides the Src kinase SmTK3 we identified a new Src kinase (SmTK6) acting upstream of SmTK4 and a MAPK-activating protein, as well as mapmodulin acting downstream. Their identities and colocalization studies pointed to a role of SmTK4 in a signaling cascade regulating the proliferation and/or differentiation of cells in the gonads of schistosomes. To confirm this decisive role we performed biochemical and molecular approaches to knock down SmTK4 combined with a novel protocol for confocal laser scanning microscopy for morphological analyses. Using the Syk kinase-specific inhibitor Piceatannol or by RNAi treatment of adult schistosomes in vitro, corresponding phenotypes were detected in the testes and ovary. In the Xenopus oocyte system it was finally confirmed that Piceatannol suppressed the activity of the catalytic kinase domain of SmTK4. Our findings demonstrate a pivotal role of SmTK4 in gametogenesis, a new function for Syk kinases in eukaryotes

Gastrointestinalhelminthosen bei Legehennen in deutschen Öko-Betrieben

Ziel einer Querschnittsstudie war es, die bislang spärlichen Erkenntnisse über den Endoparasitenbefall bei Legehennen in ökologisch geführten Betrieben Deutschlands zu erweitern. Die Studiendurchführung erfolgte im Rahmen des europäischen Forschungsprojekts “HealthyHens“. In sieben Öko-Betrieben wurden jeweils 15 Legehennen (Lohmann Brown oder TetraSL) kurz vor Ende ihrer Legeperiode (ca. 70. Lebenswoche) mittels helminthologischer Teilsektion (Magen-Darm-Trakt) untersucht; Zestodenexemplare wurden zusätzlich molekularbiologisch (rRNA-Gene) charakterisiert. Die über Deutschland verteilten Betriebe unterschieden sich u.a. in Haltungsform (Boden- oder Volierenhaltung, jeweils mit Grünlandauslauf), Größe (500–30.000 Legehennenplätze, z.T. an mehreren Standorten) und Besatzdichte in Stalleinheiten (4,3–6,8 Hühner/m2); als “Wurmmittel“ wurden überwiegend nur pflanzliche Stoffe (Kräutermischungen, Oregano) verwendet, auf einem Betrieb kam Flubendazol zum Einsatz. Infektionen mit Ascaridia galli und Heterakis gallinarum waren in allen Betrieben (innerbetriebliche Prävalenz (IP): 87–93 % bzw. 60–100 %;), mit Capillaria spp. und Raillietina cesticillus in sechs (IP: 53–100 %) bzw. vier Betrieben (IP: 7–73 %) nachzuweisen; weitere Gastrointestinalhelminthenarten wurden nicht festgestellt. Die Stärke des Ascaridia-Befalls (innerbetriebliche mittlere Abundanz (IMA): 3,3±2,7–55,7±70,0 Würmer) unterschied sich zwischen den Betrieben nicht signifikant, jedoch gab es zwischen den Betrieben signifikante Unterschiede (Kruskal-Wallis-Test mit nachfolgenden multiplen Vergleichen nach Dunn) in der Stärke des Heterakis-Befalls (IMA: 30,0±27,5–121,4±84,6 Würmer). Die Ergebnisse legen nahe, dass Helminthenbefall in ökologisch geführten Legehennenbetrieben zu erwarten ist, seine artliche Zusammensetzung sowie die Befallshäufigkeit und -stärke aber variiert. Daher sollte eine Helminthenbekämpfung zur Verbesserung der Tiergesundheit nicht pauschalen Empfehlungen folgen, sondern vielmehr auf die jeweilige Betriebssituation bezogen durchgeführt werden. Wir bedanken uns für die Förderung des Projekts durch die BLE im Rahmen des CORE Organic II Programms (www.coreorganic2.org)

Occurrence of endoparasites in wild Antillean manatees (Trichechus manatus manatus) in Colombia

The recognized impact of parasites in wildlife populations demands surveillance of endangered species like the Antillean manatees (Trichechus manatus manatus) in Colombia. We conducted a parasitological survey in four rescued sea cows in order to document the parasite diversity of this sirenian in the Caribbean wetland of Colombia and contribute to the molecular characterization of its trematodes. The flukes Chiorchis fabaceus, Nudacotyle undicola and the protozoans Eimeria manatus and E. nodulosa were identified in analysed faecal samples. For C. fabaceus and N. undicola, partial regions of ribosomal RNA genes were amplified and sequenced in order to infer their phylogenetic relations. The current study constitutes a new sirenian host (T. manatus manatus) record for the genus Eimeria and the trematode N. undicola. Keywords: Antillean manatee, Trematodes, Eimeria, Coprological survey, Phylogen

Subcutaneous merocercoids of Clistobothrium sp. in two Cape fur seals (Arctocephalus pusillus pusillus)

Fur seals represent intermediate hosts of the cestode Clistobothrium. Large sharks are definitive hosts for these parasites. Two female, 25– and 27-year-old fur seals, caught in the 1980s at the South African coast, were examined pathomorphologically. Both animals showed multifocal, up to 1 cm in diameter large cavities of the thoracic and abdominal subcutaneous adipose tissue containing intraluminal metacestodes of tapeworms, which were surrounded by a locally extensive, pyogranulomatous panniculitis. The metacestodes (merocercoids) of one fur seal were isolated from the subcutaneous adipose tissue and characterized morphologically and for the first time from this host by molecular techniques. The morphometric data corresponded with ‘delphini'-morphotype merocercoids, but the sequence of the partial 28S ribosomal RNA gene identified them as conspecific with merocercoids of the morphotype ‘grimaldii’. These merocercoid types are morphologically Type XV metacestodes of marine tapeworms and represent different species of Clistobothrium. Sequence data were generated for 18S, ITS1, 5.8S, ITS2, partial 28S ribosomal DNA and partial mitochondrial cox1 gene and phylogenetic analysis of 18S rRNA and partial 28S rRNA genes identified the fur seal merocercoids as Clistobothrium species. However, it cannot yet be assigned to species level because of limited molecular data from adult stages. Most likely, both fur seals were infected as juveniles in their original habitat, the coastal regions of South Africa. The metacestode infection is probably an incidental finding, however, there is a chronic inflammatory reaction next to the subcutaneous merocercoids. It is noteworthy, that the merocercoids remain in a potentially infective stage even after more than 20 years. Keywords: Arctocephalus pusillus pusillus, Clistobothrium sp., Fur seal, Cestode, Subcutaneous parasites, Monorygma grimaldi