Review of metazoan parasites of the northern fur seal (Callorhinus ursinus) and the analysis of the gastrointestinal helminth community of the population on St. Paul Island, Alaska.

The northern fur seal (NFS), Callorhinus ursinus (Mammalia: Otariidae), is a marine mammal species included into the IUCN Red List as the vulnerable species which population is dramatically declining. A significant amount of parasitological data collected previously and our recent data allowed us to clarify the list of NFS metazoan parasites and to perform a comprehensive analysis of the gastrointestinal helminth community. Gastrointestinal tracts from 756 NFSs (3- to 4-year-old males) were collected during the annual Aleut subsistence harvests in July-August of 2011-2014 from five separate rookeries on St. Paul Island, Alaska. Totally, 27,625 specimens of helminths and approximately 1000 nasal mites were collected and identified. Detailed analysis of the previously published and newly obtained data revealed 32 species of metazoan parasites, including trematodes (6 species), cestodes (4), nematodes (9), acanthocephalans (9) and arthropods (4). The gastrointestinal helminth community of newly studied NFSs comprised 19 species including trematodes (4), cestodes (3), nematodes (5) and acanthocephalans (7). Temporal changes in the helminth community structure were small but statistically significant. Gastrointestinal helminth infracommunities comprised from 1 to 10 species (average of 4). Small but significant correlation was found between the abundances of acanthocephalans (Corynosoma similis and C. strumosum), nematodes (Contracaecum osculatum, Pseudoterranova spp.) and cestode Diphyllobothrium tetrapterum.

Mitochondria in Obliquely Striated Muscles of the Horsehair Worm Gordionus alpestris (Nematomorpha, Gordioidea) with Structural Organization Typical of Cells with Energy-Intensive Processes.

The ultrastructure of mitochondria in the flattened circomyarian fibers of the horsehair worm Gordionus alpestris (Nemathelminthes) was examined. In contrast to the previously published data, we showed these mitochondria to be giant elongated organelles that densely fill the central cytoplasmic space of the ribbon-like muscle fibers. No fundamental differences were found in the ultrastructure of the muscle tissue mitochondria in actively moving free-living and parasitic G. alpestris worms. The functional significance of the observed ultrastructural organization of mitochondria is discussed in connection with the necessity for an extended mitochondrial membrane system for a uniform supply of active muscle tissue with energy.

Intestinal Parasites in an Ottoman Period Latrine from Acre (Israel) Dating to the Early 1800s CE.

The aim of this study is to determine the species of parasites that affected the inhabitants of the city of Acre on the coast of the eastern Mediterranean during the Ottoman Period. This is the first archaeological study of parasites in the Ottoman Empire. We analysed sediment from a latrine dating to the early 1800s for the presence of helminth eggs and protozoan parasites which caused dysentery. The samples were examined using light microscopy and enzyme-linked immunosorbent assay (ELISA) kits. We found evidence for roundworm (Ascaris lumbricoides), whipworm (Trichuris trichiura), fish tapeworm (Dibothriocephalus sp.), Taenia tapeworm (Taenia sp.), lancet liver fluke (Dicrocoelium dendriticum), and the protozoa Giardia duodenalis and Entamoeba histolytica. The parasite taxa recovered demonstrate the breadth of species present in this coastal city. We consider the effect of Ottoman Period diet, culture, trade and sanitation upon risk of parasitism in this community living 200 years ago.

Comparison of Helminth Infection among the Native Populations of the Arctic and Subarctic Areas in Western Siberia Throughout History: Parasitological Researches on Contemporary and the Archaeological Resources.

The aim of this parasitological study is examining contemporary (the late 20th century) specimens of the arctic or subarctic areas in Western Siberia and comparing them with the information acquired from archaeological samples from the same area. In the contemporary specimens, we observed the parasite eggs of 3 different species: Opisthochis felineus, Ascaris lumbricoides, and Enterobius vermicularis. Meanwhile, in archaeoparasitological results of Vesakoyakha, Kikki-Akki, and Nyamboyto I burial grounds, the eggs of Diphyllobothrium and Taenia spp. were found while no nematode (soil-transmitted) eggs were observed in the same samples. In this study, we concluded helminth infection pattern among the arctic and subarctic peoples of Western Siberia throughout history as follows: the raw fish-eating tradition did not undergo radical change in the area at least since the 18th century; and A. lumbricoides or E. vermicularis did not infect the inhabitants of this area before 20th century. With respect to the Western Siberia, we caught glimpse of the parasite infection pattern prevalent therein via investigations on contemporary and archaeoparasitological specimens.

Reconsideration of Dr. Allen's Report about Hemoptysis Patients from High Prevalence of Archaeoparasitological Paragonimiasis in Korea.

Horace N. Allen, an American physician, was a Presbyterian missionary to Korea. In 1886, he wrote the annual report of the Korean government hospital, summarizing patient statistics according to outpatient and inpatient classification for the first ever in Korean history. In the report, he speculated that hemoptysis cases of outpatient might have been mainly caused by distoma. Allen's conjecture was noteworthy because only a few years lapsed since the first scientific report of paragonimiasis. However, he was not sure of his assumption either because it was not evidently supported by proper microscopic or post-mortem examinations. In this letter, we thus revisit his assumption with our parasitological data recently obtained from Joseon period mummies.

Exosome-transported microRNAs of helminth origin: new tools for allergic and autoimmune diseases therapy?

Chronic diseases associated with inflammation show fast annual increase in their incidence. This has been associated with excessive hygiene habits that limit contacts between the immune system and helminth parasites. Helminthic infections induce regulation and expansion of regulatory T cells (Treg) leading to atypical Th2 type immune responses, with downregulation of the inflammatory component usually associated with these type of responses. Many cells, including those of the immune system, produce extracellular vesicles called exosomes which mediate either immune stimulation (DCs) or immune modulation (T cells). The transfer of miRNAs contained in T-cell exosomes has been shown to contribute to downregulate the production of inflammatory mediators. It has been recently described the delivery to the host-parasite interface of exosomes containing miRNAs by helminths and its internalization by host cells. In this sense, helminth microRNAs transported in exosomes and internalized by immune host cells exert an important role in the expansion of Treg cells, resulting in the control of inflammation. We here provide relevant information obtained in the field of exosomes, cell-cell communication and miRNAs, showing the high potential of helminth miRNAs delivered in exosomes to host cells as new therapeutic tools against diseases associated with exacerbated inflammatory responses.

Mini-FLOTAC and Kato-Katz: helminth eggs watching on the shore of Lake Victoria.

BACKGROUND: One of the challenges for monitoring helminth control programmes based on preventive chemotherapy is the lack of a copro-parasitological gold-standard method that combines good sensitivity with quantitative performance, low cost, and easy-to-learn technique.The aim of our study was to evaluate and compare, the WHO recommended quantitative diagnostic technique (Kato-Katz) and the Mini-FLOTAC. METHODS: Mini-FLOTAC is an innovative method based on floatation of helminths eggs with two different solutions (FS2 and FS7) using a close system (Fill-FLOTAC) with 5% fixative. Kato-Katz was performed following WHO recommendation. The study was carried out in a rural part of Tanzania, close to Lake Victoria, where the laboratory facilities are fairly scarce, and the basic technique used in the local laboratory (direct smear) was taken as reference standard. RESULTS: 201 children were screened for intestinal helminths and 91% of them were found to be positive. The agreement among the three techniques was calculated with k Cohen coefficient and was fairly good (k = 0.4), although the Mini-FLOTAC results were more sensitive for hookworm (98%) with FS2, and for S.mansoni (90%) with FS7 followed by Kato-Katz (91% and 60% respectively) and direct smear (30% and 10% respectively). A good agreement was found between Mini-FLOTAC and Kato-Katz (k = 0.81) with FS7 (k = 0.76) for hookworm diagnosis and a fairly good one for S.mansoni diagnosis (k = 0.5). For both infections we had a poor agreement between the two quantitative techniques and the direct smear (k<0.3). Kato-Katz diagnosed a higher number of eggs (calculated by arithmetic mean) both for hookworm (455 vs 424 EPG) and for S.mansoni (71 vs 58 EPG) compared with the Mini-FLOTAC, but the differences were not significant (p = 0.4). CONCLUSIONS: Mini-FLOTAC is a promising technique, comparable and as sensitive as the Kato-Katz, which is the recommended method in intestinal helminthology for monitoring helminth control programmes. A comparative advantage of the Mini-FLOTAC is that it comprises of a closed system with preserved samples that both protects the operators and allows subsequent examination of the samples. Further studies are needed to validate the mini-FLOTAC with other quantitative techniques (McMaster) and in different settings where other soil-transmitted helminths are also endemic.

Automated imaging and other developments in whole-organism anthelmintic screening.

Helminth infections still represent a huge public health problem throughout the developing world and in the absence of vaccines control is based on periodic mass drug administration. Poor efficacy of some anthelmintics and concerns about emergence of drug resistance has highlighted the need for new drug discovery. Most current anthelmintics were discovered through in vivo screening of selected compounds in animal models but recent approaches have shifted towards screening for activity against adult or larval stages in vitro. Larvae are normally available in greater numbers than adults, can often be produced in vitro and are small enough for microplate assays. However, the manual visualization of drug effects in vitro is subjective, laborious and slow. This can be overcome by application of automated readouts including high-content imaging. Incorporated into robotically controlled HTS platforms such methods allow the very large compound collections being made available by the pharmaceutical industry or academic organizations to be screened against helminths for the first time, invigorating the drug discovery pipeline. Here, we review the status of whole-organism screens based on in vitro activity against living worms and highlight the recent progress towards automated image-based readouts.

Cell polarity: models and mechanisms from yeast, worms and flies.

Determinants of cell polarity orient the behaviour of many cell types during development. Pioneering genetic screens in yeast, worms and flies have identified key polarity determinants that are evolutionarily conserved across the animal kingdom. Recent work in these three model organisms has combined computer modelling with experimental analysis to reveal the molecular mechanisms that drive the polarisation of determinants. Two key principles have emerged: the first is the requirement for a positive-feedback loop to drive self-recruitment of determinants to the plasma membrane; the second is the requirement for mutual antagonism between determinants that localise to opposite ends of the cell.

Cross-disciplinary approaches for measuring parasitic helminth viability and phenotype.

Parasitic worms (helminths) within the Phyla Nematoda and Platyhelminthes are responsible for some of the most debilitating and chronic infectious diseases of human and animal populations across the globe. As no subunit vaccine for any parasitic helminth is close to being developed, the frontline strategy for intervention is administration of therapeutic, anthelmintic drugs. Worryingly, and unsurprising due to co-evolutionary mechanisms, many of these worms are developing resistance to the limited compound classes currently being used. This unfortunate reality has led to a renaissance in next generation anthelmintic discovery within both academic and industrial sectors. However, a major bottleneck in this process is the lack of quantitative methods for screening large numbers of small molecules for their effects on the whole organism. Development of methodologies that can objectively and rapidly distinguish helminth viability or phenotype would be an invaluable tool in the anthelmintic discovery pipeline. Towards this end, we describe how several basic techniques currently used to assess single cell eukaryote viability have been successfully applied to parasitic helminths. We additionally demonstrate how some of these methodologies have been adopted for high-throughput use and further modified for assessing worm phenotype. Continued development in this area is aimed at increasing the rate by which novel anthelmintics are identified and subsequently translated into everyday, practical applications.

Cross-disciplinary approaches for measuring parasitic helminth viability and phenotype

Parasitic worms (helminths) within the Phyla Nematoda and Platyhelminthes are responsible for some of the most debilitating and chronic infectious diseases of human and animal populations across the globe. As no subunit vaccine for any parasitic helminth is close to being developed, the frontline strategy for intervention is administration of therapeutic, anthelmintic drugs. Worryingly, and unsurprising due to co-evolutionary mechanisms, many of these worms are developing resistance to the limited compound classes currently being used. This unfortunate reality has led to a renaissance in next generation anthelmintic discovery within both academic and industrial sectors. However, a major bottleneck in this process is the lack of quantitative methods for screening large numbers of small molecules for their effects on the whole organism. Development of methodologies that can objectively and rapidly distinguish helminth viability or phenotype would be an invaluable tool in the anthelmintic discovery pipeline. Towards this end, we describe how several basic techniques currently used to assess single cell eukaryote viability have been successfully applied to parasitic helminths. We additionally demonstrate how some of these methodologies have been adopted for high-throughput use and further modified for assessing worm phenotype. Continued development in this area is aimed at increasing the rate by which novel anthelmintics are identified and subsequently translated into everyday, practical applications.

Vermes parasíticos (helmintos) dos filos Nematoda e Platelmintos são responsáveis por algumas das doenças infecciosas crônicas e mais debilitantes das populações humana e animal em todo o globo. Já que nenhuma vacina está prestes a ser desenvolvida para nenhum parasita helmíntico, a frente estratégica de intervenção é a administração de drogas terapêuticas anti-helmínticas. De maneira preocupante, e não surpreendente devido a mecanismos coevolutivos, muitos destes vermes estão desenvolvendo resistência às limitadas classes de compostos que têm sido usados no momento. Esta infeliz realidade levou a um renascimento na descoberta de uma nova geração de anti-helmínticos tanto no setor acadêmico quanto no industrial. Contudo, um importante gargalo neste processo é a falta de métodos quantitativos para testar um grande número de pequenas moléculas em relação aos efeitos sobre o organismo inteiro. O desenvolvimento de metodologias que possam distinguir objetiva e rapidamente a viabilidade dos helmintos ou o fenótipo seria uma ferramenta valiosa para canalizar a descoberta de anti-helmínticos. Para este fim, descrevemos aqui como muitas técnicas básicas, correntemente usadas para avaliar a viabilidade de células únicas de eucariotos, têm sido aplicadas com sucesso para helmintos parasíticos. Adicionalmente demonstramos como algumas destas metodologias foram adotadas para uso em larga escala e além disso modificadas para avaliar o fenótipo de vermes. O desenvolvimento contínuo nesta área está voltado para aumentar a taxa com que novos anti-helmínticos são identificados e subsequentemente traduzidos em aplicações práticas cotidianas.

The invention of the pilidium larva in an otherwise perfectly good spiralian phylum Nemertea.

One of the most remarkable larval types among spiralians, and invertebrates in general, is the planktotrophic pilidium. The pilidium is found in a single clade of nemerteans, called the Pilidiophora, and appears to be an innovation of this group. All other nemerteans have either planktotrophic or lecithotrophic juvenile-like planuliform larvae or have direct development. The invention of the pilidium larva is associated with the formation of an extensive blastocoel that supports the delicate larval frame and elaborate ciliary band. Perhaps the most striking characteristic of the pilidium is the way the juvenile worm develops inside the larva from a series of isolated rudiments, called the imaginal discs. The paired cephalic discs, cerebral organ discs, and trunk discs originate as invaginations of larval epidermis and subsequently grow and fuse around the larval gut to form the juvenile. The fully formed juvenile ruptures the larval body and, more often than not, devours the larva during catastrophic metamorphosis. This review is an attempt to examine the pilidium in the context of recent data on development of non-pilidiophoran nemerteans, and speculate about the evolution of pilidial larval development. The author emphasizes the difference between the planuliform larvae of Palaeonemerteans and Hoplonemerteans, and suggest a new name for the hoplonemertean larvae--the decidula.

[Usage of the Hoyer's medium for diagnostics and morphological studies of some parasites]. / Zastosowanie plynu Hoyera do diagnostyki i badan morfologicznych niektórych pasozytów.

Various modification of the mixture of gum arabic with chloral hydrate can be used for mounting of small arthropods as well as some helminths. However, in diagnostic laboratories in Poland they seem to remain unknown. The authors of current work present examples of the Hoyer's medium application. The medium has been composed according to the initial, given by Hoyer, hundred-years-old recipe, which was the root of all other, later used recipes. Hoyer's medium is universal in action and very comfortable to use in slides for microscope investigation. It gives the immediate light-through effect, so it can be helpful in fast diagnostics. At the same time it allows to store the slides for a relatively long time e.g. with education purpose.

[The application of acetic acid and benzyl alcohol in parasitological preparations--advantages and disadvantages]. / Zastosowanie kwasu octowego i alkoholu benzylowego w preparatyce parazytologicznej--zalety i wady.

This work presents the application of acetic acid as a fixing and dehydrating agent and benzyl alcohol as the clearing medium for helminths of the Monogenea, Digenea, Cestoda, Nematoda and Acanthocephala, and mounting them in Canada balsam or other resins.

Worm-like artifacts in exfoliative cytology.

The presence of worms in cytological smears is occasionally reported, although various other structures exist that may be confused with such parasites. We present eight worm-like artifacts observed in routine Papanicolaou smears. Recognition of these structures is important to avoid overvaluation or confusion with true worms.