Intestinal cell diversity and treatment responses in a parasitic nematode at single cell resolution.

Background: Parasitic nematodes, significant pathogens for humans, animals, and plants, depend on diverse organ systems for intra-host survival. Understanding the cellular diversity and molecular variations underlying these functions holds promise for developing novel therapeutics, with specific emphasis on the neuromuscular system's functional diversity. The nematode intestine, crucial for anthelmintic therapies, exhibits diverse cellular phenotypes, and unraveling this diversity at the single-cell level is essential for advancing knowledge in anthelmintic research across various organ systems.

Cell Death and Transcriptional Responses Induced in Larvae of the Nematode by Toxins/Toxicants with Broad Phylogenetic Efficacy.

Establishing methods to investigate treatments that induce cell death in parasitic nematodes will promote experimental approaches to elucidate mechanisms and to identify prospective anthelmintics capable of inducing this outcome. Here, we extended recent progress on a method to monitor cell death and to identify small molecule inhibitors in to , a phylogenetically distant parasitic nematode of significance for both human and agricultural animal health. We utilized a diverse group of small molecule inhibitors referred to as nematode intestinal toxins/toxicants (NITs) coupled with motility, cytological and cell death assays to resolve gross effects on motility and individual cells and organ systems of two larval stages in culture.

Rapid determination of nematode cell and organ susceptibility to toxic treatments.

In research focused on the intestine of parasitic nematodes, we recently identified small molecule inhibitors toxic to intestinal cells of larval Ascaris suum (nematode intestinal toxins/toxicants; "NITs"). Some NITs had anthelmintic activity across the phylogenetic diversity of the Nematoda. The whole-worm motility inhibition assay quantified anthelmintic activity, but worm responses to NITs in relation to pathology or affected molecular pathways was not acquired.

De novo identification of toxicants that cause irreparable damage to parasitic nematode intestinal cells.

Efforts to identify new drugs for therapeutic and preventive treatments against parasitic nematodes have gained increasing interest with expanding pathogen omics databases and drug databases from which new anthelmintic compounds might be identified. Here, a novel approach focused on integrating a pan-Nematoda multi-omics data targeted to a specific nematode organ system (the intestinal tract) with evidence-based filtering and chemogenomic screening was undertaken. Based on de novo computational target prioritization of the 3,564 conserved intestine genes in A.

Omics Driven Understanding of the Intestines of Parasitic Nematodes.

The biological and molecular complexity of nematodes has impeded research on development of new therapies for treatment and control. We have focused on the versatility of the nematode intestine as a target for new therapies. To that end, it is desirable to establish a broad and deep understanding of the molecular architecture underlying intestinal cell functions at the pan-Nematoda level.

Direct experimental manipulation of intestinal cells in Ascaris suum, with minor influences on the global transcriptome.

Ascaris suum provides a powerful model for studying parasitic nematodes, including individual tissues such as the intestine, an established target for anthelmintic treatments. Here, we add a valuable experimental component to our existing functional, proteomic, transcriptomic and phylogenomic studies of the Ascaris suum intestine, by developing a method to manipulate intestinal cell functions via direct delivery of experimental treatments (in this case, double-stranded (ds)RNA) to the apical intestinal membrane. We developed an intestinal perfusion method for direct, controlled delivery of dsRNA/heterogeneous small interfering (hsi) RNA into the intestinal lumen for experimentation.

Compartmentalization of functions and predicted miRNA regulation among contiguous regions of the nematode intestine.

The intestine of parasitic nematodes has proven an important target for therapies aimed at prevention and treatment of diseases caused by these pathogens in humans, animals and plants. We have developed a unique research model with the intestine of Ascaris suum, the large round worm of swine and humans, that will enhance biological research on this tissue. To expand utility of this model, we quantitatively compared expression of 15,382 coding RNAs and 277 noncoding, micro RNAs (miRNAs) among 3 contiguous regions of the adult A.

Pan-Nematoda Transcriptomic Elucidation of Essential Intestinal Functions and Therapeutic Targets With Broad Potential.

The nematode intestine is continuous with the outside environment, making it easily accessible to anthelmintics for parasite control, but the development of new therapeutics is impeded by limited knowledge of nematode intestinal cell biology. We established the most comprehensive nematode intestinal functional database to date by generating transcriptional data from the dissected intestines of three parasitic nematodes spanning the phylum, and integrating the results with the whole proteomes of 10 nematodes (including 9 pathogens of humans or animals) and 3 host species and 2 outgroup species. We resolved 10,772 predicted nematode intestinal protein families (IntFams), and studied their presence and absence within the different lineages (births and deaths) among nematodes.

Functional and phylogenetic characterization of proteins detected in various nematode intestinal compartments.

The parasitic nematode intestine is responsible for nutrient digestion and absorption, and many other processes essential for reproduction and survival, making it a valuable target for anthelmintic drug treatment. However, nematodes display extreme biological diversity (including occupying distinct trophic habitats), resulting in limited knowledge of intestinal cell/protein functions of fundamental or adaptive significance. We developed a perfusion model for isolating intestinal proteins in Ascaris suum (a parasite of humans and swine), allowing for the identification of over 1000 intestinal A.

Peptidases compartmentalized to the Ascaris suum intestinal lumen and apical intestinal membrane.

The nematode intestine is a tissue of interest for developing new methods of therapy and control of parasitic nematodes. However, biological details of intestinal cell functions remain obscure, as do the proteins and molecular functions located on the apical intestinal membrane (AIM), and within the intestinal lumen (IL) of nematodes. Accordingly, methods were developed to gain a comprehensive identification of peptidases that function in the intestinal tract of adult female Ascaris suum.

Genome-wide tissue-specific gene expression, co-expression and regulation of co-expressed genes in adult nematode Ascaris suum.

Background: Caenorhabditis elegans has traditionally been used as a model for studying nematode biology, but its small size limits the ability for researchers to perform some experiments such as high-throughput tissue-specific gene expression studies. However, the dissection of individual tissues is possible in the parasitic nematode Ascaris suum due to its relatively large size. Here, we take advantage of the recent genome sequencing of Ascaris suum and the ability to physically dissect its separate tissues to produce a wide-scale tissue-specific nematode RNA-seq datasets, including data on three non-reproductive tissues (head, pharynx, and intestine) in both male and female worms, as well as four reproductive tissues (testis, seminal vesicle, ovary, and uterus).

Gene expression analysis distinguishes tissue-specific and gender-related functions among adult Ascaris suum tissues.

Over a billion people are infected by Ascaris spp. intestinal parasites. To clarify functional differences among tissues of adult A.

Effect of heat treatment on viability of Taenia hydatigena eggs.

Effects of heat treatments on activation and infectivity of Taenia hydatigena eggs were assessed. Eggs containing oncospheres were used for in vitro and in vivo studies to determine the response to 5min of heat treatment, ranging from room temperature (22°C) to 60°C. The study demonstrated 99.

Effect of ensilation of potato on viability of Taenia hydatigena eggs.

A Taenia hydatigena model was used to assess the effect 0, 7, 14, 21, and 28 days of ensilation of minced potato on viability of tapeworm eggs. For infection of lambs, 2,000 T. hydatigena eggs were ensiled for 0, 7, 14, 21 and 28 days in minced potato at 22°C and fed to recently weaned lambs (29.

The draft genome of the parasitic nematode Trichinella spiralis.

Genome evolution studies for the phylum Nematoda have been limited by focusing on comparisons involving Caenorhabditis elegans. We report a draft genome sequence of Trichinella spiralis, a food-borne zoonotic parasite, which is the most common cause of human trichinellosis. This parasitic nematode is an extant member of a clade that diverged early in the evolution of the phylum, enabling identification of archetypical genes and molecular signatures exclusive to nematodes.

Direct immunization of the abomasum or rectum of goats induces local lymph node responses against Haemonchus contortus mucosal antigens.

We investigated several methods to immunize the abomasum (fourth and gastric stomach) of kid goats by direct (abomasal) or distal (rectal or nasal) routes utilizing mucosal antigens isolated from the abomasal parasite, Haemonchus contortus. Direct (ultrasound guided), immunization of the abomasal mucosa together with rectal immunization established lymphocyte proliferation responses in abomasal lymph nodes (ALNs), while distal methods, alone, produced equivocal results. The differential responses (cellular and antibody) induced by alternative immunization methods demonstrated an experimental system that can facilitate advances in mucosal immunization against H.

Trichinella spiralis: genomic application to control a zoonotic nematode.

The nematode Trichinella spiralis and related species are zoonotic food-borne pathogens of humans. The muscle larval stage of this parasite establishes a chronic infection in skeletal muscle cells of humans who acquire trichinellosis. Muscle larvae also reside in skeletal muscles of animals, swine in particular, and other food animals, including game animals.

Intestinal transcriptomes of nematodes: comparison of the parasites Ascaris suum and Haemonchus contortus with the free-living Caenorhabditis elegans.

Background: The nematode intestine is a major organ responsible for nutrient digestion and absorption; it is also involved in many other processes, such as reproduction, innate immunity, stress responses, and aging. The importance of the intestine as a target for the control of parasitic nematodes has been demonstrated. However, the lack of detailed knowledge on the molecular and cellular functions of the intestine and the level of its conservation across nematodes has impeded breakthroughs in this application.

Advances in the sequencing of the genome of the adenophorean nematode Trichinella spiralis.

The adenophorean nematodes are evolutionarily distant from other species in the phylum Nematoda. Interspecific comparisons of predicted proteins have supported such an ancient divergence. Accordingly, Trichinella spiralis represents a basal nematode representative for genome sequencing focused on gaining a deeper insight into the evolutionary biology of nematodes.

Taenia taeniaeformis: effectiveness of staining oncospheres is related to both temperature of treatment and molecular weight of dyes utilized.

Methods to determine viability of taeniid oncospheres following treatments with potential lethality have practical application in efforts to control transmission. Here we investigated several methods, in lieu of infectivity studies, to assess oncosphere viability and determine lethal temperature treatment regimens. In the first experiment, a standard treatment to exshell oncospheres with 0.

Biology and genome of Trichinella spiralis.

Clade I nematode species in the genus Trichinella can cause infections in humans that lead to mortality and serious morbidity. There are currently eight recognized species or genotypes that comprise this genus. The species display diverse biological characteristics, the evolutionary significance of which recently has been extensively clarified.

Parasitic nematodes - from genomes to control.

The diseases caused by parasitic nematodes in domestic and companion animals are major factors that decrease production and quality of the agricultural products. Methods available for the control of the parasitic nematode infections are mainly based on chemical treatment, non-chemical management practices, immune modulation and biological control. However, even with integrated pest management that frequently combines these approaches, the effective and long-lasting control strategies are hampered by the persistent exposure of host animals to environmental stages of parasites, the incomplete protective response of the host and acquisition of anthelmintic resistance by an increasing number of parasitic nematodes.

Localization of a TNF-activated transcription site and interactions with the gamma activated site within the CAEV U3 70 base pair repeat.

The cytokines TNFalpha and IFNgamma have previously been shown to activate caprine arthritis encephalitis virus (CAEV) transcription. Increased viral titers correlate with increased lesion severity. Therefore, TNFalpha and IFNgamma may augment the caprine arthritis lesion by increasing viral titers.

Haemonchus contortus intestine: a prominent source of mucosal antigens.

We sought to identify antigens from Haemonchus contortus, an abomasal nematode of small ruminants, that stimulate local (abomasal lymph node, ALN) CD4+ T lymphocyte responses during a primary infection. Results led to a focus on antigens from the parasite intestine. The H.

TNFalpha and GM-CSF-induced activation of the CAEV promoter is independent of AP-1.

Caprine arthritis encephalitis virus transcription is under the control of the viral promoter within the long terminal repeat. Previous studies with the closely related maedi visna lentivirus have indicated that viral transcription is dependent upon the AP-1 transcription factor. Other studies have indicated a potential role for the cytokines TNFalpha and GM-CSF in CAEV pathogenesis by increasing viral loads in infected tissues.