Multiple Worms Research Articles

Parasitic Zoo of Fasciolopsis buski, Gastrodiscoides hominis, Giardia intestinalis and Entamoeba histolytica

Gastrodiscoides hominis) has never been reported. A young lady presented to us with bilateral pitting pedal edema of 2 months duration and laboratory evaluation showed microcytic anemia, low total protein and albumin. Endoscopy and colonoscopy revealed multiple motile worms which were retrieved for species identification. Stool examination revealed polyparasites - Fasciola, Gastrodiscoides, Giardia and Entamoeba. She improved with praziquantel, albendazole and metronidazole. Factors favouring the high endemicity are poor hygiene, consumption of undercooked food, the climatic compatibility for the parasites, swine population and usage of fecal manures for farming. A simple use of anti-helminthic or antiprotozoals in areas of high endemicity can prevent poor outcome among these patients.

A multi-channel device for high-density target-selective stimulation and long-term monitoring of cells and subcellular features in C. elegans.

Selective cell ablation can be used to identify neuronal functions in multicellular model organisms such as Caenorhabditis elegans. The optogenetic tool KillerRed facilitates selective ablation by enabling light-activated damage of cell or subcellular components in a temporally and spatially precise manner. However, the use of KillerRed requires stimulating (5 min-1 h), culturing (~24 h) and imaging (often repeatedly) a large number of individual animals. Current manual manipulation methods are limited by their time-consuming, labor-intensive nature, and their usage of anesthetics. To facilitate large-scale selective ablation, culturing, and repetitive imaging, we developed a densely-packed multi-channel device and used it to perform high-throughput neuronal ablation on KillerRed-expressing animals. The ability to load worms in identical locations with high loading efficiency allows us to ablate selected neurons in multiple worms simultaneously. Our device also enables continuous observation of animals for 24 h following KillerRed activation, and allows the animals to be recovered for behavioural assays. We expect this multi-channel device to facilitate a broad range of long-term imaging and selective illumination experiments in neuroscience.

Multi-well imaging of development and behavior in Caenorhabditis elegans

BackgroundThe nematode Caenorhabditis elegans is widely used as a model for understanding the neuronal and genetic bases of behavior. Recent studies have required longitudinal assessment of individual animal's behavior over extended periods. New methodHere we present a technique for automated monitoring of multiple worms for several days. Our method uses an array of plano-concave glass wells containing standard agar media. The concave well geometry allows worms to be imaged even at the edge of the agar surface and prevents them from burrowing under the agar. We transfer one worm or embryo into each well, and perform imaging of the array of wells using a single camera. Machine vision software is used to quantify size, activity, and/or fluorescence of each worm over time. ResultsWe demonstrate the utility of our method in two applications: (1) quantifying behavioral quiescence and developmental rate in wild-type and mutant animals, and (2) characterizing differences in mating behavior between two C. elegans strains. Comparison with existing method(s)Current techniques for tracking behavior in identified worms are generally restricted to imaging either single animals or have not been shown to work with arbitrary developmental stages; many are also technically complex. Our system works with up to 24 animals of any stages and is technically simple. ConclusionsOur multi-well imaging method is a powerful tool for quantification of long-term behavioral phenotypes in C. elegans.

Protein-losing enteropathy—vomiting the diagnosis

Protein-losing enteropathy is uncommonly due to intestinal worm infestations. The 20-year-old lady reported here presented with bilateral progressive pedal edema for 2 months without abdominal distension, facial puffiness, or other symptoms. Physical examination was unremarkable except for pedal edema. Her body mass index was 17.1 kg/m. Laboratory evaluation revealed microcytic anemia (hemoglobin 10.8 g/dL) and low levels of total protein (4.2 g/dL) and albumin (1.8 g/dL). Cardiac, thyroid, and renal functions were normal and there was no eosinophilia. Ultrasound abdomen revealed mild ascites. Before the malabsorption work up was done, while hospitalized, she vomited pink worms that gave us the diagnosis! Upper gastrointestinal endoscopy revealed multiple motile worms in the duodenum, which range from 1 to 5 cm

Tracking Movement Behavior of Multiple Worms on Food

Neurobiological research in genetically tractable organisms relies heavily on robust assays for behavioral phenotypes. The simple body plan of the nematode Caenorhabditis elegans makes it particularly amenable to the use of automated microscopy and image analysis to describe behavioral patterns quantitatively. Forward genetic screens and screens of drug libraries require high-throughput phenotyping, a task traditionally incompatible with manual scoring of quantitatively varying behaviors. High-throughput automated analysis of C. elegans movement behavior is now possible with several different tracking software packages. The Multiworm Tracker (MWT) described here is designed for high-throughput analysis: it can record dozens of worms simultaneously at 30 frames per second for hours or days at a time. This is accomplished by performing all image analysis in real time, saving only the worm centroid, bearing, and outline data to the disk. To simplify image processing, the system focuses only on worms that have moved, and detects and discards worms that are touching rather than trying to isolate them computationally. Because the software is entirely automated, protocols can run unattended once the worms have been placed and the software has been started. The MWT does not save images for later analysis, but behavior can be validated manually with a companion analysis tool that replays recorded body postures. This protocol describes a basic basal movement assay on food using the MWT; similar protocols apply to related assays and to similar multiple animal trackers. The protocol can be extended to a variety of assays ranging from tap response to chemotaxis.

Facial wrigglies: live extralymphatic filarial infestation in subcutaneous tissues of the head and neck

We report a rare case of a 32-year-old male with live extralymphatic filarial infestation presenting as a facial subcutaneous soft-tissue swelling. To the best of our knowledge these imaging findings have not been previously reported in the head and neck region in the existing English language literature. Real-time high-resolution ultrasonography revealed a solitary well-defined subcutaneous cystic lesion over the right zygomatic arch. It showed multiple linear, echogenic, undulating structures exhibiting a persistent twirling motion during the examination. This typical ultrasonographic appearance was consistent with the filarial dance sign (FDS) of live adult filarial worms. Subsequent MRI confirmed the cystic and solitary nature of the lesion. Complete excision of the cyst was performed, which revealed intracystic straw-coloured fluid and multiple white-coloured adult worms within the lesion. Histopathological examination confirmed multiple adult filarial worms with surrounding reactive inflammatory changes. In an endemic region, identification of the FDS in any normal anatomical structure or abnormal swelling, however remote or unusual the location within the body, should strongly suggest the diagnosis of live active filarial infestation. In view of the increasing migratory trends in the global population, it is imperative for radiologists in all countries to be aware of the typical imaging findings of this disease to arrive at the correct diagnosis.

<em>In vivo</em> Laser Axotomy in <em>C. elegans</em>

Neurons communicate with other cells via axons and dendrites, slender membrane extensions that contain pre- or post-synaptic specializations. If a neuron is damaged by injury or disease, it may regenerate. Cell-intrinsic and extrinsic factors influence the ability of a neuron to regenerate and restore function. Recently, the nematode C. elegans has emerged as an excellent model organism to identify genes and signaling pathways that influence the regeneration of neurons1-6. The main way to initiate neuronal regeneration in C. elegans is laser-mediated cutting, or axotomy. During axotomy, a fluorescently-labeled neuronal process is severed using high-energy pulses. Initially, neuronal regeneration in C. elegans was examined using an amplified femtosecond laser5. However, subsequent regeneration studies have shown that a conventional pulsed laser can be used to accurately sever neurons in vivo and elicit a similar regenerative response1,3,7. We present a protocol for performing in vivo laser axotomy in the worm using a MicroPoint pulsed laser, a turnkey system that is readily available and that has been widely used for targeted cell ablation. We describe aligning the laser, mounting the worms, cutting specific neurons, and assessing subsequent regeneration. The system provides the ability to cut large numbers of neurons in multiple worms during one experiment. Thus, laser axotomy as described herein is an efficient system for initiating and analyzing the process of regeneration.

In vivo laser axotomy in C. elegans.

Neurons communicate with other cells via axons and dendrites, slender membrane extensions that contain pre- or post-synaptic specializations. If a neuron is damaged by injury or disease, it may regenerate. Cell-intrinsic and extrinsic factors influence the ability of a neuron to regenerate and restore function. Recently, the nematode C. elegans has emerged as an excellent model organism to identify genes and signaling pathways that influence the regeneration of neurons1-6. The main way to initiate neuronal regeneration in C. elegans is laser-mediated cutting, or axotomy. During axotomy, a fluorescently-labeled neuronal process is severed using high-energy pulses. Initially, neuronal regeneration in C. elegans was examined using an amplified femtosecond laser5. However, subsequent regeneration studies have shown that a conventional pulsed laser can be used to accurately sever neurons in vivo and elicit a similar regenerative response1,3,7.We present a protocol for performing in vivo laser axotomy in the worm using a MicroPoint pulsed laser, a turnkey system that is readily available and that has been widely used for targeted cell ablation. We describe aligning the laser, mounting the worms, cutting specific neurons, and assessing subsequent regeneration. The system provides the ability to cut large numbers of neurons in multiple worms during one experiment. Thus, laser axotomy as described herein is an efficient system for initiating and analyzing the process of regeneration.

Gallbladder ascariasis

Gallbladder ascariasis is rarely encountered. The causative agent is Ascaris lumbricoides. Presence of ascaris inside the gallbladder can incite acalculous cholecystitis. We aimed in this report to study gallbladder ascariasis. This is a case study of 28 patients with a diagnosis of gallbladder ascariasis from 2005 to 2010 who were seen in the Department of General Surgery, SMHS Hospital. Three patients with gallbladder ascariasis had associated worms in the common bile duct. On abdominal sonography, all patients showed worms as linear echogenic shadows or the worms showed 'belly dance' of worms when actively moving. Acute acalculous cholecystitis was present in 21 patients, and 7 had biliary colic. Dead worms in the gallbladder were seen in 4 patients, of which, one was fragmented and another had calcified form of parasite. Twenty-two patients had spontaneous exit of worms from the gallbladder with a time interval ranging from 1 hour to 144 hours from the time of a diagnosis. Six patients had cholecystectomy. Two patients were subjected to emergency cholecystectomy due to multiple impacted worms not exiting from the gallbladder after conservative treatment. Based on our experience, ultrasound diagnosis is advocated as a highly sensitive and noninvasive technique for suspected gallbladder ascariasis and it can be repeated frequently to monitor movement of worms in the gallbladder and common bile ducts.

P01-467 - Ekbom syndrome - cultural aspects from a clinical case

IntroductionEkbom syndrome is a delusional parasitosis in which the patient has the delusion of infestation by parasites. This condition might occur in concordance with senile dementia or cerebrovascular disease, but it is also present in mood disorders or schizophrenia.ObjectivesTo present a clinical description of a delusional parasitosis that appeared in a young nigerian women after she immigrated in France.AimsOur case is suggestive for showing that delusional parasitosis might develop in circumstances of social vulnerability such as the immigration and might have different cultural aspects.MethodsWe present the case of a 29 years old nigerian women that developed a delusional parasitosis 3 years after her arrival in France. The diagnosis was based on a carefully detailed clinical history, an MMSE was also applied. In order to exclude secondary causes an extensive laboratory evaluation was performed including: complete blood cell count, liver, renal and thyroid function tests, serum electrolytes and glucose levels, vitamin B12, folate, iron studies, coproanalysis, neuroimaging.ResultsThe patient had all laboratory and neuroimaging tests normal. She presented a delusional parasitosis and she described an infestation with multiple intestinal worms. She was capable of describing them and their movements under the skin and also in all the organs. She described dracunculiasis and three more different species that are commonly present in Nigeria, she never mentioned lice or mites.ConclusionsEkbom syndrome is a delusion of hallucinatory mechanism that might have different cultural presentations and could be favored by social vulnerability such as immigration.

Cases and Literature Review of Breast Sparganosis

Sparganosis is a seldom encountered disease of the breast. Based on the results of previous studies and our own experience, we investigated the clinical characteristics of breast sparganosis. Four patients who were treated for breast sparganosis at Samsung Medical Center were included in the study, and additional data were retrieved from the Index Medicus and the Science Citation Index (SCI). The Korean Medical (KM) database and the Korean Association of Medical Journal Editors (KAMJE) database were searched to include information from Korean journals that is not included in international citation indexes. Among 35 patients, 8 (32%) had a record of oral ingestion of an intermediary host. All patients presented with a breast mass, with migration in 6 cases (25%) and pain or an itching sensation in 6 cases (25%). Elongated tubular structures were found in the subcutaneous layer in 22 cases (88%) during radiologic exam. All patients underwent complete surgical excision. Twenty-seven of 29 (93.1%) patients' spargana was separated and confirmed grossly. Histologically, granulomatous inflammation was the most common feature. A single worm was extracted in 16 cases (76.2%) and 5 cases (23.8%) had multiple worms. Sparganosis tends to present as a suspicious mass in the subcutaneous layer of the breast. Complete surgical excision is the treatment of choice, with careful postoperative surveillance.

Molecular and serological survey on taeniasis and cysticercosis in Kanchanaburi Province, Thailand

A community-based field survey on taeniasis and cysticercosis was performed in two villages in Thong Pha Phum District, Kanchanaburi Province, central Thailand, where 3 Taenia species, T. solium, T. saginata and T. asiatica, are sympatrically occurring. Four (0.6%) out of 667 stool samples were egg-positive for Taenia sp. by Kato–Katz technique. Three out of those four persons and other three persons who were Taenia egg-negative but having a recent (< 1 year) history of discharging worms in stool were treated with niclosamide. One Taenia egg-positive woman was not treated because of severe ascites. After treatment, three persons expelled long strobilae with scolices and two persons expelled strobilae without scolex. One Taenia egg-positive person did not expel any worms post-treatment. Among 5 persons, four expelled a single worm, whereas one expelled multiple worms, may be 6 worms but not confirmed by detection of scolices. One scolex was armed with hooklets, whereas 2 others did not. Multiplex PCR of 10 expelled proglottids (including 6 estimated worms from one patient) revealed that one sample was T. solium, one T. saginata, and 8 T. asiatica. A total of 159 residents agreed to receive a serological test for cysticercosis. By ELISA using partially purified glycoprotein antigen, 9 cases, 5 and 4 from villages A and B respectively, were found to be sero-positive. The five and an additional sample on the border line from village A were evaluated using confirmative immunoblot using recombinant chimeric antigen. Among the six samples, four including the border line sample were confirmed to be cysticercosis by immunoblotting. One of the 4 persons had neurological symptoms with nodular lesions in the brain by computed tomography. These 4 confirmed or suspected cysticercosis cases were free of T. solium worms, but two of them including confirmed NCC case had a past (> 1 year) history of expelling proglottids in the stool.

Image Tracking of Multiple C. Elegans Worms Using Adaptive Scanning Optical Microscope (ASOM)

Long-term imaging of living biological specimens is important to infer behaviorial trends and correlate neural structure with behavior. Such study is plagued by the field of view limitation in standard optical microscopes, as the motile specimen would frequently move out of view. A novel microscope, called the adaptive scanning optical microscope (ASOM), has recently been proposed to address this limitation. Through high speed post-objective scanning with a steering mirror, and compensation for optical aberrations with a MEMS deformable mirror, simultaneous imaging and tracking of multiple Caenorhabditis elegans worms has been demonstrated. This article presents the image processing algorithm for tracking multiple worms. Since the steering mirror has to move based on the predicted worm motion, image processing and stable steering mirror motion need to be executed at higher than the composite mosaic video frame rate (in contrast to existing works on image-based worm tracking, which are predominantly based on post-processing). Particular care is placed on disambiguating the worms when they overlap, collide, or entangle, where the worm tracking algorithm may fail. Results from both real time and simulated tracking are presented.

The Parallel Worm Tracker: a platform for measuring average speed and drug-induced paralysis in nematodes.

Background Caenorhabditis elegans locomotion is a simple behavior that has been widely used to dissect genetic components of behavior, synaptic transmission, and muscle function. Many of the paradigms that have been created to study C. elegans locomotion rely on qualitative experimenter observation. Here we report the implementation of an automated tracking system developed to quantify the locomotion of multiple individual worms in parallel.Methodology/Principal FindingsOur tracking system generates a consistent measurement of locomotion that allows direct comparison of results across experiments and experimenters and provides a standard method to share data between laboratories. The tracker utilizes a video camera attached to a zoom lens and a software package implemented in MATLAB®. We demonstrate several proof-of-principle applications for the tracker including measuring speed in the absence and presence of food and in the presence of serotonin. We further use the tracker to automatically quantify the time course of paralysis of worms exposed to aldicarb and levamisole and show that tracker performance compares favorably to data generated using a hand-scored metric.Conclusions/SignficanceAlthough this is not the first automated tracking system developed to measure C. elegans locomotion, our tracking software package is freely available and provides a simple interface that includes tools for rapid data collection and analysis. By contrast with other tools, it is not dependent on a specific set of hardware. We propose that the tracker may be used for a broad range of additional worm locomotion applications including genetic and chemical screening.

A Computational Model forC. elegansLocomotory Behavior: Application to Multiworm Tracking

A computational approach is presented for modeling and quantifying the structure and dynamics of the nematode C. elegans observed by time-lapse microscopy. Worm shape and conformations are expressed in a decoupled manner. Complex worm movements are expressed in terms of three primitive patterns--peristaltic progression, deformation, and translation. The model has been incorporated into algorithms for segmentation and simultaneous tracking of multiple worms in a field, some of which may be interacting in complex ways. A recursive Bayesian filter is used for tracking. Unpredictable behaviors associated with interactions are resolved by multiple-hypothesis tracking. Our algorithm can track worms of diverse sizes and conformations (coiled/uncoiled) in the presence of imaging artifacts and clutter, even when worms are overlapping with others. A two-observer performance assessment was conducted over 16 image sequences representing wild-type and uncoordinated mutants as a function of worm size, conformation, presence of clutter, and worm entanglement. Overall detected tracking failures were 1.41%, undetected tracking failures were 0.41%, and segmentation errors were 1.11% of worm length. When worms overlap, our method reduced undetected failures from 12% to 1.75%, and segmentation error from 11% to 5%. Our method provides the basis for reliable morphometric and locomotory analysis of freely behaving worm populations.

33-Year-Old Woman With Marked Eosinophilia

33-Year-Old Woman With Marked Eosinophilia

Detection and measurement of benzimidazole resistance alleles in Haemonchus contortus using real-time PCR with locked nucleic acid Taqman probes

Benzimidazole resistance is a common problem in parasitic nematodes of ruminants and early detection is vital if its spread is to be monitored and controlled. Real time PCR offers a fast and reliable method for rapid detection and measurement of resistance allele frequencies. In Haemonchus contortus a single nucleotide polymorphism at codon 200 of the β-tubulin gene (TTC to TAC), causing a phenylalanine to tyrosine amino acid substitution, has been shown to be involved in many cases of resistance. Locked nucleic acid (LNA) Taqman probes have been used in this work to detect and measure the frequency of resistance alleles in individual and multiple H. contortus. Detection of resistant genotypes using LNA Taqman probes in individual H. contortus is simpler and more reliable than with previously described assays. Measurement of the frequency of resistant alleles in populations of H. contortus was achieved by using the cycle threshold ( C t) values and a standard curve derived from populations with known allele frequencies. Results using the LNA probes on individual and multiple worms gave similar results to the allele specific PCR. The sensitivity of the LNA assay on multiple nematodes allowed reliable detection of ≥10% resistance allele frequency. Using the final fluorescence method, it was possible to differentiate populations with ∼0, 5 and 10% resistance allele frequencies.

Nucleotide alterations in the D3 domain of the large subunit of ribosomal DNA among 21 species of equine strongyle

The expansion segments or divergent (D) domains in the large subunit (LSU) of the ribosomal DNA have been suggested as genetic markers for taxonomic and/or phylogenetic studies of parasites. In the present study, we assessed the degree of sequence variation in the D3 domain and flanking core regions of the LSU for 21 species of equine strongyles (Strongylida: Strongylidae) and determined which positions in the secondary structure of the LSU were associated with the nucleotide alterations. No intraspecific sequence variation was detected in 17 species, for which multiple individual worms were available. Mutations in sequence among species were detected at 19 nucleotide positions, most of which were located in the D3 domain. Fifteen alterations were transitions, three were transversions and one represented a site of multiple mutations. In relation to the secondary structure element of D3, 26% of these mutations were located in unpaired regions (i.e., end of loops, or in bulges of helices) and thus did not appear to alter the pairing arrangement in the helices of the secondary structure. Many of the other mutations represented partial or complete compensatory base pair changes. The magnitude of interspecific nucleotide variation in the D3 domain (0-4%) was considerably less than that recorded for some other nematode groups (enoplids and thelastomatoids), indicating that this region alone is of limited value for taxonomic and phylogenetic studies for strongyles of equids but is interesting in relation to the evolution of ribosomal DNA.

The nuclear hormone receptor DAF‐12 has opposing effects on Caenorhabditis elegans lifespan and regulates genes repressed in multiple long‐lived worms

The orphan nuclear hormone receptor gene daf-12 in Caenorhabditis elegans plays a key role in the regulation of development and determination of adult longevity. To understand the effects of daf-12 on aging we characterized the lifespan of loss-of-function and gain-of-function daf-12 alleles that have been identified on the basis of their effects on dauer development. We find that these mutations have opposing effects on longevity and resistance to oxidative and thermal stress which makes daf-12 the first gene with alleles that can extend or shorten lifespan. We find that the shortened lifespan of the loss-of-function mutation is due to accelerated aging in young adulthood rather than an adverse effect of the mutation on development. Microarray analysis of worms carrying the two alleles revealed a relatively small number of genes differentially expressed between the two genotypes. Comparison of the expression profiles with the profiles associated with dauer formation and long-lived daf-2 mutants revealed that while the profiles are largely different, there is significant overlap among the genes down-regulated, but not up-regulated, in all profiles. Several of these genes down-regulated in multiple long-lived worms have known effects on lifespan, and many of the genes belong to a family of poorly characterized genes that are strongly down-regulated in dauers, daf-2 mutants, and long-lived daf-12 mutants. Our results point to daf-12 modulating aging and stress responses in part through the repression of specific genes, and emphasize the role that the repression of genes that curtail maximal lifespan plays in lifespan determination.

FATAL AUTOCHTHONOUS EOSINOPHILIC MENINGITIS IN A JAMAICAN CHILD CAUSED BY ANGIOSTRONGYLUS CANTONENSIS

A fatal case of infection with Angiostrongylus cantonensis is reported in a 14-month-old Jamaican boy. Although infection with Angiostrongylus was not considered initially, sections of multiple worms were observed in the brain and lungs at autopsy and confirmed the infection. This is the first reported fatality due to this infection in the Western Hemisphere, and follows shortly after an outbreak of eosinophilic meningitis among a group of travelers to Jamaica. The source of infection in this case could not be determined.