Caenorhabditis elegans Multi-Tracker Based on a Modified Skeleton Algorithm

Automatic tracking of Caenorhabditis elegans (C. egans) in standard Petri dishes is challenging due to high-resolution image requirements when fully monitoring a Petri dish, but mainly due to potential losses of individual worm identity caused by aggregation of worms, overlaps and body contact. To date, trackers only automate tests for individual worm behaviors, canceling data when body contact occurs. However, essays automating contact behaviors still require solutions to this problem. In this work, we propose a solution to this difficulty using computer vision techniques. On the one hand, a skeletonization method is applied to extract skeletons in overlap and contact situations. On the other hand, new optimization methods are proposed to solve the identity problem during these situations. Experiments were performed with 70 tracks and 3779 poses (skeletons) of C. elegans. Several cost functions with different criteria have been evaluated, and the best results gave an accuracy of 99.42% in overlapping with other worms and noise on the plate using the modified skeleton algorithm and 98.73% precision using the classical skeleton algorithm.

Microfluidic Electric Egg-Laying Assay and Application to In-vivo Toxicity Screening of Microplastics using C. elegans

Environmental pollutants like microplastics are posing health concerns on aquatic animals and the ecosystem. Microplastic toxicity studies using C. elegans as a model are evolving but methodologically hindered from obtaining statistically strong data sets, detecting toxicity effects based on microplastics uptake, and correlating physiological and behavioural effects at an individual-worm level. In this paper, we report a novel microfluidic electric egg-laying assay for phenotypical assessment of multiple worms in parallel. The effects of glucose and polystyrene microplastics at various concentrations on the worms’ electric egg-laying, length, diameter, and length contraction during exposure to electric signal were studied. The device contained eight parallel worm-dwelling microchannels called electric traps, with equivalent electrical fields, in which the worms were electrically stimulated for egg deposition and fluorescently imaged for assessment of neuronal and microplastic uptake expression. A new bidirectional stimulation technique was developed, and the device design was optimized to achieve a testing efficiency of 91.25%. Exposure of worms to 100mM glucose resulted in a significant reduction in their egg-laying and size. The effects of 1μm polystyrene microparticles at concentrations of 100 and 1000 mg/L on the electric egg-laying behaviour, size, and neurodegeneration of N2 and NW1229 (expressing GFP pan-neuronally) worms were also studied. Of the two concentrations, 1000 mg/L caused severe egg-laying deficiency and growth retardation as well as neurodegeneration. Additionally, using single-worm level phenotyping, we noticed intra-population variability in microplastics uptake and correlation with the above physiological and behavioural phenotypes, which was hidden in the population-averaged results. Taken together, these results suggest the appropriateness of our microfluidic assay for toxicological studies and for assessing the phenotypical heterogeneity in response to microplastics.

Biomass measurement of living Lumbriculus variegatus with impedance spectroscopy

Impedance spectroscopy is a useful tool for non-invasive and real time measurements of cell suspensions and a variety of biological tissues. The objective of this study was the investigation of the dielectric properties of living aquatic worms (Lumbriculus variegatus) using impedance spectroscopy in a frequency range between 100 Hz and 10 MHz. We demonstrate a linear relation between the worm biomass and the phase response of the signal thereby providing a quick and precise method to determine the biomass of aquatic worms in situ. Possible applications for non-destructive online biomass monitoring of aquatic worms and other aqueous organisms are discussed. Furthermore, we show that groups of worms fed different diets can be distinguished by the method presented. These results reveal a close relationship between the nutritional composition of the worms and the measured phase response. We also demonstrate that the phase response at 90 kHz does not depend on the worm size. In contrast, the response function for the signal at 440 Hz reveals a linear correlation of average individual worm size and phase. Therefore, we conclude that the measured phase response at 90 kHz qualifies as a measure of the total amount of worm biomass present in the measuring cell, whereas the phase measurement at 440 Hz can be used to estimate the average individual worm size.

Antigenic variability in Trichuris trichiura populations

The present study examines antigenic variability for the human whipworm Trichuris trichiura. Recognition by IgG of somatic antigens of individual worms collected from 3 intensely infected children from Jamaica, West Indies has been investigated by immunoblotting. When probed with 1 plasma sample, significant differences in recognition of 2 selected antigens among worm populations and between male and female worms was observed. In addition, there was evidence for antigenic variability within worm populations at the individual worm level. Such variation may have considerable implications for the development of immunity to parasitic nematodes.

Schistosoma mansoni: stage-dependent damage after in vivo treatment with praziquantel

SummaryThe in vivo effects of a single, subcurative dose (200 mg/kg body wt of mouse) of praziquantel on the structure of the tegument and subtegumental tissues of juvenile (21-, 26- and 30-day-old) and adult Schistosoma mansoni are described. In juvenile worms praziquantel caused only moderate damage to both the tegument and subtegumental tissues although the degree of drug-induced damage was related to both the sex of the worms and to the developmental status of each individual worm. In general, male worms exhibited more extensive and longer lasting surface damage than females. However, the level and extent of the changes increased as the worms became more developmentally advanced with adult worms showing extensive damage to the tegument and vacuolization and disruption of the subtegumental tissues. The possible correlation between the stage-dependent changes in susceptibility to praziquantel and age-related changes, particularly in the phospholipid composition of the outer tegumental membrane is discussed.

The use of faecal egg counts for estimating worm burdens in sheep infected with Haemonchus contortus

SUMMARYClun forest sheep, worm-free from birth, were given a single dose of 20000 infective larvae of Haemonchus contortus. The total number of eggs/day in the faeces was recorded for 21 infections and data on population size, sex ratio and individual worm development were collected from 76 sheep. The female to male ratio was 1·28±0·07 (s.E.). The relation between increase in worm size and uterine egg content was linear. The number of eggs present in the uteri was found to be an accurate measure of eggs passed. It was shown that the daily faecal egg output is related to total parasite weight and is not a measure of the number of individuals present.

Two Species of Algal Symbiont in Naturally Occurrin G Populations of Convoluta Roscoffensis

Convoluta roscoffensis (Graff) has long been known for its symbiosis with a green flagellate definitively described from French stock as Platymonas convolutae by Parke and Manton (Parke & Manton, 1967). Provasoli subsequently discovered a species of Prasinocladus in a culture isolated from C. roscoffensis, but concluded that it was a surface contaminant of the animal (Provasoli, Yamasu & Manton, 1968).Populations of C. roscoffensis from Aberthaw, South Wales (Mettam, 1979), and eighteen sites on Guernsey have been studied. In most patches of the Aberthaw colony, and in two of the Guernsey sites, two algal symbionts have been found. One of the Guernsey, and one of the Aberthaw symbionts appear identical to Platymonas convolutae.They can be distinguished at the ultrastructural level by the presence of canaliculi which penetrate the pyrenoid, and are lined with the double-layered plastid envelope. In both cases the second flagellate shows the nucleus-pyrenoid complex typical of the genus Prasinocladus, where the nucleus penetrates the pyrenoid, and a pair of thylakoid membranes separate the pyrenoid from the starch shell (Parke & Manton, 1965). Neither symbiont has been identified specifically.It is also possible to distinguish the two symbionts in the living animals, since the cup-shaped pyrenoid of the Prasinocladus type is easily distinguished from the spherical condition in Platymonas using the light microscope. A mixed population of symbionts has never been found in an individual worm. For each patch of animals in a colony it was possible to establish the percentage of animals containing each symbiont. This ratio varies from patch to patch in the colony, but has remained remarkably constant within each patch over the six months it has been studied. The Prasinocladus type always predominates at Aberthaw, the Platymonas type being absent in places.

Chemical Prey Preference Polymorphism in Newborn Garter Snakes Thamnophis Sir Talis

Newborn, previously unfed garter snakes (Thamnophis sirtalis) respond with prey attack and tongue flicking to water based extracts prepared from the surface substances of normally eaten prey and presented on cotton swabs. The present experiments demonstrate reliable individual differences in preferred stimuli among members of the same litter. (I) Of 12 snakes tested on six different days with redworm extract, minnow extract, and distilled water, six responded reliably more to worm and one reliably more to minnow. Redworm was overall more effective than minnow. Using two ranking procedures, individual responsivity to redworm was not correlated with responsivity to minnow. Although distilled water was relatively ineffective, correlations of individual snake scores to extracts and water were high and often significant, in contrast to the low correlations between extracts. (2) Of an entire litter of 13 snakes tested on seven days with extracts from two species of earthworms and two species of fish, 11 responded significantly more to worm extracts and one significantly more to fish extracts. The two worm extracts gave almost identical overall scores and the same occurred for the two fish extracts, with the worm being more effective. While individual responsivities to the two worm or two fish extracts were highly correlated, responsivities across worms and fish were not. A few snakes discriminated between individual worm or fish extracts. (3) In 15 sibling newborn snakes tested on three concentrations (100%, 10%, 1%) of earthworm and fish extracts, most responded more to worm regardless of concentration, a 1% worm extract being more effective than 100% fish. The method of limits was used in both ascending and descending sequences. (4) In all three litters, the snakes could be divided into a few discrete groups based on the relative preference for fish or worm extracts. However, there was wide individual variation in attack frequency, attack latency, and tongue flicking in absence of prey attack. (5) The results are discussed in terms of a genetically based perceptual polymorphism. The phenomenon's possible role in the natural history and evolution of snakes, especially the interaction with feeding experience, is elaborated.