Nematode Behavior Research Articles

Food familiarity does not change nematode feeding behavior

Ability to identify familiar foods is a major driver of animal feeding behavior. Animals show preference for familiar foods over novel ones, presumably to avoid poisonous substances and better support their growth and reproduction. However, we do not know how their feeding behavior is affected when the familiar food is inferior to a novel food with regards to its palatability and nutritional properties. We tested the hypothesis that feeding preference is determined by specific food characteristics rather than food familiarity with fast-growing soil-dwelling nematodes. Two sequential experiments were designed to test the feeding behavior of soil bacterial-feeding nematodes (Mesorhabditis sp. and Acrobeloides sp.) resulting from familiar food recognition, including a 6-months training period to develop their food familiarity and a feeding experiment to test their food preference for five bacterial species. Nematodes showed clear preference for specific food sources, which were not necessarily the familiar food. Their feeding behavior was not affected by food familiarity, but correlated highly with bacterial traits such as cell size, growth rates, and water content. We conclude that nematode feeding behavior was controlled by food specificity rather than familiarity. We further suggest that physiological factors, possibly the physical size and presence of chemical receptors in the buccal cavity, allow nematodes to rapidly assess their food supply and switch from a familiar food to a preferred, novel food that is easier to ingest and better suited to support their growth and reproduction.

Assessment of the behaviour and survival of nematodes under low oxygen concentrations.

Oxygen is required for the completion of almost all known metazoan lifecycles, but many metazoans harbour abilities to withstand varying degrees and periods of hypoxia. Caenorhabditis elegans, one of the most popular model organism is extensively used as a model for the study of hypoxia and anoxia biology and it has been found that this nematode is capable of tolerance to varying degrees of hypoxia. Considering the extremely high diversity of nematodes, the effects of low oxygen concentration and mechanisms of adaptation to oxygen depletion differ among species. In this study, we used a simple assay to examine anoxia tolerance in four nematode species, including three free-living and one plant parasitic nematode. We found that the plant parasitic nematode Bursaphelenchus xylophilus can survive more than 14 days under anoxic conditions. Comparisons of behaviour during anoxia induction and the repertoire of oxygen sensation genes among the tested species suggested the existence of different oxygen sensation systems between B. xylophilus and C. elegans, which quickly introduce suspended animation in response to oxygen depletion to survive long-term anoxia.

Toxicity evaluation of Wanzhou watershed of Yangtze Three Gorges Reservoir in the flood season in Caenorhabditis elegans

Three Gorges Reservoir (TGR) in the upper stream of Yangtze River in China is a reservoir with the largest and the longest yearly water-level drop. Considering the fact that most of safety assessments of water samples collected from TGR region were based on chemical analysis, we here employed Caenorhabditis elegans to perform in vivo safety assessment of original surface water samples collected from TGR region in the flood season in Wanzhou, Chongqing. Among the examined five original surface water samples, only exposure to original surface water sample collected from backwater area could induce the significant intestinal ROS production, enhance the intestinal permeability, and decrease the locomotion behavior. Additionally, exposure to original surface water sample collected from backwater area altered the expressions of sod-2, sod-5, clk-1, and mev-1. Moreover, mutation of sod-2 or sod-5 was susceptible to the potential toxicity of original surface water sample collected from backwater area on nematodes. Together, our results imply that exposure to surface water sample from the backwater area may at least cause the adverse effects on intestinal function and locomotion behavior in nematodes.

Identification of interneurons required for the aversive response of Caenorhabditis elegans to graphene oxide

BackgroundSo far, how the animals evade the environmental nanomaterials is still largely unclear. In this study, we employed in vivo assay system of Caenorhabditis elegans to investigate the aversive behavior of nematodes to graphene oxide (GO) and the underlying neuronal basis.ResultsIn this assay model, we detected the significant aversive behavior of nematodes to GO at concentrations more than 50 mg/L. Loss-of-function mutation of nlg-1 encoding a neuroligin with the function in connecting pre- and post-synaptic neurons suppressed the aversive behavior of nematodes to GO. Moreover, based on the neuron-specific activity assay, we found that the NLG-1 activity in AIY or AIB interneurons was required for the regulation of aversive behavior to GO. The neuron-specific activities of NLG-1 in AIY or AIB interneurons were also required for the regulation of GO toxicity.ConclusionsUsing nlg-1 mutant as a genetic tool, we identified the AIY and AIB interneurons required for the regulation of aversive behavior to GO. Our results provide an important neuronal basis for the aversive response of animals to environmental nanomaterials.

Laboratory Activity Using Accessible Microfluidics to Study Nematode Behavior in an Electrical Field.

Microfluidic devices are used in a broad range of technological applications, from creating ingredients for cosmetics to discovering new medicines. The small size of microfluidic channels makes it possible to isolate individual cells, collections of cells, and multicellular organisms and study their biology, ecology, and behavior. Microfluidics is particularly well suited to teaching students concepts from different fields of science. A challenge with conventional microfluidic devices is that they are difficult and expensive to make, which has been a barrier for their entry into curricula and classrooms. We describe a simple and low-cost method for creating microfluidic devices and use them to study the behavior of nematodes in an electrical field. Nematodes are ecologically and agriculturally important organisms that respond robustly to various environmental cues. In this activity, we demonstrate that nematodes swim through liquid in microfluidic channels in response to an applied electric field and describe student responses to this activity.

Movement and Motion of Soybean Cyst Nematode Heterodera glycines Populations and Individuals in Response to Abamectin.

Two new in vitro methods were developed to analyze plant-parasitic nematode behavior, at the population and the individual organism levels, through time-lapse image analysis. The first method employed a high-resolution flatbed scanner to monitor the movement of a population of nematodes over a 24-h period at 25°C. The second method tracked multiple motion parameters of individual nematodes on a microscopic scale, using a high-speed camera. Changes in movement and motion of second-stage juveniles (J2) of the soybean cyst nematode Heterodera glycines Ichinohe were measured after exposure to a serial dilution of abamectin (0.1 to 100 μg/ml). Movement and motion of H. glycines were significantly reduced as the concentration of abamectin increased. The effective range of abamectin to inhibit movement and motion of H. glycines J2 was between 1.0 and 10 μg/ml. Proof-of-concept experiments for both methods produced one of the first in vitro sensitivity studies of H. glycines to abamectin. The two methods developed allow for higher-throughput analysis of nematode movement and motion and provide objective and data-rich measurements that are difficult to achieve from conventional microscopic laboratory methods.

A Technique for Thermal Desorption Analyses Suitable for Thermally-Labile, Volatile Compounds.

Many plant and insect interactions are governed by odors released by the plants or insects and there exists a continual need for new or improved methods to collect and identify these odors. Our group has for some time studied below-ground, plant-produced volatile signals affecting nematode and insect behavior. The research requires repeated sampling of volatiles of intact plant/soil systems in the laboratory as well as the field with the help of probes to minimize unwanted effects on the systems we are studying. After evaluating solid adsorbent filters with solvent extraction or solid phase micro extraction fiber sample collection, we found dynamic sampling of small air volumes on Tenax TA filters followed by thermal desorption sample introduction to be the most suitable analytical technique for our applications. Here we present the development and evaluation of a low-cost and relatively simple thermal desorption technique where a cold trap cooled with liquid carbon dioxide is added as an integral part of a splitless injector. Temperature gradient-based focusing and low thermal mass minimizes aerosol formation and eliminates the need for flash heating, resulting in low sample degradation comparable to solvent-based on-column injections. Additionally, since the presence of the cold trap does not affect normal splitless injections, on-the-fly switching between splitless and thermal desorption modes can be used for external standard quantification.

Parameters affecting plant defense pathway mediated recruitment of entomopathogenic nematodes

ABSTRACTEntomopathogenic nematodes are natural enemies and effective biological control agents of subterranean insect herbivores. Interactions between herbivores, plants, and entomopathogenic nematodes are mediated by plant defense pathways. These pathways can induce release of volatiles and recruit entomopathogenic nematodes. Stimulation of these plant defense pathways for induced defense against belowground herbivory may enhance biological control in the field. Knowledge of the factors affecting entomopathogenic nematode behaviour belowground is needed to effectively implement such strategies. To that end, we explore the effect of elicitor, elicitor dose, mechanical damage, and entomopathogenic nematode release distance on recruitment of entomopathogenic nematode infective juveniles to corn seedlings. Increasing doses of methyl jasmonate and methyl salicylate elicitors recruited more entomopathogenic nematodes as did mechanical damage. Recruitment of entomopathogenic nematodes was higher at greater release distances. These results suggest entomopathogenic nematodes are highly tuned to plant status and present a strategy for enhancing biological control using elicitor-stimulated recruitment of entomopathogenic nematodes.

Neuromodulation: The Fevered Mind of the Worm

A landmark study has revealed that an interleukin-17-like signaling system modulates a neural circuit that controls the aggregation behavior of nematodes.

Molecular characterization of FMRFamide-like peptides in Meloidogyne graminicola and analysis of their knockdown effect on nematode infectivity

The rice root-knot nematode, Meloidogyne graminicola, seriously impairs the growth and yield of rice which is an important staple food worldwide. The disruption of neuropeptide signalling leading to attenuation in nematode behaviour and thereby perturbed infection, offers an attractive alternative to control nematodes. In this direction, the present study was aimed at mining of putative FMRFamide-like peptides (FLPs) from the transcriptomic dataset of M. graminicola followed by characterization of those FLPs via sequencing of PCR products, qRT-PCR and Southern hybridization analysis. We have characterized nine flp genes (flp-1, flp-3, flp-6, flp-7, flp-11, flp-12, flp-14, flp-16 and flp-18) and a partial neuropeptide receptor gene (flp-18 GPCR) from M. graminicola in the present study. In addition, in situ localization revealed the expression of flp-1 and flp-7 in neurons posterior to the circumpharyngeal nerve ring of M. graminicola. In vitro silencing of nine flp genes and flp-18 GPCR in M. graminicola J2 and their subsequent infection in rice and wheat roots demonstrated the reduced penetration ability of FLP silenced worms which underscores the potential of the FLPergic system as a broad-spectrum target to manage the root-knot nematode problem in rice-wheat cropping system.

Arabidopsis thaliana as a suitable model host for research on interactions between plant and foliar nematodes, parasites of plant shoot.

The rice white tip nematode (RWTN), Aphelenchoides besseyi and the chrysanthemum foliar nematode (CFN), Aphelenchoides ritzemabosi are migratory plant parasitic nematodes that infect the aboveground parts of plants. In this research, Arabidopsis thaliana was infected by RWTN and CFN under indoor aseptic cultivation, and the nematodes caused recognizable symptoms in the leaves. Furthermore, RWTN and CFN completed their life cycles and proliferated. Therefore, A. thaliana was identified as a new host of RWTN and CFN. The optimum inoculum concentration for RWTN and CFN was 100 nematodes/plantlet, and the optimum inoculum times were 21 and 24 days, respectively. For different RWTN populations, the pathogenicity and reproduction rates were different in the A. thaliana Col-0 ecotype and were positively correlated. The optimum A. thaliana ecotypes were Col-0 and WS, which were the most susceptible to RWTN and CFN, respectively. Additionally, RWTN was ectoparasitic and CFN was ecto- and endoparasitic in A. thaliana. The RWTN and CFN migrated from inoculated leaves to the entire plantlet, and the number of nematodes in different parts of A. thaliana was not correlated with distance from the inoculum point. This is a detailed study of the behavior and infection process of foliar nematodes on A. thaliana.

Influence of temperature and storage conditions on the hatching behavior of cereal cyst nematodes (Heterodera avenae Wollenweber) from Egypt

The cereal cyst nematode Heterodera avenae has been reported in wheat fields in Egypt; however no information is available on the hatch of second-stage juveniles (J2) from such populations of H. avenae. The aim of this study was to subject five Egyptian populations of H. avenae that were obtained from different regions in the Ismailia Province to different temperatures and storage durations to determine the effects on J2 emergence. The Egyptian populations of H. avenae were compared to a population originating from a different climatic zone in Germany (Grafenreuth, Bavaria). Results showed that the J2 hatching pattern of the Egyptian H. avenae populations did not differ from each other. Although the Egyptian and the German populations of H. avenae hatched primarily between 5 and 20 °C, significant differences in J2 hatching have been observed among them. Highest emergence of J2 of the Egyptian populations was observed between 10 and 15 °C, whereas the highest level of J2 emergence of the German population was between 5 and 10 °C. Storing cysts to 5 °C before incubation at 10 °C stimulated the hatch of the German population of H. avenae significantly compared to the control. Whereas, storing cysts at 20 °C, before incubation at 10 °C, significantly stimulated J2 hatching of the Egyptian populations compared to the control. The J2 hatching behavior of the Egyptian populations of H. avenae was in agreement with reports for Mediterranean ecotypes which have winter activity, while the hatching pattern of the German population was similar to those of the Northern ecotypes with spring activity.

Chip Technologies for Screening Chemical and Biological Agents Against Plant-Parasitic Nematodes.

Plant-parasitic nematodes cause substantial damage to agricultural crops worldwide. Long-term management of these pests requires novel strategies to reduce infection of host plants. Disruption of nematode chemotaxis to root systems has been proposed as a potential management approach, and novel assays are needed to test the chemotactic behavior of nematodes against a wide range of synthetic chemicals and root exudates. Two microfluidic chips were developed that measure the attraction or repulsion of nematodes to chemicals ("chemical chip") and young plant roots ("root chip"). The chip designs allowed for chemical concentration gradients to be maintained up to 24 h, the nematodes to remain physically separate from the chemical reservoirs, and for images of nematode populations to be captured using either a microscope or a flatbed scanner. In the experiments using the chemical chips, seven ionic solutions were tested on second-stage juveniles (J2s) of Meloidogyne incognita and Heterodera glycines. Results were consistent with previous reports of repellency of M. incognita to a majority of the ionic solutions, including NH4NO3, KNO3, KCl, MgCl2, and CaCl2. H. glycines was found to be attracted to both NH4NO3 and KNO3, which has not been reported previously. A software program was written to aid in monitoring the location of nematodes at regular time intervals using the root chip. In experiments with the root chip, H. glycines J2s were attracted to roots of 3-day-old, susceptible (cultivar Williams 82) soybean seedlings, and attraction of H. glycines to susceptible soybean was similar across the length of the root. Attraction to resistant (cultivar Jack) soybean seedlings relative to the water only control was inconsistent across runs, and H. glycines J2s were not preferentially attracted to the roots of resistant or susceptible cultivars when both were placed on opposite sides of the same root chip. The chips developed allow for direct tests of plant-parasitic nematode chemotaxis to chemicals and roots with minimal human intervention.

Mechanisms Involved in Nematode Control by Endophytic Fungi.

Colonization of plants by particular endophytic fungi can provide plants with improved defenses toward nematodes. Evidently, such endophytes can be important in developing more sustainable agricultural practices. The mechanisms playing a role in this quantitative antagonism are poorly understood but most likely multifactorial. This knowledge gap obstructs the progress regarding the development of endophytes or endophyte-derived constituents into biocontrol agents. In part, this may be caused by the fact that endophytic fungi form a rather heterogeneous group. By combining the knowledge of the currently characterized antagonistic endophytic fungi and their effects on nematode behavior and biology with the knowledge of microbial competition and induced plant defenses, the various mechanisms by which this nematode antagonism operates or may operate are discussed. Now that new technologies are becoming available and more accessible, the currently unresolved mechanisms can be studied in greater detail than ever before.

Foraging behavior and virulence of some entomopathogenic nematodes

At present the biological control as a pest control technology is becoming more desirable. Biological formulations on basis of entomopathogenic nematodes are one of the effective means for the protection of agricultural and forest plants from harmful insects. Nowadays, the use of entomopathogenic nematodes as biological control agents is a key component in IPM system. The foraging strategies of entomopathogenic nematodes (EPNs) vary between species. This variation is consistent with use of different foraging strategies between ambush, cruise and intermediate to find their host insects. In order to ambush prey, some species of EPNs nictate, or raise their bodies of the soil surface so they are better poised to attach passing insects, other species adopt a cruising strategy and rarely nictate. Some species adopt an intermediate strategy between ambush and cruise. We compared in laboratory the foraging strategies of the entomopathogenic nematode species: Steinernema carpocapsae, Heterorhabditis bacteriophora and the recently described species Steinernema tbilisiensis and assessed their virulence against mealworm beetle, Tenebrio molitor L. (Coleoptera: Tenebrionidae). The tests showed that S. tbilisiensis adopts both foraging strategies.

NemaCount: quantification of nematode chemotaxis behavior in a browser.

Nematodes such as Caenorhabditis elegans offer a very effective and tractable system to probe the underlying mechanisms of diverse sensory behaviors. Numerous platforms exist for quantifying nematode behavior and often require separate dependencies or software. Here I describe a novel and simple tool called NemaCount that provides a versatile solution for the quantification of nematode chemotaxis behavior. The ease of installation and user-friendly interface makes NemaCount a practical tool for measuring diverse behaviors and image features of nematodes such as C. elegans. The main advantage of NemaCount is that it operates from within a modern browser such as Google Chrome or Apple Safari. Any features that change in total number, size, shape, or angular distance between control and experimental preparations are suited to NemaCount for image analysis, while commonly used chemotaxis assays can be quantified, and statistically analyzed using a suite of functions from within NemaCount. NemaCount also offers image filtering options that allow the user to improve object detection and measurements. NemaCount was validated by examining nematode chemotaxis behavior; angular distances of locomotory tracks in C. elegans; and body lengths of Heterorhabditis bacteriophora nematodes. Apart from a modern browser, no additional software is required to operate NemaCount, making NemaCount a cheap, simple option for the analysis of nematode images and chemotaxis behavior.

Method for the assessment of neuromuscular integrity and burrowing choice in vermiform animals

BackgroundThe study of locomotion in vermiform animals has largely been restricted to animals crawling on agar surfaces. While this has been fruitful in the study of neuronal basis of disease and behavior, the reduced physical challenge posed by these environments has prevented these organisms from being equally successful in the study of neuromuscular diseases. Our burrowing assay allowed us to study the effects of muscular exertion on locomotion and muscle degeneration during disease (Beron et al., 2015), as well as the natural burrowing preference of diverse Caenorhabditis elegans strains (Vidal-Gadea et al., 2015). New methodWe describe a simple, rapid, and affordable set of assays to study the burrowing behavior of nematodes and other vermiform organisms which permits the titration of muscular exertion in test animals. ResultsWe show that our burrowing assay design is versatile and can be adapted for use in widely different experimental paradigms. Comparison with existing method(s)Previous assays for the study of neuromuscular integrity in nematodes relied on movement through facile and homogeneous environments. The ability of modulating substrate density allows our burrowing assay to be used to separate animal populations where muscular fitness or health are not visible differentiable by standard techniques. ConclusionThe simplicity, versatility, and potential for greatly facilitating the study of previously challenging neuromuscular disorders makes this assay a valuable addition that overcomes many of the limitations inherent to traditional behavioral tests of vermiform locomotion.

Lactic Acid Bacteria Protects Caenorhabditis elegans from Toxicity of Graphene Oxide by Maintaining Normal Intestinal Permeability under different Genetic Backgrounds.

Lactic acid bacteria (LAB) is safe and useful for food and feed fermentation. We employed Caenorhabditis elegans to investigate the possible beneficial effect of LAB (Lactobacillus bulgaricus) pretreatment against toxicity of graphene oxide (GO) and the underlying mechanisms. LAB prevented GO toxicity on the functions of both primary and secondary targeted organs in wild-type nematodes. LAB blocked translocation of GO into secondary targeted organs through intestinal barrier by maintaining normal intestinal permeability in wild-type nematodes. Moreover, LAB prevented GO damage on the functions of both primary and secondary targeted organs in exposed nematodes with mutations of susceptible genes (sod-2, sod-3, gas-1, and aak-2) to GO toxicity by sustaining normal intestinal permeability. LAB also sustained the normal defecation behavior in both wild-type nematodes and nematodes with mutations of susceptible genes. Therefore, the beneficial role of LAB against GO toxicity under different genetic backgrounds may be due to the combinational effects on intestinal permeability and defecation behavior. Moreover, the beneficial effects of LAB against GO toxicity was dependent on the function of ACS-22, homologous to mammalian FATP4 to mammalian FATP4. Our study provides highlight on establishment of pharmacological strategy to protect intestinal barrier from toxicity of GO.

Lateral Dispersal and Foraging Behavior of Entomopathogenic Nematodes in the Absence and Presence of Mobile and Non-Mobile Hosts.

Entomopathogenic nematodes have been classified into cruisers (active searchers) and ambushers (sit and wait foragers). However, little is known about their dispersal and foraging behavior at population level in soil. We studied lateral dispersal of the ambush foraging Steinernema carpocapsae (ALL strain) and cruise foraging Heterorhabditis bacteriophora (GPS11 strain) from infected host cadavers in microcosms (0.05 m2) containing Wooster silt-loam soil (Oxyaquic fragiudalf) and vegetation in the presence or absence of non-mobile and mobile hosts. Results showed that the presence of a non-mobile host (Galleria mellonella larva in a wire mesh cage) enhanced H. bacteriophora dispersal for up to 24 hr compared with no-host treatment, but had no impact on S. carpocapsae dispersal. In contrast, presence of a mobile host (G. mellonella larvae) increased dispersal of S. carpocapsae compared with no host treatment, but had no effect on H. bacteriophora dispersal. Also H. bacteriophora was better at infecting non-mobile than mobile hosts released into the microcosms and S. carpocapsae was better at infecting mobile than non-mobile hosts, thus affirming the established cruiser-ambusher theory. However, results also revealed that a large proportion of infective juveniles (IJs) of both species stayed near (≤ 3.8 cm) the source cadaver (88-96% S. carpocapsae; 67–79% H. bacteriophora), and the proportion of IJs reaching the farthest distance (11.4 cm) was significantly higher for S. carpocapsae (1.4%) than H. bacteriophora (0.4%) in the presence of mobile hosts. S. carpocapsae also had higher average population displacement than H. bacteriophora in the presence of both the non-mobile (5.07 vs. 3.6 cm/day) and mobile (8.06 vs. 5.3 cm/day) hosts. We conclude that the two species differ in their dispersal and foraging behavior at the population level and this behavior is affected by both the presence and absence of hosts and by their mobility.

GLOWORM-FL: A simulation model of the effects of climate and climate change on the free-living stages of gastro-intestinal nematode parasites of ruminants

Gastrointestinal nematodes are important parasites of livestock and wildlife worldwide, causing mortality and morbidity, regulating host populations and threatening food security through reduced productivity of ruminant livestock. A significant part of the life-cycle of most GINs is completed outside of the host. GINs are therefore susceptible to changes in climate, and evidence of climate-driven changes in the phenology of GINs and the seasonal incidence of disease already exists. A modelling framework, GLOWORM-FL was developed to predict changes in the seasonal dynamics of the free-living stages of trichostrongylid GINs on pasture as a first step towards evaluating potential mitigation strategies. The general model framework was parameterised and validated for three GIN species that infect a range of ruminants worldwide: Haemonchus contortus, Teladorsagia circumcincta and Ostertagia ostertagi. The model builds significantly on previous models of GIN population dynamics by incorporating the behaviour of nematodes in response to climate variability, facilitated by recent advances in our understanding of the ecology of GINs. Simulations using historical and predicted future climatic data for a temperate region reveal the potential for an increase in annual infection pressure of H. contortus and T. circumcincta in small ruminants as increasing temperatures accelerate development and remove constraints on the development of H. contortus during the winter months. In contrast, a significant decrease in annual infection pressure is predicted for O. ostertagi in cattle due to accelerated development being offset by rapid mortality at higher temperatures. A similar trade-off is predicted during the summer months for H. contortus and T. circumcincta resulting in complex seasonal dynamics of the availability of infective stages on pasture. These changes could have significant impacts on the seasonal incidence and pathology of infection by GINs. GLOWORM-FL therefore provides an important tool to predict the seasonal risk of transmission of GINs and will aid in the design of climate-driven, risk-based GIN control strategies.