Ascaris lumbricoides, the roundworm, and Trichuris trichiura, the whipworm, are human intestinal nematode parasites; both are soil-transmitted helminths, are often placed together in an epidemiological context and both remain neglected despite high prevalence. Our understanding of parasitic disease continues to be enhanced through animal models. Despite the similarities between whipworm and roundworm, there are key differences between the two species and these have influenced the application of their respective animal models. In the case of T. trichiura, the fact that a murine equivalent, T. muris completes its life cycle in a mouse model has greatly enhanced our knowledge of whipworm biology, pathogenicity and immunology. In contrast, A. lumbricoides and its porcine equivalent, Ascaris suum, lack a rodent model in which the life cycle is completed. However, evidence continues to accumulate demonstrating that mice represent useful models of early Ascaris infection, a key stage of the life cycle. The use of mouse models for both Ascaris and Trichuris has a long history with early pioneers discovering fundamental aspects of each parasite's biology. Novel technologies and perspectives, as outlined in this special issue, demonstrate how through the prism of mouse models, we can continue to explore the similarities and differences between roundworms and whipworms.

Human ascariasis is the most common and prevalent neglected tropical disease and is estimated that ~819 million people are infected around the globe, accounting for 0.861 million years of disability-adjusted life years in 2017. Even with the existence of highly effective drugs, the constant presence of infective parasite eggs in the environment contribute to a high reinfection rate after treatment. Due to its high prevalence and broad geographic distribution Ascaris infection is associated with a variety of co-morbidities and co-infections. Here, we provide data from both experimental models and humans studies that illustrate how complex is the interaction of Ascaris with the host immune system, especially, in the context of reinfections, co-infections and associated co-morbidities.

Helminth infections such as ascariasis elicit a type 2 immune response resembling that involved in allergic inflammation, but differing to allergy, they are also accompanied with strong immunomodulation. This has stimulated an increasing number of investigations, not only to better understand the mechanisms of allergy and helminth immunity but to find parasite-derived anti-inflammatory products that could improve the current treatments of chronic non-communicable inflammatory diseases such as asthma. A great number of helminth-derived immunomodulators have been discovered and some of them extensively analysed, showing their potential use as anti-inflammatory drugs in clinical settings. Since Ascaris lumbricoides is one of the most successful parasites, several groups have focused on the immunomodulatory properties of this helminth. As a result, several excretory/secretory components and purified molecules have been analysed, revealing interesting anti-inflammatory activities potentially useful as therapeutic tools. One of these molecules is A. lumbricoides cystatin, whose genomic, cellular, molecular, and immunomodulatory properties are described in this review.

The aim of this study is to determine the species of parasite that infected the population of Brussels during the Medieval and Renaissance periods, and determine if there was notable variation between different households within the city. We compared multiple sediment layers from cesspits beneath three different latrines dating from the 14th–17th centuries. Helminths and protozoa were detected using microscopy and enzyme-linked immunosorbent assay (ELISA). We identified Ascaris sp., Capillaria sp., Dicrocoelium dendriticum, Entamoeba histolytica, Fasciola hepatica, Giardia duodenalis, Taenia sp. and Trichuris sp. in Medieval samples, and continuing presence of Ascaris sp., D. dendriticum, F. hepatica, G. duodenalis and Trichuris sp. into the Renaissance. While some variation existed between households, there was a broadly consistent pattern with the domination of species spread by fecal contamination of food and drink (whipworm, roundworm and protozoa that cause dysentery). These data allow us to explore diet and hygiene, together with routes for the spread of fecal–oral parasites. Key factors explaining our findings are manuring practices with human excrement in market gardens, and flooding of the polluted River Senne during the 14th–17th centuries.

This longitudinal study explored whether aspects of subsistence agriculture were associated with presence and intensity of Ascaris and hookworm in preschool children in rural Panama. Questionnaires were used to collect data on household socio-demographics, child exposure to agriculture and household agricultural practices. Stool samples were collected from children (6 months–5 years) at 3 time points, with albendazole administered after each to clear infections, resulting in 1 baseline and 2 reinfection measures. A novel Agricultural Activity Index (AAI) was developed using principal components analysis to measure the intensity of household agricultural practices. Zero-inflated negative binomial regression models revealed baseline hookworm egg counts were higher if children went to the agricultural plot and if the plot was smaller. Baseline and reinfection Ascaris egg counts were higher if children went to the plot and households had higher AAI, and higher at baseline if the plot was smaller. Caregiver time in the plot was negatively associated with baseline Ascaris egg counts, but positively associated with baseline hookworm and Ascaris reinfection egg counts. Children who spent more time playing around the home were less likely to be infected with Ascaris at baseline. We conclude that preschool child exposure to subsistence agriculture increased Ascaris and hookworm intensity.

This study explored whether the yard environment and child hygiene and play behaviours were associated with presence and intensity of Ascaris and hookworm in preschool children and with eggs and larvae in soil. Data were collected using questionnaires, a visual survey of the yard, soil samples and fecal samples collected at baseline and following re-infection. The presence of eggs/larvae in soil was associated negatively with water storage (eggs) but positively with dogs (eggs) and distance from home to latrine (larvae). Baseline and re-infection prevalences were: hookworm (28·0%, 3·4%); Ascaris (16·9%, 9·5%); Trichuris (0·9%, 0·7%). Zero-inflated negative binomial regression models revealed a higher baseline hookworm infection if yards had eggs or larvae, more vegetation or garbage, and if the child played with soil. Baseline Ascaris was associated with dirt floor, dogs, exposed soil in yard, open defecation and with less time playing outdoors, whereas Ascaris re-infection was associated with water storage, vegetation cover and garbage near the home and not playing with animals. Our results show complex interactions between infection, the yard environment and child behaviours, and indicate that transmission would be reduced if latrines were closer to the home, and if open defecation and water spillage were reduced.

The somatic muscle cells of the parasitic nematode Ascaris suum possess GABA receptors that gate chloride conductances in a similar fashion to the mammalian GABAA receptor subtype. These receptors mediate muscle relaxation and are the site of action of the anthelmintic piperazine. The properties of this receptor differ from the properties of the GABA-gated chloride receptors in the mammalian host, in particular they are not as sensitive to mammalian GABA receptor antagonists such as bicuculline and picrotoxin. Using two-electrode intracellular electrophysiological recording techniques from Ascaris muscle cells, we have tested the potency of a series of azole derivatives for their ability to block the chloride-dependent GABA response. The lead compound, SN606078, 2-(2,6-dichloro-4-trifluromethylphenyl)-4-(4,5-dicyano-lH-imidazol-2-yl)-2H-l,2,3-triazoIe, and 4 structurally related compounds reversibly blocked the conductance increase elicited by 30 μM GABA with IC50s of less than 10 μM. SN606078 (10 μM) decreased the slope of the dose-response curve for GABA, suggesting a non-competitive mechanism of action. In two-electrode voltage clamp experiments, 10μM SN606078 blocked the outward current elicited by 20 μM GABA in a voltage-dependent manner with 72 ± 2% inhibition at −20 mV and 49±6% inhibition at −40 mV. These observations indicate that SN606078 may act as an open-channel blocker of the GABA-gated chloride channel in A. suum.

PF1 (SDPNFLRFamide) is a FMRFamide-like peptide extracted from the free-living nematode Panagrellus redivivus. Here we show that this peptide causes a hyperpolarization of somatic muscle cells of the parasitic nematode Ascaris suum and a relaxation of the somatic muscle strip preparation. We have assessed whether or not the relaxation of Ascaris dorsal muscle strip by PF1 is due to (i) inhibition of the release of the excitatory neuromuscular junction transmitter acetylcholine (ACh), (ii) potentiation of the release of the inhibitory neuromuscular junction transmitter γ-aminobutyric acid (GABA) or (iii) a direct inhibitory action of the peptide on the muscle cells. Under the experimental conditions described here, tonic ACh release does not seem to be involved in determining the resting membrane potential or resting tone of the Ascaris dorsal muscle strip and thus inhibition of tonic ACh release is unlikely to explain the relaxation elicited by the peptide. Furthermore, PF1 (100 nM–1 μM) inhibited the contraction of the muscle strip elicited by bath application of ACh, suggesting either a direct inhibitory action of the peptide on the muscle cells or a potentiation of GABA release. In electrophysiological experiments, the reversal potential for the PF1 hyperpolarization was not the same as that for GABA. Thus, PF1 hyperpolarizes Ascaris muscle by a mechanism that does not involve stimulation of GABA release from inhibitory pre-synaptic terminals.

Cases of human Ascaris infection occur sporadically in areas such as N. America and Western Europe, where this parasite is thought to be non-endemic. Clinical case histories suggest that many of these cases may be cross-infections from pigs. I describe patterns of variation in the ribosomal DNA of Ascaris from 9 such cases. For comparative purposes, I also describe patterns of variation in parasites obtained from pigs and humans from worldwide locations. A Hae III restriction site distinguishes two classes of rDNA repeats; repeats bearing this restriction site were found in > 96% of parasites from pig populations worldwide and in all 9 worms from humans in N. America. In contrast, repeats bearing this restriction site were detected in < 2% of parasites from humans in endemic areas. The molecular data clearly incriminate pigs as the source of infection in the N. American cases. I discuss evolutionary and public health implications of incomplete host fidelity in Ascaris.

Analysis of the electrical properties of neurons in the motornervous system of Ascaris suum suggests that it is largely an analogue system. The motorneurons do not conduct action potentials and they release transmitter tonically at their normal resting potential; transmitter release is increased or decreased as a continuous function of membrane potential. Despite extensive physiological descriptions of the electrical properties of the neurons and their synapses, as well as morphological descriptions of the synaptic circuitry of the system, the predicted activities of the neurons in the circuit differ from those observed by direct recording in semi-intact behaving animals. We conclude that the description of the circuit is incomplete. There are several possibilities for the missing elements, including chemical, proprioceptive, and additional neuronal components. Recently, attention has been focussed most heavily on the intercellular chemical signalling systems; in addition to those mediated by classical neurotransmitters, a surprisingly complex array of neuropeptides has been identified. One family of these peptides, the AF peptides, has been analyzed in detail. It comprises at least 20 peptides, and they fall into sequence-related subfamilies. One of these subfamilies, containing 6 peptides, is encoded by a single transcript, suggesting that the AF peptides are under multiple genetic control. All AF peptides tested have potent activity on the motornervous system and or on muscle. There are multiple physiological activities, and cellular localization studies show multiple patterns of cellular expression. Studies on Panagrellus and Caenorhabditis emphasize the diversity of this family and its genetic control.

The mechanism underlying the ability of the anthelmintic avermectin to paralyse the nematode Ascaris is not yet fully understood. Using conventional two-electrode electrophysiological recording techniques we have demonstrated that micromolar concentrations of ivermectin block the inhibitory GABA response on the muscle cells of the parasitic nematode Ascaris. The ability of a number of avermectin derivatives to act as receptor antagonists for the Ascaris muscle GABA receptor has been determined. This provides useful information to compare with the in vivo anthelmintic potency of these compounds. Abamectin, the most potent anthelmintic, was the most potent compound at inhibiting the GABA response whilst octahydroavermectin, a compound which lacks anthelmintic activity, did not block the GABA receptor. This is consistent with the notion that the GABA receptor antagonist properties of the avermectins could contribute to their anthelmintic action.

The action of two peptides isolated from the nematode Panagrellus redivivus, PF1 (SDPNFLRFamide) and PF2 (SADPNFLRFamide) have been studied on synaptic transmission in the motornervous system of the parasitic nematode Ascaris suum. Intracellular recordings were made from Ascaris somatic muscle cells and excitatory junction potentials (EJPs) elicited by stimulation of the ventral nerve cord. The EJPs were cholinergic as they were blocked by the Ascaris nicotinic receptor antagonist, benzoquinonium. PF1 caused a slow hyperpolarization, similar to the action of this peptide first reported by Bowman, Geary & Thompson (1990) and further characterized by Franks et al. (1994). The hyper-polarization was accompanied by a marked decrease in the amplitude of the EJPs with an EC50 of 311 ± 30 nM (n = 5). This inhibition is unlikely to be due to a post-synaptic site of action of the peptide as the muscle cell input conductance was not significantly altered by PF1 and furthermore the response to bath-applied acetylcholine was not inhibited by PF1 at concentrations up to 10μM (n = 6). PF2 also inhibited the EJPs in a similar manner to PF1. These studies indicate that both of the peptides isolated from the free-living nematode Panagrellus redivivus have biological activity in the parasitic nematode Ascaris suum. PF1 and PF2 have inhibitory actions in contrast to the predominantly excitatory actions of the Ascaris endogenous peptides AF1 (KNEFIRFamide) and AF2 (KHEYLRFamide). The potent actions of the Panagrellus neuropeptides PF1 and PF2 in Ascaris suggest that peptides with a similar or identical sequence may also occur in Ascarisand have an inhibitory role in the motornervous system.

In Guatemalan villages people commonly rear pigs, and both hosts may be infected with Ascaris. This study was designed to ask whether both humans and pigs are potential hosts in a single parasite transmission cycle in such villages, or alternatively, if there are two separate transmission cycles, one involving pigs and one involving human hosts. Parasites were collected from both host species from locations in the north and south of Guatemala. Allelic variation in the nuclear genome of Ascaris was measured using enzyme electrophoresis, while mitochondrial DNA (mtDNA) sequence variation was quantified using restriction mapping. Low levels of enzyme polymorphism were found in Ascaris, but allele frequencies at two loci, mannose phosphate isomerase and esterase, suggest that there is little gene exchange between parasite populations from humans and pigs. MtDNA haplotypes fall into two distinct clusters which differ in sequence by 3–4%; the two clusters broadly correspond to worms collected from humans and those collected from pigs. However, some parasites collected from humans have mtDNA characteristic of the ‘pig Ascaris’ haplotype cluster, while some parasites collected from pigs have mtDNA characteristic of the ’ haplotype cluster. These shared haplotypes are unlikely to represent contemporary cross-infection events. Patterns of phylogenetic similarity and geographical distribution of these haplotypes suggest, instead, that they are the result of two historical introgressions of mtDNA between the two host-associated Ascaris populations. The results clearly demonstrate that Ascaris from humans and pigs are involved in separate transmission cycles in Guatemala.

Patterns of genetic subdivision in parasite populations can provide important insights into transmission processes and complement information obtained using traditional epidemiological techniques. We describe mitochondrial sequence variation in 265 Ascaris collected from 62 individual hosts (humans and pigs) from 35 households in 3 Guatemalan locations. Restriction mapping of individual worms revealed 42 distinct mitochondrial genotypes. We ask whether the mitochondrial genotypes found in worms from individual hosts, from families of hosts and from villages represent random samples from the total Ascaris population. Patterns of genetic subdivision were quantified using F-statistics, while deviations from the null hypothesis of randomness were evaluated by a simple resampling procedure. The analysis revealed significant deviations from panmixia. Parasite populations were strongly structured at the level of the individual host in both humans and pigs: parasites bearing the same mitochondrial genotype were found more frequently than would be expected by chance within hosts. Significant heterogeneity was also observed among populations from different villages, but not from different families within a village. The clustering of related parasites within hosts suggests a similar clustering of related infective stages in the environment and may explain why sex ratios in Ascaris are female-biased. We discuss aspects of Ascaris biology which may lead to the observed patterns.

The DNA from a number of free-living and parasitic nematode species was examined to determine the genomic number and distribution of DNA sequences encoding two evolutionary conserved proteins; the major sperm protein (MSP) and nematode actin. Ascaris and Caenorhabditis MSP cDNA sequences and Ascaris genomic actin sequences were used to probe Southern blots of Eco RI and Hin d III digested nematode DNA. The number of MSP genes varied widely between the 1 MSP gene in Ascaris and the 60 MSP genes in Caenorhabditis. Filarial nematodes appeared to have 1—4 MSP genes while the plant and insect parasitic species showed from 5–12 MSP-hybridizing restriction fragments. Mammalian intestinal parasites showed between 1 and 13 bands hybridizing with the MSP probes. Blots probed to estimate the number of actin genes showed that, with the exception of Ascaris which contains more than 20 germ line sequences that encode actin, all of the nematodes tested had between 3 and 9 bands that hybridized to the Ascaris genomic actin probe. The possible use of highly conserved sequences such as MSP and actin to differentiate between nematode species in diagnostic and taxonomic studies is discussed.

This study examines the persistence of predisposition to Ascaris lumbricoides and Trichuris trichiura during repeated chemotherapy in an urban community in Kuala Lumpur, Malaysia. Significant predisposition was observed over 2 periods of reinfection with and without age-standardization of data. Analysis of different age groups indicated that predisposition was most strongly detectable in the younger age classes. The intensities of infection with both parasites were strongly correlated at each cycle of intervention, suggesting that individuals were similarly predisposed to both species.

The genotypes of both host and parasite may influence the outcome of parasitic infections, but few attempts have been made to quantify the effect of parasite genotype on macroparasite infections of socio-economic importance. We examined variation in particular traits during the infection in pigs with the parasitic nematode Ascaris suum. We infected 26 pigs with mixtures of equal proportions of embryonated eggs from 4 single female worms each with a unique mtDNA haplotype – the eggs from each female worm were a mixture of siblings and half-siblings. Pigs were necropsied on days 14, 17 and 28 following inoculation, which corresponded to time-points before, during and after the main immune responses against the nematode. A total of approximately 11 000 worms were recovered at necropsy. The location in the small intestine was recorded for all worms and the length and mtDNA haplotype were determined for about 4200 individual worms. There were significant differences in the distribution and abundance of the 4 individual haplotypes among individual pigs demonstrating strong interactions between parasite and host. We found significant differences in the abundance and position in the small intestine as well as the size of worms among haplotypes. We conclude that both parasite and host effects as well as the interplay between them play important roles in determining the characteristics and outcome of infection.

Soil-transmitted helminthiasis (STH) is a scourge to the health and well-being of infants and pre-schoolchildren throughout many parts of sub-Saharan Africa. To improve maternal and child health, regular de-worming is recommended and often delivered from mother and child health (MCH) clinics, yet there have been few studies monitoring the progress and impact of interventions on local levels of disease. A cross-sectional parasitological survey, supplemented with questionnaires, was therefore conducted across 10 Ungujan villages examining mothers (n=322) and their pre-school children (n=359). Within children, mean prevalence of ascariasis, trichuriasis and hookworm was 8·6% (95% CI 5·5–11·8), 18·9% (95% CI 14·5–23·4) and 1·7% (95% CI 0·2–3·5) while in mothers mean prevalence was 6·7% (95% CI 3·7–9·7), 11·9% (95% CI 8·0–15·8) and 1·9% (95% CI 0·2–3·5), respectively. There was, however, significant spatial heterogeneity of STH by village, 2 villages having much elevated levels of infection, although general access to anthelminthics and utilization of village MCH clinics was good. Levels of parasite aggregation (k) were determined and a multilevel logistic regression model identified access to a household latrine [OR=0·56 (95% CI 0·32–0·99)] and having an infected household member [OR=3·72 (95% CI 2·22–6·26)] as observed risk factors. To further investigate worm burdens of Ascaris lumbricoides, adult worms were expelled using Combantrin® and measured. A negative relationship between mean worm burden and mean worm mass was found. Villages in the north of Unguja represent locations where there is elevated prevalence of both ascariasis and trichuriasis and it appears that local factors are particularly favourable for transmission of these helminths. From a perspective of control, in such locations, intervention efforts should be stepped up and greater efforts placed upon improving household sanitation.

Ascaris is a large parasitic roundworm (nematode) of the small intestine of humans and pigs, which causes the socio-economically important disease, ascariasis. To better understand the relationship of Ascaris between the 2 host species, recent studies in China have focused on investigating the genetics and epidemiology of Ascaris from humans and pigs using a mutation scanning-based approach. Findings provided support for a low level of gene flow between the human and porcine Ascaris populations. Extending the studies of genotypic variability within Ascaris from humans and pigs, experimental infections of mice and pigs with selected genotypes of Ascaris were carried out. Initial results indicate that there is a significant difference in the ability of Ascaris eggs of genotype G1 (derived from human) and G3 (derived from pig) to infect and establish as adults in pigs, supporting the difference in the frequencies of these genotypes in natural Ascaris populations between pigs and humans in China. Taken together, current information supports that there is limited cross-infection of Ascaris between humans and pigs in endemic regions and that pigs are not a significant reservoir of human infection with the adult nematode in such areas.

Extending a previous investigation of the genotypic variability within Ascaris from humans and pigs, experimental infections of mice and pigs with selected genotypes of Ascaris were performed in the present study to explore possible host affiliation. Initial findings indicate that there is a significant difference in the ability of Ascaris eggs of genotype G1 (derived from human) and G3 (derived from pig) to infect and establish as larvae in mice and as adults in pigs, supporting the difference in the frequencies of the genotypes reported previously in natural Ascaris populations in pigs and humans in areas in China endemic for ascariasis.