Application of test-tube cultivation method on the survey of bookworm and related human nematodes infection.
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A field study was conducted between May and October 1995 to examine the efficacy of 0.5 mg/kg ivermectin pour-on on parasitic gastroenteritis of set-stocked calves at turnout and 6 weeks later. A treated group of 10 calves was compared with a separately grazed group of 10 non-treated calves; 14 calves were used as tracer animals. Initial infection levels appeared to be very low in both groups and faecal egg counts, pasture larval counts, serum pepsinogen levels and optical density (OD) values of an ELISA with a specific recombinant antigen for Cooperia oncophora remained low for 4 months. Thereafter, a rapid build up of gastrointestinal nematode infections occurred in both groups.
A longitudinal study of faecal nematode egg counts was made in naturally infected Scottish Blackface sheep over two grazing seasons to 75 weeks of age. Although egg counts were lower in the second grazing season the variation among animals was greater. Egg counts were repeatable from 3 months of age. The repeatability of faecal egg counts within and between grazing seasons was about 0.3. Animals with lower than average egg counts in the first grazing season tended to have lower than average egg counts in the second grazing season. Therefore lambs with relatively low faecal egg counts after 3 months of age are likely to retain their advantage in the following year.
Herbage sampling has been used to ascertain the contamination and epidemiology of cattle nematode infective larvae in large extensive pastures situated in the centre of France, where heifers graze for four months on a total area of one ha/heifer in mountains virtual "hectares", tufts of grass being picked both close to faecal pats or "refusals", and far from these pats. Ostertagia was the predominant parasite and the occurrence of Dictyocaulus resulted from silent-carrier heifers. Just after the thawing of the snow, when the heifers arrived in the mountains, the contamination was very high: 8000-9000 L3 and 45.00 to 63.00 L3 kg-1 dry herbage, respectively, far from and close to faecal pats, but this contamination decreases regularly during the season. The sampling of four areas (four "hectares") in each paddock was found to be a very valuable method. The variation of the mean was low and found mainly when the number of larvae was high (6-19% only for the spring sampling). This technique could have some merit in parallel or concurrently with tracer calves which are always difficult and expensive to use.
Research on the complex interactions among host nutritional status, parasitic infection and immune responsiveness has focused on the detrimental consequences of parasitic infections on host nutritional status and on mechanisms by which malnutrition impairs immunocompetence. Curiously, relatively few studies have examined the effects of malnutrition on the immune response in the parasite-infected host, and even fewer have considered the events occurring at the intestinal level, where absorption of nutrients occurs, intestinal parasites reside, and the gastrointestinal-associated lymphoid tissues play a role in directing both the local and the more systemic immune responses. Our work using a zinc-deficient nematode-infected mouse model reveals that parasites are better able to survive in the zinc-deficient hosts than in well-nourished hosts; that the production of interleukin-4 in the spleen of zinc-deficient mice is depressed, leading to depressed levels of IgE, IgG(1) and eosinophils; and that the function of T cells and antigen-presenting cells is impaired by zinc deficiency as well as by energy restriction. Given the paramount role of the gastrointestinal-associated lymphoid tissues in inducing and regulating immune responses to intestinal parasites and in orchestrating responses in the spleen and peripheral circulation, we conclude that zinc deficiency (in association with energy restriction) exerts profound effects on the gut mucosal immune system, leading to changes in systemically disseminated immune responses and, importantly, to prolonged parasite survival.
The major gastrointestinal nematode parasites of ruminants all belong to the Order Strongylida and the family Trichostrongyloidea. Despite this close evolutionary relationship, distinct differences exist in the microenvironmental niches occupied by the developmental stages of the various parasites, which may account for the variable susceptibility of the different parasite species to the immune effector mechanisms generated by the host. In addition, different manifestations of resistance have been observed against the adult and larval stages of the same parasite species, and even against the same parasite stage. In particular, both rapid and delayed rejection of infective larval stages of gastrointestinal nematode parasites has been documented. This review will give an overview of the various manifestations of resistance to gastrointestinal nematode parasites of ruminants, as well as the immune mechanisms and antigens associated with the generation of immunity by the ruminant hosts to these parasites. In addition, a working model is provided aimed at reconciling most of the present knowledge on the different immune responses generated during infection with the various parasite rejection profiles. Extrapolation of these results to field conditions will need to take into account the variability imposed by seasonal changes and management practices, as well as the individual variability in immune responsiveness present in outbred animal populations.
In two experiments groups of calves were exposed to different levels and patterns of infection with Ostertagia spp. and Cooperia spp. The experimental design simulated the stereotypic pattern of herbage infestation, including a normal or a delayed midsummer increase, under conditions of set-stocking. The purpose of the experiments was to investigate the accuracy of egg counts, pepsinogen and gastrin values and antibody titres as estimators of the level of exposure to infection. Faecal egg counts significantly reflected levels of exposure during the first half of the simulated grazing season. Antibody titres and pepsinogen values reflected levels of exposure best during August and September, partly depending on the pattern and range of levels of exposure. Antibody titres against Cooperia spp. were particularly useful when levels of exposure to gastrointestinal nematode infection were low. Gastrin values were elevated only at high levels of exposure, which caused large weight gain reductions, in the later part of the simulated first grazing season. It is suggested that antibody titres and pepsinogen values can be used for prognostic diagnosis, indicating whether or not control measures should be taken. Both estimators of infection correlated significantly with the realised weight gain at the end of the simulated grazing season. Egg counts in the second month after the initial infection (turnout) also may be of significant value to support decisions concerning control measures. Comparisons with data from field trials and experiments conducted by others under various conditions suggested that the conclusions of the present experiments are also valid under field conditions. Furthermore, the results supported the conclusions drawn from previous field work, that levels of exposure are often very low on commercial farms in the Netherlands.
Infestation of the gastrointestinal tract by parasitic nematodes is invariably associated with mucosal mastocytosis, which is a thymus-dependent phenomenon in parasitized rats, and is adoptively transferable with a T cell-enriched population of thoracic duct lymphocytes. When derived by in vitro culture, mucosal mast cells (MMC) arise from a bone marrow precursor after stimulation by T cell-derived factors. In rats infected with the nematode Trichinella spiralis, mucosal mastocytosis is temporally associated with the immune expulsion of the adult worms whereas in the case of Nippostrongylus brasiliensis, mastocytosis is frequently observed to occur after worm expulsion has been completed. Consequently, there has been doubt as to whether MMC are active and serve a functional role in the expulsion of rat intestinal nematodes. MMC contain and secrete a neutral proteinase, rat mast cell protease II (RMCP II); detection and assay of secreted RMCP II therefore provides a direct measurement of MMC activity. Here we describe the release of this enzyme into the blood of rats infected with N. brasiliensis or T. spiralis. Our results show that the systemic secretion of RMCP II coincides with the immune expulsion of these nematodes, demonstrating clearly for the first time that rat MMC are functionally active during the immune elimination of primary nematode infections.
The relative efficacy of a single 400 mg dose of albendazole or mebendazole in the treatment of nematode infections was assessed in 2- to 9-year-old children living in two different Bolivian rural communities. Both agents were equally very effective (100% cure rate) in treating ascariasis. Albendazole was clearly more active than mebendazole against hookworm infections, both in terms of egg reduction rate (92.8% vs. 62.4%) and cure rate (81.8% vs. 17.2%). As far as trichuriasis is concerned, albendazole produced a higher egg reduction rate than mebendazole (45.7% vs 15%), but a lower cure rate (33.3% vs 60%). Both drugs were well tolerated.
Persistent anthelmintic efficacy of topical formulations (all at a dosage of 500 microg/kg) of doramectin (DOR), ivermectin (IVM), eprinomectin (EPR) and moxidectin (MOX), in comparison with untreated control cattle (CONT), was observed in stocker beef calves during a 112-day winter-spring grazing trial. Five groups of 15 calves per group were grazed on 15 separate 2 ha pastures following random assignment of animals to specific pastures and then to treatment groups. All of the 5 treatments were represented in each of the 15 pastures. All cattle were weighed on study Days 1, 0, 28, 56, 84, 111 and 112. Fecal samples for nematode egg counts were collected on Days 7, 0, at 7 day intervals through Day 56 and at 14 day intervals to Day 1 12. Pooled group fecal cultures for determining generic composition of nematode infections were prepared at 14 day intervals throughout the study. As based on fecal egg counts, anthelmintic activity of EPR and MOX was greater (p < 0.05) than values for IVM or CONT through Day 28. Activity of DOR was greater (p < 0.05) than that of IVM on Days 7 and 14 only. Although significance levels varied little among treated groups from Day 42 to the end of the study, egg counts and percent reduction values of EPR and MOX remained consistently lower numerically than egg counts and higher than reduction values respectively, of DOR and IVM through Day 70. From Day 70 on, IVM counts were numerically, but not significantly higher than values of CONT. Based on larval culture, Cooperia predominated from Day 0 through 28 and again from Days 70 to 98; Ostertagia was second in prevalence with highest percentages, which exceeded those of Cooperia, between Days 42 and 70. Bodyweights of all treated groups, with exception of IVM, were always significantly greater (p < 0.05) than weights of CONT. Weights of IVM were numerically greater, but not significantly greater than CONT only on Days 84 and 112. From Day 56 on, there were no significant differences between weights of DOR, EPR and MOX, however, numerical values for MOX were consistently higher than values for the other two. Final average total bodyweight gains were: 153.7 kg for MOX, 148.5 kg for EPR, 146.9 kg for DOR, 139.7 kg for IVM and 127.7 kg for CONT.
Attempts to control a summer diarrhoea in grazing Finnish landrace lambs which had been unresponsive to anthelmintics and coccidiostats were made by supplementing them with cupric oxide particles and withdrawing a magnesium-rich mineral, while maintaining parasite control measures. The diarrhoea persisted from July to September and plasma pepsinogen activities were raised, suggesting that the anthelmintic did not prevent abomasal damage; the jejunum of an affected lamb showed lesions of parasitic gastroenteritis. Small responses to cupric oxide particles and larger responses to the withdrawal of magnesium were deceptive, possibly being confounded by differences in parasite challenge. In another experiment Finnish landrace lambs were more susceptible to diarrhoea than Suffolk cross lambs in autumn. The susceptibility was then linked to a strong inhibition of worm egg output and may have been caused by a hypersensitive mucosal response to the larval challenge. Plasma pepsinogen concentrations were again raised in the Finnish landrace lambs and did not decline after treatment with anthelmintic, whereas the concentrations increased later in the Suffolk cross lambs, and were apparently responsive to anthelmintic. The cases of diarrhoea were similar to 'July disease' and may have been caused by continuous nematode infections which were only briefly controlled by drenches. Anthelmintic-unresponsive diarrhoea is the term proposed for the disorder, which may be controllable by devices releasing anthelmintic continuously or by a move to less infected pasture. Faecal egg counts remained low in the condition and were diagnostically misleading.
We illustrate the importance of stochastic effects in population models of biological systems and demonstrate a number of analytic and simulation-based approaches that can usefully be applied to such models. In so doing, we compare the stochastic approach to the more usual deterministic one. The model studied represents the gastrointestinal infection of ruminants by nematodes when the hosts maintain a fixed density. The incorporation of a feedback mechanism, which accounts for the immune response of the infected animals, results in a highly nonlinear model; similar forms of nonlinearity are a feature of many plausible models in population biology. In the absence of an analytic solution to the full stochastic model we explore a number of approximations and compare them to simulations of the full stochastic process. We explore three modes of behaviour of the system. In the endemic regime the stochastic system fluctuates widely around the non-zero fixed points of the deterministic model. In the managed regime, where the system is subject to external periodic perturbation, stochastic effects are negligible. Finally, we find that in a regime in which the deterministic model predicts the long-term persistence of oscillations the stochastic model shows that extinction can occur. Of the approximation procedures we consider, the Normal approximation to the full stochastic process is the most generally applicable, and it is also the most accurate in the light of simulation results. Local linearization provides reasonably accurate prediction of the variance-covariance structure, and a transfer function approach allows calculation of the time-lagged auto- and cross-correlations in the endemic regime. Linearization of the stochastic updates themselves results in poor prediction of the population variances.
The cytokine microenvironment is thought to play an important role in the generation of immunoregulatory cells. Nematode infections are commonly associated with Th2 cytokines and hyporesponsive T cells. Here we show that IL-4-dependent macrophages recruited in vivo by the nematode parasite Brugia malayi actively suppress the proliferation of lymphocytes on co-culture in vitro. These alternatively activated macrophages block proliferation by cell-to-cell contact, implicating a receptor-mediated mechanism. Further, the proliferative block is reversible and is not a result of apoptosis. Suppressed cells accumulate in the G1 and G2/M phase of the cell cycle. Interestingly, the G1 and G2/M block correlates with increased levels of Ki-67 protein, suggesting a mechanism that affects degradation of cell cycle proteins. We also show that, in addition to lymphocyte cell lines of murine origin, these suppressive cells can inhibit proliferation of a wide range of transformed human carcinoma lines. Our data reveal a novel mechanism of proliferative suppression induced by a parasitic nematode that acts via IL-4-dependent macrophages. These macrophages may function as important immune regulatory cells in both infectious and noninfectious disease contexts.
Treatment of soil-transmitted nematodes with mebendazole was carried out in 137 children aged 6--15 years in Thailand. There were 100 cases of hookworm infection, 37 of ascariasis, 16 of trichuriasis and 32 of strongyloidiasis. Mebendazole was given at 100 mg twice daily for three consecutive days irrespective of age or weight. The reduction rates as seen by mean egg counts four weeks after treatment were 94.9% for hookworm infection, 100% for ascariasis and 93.9% for trichuriasis; the reduction rates of the mean larval count in 15 cases of strongyloidiasis was 82.1%. No side-effects were observed. Mebendazole was thus confirmed as effective and safe in the treatment of soil-transmitted nematode infections in children in Thailand.
Gastrointestinal parasitism of Zebu crossbred (Z), Hereford (H), and Hereford X Brahman (HB) weaned steer calves was observed from March 1985 to May 1986. Three groups of 60 calves were randomised and in each group the three breeds were equally present. Calves in GI were untreated, in GII were treated strategically and in GIII treated monthly with anthelmintic. The effects of anthelmintic treatments on the growth of cattle were assessed by comparing mean live-weight gain (MLG) responses of treated and untreated calves grazing the same permanent pasture. Worm recovery and egg counts increased from autumn to winter. After abundant July rainfalls, herbage infective larvae (L3) and worm burdens (mainly Ostertagia) reached the highest levels and Z calves presented symptoms of clinical parasitism. After high availability of L3 in spring, L3 and egg counts decreased to the end of the study. From the end of winter until summer a high percent of inhibited Ostertagia EL4 was seen. Blood samples showed reduced serum copper lower levels for GI. MLG responses of Z treated calves were significantly higher during the autumn-spring and summer-autumn periods. MLG responses of treated H calves were significant only during the winter-spring period. For HB calves there were no significant MLG responses during any period. The highest monthly treatment response was observed during winter and early spring. All strategic treatments gave significant MLG responses in spring and late summer. Initial effects of parasitism, from autumn until early spring had the greatest consequences in current and later productivity. For Z calves herbage L3, worm burdens, egg counts and MLG responses were higher than for H and HB calves. The Z breed were significantly heavier (+22.7%) at the end of the experiment in GIII than in GI. Similar trends were recorded, significantly, for H breed (+17.7%) and not statistically significant for HB (+12.7%). Under our temperate conditions it seems that H and HB calves showed greater capacity to resist nematode infection than Z calves.
The immune expulsion of gastrointestinal nematode parasites is usually associated with T helper type 2 (Th2) responses, but the effector mechanisms directly responsible for parasite loss have not been elucidated. The intestinal inflammatory response accompanying infection with gastrointestinal helminths is thought to be a contributory factor leading to the expulsion of the parasite. However, we have shown that the intestinal inflammation, which is controlled by interleukin (IL)-4, is not required for parasite expulsion. OX40-OX40 ligand (L) signals have been shown to be important for the development of Th2 immune responses but are also involved in a number of inflammatory diseases including those of the intestine. Here, we have investigated the effect of OX40 and OX40L fusion protein treatment on the induction of protective Th2 responses and enteropathy following infection with the gastrointestinal nematode Trichinella spiralis. Treatment with an OX40-immunoglobulin (Ig) blocking fusion protein resulted in enhanced expulsion of the parasite and an increase in the accompanying mastocytosis, despite unaltered levels of Th2 cytokines. Furthermore, there was a delay in the villus atrophy and crypt hyperplasia usually associated with this infection. In contrast, levels of Th2 cytokines were greatly up-regulated in mice treated with an OX40L-Ig activating fusion protein, yet the expulsion of the parasite and the enteropathy were unaffected. Therefore, OX40 ligation potentiates the Th2 response without enhancing host protective immune responses, whereas blocking the OX40-OX40L interaction enhances host protection without promoting Th2 cytokine responses during Trichinella spiralis infection.
In a 30 months' longitudinal study, primary health care intervention was effective in reducing the prevalence of three common intestinal nematode infections (Ascaris lumbricoides, Trichuris trichiura and Necator americanus) in three communities. This was achieved by training school leavers and auxiliary aides as microscopists, health inspectors and field assistants and deploying them to provide screening, surveillance, environmental sanitation, and mass-expulsion chemotherapy (MEC). Post-control surveillance confirmed that the prevalence of these infections had been greatly reduced. The prevalence of Ascaris lumbricoides had declined from 49.3% (pre-intervention) to 10.5% (post-intervention). Hookworm had fallen from 31.4% (pre-intervention) to 4.1% (post-intervention) and whipworm from 40.7% (pre-intervention) to 6.5% (post-intervention). Overall percentage decreases of 78.7%, 86.9% and 84.0% were recorded for Ascaris lumbricoides, Necator americanus and Trichuris trichiura respectively. The initial decline in prevalence was due to the MEC campaign, but the improved sanitation and health education presumably reduced the reinfection rate. If the entire population participated, periodic repetition of the mass expulsion therapy campaign at appropriate intervals combined with continued attention to environmental hygiene and prolonged health education could bring these infections under control within a few years.
Intestinal infections often trigger functional bowel disorders. The nematode Nippostrongylus brasiliensis induces post-infective alterations mainly consisting in an intestinal mast cell hyperplasia. Mast cells contact vagal afferent nerve fibres. Therefore, it is possible that the anatomical sequels of intestinal nematode infection induce long term alterations in the mediation of afferent signals from the gut to the brain. To test this hypothesis, we examined hindbrain expression of Fos immunoreactivity following systemic cholecystokinin (CCK) administration in control rats and 35 days after N. brasiliensis infection. In controls, Fos was expressed in the area postrema and the nucleus of solitary tract. After infection, this expression was increased by 262 and 157%, respectively. We conclude that an intestinal infection, at least in this model, is followed by an enhancement of the activation of hindbrain sites by CCK.