Effect of size of Trichinella spiralis (Nematode) infections on glucose and ion transport in the rat intestine

An in vivo perfusion technique, using 3 intestinal loops representing the anterior, mid and posterior regions of the rat small intestine, was used to determine intestinal glucose uptake 5 days after infection with Trichinella spiralis. At high levels of infection (3,000 and 6,000 larvae/rat) net glucose absorption by the intestinal mucosa was significantly impaired in all regions of the small intestine when compared to uninfected controls. At low levels of infection (50 larvae/rat) glucose uptake by the mucosa was significantly enhanced in all 3 regions of the small intestine. Intermediate levels of infections (200-1,000 larvae/rat) also enhanced glucose uptake, but only in the anterior regions of the small intestine. When washings from the small intestine of rats infected with 50 larvae/rat were added to the perfusion fluid used on uninfected rats, glucose uptake was also significantly enhanced. These results suggest that at low levels of infection the intestinal lumen contains a metabolite which may affect the mucosal transport of glucose and the related fluxes of H2O, Na+, Cl-, and K+, in the rat intestine. Luminal [H+] and pCO2 decreased from the proximal to distal regions of the small intestine following perfusion; pO2 was significantly decreased in the proximal and distal regions.     

Effect of level of infection with Nippostrongylus brasiliensis on intestinal absorption of hexoses in rats

Transfer of glucose and galactose in vitro from the mucosal to serosal side of the entire small intestine was significantly reduced in male rats 10 days after infection with 100 or more larvae. Transfer of hexoses across the mucosa into the gut tissue by the entire intestine was generally not significantly affected by infection but transfer from the tissue to serosal fluid was significantly reduced, the reductions occurring throughout the intestine, and the metabolism of glucose significantly increased, the increases occurring in the distal two-thirds of the intestine. Females showed a reduction in serosal glucose transfer at an infection level of 50 larvae whereas males did not and the uninfected distal third of the intestine showed the same response to infection as the infected proximal third. The length of the intestine increased significantly at infection levels of 100 or more larvae.     

The responses of a tropical breed of domestic rabbit, Oryctolagus cuniculus, to experimental infection with Trichostrongylus colubriformis

Clinical, parasitological and pathological responses of a tropical out-bred domestic rabbit to experimental Trichostrongylus colubriformis infection were used to evaluate its suitability as a laboratory host and model for studying the host-parasite relationships of T. colubriformis. In the first experiment, three groups each of 16, predominantly juvenile male, 8- to 10-week-old rabbits were given a single pulse infection with 500, 5000 or 25000 infective larvae (L3) of T. colubriformis, to represent low, medium and high levels of infection, respectively. A fourth group of 16 rabbits of similar age formed the uninfected controls. In the second experiment, two groups of 10 juvenile (8- to 10-week-old) and 10 adult (8- to 10-month-old) rabbits were similarly infected with 20000 L3, with appropriate naive controls. Prepatency was 14 and 16 days and peak faecal egg counts occurred on days 24 and 20 after infection in young and adult rabbits respectively. Peak worm counts occurred on day 14 in both age groups and at all levels of infection. Subsequently, parasite burdens declined in a highly significantly dose- and age-dependent manner. At low and moderate levels of infection, approximately 83-98% of worms were recovered from the first 60 cm of the small intestine. Worm fecundity was also significantly influenced by host age and larval dose. Host age also had a significant effect on worm length. Infections with T. colubriformis were associated with a highly significant loss of body weight, accompanied by anorexia, diarrhoea and 25% mortality at high dose levels during the patent period of infection. There were no significant changes in packed cell volume and eosinophil counts at all ages and levels of infection but significant lymphocytosis occurred at the high dose level between days 7 and 21. Parasite-specific serum IgG responses were not related to worm burden. Overall, data showed that this miniature, docile and relatively inexpensive breed of rabbit is a potentially valuable laboratory host for studying T. colubriformis infections. The larval dose, duration of infection and host age were major determinants of host responsiveness to primary infections in this rabbit genotype.     

Studies on the role of mucus and mucosal hypersensitivity reactions during rejection of Trichostrongylus colubriformis from the intestine of immune sheep using an experimental challenge model.

Nematode-naive sheep and sheep immunised by truncated infections with Trichostrongylus colubriformis were fitted with intestinal cannulae to allow administration of challenge infection and collection of intestinal fluids. Sheep were slaughtered at various times after challenge and the distribution of larvae along the small intestine was determined. Results showed that immune sheep had significantly fewer larvae in their intestines and that some sheep could expel the challenge infection within 2 h. Mucus samples from immune sheep contained increased parasite-specific antibody, histamine and anti-parasite activity as measured by larval migration inhibition assay. Higher levels of antibody and histamine were seen in intestinal fluids of immune sheep after challenge. Immunisation of sheep by truncated infections stimulated serum IgE and resulted in significantly higher numbers of IgE-positive cells in gut tissue sections before challenge and at 2 h and 24 h after challenge. Immune sheep also had greater numbers of mucosal mast cells and globule leucocytes after challenge, compared with naive sheep. When challenge larvae were mixed with mucus from immune sheep and infused back into naive recipient sheep, there was a distinct displacement of the larval population towards the distal part of the intestine, compared with the profile of larval establishment after infusion with mucus from naive sheep. These results are further evidence for an immediate hypersensitivity reaction in the intestine of immune sheep, where challenge larvae are expelled within 2 h and confirm the direct anti-larval properties of mucus. The cannulated-sheep challenge model described here will be a useful tool to unravel the mechanism of larval rejection from immune sheep and could lead to novel therapies.     

Nippostrongylus brasiliensis: local humoral responses in the lungs and intestines of rats following vaccination with irradiated larvae

Groups of rats were infected with 2000 normal larvae of Nippostrongylus brasiliensis or larvae irradiated with 10 to 120 kR. On Day 10 after infection half the animals from each group were autopsied. The remainder were challenged with 5000 unirradiated larvae on Day 15 and killed ten days later. During the experiment enteric antibody levels were estimated by coproantibody measurement. At autopsy the worm burdens were determined and worm-specific antibodies evaluated in lung extracts and serum. It was found that the levels of coproantibody detected with adult worm metabolites were positively correlated with the number of adult nematodes recovered from the intestine after primary infection. The challenge induced a similar increase of these antibodies in all immunised rats which reflected a high immunity to reinfection of vaccinated animals. Preliminary immunochemical studies suggested that the coproantibodies had SIgA properties. In lung extracts of rats immunised with larvae irradiated at 40, 80, or 120 kR and in all animals after challenge, antibodies reacting with infective larval antigens were found. Their titres were negatively correlated with serum antibody levels. The significance of bronchial and enteric antibodies in conferring protection against challenge remains to be elucidated.     

Alterations in Ascaris suum larval burdens, eosinophil numbers, and lysophospholipase activity associated with low levels of dietary iron

The effect of various amounts of dietary iron on the immune response was investigated using BALB/cAnNCr/BR mice infected with Ascaris suum. Changes in numbers of larvae, numbers of eosinophils, and levels of lysophospholipase (LPL) activity in lung or liver tissues were analyzed from nonimmune and immunized mice at 2 and 7 days postinfection (PI). Various iron diets did not influence the numbers of tissue larvae, eosinophils, or the LPL activity in lungs or livers of nonimmunized mice at various times after infection. Lung and liver LPL activity was reduced in immunized mice without significant changes in larval numbers at 2 days PI. At 7 days PI, lung and liver LPL activity, eosinophil numbers, and numbers of larvae were increased in immunized mice receiving low iron diets. Results confirm that low iron diets affect the host response to A. suum.     

Observations on Toxocara pteropodis infections in mice

Infection in mice with Toxocara pteropodis was investigated. In mice fed infective eggs, third-stage larvae hatched out and penetrated the mucosa, predominantly that of the lower intestine. They travelled via the portal vein to the liver, where they remained at least 14 months. They grew in length from 430 +/- 15 micron, at three days post infection (p.i.), to 600 +/- 50 micron, at six to nine weeks p.i., after which time growth ceased. Blood eosinophilia appeared at 28 days p.i., and eosinophil levels continued to rise gradually beyond this time. In female mice the larvae did not migrate from the liver in response to pregnancy or lactation. When infective eggs were inoculated subcutaneously or intra-peritoneally, larvae hatched out and ultimately appeared in the liver in larger numbers than seen with oral infections.     

Suppression of peripheral eosinophilia by the coccidium Eimeria nieschulzi (Apicomplexa: Eimeriidae) in experimentally infected rats

Four groups of 60 rats each were used to examine interspecific interactions between Eimeria nieschulzi and Nippostrongylus brasiliensis. Rats in group 1 served as uninoculated controls. Group 2 rats were each injected subcutaneously with 2.0 X 10(3) L3 larvae of N. brasiliensis. Group 3 rats were each inoculated per os with 2.5 X 10(5) sporulated oocysts of E. nieschulzi. Rats in group 4 were first infected with 2.0 X 10(3) larvae of N. brasiliensis and, at 8 days postinoculation, with 2.5 X 10(5) oocysts of E. nieschulzi. Ten animals from groups 1-3 were sacrificed at 4-day intervals postinoculation and group 4 rats were sacrificed at 4-day intervals beginning after the secondary infection. Blood smears were prepared from each animal to determine differential blood cell counts, bone marrow was examined at the times of peak infection for absolute and relative numbers of eosinophils, portions of the duodenum and jejunum were examined histologically for mast cells, and feces obtained from the cecum and large intestine were examined for ova/gram of feces. Results revealed that relative numbers of peripheral neutrophils and monocytes became elevated during the course of infection for all infected animals, and rats infected with the helminth only also had elevated eosinophil levels. However, rats infected singly with E. nieschulzi, or concurrently with the coccidium and helminth, had peripheral eosinophil levels that were not significantly different from controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Kinetics of primary and secondary infections with Strongyloides ratti in mice

Dawkins H. J. S. and Grove D. I. 1981 Kinetics of primary and secondary infections with Strongyloides ratti in mice. International journal for Parasitology11: 89–96. The kinetics of infection with S. ratti were quantitated in normal and previously exposed C57B1 /6 mice. In primary infections, larvae penetrated the skin rapidly and were seen in peak numbers 12 h after infection. By 24 h after infection, larval numbers had declined appreciably and there was a slow decrease in numbers thereafter. Larvae were first observed in the lungs at 24 h and maximal recovery occurred at 48 h. It is thought that larval migration through the lungs is rapid. Worms were first seen in the intestines two days after infection. Maximum numbers were seen on the fifth day and worm expulsion was complete by day 10. Two moults took place in the small intestine during days 3 and 4 after infection. Rhabditiform larvae were first noted on the fourth day after infection. Mice exposed to S. ratti four weeks previously had significantly less larvae in the skin 4 and 12 h after infection but by 24 h there was no difference when compared with mice with primary infections. Peak recovery of larvae from the lungs occurred 24 h after infection; significantly less larvae were recovered on days 2 and 3 when compared with normal mice. There was a marked reduction in the adult worm burden in the gut; the number of worms recovered was less than one fifth of that seen in primary infections. Those worms which did mature were less fecund and were expelled from the intestines within 7 days of infection. It is suggested that in previously exposed animals, the migration of larvae from the skin is hastened, many of these larvae are destroyed in the lungs and that expulsion of worms which do mature in the intestines is accelerated.     

Heligmosomoides polygyrus: inoculum size and glucose, ion, and gas fluxes in the small intestine of mice

Female CDI mice were inoculated with 10, 50, 100, 250, or 500 larvae of Heligmosomoides polygyrus. At Days 7, 9, and 12 after infection, the anterior third of the small intestine was perfused using an in vivo technique. The distribution of worms in the mouse intestine was determined after 7, 9, and 12 days. All worms that were recovered were from the proximal half of the small intestine. When compared to uninfected controls, there was a significant increase (+56%) in glucose absorption of the small intestine at Day 7 after infection with inocula of 50 and 100 larvae; at Day 9, glucose absorption was significantly increased with a 10-larvae inoculum. A decrease in glucose absorption occurred at Days 7 and 9 after infection with a 500-larvae inoculum. Net water absorption was significantly increased (+183%) with the 50- and 100-larvae inocula at Day 7, but was significantly reduced at Day 9 after infection with the 50-, 100-, 250-, and 500-larvae inocula. Both Cl- and Na+ absorption were significantly increased with the 50-, 100-, and 250-larvae inocula at Day 7 after infection; at 9 and 12 days, there was significant net secretion of both ions. In control mice, there was net secretion of K+, while with the 50-, 100-, and 250-larvae inocula on Day 7 there was significant net absorption of K+ ions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Angiostrongylus cantonensis: role of eosinophils in the neurotoxic syndrome (Gordon-like phenomenon)

The role of eosinophils in the pathophysiology of Angiostrongylus cantonensis infections was investigated in nonpermissive (guinea pig) and permissive (rat) hosts. Neurological symptoms similar to the Gordon phenomenon (ataxia, tremor, paralysis) together with a loss of Purkinje cells in the cerebellum were observed after intracraneal injection of human eosinophil extracts or after infection with A. cantonensis, only in guinea pigs and not in rats. Blood eosinophilia as well as eosinophil numbers present in the cerebellum and in the cerebrospinal fluid were higher in guinea pigs than in rats, at all times after infection with A. cantonensis. Increased levels of cytotoxicity toward L3 larvae in vitro were obtained in the presence of guinea pig eosinophils and IgE antibodies, rather than with the corresponding rat effector system. The detection of one eosinophil granule component, the eosinophil peroxidase, in the cerebrospinal fluid from infected guinea pigs but not from rats suggested that in nonpermissive hosts, neurological disorders, similar to the previously described Gordon phenomenon, might be due to eosinophil neurotoxins released after interaction of eosinophils with the parasites.     

Trichinella spiralis: phospholipase in challenged mice and rats

Mice challenged with 400 larvae of T. spiralis showed greatly elevated phospholipase levels in tissues of the small intestine from 5 to 29 days after infection, after which the levels returned to those of the uninfected controls. Related to these rises in enzyme levels were greatly increased numbers of eosinophils in the bone marrow. Increases strikingly above those in the controls were noted from 8 to 20 days after infection. In rats challenged with 3000 larvae, the enzyme levels were elevated from 4 to 18 days, and the eosinophils were increased during the same periods. A working hypothesis is proposed to explain these phenomena.     

IgE enhances parasite clearance and regulates mast cell responses in mice infected with Trichinella spiralis

Trichinella spiralis infection elicits a vigorous IgE response and pronounced intestinal and splenic mastocytosis in mice. Since IgE both activates mast cells (MC) and promotes their survival in culture, we examined its role in MC responses and parasite elimination in T. spiralis-infected mice. During primary infection, wild-type but not IgE-deficient (IgE(-/-)) BALB/c mice mounted a strong IgE response peaking 14 days into infection. The splenic mastocytosis observed in BALB/c mice following infection with T. spiralis was significantly diminished in IgE(-/-) mice while eosinophil responses were not diminished in either the blood or jejunum. Similar levels of peripheral blood eosinophilia and jejunal mastocytosis occurred in wild-type and IgE-deficient animals. Despite the normal MC response in the small intestine, serum levels of mouse MC protease-1 also were lower in parasite-infected IgE(-/-) animals and these animals were slower to eliminate the adult worms from the small intestine. The number of T. spiralis larvae present in the skeletal muscle of IgE(-/-) mice 28 days after primary infection was about twice that in BALB/c controls, and the fraction of larvae that was necrotic was reduced in the IgE-deficient animals. An intense deposition of IgE in and around the muscle larvae was observed in wild-type but not in IgE null mice. We conclude that IgE promotes parasite expulsion from the gut following T. spiralis infection and participates in the response to larval stages of the parasite. Furthermore, our observations support a role for IgE in the regulation of MC homeostasis in vivo.     

Rapidly Boosted Plasma IL-5 Induced by Treatment of Human Schistosomiasis haematobium Is Dependent on Antigen Dose,IgE and Eosinophils

Background

IgE specific to worm antigen (SWA) and pre-treatment eosinophil number, are associated with human immunity to re-infection with schistosomes after chemotherapeutic treatment. Treatment significantly elevates circulating IL-5 24-hr post-treatment of Schistosoma mansoni. Here we investigate if praziquantel treatment of human schistosomiasis haematobium also boosts circulating IL-5, the immunological and parasitological factors that predispose to this, and the relationship between these and subsequent immunity to post-treatment re-infection.

Methodology/Principle Findings

The relationship between pre-treatment SWA-IgE, eosinophil number and infection intensity and the 24-hr post-treatment IL-5 boost was investigated in a Malian cohort (aged 5–40 yrs), exposed to S. haematobium. Eotaxin levels were measured at 24-hr post-treatment as a proxy of eosinophil migration. The relationship between the 24-hr post-treatment IL-5 boost and later eosinophil numbers and SWA-IgE levels (9-wk post-treatment) was examined, then investigated in the context of subsequent levels of re-infection (2-yr post-treatment). Circulating IL-5 levels increased 24-hr post-treatment and were associated with pre-treatment infection intensity, SWA-IgE levels, eosinophil number, as well as 24-hr post-treatment eotaxin levels. 24-hr IL-5 levels were, in turn, significantly associated with eosinophil number and elevated SWA-IgE 9-wk later. These SWA-IgE levels were significantly associated with immunity to re-infection.

Conclusions/Significance

Early IL-5 production after treatment-induced exposure to S. haematobium worm antigen is positively associated with antigen dose (infection intensity), IgE availability for arming of effector cells at time of treatment and subsequent eosinophil migration response (as indicated by eotaxin levels). The IL-5 produced is positively associated with increased downstream eosinophil number and increases in specific IgE levels, implicating this cytokine boost and its down-stream consequences in the production and maintenance of IgE, and subsequent re-infection immunity.     

Visceral larva migrans: migratory pattern of Toxocara canis in pigs.

The migratory pattern of Toxocara canis was investigated following infection of pigs with 60000 infective eggs. Groups of six pigs were slaughtered at 7, 14 and 28 days after infection (p.i.), and the number of larvae in selected organs and muscles was determined by digestion. A group of uninfected pigs was used as negative controls for blood parameters and weight gain. Toxocara canis migrated well in the pig, although the relative numbers of larvae recovered decreased significantly during the experiment. On day 7 p.i., high numbers of larvae were recovered from the lymph nodes around the small intestine and to some extent also from the lymph nodes around the large intestine, and from the lungs and the liver. On day 14, the majority of larvae were recovered from the lungs and the lymph nodes around the small intestine, and by day 28 p.i. most larvae were found in the lungs. Larvae were recovered from the brain on days 14 and 21, with a maximum on day 14 p.i. No larvae were found in the eyes. Severe pathological changes were observed in the liver and lungs, especially on day 14 p.i.; also, development of granulomas was observed in the kidneys. Finally, a strong specific antibody response towards T. canis L2/L3 ES products was observed from day 14 p.i. until termination of the experiment, and the maximum eosinophil response was observed 14 days p.i. The pig is a useful non-primate model for human visceral larva migrans, since T. canis migrate well and induce a strong immunological response in the pig. However, the importance of the pig as a paratenic host is probably minor, because of the relatively early death of most of the larvae.     

Effect of dietary magnesium on Ascaris suum infections of mice

Low amounts of dietary magnesium affected the inflammatory tissue response in nonimmunized mice differently than in immunized mice. Eosinophil numbers and LPL activity in lung tissue following infection with A. suum larvae were altered by the level of magnesium in the diets of mice. Average or higher dietary levels of magnesium resulted in decreased numbers of lung larvae indicating an overall protective effect. Increases in eosinophil numbers or LPL activity were not directly related to the numbers of larvae/lungs. Larvae/livers, eosinophil numbers, and LPL activity were affected by the types of magnesium diets that mice received. Nonimmunized mice had differences in larvae/liver (at 2 days and 7 days pi) and LPL activity (at 2 days pi). Immunized mice had varying findings at 2 days pi but a direct relationship between dietary magnesium and numbers of larvae, numbers of eosinophils, and liver LPL activity at 7 days pi.     

Observations on the self-cure reaction and other forms of immunological responsiveness against Haemonchus contortus in sheep

Self-cure reactions and immunological responses preventing establishment of Haemonchus contortus in sheep may operate as separate entities. In one experiment, self-cure occurred when challenge infection with 5000 larvae was superimposed on an infection with 5000 larvae given to worm-free sheep 6 weeks previously. Resident worms were rejected and establishment of infection by incoming larvae was impeded. The latter effect was not observed in sheep treated similarly but with resident parasites removed by treatment with oxfendazole before challenge. In another experiment, younger worm-free sheep primed by three infections with 2000 larvae at intervals of 2 weeks or a single infection with 6000 larvae were challenged with 10,000 larvae 6 weeks after the first priming infection. Self-cure was not incited but establishment of infection was impeded in sheep primed with three divided doses of larvae whether or not priming infections had been removed by oxfendazole. Infection regimes used for priming did not influence numbers of arrested fourth-stage larvae derived from challenge infection. However, more arrested larvae were present when challenge was superimposed on extant infections, indicating that resident worms or a factor activated by their presence induced developmental arrest. In a third experiment, large burdens with H. contortus were established in sheep immunosuppressed with the corticosteroid, dexamethasone, at the time of infection. Self-cure was not triggered by a challenge infection given 32 days later either in these sheep, or in sheep with a smaller worm burden derived from infection given without immunosuppression. Faecal egg counts, however, indicated that development of the challenge infection was prevented in both groups of sheep.

Investigation of self-cure is restricted by lack of a predictable system for reproducing the phenomenon. Self-cure was induced by a single infection with 5000 larvae in mature sheep but not with 6000 larvae in immature sheep. Three infections with 3000 larvae given at intervals of 2 weeks to mature sheep did not prime for self-cure. Procedures aimed at heightening immediate hypersensitivity, i.e. treatment with pertussis vaccine or concurrent infections with Ostertagia circumcincta, did not promote self-cure reactivity in the latter situation.     

The effect of food limitation on immunity factors and disease resistance in the western tent caterpillar

Epizootics of nucleopolyhedrovirus characterize declines of cyclic populations of western tent caterpillars, Malacosoma pluviale californicum. In field populations, infection can be apparently lacking in one generation and high in the next. This may suggest an increase in the susceptibility to infection of larvae at peak density or the triggering of a vertically transmitted virus. Here, we test the hypothesis that reduced food availability, as may occur during population outbreaks of tent caterpillars, influences the immunocompetence of larvae and increases their susceptibility to viral infection. We compared immunity factors, hemolymph phenoloxidase and hemocyte numbers, and the susceptibility to nucleopolyhedroviral infection of fifth instar larvae that were fully or partially fed as fourth instars. To determine if maternal or transgenerational influences occurred, we also determined the susceptibility of the offspring of the treated parents to viral infection. Food limitation significantly reduced larval survival, development rate, larval and pupal mass, moth fecundity and levels of hemolymph phenoloxidase, but not the numbers of hemocytes. Neither the food-reduced larvae nor their offspring were more susceptible to viral infection and were possibly even less susceptible at intermediate viral doses. Food reduction did not activate latent or covert viral infection of larvae as might be expected as a response to stress. We conclude that reducing the food intake of fourth instar larvae to an extent that had measurable and realistic impacts on their life history characteristics was not translated into increased susceptibility to viral infection.     

Experimental infection with Ancylostoma caninum larvae in mice. V. Serum protein pattern during infection with various doses.

Serum protein patterns of Swiss albino mice during Ancylostoma caninum infection with varying dose levels were studied electrophoretically. There was a significant decrease in albumin associated with a corresponding increase in beta globulin in all the infected groups. Gamma globulin decreased significantly in all groups except the one infected with a challenge dose of 4000 larvae. Maximum changes occurred on day 31, 15, 9, 15 and 6 when mice were infected with 250, 500, 1000, 2000 and 4000 larvae, respectively. There was a significant negative correlation between albumin and the infective doses and a positive correlation between beta globulin and the infective doses and among the changes caused by different infective doses.