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Use of secondarily revised VH genes in IgE antibodies produced in mice infected with the nematode Nippostrongylus brasiliensis.

Although a high level of IgE is produced after primary infection with Nippostrongylus brasiliensis (Nb), most of the IgE antibodies (Abs) are not specific to the worm. Analyses with Western blotting and enzyme-linked immunosorbent assay (ELISA) revealed that the IgE Abs from Nb-infected BALB/c mice did not show reactivity with Nb-derived excretory-secretory proteins (NES) and antigens present in the cell-free extracts of the worm. Monoclonal IgE Abs obtained from the Nb-infected mice were not reactive with these Nb antigen either. To characterize Nb-induced IgE response, we used (QM x C57BL/6)F1 (QBF1) mice that bear the knock-in 17.2.25 VHDJH segment (VHT) encoding a VH region specific to 4-hydroxy-3-nitrophenylacetyl hapten, and express VHT-encoded antigen receptors on 80-85% of their B cells. Consistent with the frequency of VHT-positive B cells, more than 80% of IgE Abs induced in QBF1 B cells that were cultured with LPS plus IL-4 were found to bear VHT-encoded H chains. In contrast, when QBF1 mice were infected with Nb, less than 10% of Nb-induced IgE Abs were found to use VHT. The QBF1-derived IgE did not react with Nb antigens either. Taken together, data suggest that Nb-induced IgE response in mice is not merely the result of polyclonal activation of B cells, but may involve a mechanism that revises Ig genes secondarily.

Animals↗

Reduced spatial learning in mice infected with the nematode, Heligmosomoides polygyrus.

Parasite modification of host behaviour influences a number of critical responses, but little is known about the effects on host spatial abilities. This study examined the effects of infection with the intestinal trichostrongylid nematode, Heligmosomoides polygyrus, on spatial water maze learning by male laboratory mice, Mus musculus. In this task individual mice had to learn the spatial location of a submerged hidden platform using extramaze visual cues. Determinations of spatial performance were made on day 19 post-infection with mice that had been administered either 50 or 200 infective larvae of H. polygyrus. The infected mice displayed over 1 day of testing (6 blocks of 4 trials) significantly poorer acquisition and retention of the water maze task than either sham-infected or control mice, with mice that had received 200 infective larvae displaying significantly poorer spatial performance than individuals receiving 50 larvae. The decrease in spatial learning occurred in the absence of either any symptoms of illness and malaise, or any evident motor, visual and motivational impairments. It is suggested that in this single host system the parasitic infection-induced decrease in spatial learning arises as a side-effect of the host's immunological and neuromodulatory responses and represents a fitness cost of response to infection.

Analysis of Variance↗

Activation of alveolar macrophages following infection with the parasitic nematode Nippostrongylus brasiliensis.

Alveolar macrophages (AM) of rats infected with 3000 Nippostrongylus brasiliensis infective larvae for 2, 8 or 32 days (D2, D8 or D32 AM) quantitatively surpassed AM from uninfected rats in one or more of IgG- or C3-dependent phagocytosis indices, beta-D-glucuronidase release, or spontaneous release of thymocyte activating factor (interleukin-1, IL-1) and hepatocyte stimulation factor (HSF). These observations suggest that N. brasiliensis infection results in the activation of AM. We have reported previously that a greater proportion of AM from infected rats expressed C3 receptors and were helminthocidal in vitro in the presence of complement than normal AM which were not helminthocidal. The acquisition of the activated state by AM during infection may play a role in vivo lung resistance against migrating helminth parasites.

Animals↗

A novel antimicrobial epoxide isolated from larval Galleria mellonella infected by the nematode symbiont, Photorhabdus luminescens (Enterobacteriaceae).

A novel antimicrobial epoxide, 2-isopropyl-5-(3-phenyl-oxiranyl)-benzene-1,3-diol (1), was identified from larval Galleria mellonella infected by a symbiotically associated bacterium-nematode complex (Photorhabdus luminescens C9-Heterorhabditis megidis 90). Its structure was determined with spectroscopic analysis and confirmed by chemical synthesis starting from a known antibiotic, 2-isopropyl-5-(2-phenylethenyl)-benzene-1,3-diol (2). Epoxide 1 was active against Bacillus subtilis, Escherichia coli, Streptococcus pyogenes, and a drug-resistant, clinical strain of Staphylococcus aureus (RN4220) with minimum inhibitory concentrations in the range of 6.25-12.5 microg/ml. Epoxide 1 was cytotoxic against human cancer cell lines, MCF-7 wt, H460, and Jurkat, with GI(50) of 2.14, 0.63, and 0.42 microM, respectively, but was less toxic on normal, mouse splenic lymphocytes with a GI(50) of 45.00 microM.

Animals↗

The effect of fenbendazole on nematode parasites in experimentally infected lambs.

Fenbendazole given orally to experimentally infected lambs at a dose rate of 5 mg per kg was found to be 100 per cent effective against three and 10-day larave and also against 20-day adults of H contortus, O circumcincta, N battus and T colubriformis. The same dose was also 100 per cent effective against 10-day, 17-day larvae and 27-day adult D filaria.

Animals↗

DNA analysis in the diagnosis of infection and in the speciation of nematode parasites.

DNA analysis is playing an increasingly important role in characterising and classifying nematode parasites. Though less emphasis has been placed on utilising DNA elements to study nematodes of veterinary importance, correct diagnosis is, nevertheless, critical to proper treatment and control. The genus Trichinella presents a particularly interesting problem since the level of classification within this genus remains unclear. Herein we discuss the application of DNA analysis to the diagnosis and speciation of the parasitic nematode Trichinella spiralis, where the study of cloned repetitive DNA elements and ribosomal RNA genes has led to significant advances in the understanding of the species-level systematics of this nematode. Despite advances in the recognition of multiple gene pools within the genus Trichinella, their taxonomic level is still uncertain because of insufficient knowledge regarding intraspecific variation within this genus. The utility of mitochondrial DNA and ribosomal DNA sequence analysis in studying the phylogeny and evolutionary history of parasites within this genus is also discussed.

Animals↗

Effects of preventive anthelmintic treatment on acquired resistance to gastrointestinal nematodes in naturally infected cattle.

The objective of this study was to investigate the influence of different types of chemoprophylaxis in first season grazing calves on their resistance against a natural reinfection with Ostertagia ostertagi and Cooperia oncophora in the second grazing season. Thirty helminth-naive crossbred calves were randomly divided in three groups of 10 animals. The animals of group B received an ivermectin sustained release bolus on day 0. The calves of group D were treated on days 0 and 56 with a subcutaneous injection of doramectin (0.2 mg kg(-1) BW). Group C was the untreated control group ('immune' controls). Although exposure to gastrointestinal nematodes in the first grazing season was only limited, the chemoprophylactic treatments in groups B and D resulted in three distinctly different infection levels (group C > group D > group B). At the start of the second grazing season, six helminth-naive steers (group N, 'susceptible' controls) were turned out together with the second season animals. After 3 weeks of grazing, the 'susceptible' controls were slaughtered, together with four animals from each other group. Parasitological and immunological parameters indicated that resistance to reinfection with Ostertagia was reduced in the chemoprophylactic treated animals, and was negatively related to the degree of suppression of host-parasite contact in the first grazing season (group C > group D > group B > group N). None of the groups had developed a complete resistance against Cooperia yet. A negative relationship was observed between reduction of first grazing season exposure, and weight gains early in the second grazing season. The remaining animals stayed on pasture until the beginning of November. At the end of the second grazing season, levels of acquired resistance against Ostertagia infection were similar in all groups, and all animals had become immune against Cooperia. No effect of first year chemoprophylaxis on total weight gains could be demonstrated. Because of discrepancy between pasture larval counts and tracer worm counts, it was not possible to draw firm conclusions on the effect of chemoprophylaxis on pasture infestation levels in the second year.

Abomasum↗

Heat shock enhances thermotolerance of infective juvenile insect-parasitic nematodes Heterorhabditis bacteriophora (Rhabditida: Heterorhabditidae).

Insect-parasitic nematodes possess many of the attributes of ideal biological control agents, but intolerance to extreme temperatures can restrict their use. We examined whether heat-shock treatments could improve nematode survival and infectivity at temperatures that normally inhibit their activity (35 and 40 degrees C). Nematodes exposed to a sub-lethal temperature (35 degrees C) for 3 h with a latency period of 1-2 h at 25 degrees C killed insects at 35 and 40 degrees C. Correlative evidence was obtained between increased thermotolerance and the synthesis of 70-kDa heat-shock proteins (hsps). These results provide the first evidence of hsp synthesis in the development of thermotolerance and biological activity in the non-feeding, developmentally arrested, infective juvenile nematodes.

Animals↗

Neutral lipids and the assessment of infectivity in entomopathogenic nematodes: observations on four Steinernema species.

An 8-point visual index was developed for Oil Red O staining of neutral lipids in infective juveniles (IJs) of Steinernema carpocapsae (A11), S. riobravis (Biosys 355), S. feltiae (UK76) and S. glaseri (NC). The visual index was found to be a reliable and rapid method for determining the relative neutral lipid content of individual IJs and was validated quantitatively by gas chromatography. The relationship between neutral lipid utilization and infectivity of IJs stored in distilled water at 25 degrees C was also investigated and the first quantitative results on neutral lipid utilization in entomopathogenic nematodes are reported. Neutral lipid contents of freshly harvested IJs of S. carpocapsae, S. riobravis. S. feltiae and S. glaseri were 31, 31, 24 and 26% dry wt, respectively. Steinernema carpocapsae showed a sigmoidal pattern for neutral lipid utilization while S. riobravis used neutral lipids at an almost constant rate. Survivorship of these two species ranged between 120 and 135 days, whereas S. feltiae and S. glaseri lived > 450 days and had a slower rate of lipid utilization during a 260 day storage period. Oil Red O staining showed that individual IJs in each population utilized lipids at different rates, even though they had the same initial lipid index. The infectivity of S. riobravis, S. feltiae and S. glaseri declined with lipid utilization. In contrast, S. carpocapsae maintained a high level of infectivity even at relatively low lipid levels. Therefore, neutral lipid content was found to be a suitable indicator of infectivity for S. riobravis, S. feltiae and S. glaseri but not for S. carpocapsae.

Animals↗

Influence of protein supplementation on the resistance and resilience on young hair sheep naturally infected with gastrointestinal nematodes during rainy and dry seasons.

Thirty, 4-month-old entire Santa Ines lambs were grazed on an Andropogon gayanus pasture, during a 34-week period (rainy season weeks 0-20 and dry season weeks 21-34) and allocated in two treatment groups (n = 15) each with different protein supplementation: high protein (HP-19% CP) and low protein (LP-11% CP). These were subdivided into those receiving anthelmintic treatment (c) (n = 7) and without anthelmintic treatment (i) (n = 8). The objective was to evaluate the effects of supplementation with protein on resistance and resilience to natural helminth infection of hair breed lambs. Lamb weight, blood collection and faecal egg counts (FEC) were carried out monthly. The lambs were slaughtered after 34 weeks, when worm burdens, worm length and eosinophil cell counts were taken. The sheep on treatments HPc and HPi were heavier in live weight than those from LPi and LPc (P < 0.05) at the end of the rainy period. The HPc group finished heavier (P < 0.05) than the other groups in the dry season, which had no significant differences between them. The predominant species of nematode found was T. colubriformis followed by H. contortus, Trichuris globulosa and Moniezia expansa. Animals on HPi had lower FEC than LPi (P < 0.05). The number of worms was lower for both HP groups (P < 0.05) with worm length shorter in the HPc group (P < 0.05) compared with all other groups. The number of eosinophils was higher in animals in the LPi group, which also showed anaemia and lower plasma urea at the end of the dry season. Diet supplementation with high protein was able to improve resilience and resistance to natural infection by endoparasites during the rainy season. In the dry season there was a decrease in both of these traits, which were intimately linked to the quality of available forage under tropical conditions.

Animal Feed↗

New oral linguiform projections and their associated neurons in the third-stage infective larva of the parasitic nematode Oesophagostomum dentatum.

The infective larvae (L3i) of the nematode parasite of swine, Oesophagostomum dentatum, are passively ingested by their hosts. The L3i exhibit certain behaviors that are probably selected to increase the likelihood of ingestion, by strategic positioning in the environment. The larvae show positive geotactic behavior and respond to temperature variations in their environment, as shown by their behavior on a thermal gradient. To investigate neuronal control of this behavior, we initiated a study of the structure of the amphidial neurons of this parasite. The same number and types of neuronal dendritic processes are found in the amphids of the O. dentatum L3i as in those of its close relatives Haemonchus contortus and Ancylostoma caninum. Well-developed dendritic processes of wing cells are located in the amphidial sheath cells, these being similar to wing cells AWA in the free-living nematode Caenorhabditis elegans but actually more extensive. Similar to its close relatives just mentioned, and C. elegans as well, O. dentatum L3i has prominent finger cell processes, the finger cell neurons being the thermoreceptors in all 3 of the preceding species. However, unlike the arrangement seen in H. contortus and A. caninum, where the microvilli-like "fingers" of these neurons lie dorsal to the amphidial channel and occupy a very large portion (>50%) of the anterior end of the larva, the dendritic process of the finger cells in O. dentatum extends into unusual linguiform projections that, in turn, extend into the lumen of the mouth tube, a complex structural arrangement that has not been described for any other nematode.

Animals↗

Lipopolysaccharides of Rhizobium etli strain G12 act in potato roots as an inducing agent of systemic resistance to infection by the cyst nematode Globodera pallida.

Recent studies have shown that living and heat-killed cells of the rhizobacterium Rhizobium etli strain G12 induce in potato roots systemic resistance to infection by the potato cyst nematode Globodera pallida. To better understand the mechanisms of induced resistance, we focused on identifying the inducing agent. Since heat-stable bacterial surface carbohydrates such as exopolysaccharides (EPS) and lipopolysaccharides (LPS) are essential for recognition in the symbiotic interaction between Rhizobium and legumes, their role in the R. etli-potato interaction was studied. EPS and LPS were extracted from bacterial cultures, applied to potato roots, and tested for activity as an inducer of plant resistance to the plant-parasitic nematode. Whereas EPS did not affect G. pallida infection, LPS reduced nematode infection significantly in concentrations as low as 1 and 0.1 mg ml(-1). Split-root experiments, guaranteeing a spatial separation of inducing agent and challenging pathogen, showed that soil treatments of one half of the root system with LPS resulted in a highly significant (up to 37%) systemic induced reduction of G. pallida infection of potato roots in the other half. The results clearly showed that LPS of R. etli G12 act as the inducing agent of systemic resistance in potato roots.

Animals↗

Natural infection of gastrointestinal nematodes in long-nosed armadillos Dasypus novemcinctus Linnaeus, 1758 from Pantanal wetlands, Aquidauana sub-region, Mato Grosso do Sul State, with the description of Hadrostrongylus speciosum n. gen. et n. sp. (Molineidae: Anoplostrongylinae).

This study evaluated the gastrointestinal helminth fauna of long-nosed armadillos, Dasypus novemcinctus, from the Pantanal wetlands, Aquidauana sub-region, Aquidauana County, Mato Grosso do Sul State, Brazil. Thirteen species of nematodes, comprising seven genera and four families, were recovered from their gastrointestinal tracts. The following descriptors of infection were determined: prevalence, variation of intensity, average intensity and abundance. Hadrostrongylus speciosum n. gen. et n. sp. is first described here.

Animals↗