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Nematode sperm.

Parasitic nematode infections remain a major public health problem in many parts of the world. Because most of the current strategies aimed at controlling parasitic nematode infections have met with only limited success, it may be time to consider alternative approaches. An aspect of nematode biology that has drawn little attention as a target for control is the reproductive process. Although there are numerous facets of the overall reproductive biology of nematodes that hold potential as targets for intervention, Alan Scott here focuses on the male reproductive system, and outlines some of the known unique processes and characteristics of sperm formation and sperm function that could be exploited to block fertilization.

Journal Article↗

Effect of repeated anthelmintic treatment on ability to detect predisposition of mice to Heligmosomoides polygyrus and Aspiculuris tetraptera (Nematoda) infections.

Several recent studies have demonstrated predisposition of certain humans to heavy gastro-intestinal nematode infections. This finding has been used to support the concept of community control of nematodes based on selective treatment of the few heavily infected individuals in the community. A mouse model has been used in this study to determine whether those individuals found to be 'predisposed' to heavy infection continue to regain heavy nematode infections following repeated anthelmintic treatments. Re-infection was examined over 3 intervals of 8 weeks in 52 mice and individual worm burdens attained following drug-induced expulsion were analysed. In the case of Heligmosomoides polygyrus, parasite numbers obtained after first treatment were only significantly correlated with those obtained after the second treatment. Re-infection levels in mice during the second and third re-infection periods did not correlate with parasite loads at the beginning of the study. In the case of Aspiculuris tetraptera, correlations after each re-infection period remained significant, although the magnitude of the correlation coefficient decreased with successive re-infection intervals. The data suggest that a selective control programme based on repeated screening to identify heavily infected individuals prior to each treatment would reduce the number of H. polygyrus in the host population 17.6% more than a single screening at the beginning of the control programme, and would reduce the number of A. tetraptera by 14% more than a single screening. Re-infection rates for H. polygyrus were shown to decrease with increasing mouse age whereas re-infection rates for A. tetraptera increased with increasing mouse age.

Age Factors↗

Arabidopsis thaliana mutants with an altered susceptibility to the root-knot nematode Meloidogyne incognita.

Sedentary endoparasitic nematodes of plants are obligate parasites of roots in which they complete their sophisticated life cycle. To study the plant/root-knot nematode interaction, a screening was performed for mutants of Arabidopsis thaliana that were less susceptible to nematode infection. Ethyl methanesulfonate-mutagenized A. thaliana M2 seeds (13,000) were germinated and the seedlings were screened in vitro for nematode susceptibility. Another 5,000 plants were screened in a sand mixture under glasshouse conditions. Ten mutants that were at least a 2-fold less susceptible than the wild-type were retained and analyzed in more detail in vitro as well as in the greenhouse. All mutants with a fairly normal morphology had only a slightly, but reproducibly, decreased susceptibility to nematodes. One mutant (AMi 2) with a 17-fold lower susceptibility to nematode infection and an aberrant phenotype could be rescued by tryptophan.

Animals↗

Effects of various hormones on the sexual maturity of European eel (Anguilla anguilla L.) females from farm and lakes.

Previously described and alternative methods of the induction of sexual maturation in the European eel were investigated. Weekly administrations of a gonadoliberin agonist (nnRH-A=D-Phe6-GnRH-Ea) did not induce statistically significant effect on the gonads of treated eels in none of the dosages used (0.1 microg and 10 microg/fish). Carp pituitary extract and carp pituitary extract together with a dopamine antagonist caused considerable external changes (increase in eye size) and significant gonadal development in two treatment groups: wild and cultivated stocks. The induction of the ovulation by double amount of CP and gonadoliberin agonist with dopamine antagonist mixture was not successful in a wild stock. Fertilisation of stripped eggs of farm eel was attempted unsuccessfully in, due to low egg quality. An advanced phase of the sexual maturation process could be induced in specimen infected by Anguillicola crassus indicating, that nematode infection is not a limiting factor in the artificial propagation of the European eel.

Air Sacs↗

Genes induced during early response to Meloidogyne incognita in roots of resistant and susceptible alfalfa cultivars.

A cDNA library made to RNA from roots of Meloidogyne incognita (root-knot nematode) susceptible alfalfa cv. Lahontan seedlings 72 h after root-knot nematode inoculation was differentially screened with cDNA made from uninoculated control and M. incognita infested (72 h) root RNA. Of the six cDNAs isolated, the deduced amino acid sequences of four showed significant homology to sequences present in the databank, while two were pioneer sequences. The four cDNAs with matches to known sequences include those for glycine-rich protein, the gluconeogenic pathway enzyme phosphoenolpyruvate carboxykinase, an isoflavone reductase-like protein, and metallothionein. We have followed the expression of these genes during the course of nematode infection in both the susceptible and resistant host and also in different plant organs. Based on these analyses, the genes induced early in nematode infection are related either to metabolic pathways or to stress/defense.

Journal Article↗

An extracellular protease from Brevibacillus laterosporus G4 without parasporal crystals can serve as a pathogenic factor in infection of nematodes.

Brevibacillus laterosporus is an aerobic spore-forming bacterium with the ability to produce canoe-shaped lamellar parasporal inclusions adjacent to spores. An isolate named G4 was identified as a B. laterosporus which does not produce parasporal crystals and shows significant toxic activity toward nematodes. Crude extracellular protein extract from culture supernatant of B. laterosporus G4 killed the nematodes within 12 h and finally destroyed the targets within 24 h, which suggested possible proteinaceous pathogeny. A homogeneous extracellular protease with nematicidal activities, purified by chromatography, confirmed the hypothesis that it might serve as a pathogenic factor during infection of the G4 strain. Characterization of the purified protease revealed a molecular mass of 30 kDa and optimum activity at pH 10 and 50 degrees C. The protease hydrolyzed relatively broad substrates including collagen and the cuticle of nematodes, and histopathological observations demonstrated the resulting destroyed nematode cuticle upon treatment by purified protease. Our present study reveals that extracellular protease, but not previously reported parasporal crystals, can be employed in infection against invertebrates by the B. laterosporus G4 strain.

Animals↗

Efficacy of moxidectin, ivermectin and albendazole oral drenches for suppression of periparturient rise in ewe worm egg output and reduction of anthelmintic treatment for lambs.

Sixty multiparous crossbred ewes which had lambed within three days in the first week of April 1996, were divided into four groups. Each group consisted of 15 ewes plus 12 pairs of twins and three single lambs. Group 1 was left untreated, group 2 was treated with albendazole 2.5 per cent drench, group 3 received moxidectin 0.1 per cent drench and group 4 received ivermectin 0.08 per cent drench. The ewes in each group were dosed with their anthelmintic on April 4 (day 0) before being turned out to separate equal-sized paddocks within the same field on the following morning. The field had been used for grazing sheep annually for many years and was considered to be contaminated with infective larvae of the common gastrointestinal nematodes infecting sheep in the region. Faecal samples were collected every two weeks from the ewes and lambs until July 25 (day 112). The lambs in each group were dosed with the anthelmintic used for their dams on day 42, and the dose was repeated when more than 50 per cent of the lambs in any group had a faecal egg count of more than 200 eggs per gram (epg). The total faecal egg output of the treated ewes over days 14 to 70, compared with that of the untreated control group, was reduced by 78.9 per cent by the moxidectin drench, by 47.6 per cent by ivermectin, and by 21.5 per cent by albendazole. The lambs in the groups treated with moxidectin and ivermectin required only one treatment on day 42 before reaching finishing weight; those in the albendazole-treated group were treated twice and the control group once. The faecal egg outputs of the lambs from day 42 until the end of the experiment on day 112 were reduced by 75 per cent by the moxidectin drench, by 48.5 per cent by ivermectin, and by 9 per cent by albendazole. There were no significant differences between the rates of weight change of either ewes or lambs in any of the groups.

Albendazole↗

Epidemiological study and comparison of pyrantel and levamisole in the treatment of roundworm and hookworm infestations.

Stool examined for presence of nematodes from 467 school students in Dacca city showed 48% ova positive cases with 22% among them cases of mixed nematode infection. Highest prevalence of 58% was observed with roundworm and 5% was hookworm infestations. Among hookworm cases, Necator americana (68%) appears to be the predominant species. Two anthelmintics-pyrantel and levamisole achieved similar high cure rate (around 97%) and Egg reduction rate of 99% in roundworm infection. However, pyrantel was observed to be more effective in hookworm infestation (87% against 60%) particularly in Necator americana cases.

Adolescent↗

Mebendazole in the treatment of helminthiasis.

Mebendazole, a new broad-spectrum anthelmintic, was used to treat patients with nematode infections--ascariasis, trichuriasis and hookworm. The dosage for adults was 100 mg twice daily for 3 days and for children, 50 mg twice daily for 3 days. Pretreatment and post-treatment egg counts on stool specimens showed that after mebendazole there was a reduction of over 99% in egg count per gram of stool in all three types of infection. The overall cure rates for the infections were as follows: Ascaris lumbricoides, 86.8% (59/68); Trichuris trichiura, 86.0% (37/43); and hookworm, 85.7% (24/28). The drug was equally effective in light and heavy infections. No important side effect was noted with this drug. It is suggested that mebendazole is the drug of choice for trichuriasis and mixed nematode infection.

Adult↗

Environmental interventions and the pattern of geohelminth infections in Salvador, Brazil.

This paper reports a longitudinal study, conducted in 1989/90, of 1893 children aged 5 to 14 years in 9 poor urban areas of the city of Salvador (population 2.44 million), capital of Bahia State in northeast Brazil. Stool examinations were performed to measure nematode infection and reinfection 9 months after treatment, and an extensive questionnaire was applied to collect information on each child and on the conditions of the household. Comparison of areas with different levels of infrastructure showed the following trends as the level of community sanitation improved: clustering of cases by household became more significant, predisposition of individuals to reinfection and to heavy infection became more marked, and infections with different species were increasingly aggregated in the same individuals. These results suggest that sewerage and drainage can significantly reduce transmission of intestinal nematode infections in the public domain, but that other measures are required to control transmission within the household.

Adolescent↗

Role of the bovine immune system and genome in resistance to gastrointestinal nematodes.

Gastrointestinal nematode infections of cattle remain a constraint on the efficient raising of cattle on pasture throughout the world. Most of the common genera of parasites found in cattle stimulate an effective level of protective immunity in most animals within the herd after the animals have been on pasture for several months. In contrast, cattle remain susceptible to infection by Ostertagia for many months, and immunity that actually reduces the development of newly acquired larvae is usually not evident until the animals are more than 2 years old. This prolonged susceptibility to reinfection is a major reason that this parasite remains the most economically important GI nematode in temperate regions of the world. Although, animals remain susceptible to reinfection for a prolonged period of time, there are a number of manifestations of the immune response that result in an enhanced level of herd immunity. These include a delay in the development time of the parasites, an increase in the number of larvae that undergo an inhibition in development, morphological changes in the worms, stunting of newly acquired worms, and most importantly a reduction in the number of eggs produced by the female worms. The overall result of these manifestations of immunity is a reduction in parasite transmission within the cattle herd. The immune mechanisms responsible for these different types of functional immunity remain to be defined. In general, GI nematode infections in mammals elicit very strong Th2-like responses characterized by high levels of Interleukin 4 (IL4), high levels of IgG1 and IgE antibodies, and large numbers of mast cells. In cattle, the most extensively studied GI nematode, in regards to host immune responses, is Ostertagia ostertagi. In Ostertagia infections, antigens are presented to the host in the draining lymph nodes very soon after infection, and within the first 3-4 days of infection these cells have left the nodes, entered the peripheral circulation, and have homed to tissues immediately surrounding the parasite where they become established. The immune response seen in the abomasum is in many ways are similar to that seen other mammalian hosts, with high levels of expression of IL4 in the draining lymph nodes and in lymphocytes isolated from the mucosa. But unlike a number of other systems, lymphocyte populations taken from Ostertagia infected cattle seem to be up-regulated for a number of other cytokines, most notably Interferon (IFN, implying that in Ostertagia infections, the immune response elicit is not simply a stereotypic Th2 response. In addition, effector cell populations in the tissues surrounding the parasites, are not typical, inferring the Ostertagia has evolved means to suppress or evade protective immune mechanisms. Studies have also demonstrated that the number of nematode eggs/gram (EPG) in feces of pastured cattle is strongly influenced by host genetics and that the heritability of this trait is approximately 0.30. In addition, EPG values are not "normally" distributed and a small percentage of a herd is responsible for the majority of parasite transmission. This suggests that genetic management of a small percentage of the herd can considerably reduce overall parasite transmission. A selective breeding program has been initiated to identify the host genes controlling resistance/susceptibility to the parasites. The best indicator of the number of Cooperia infecting a host is the EPG value, while Ostertagia is best measured by serum pepsinogen levels, weight gain, and measures of anemia. Other phenotypic measures are either not significantly associated with parasite numbers or are very weakly correlated. In addition, calves can be separated into three types: (1) Type I which never demonstrates high EPG values, (2) Type II which shows rises in EPG values through the first 2 months on pasture which then fall and remain at levels associated with Type I calves, and (3) Type III calves which maintain high EPG levels. The approximate percentage of these calves is 25:50:25 respectively. Because these cattle are segregating for traits involved in resistance and susceptibility to GI nematodes, this resource population is being used to effectively detect the genomic locations of these Economic Trait Loci (ETL). For relational analysis between phenotype and genome location, over 80,000 genotypes have been generated by PCR amplification, and marker genotypes have been scored to produce inheritance data. The marker allele inheritance data is currently being statistically analyzed to detect patterns of co-segregation between allele haplotype and EPG phenotypes. Statistical power of this genome-wide scan has been strengthened by including genotypic data from the historic pedigree. In our herd, paternal half-sib families range from 5-13 progeny/sire, and extensive marker genotypes are available from ancestors of the population most of which are paternally descended from a single founding sire. Once ETL have been identified the next will be to refine ETL map resolution in attempt to discover the genes underlying disease phenotypes. Accurate identification of genes controlling resistance will offer the producer several alternatives for disease control. For a non-organic producer, the small percentage of susceptible animals can be targeted for drug administration. This approach would reduce both the cost of anthelmintics used and the odds for selection of drug resistant mutants, because the selective agent (drug) would not be applied over the entire parasite population. A second treatment option would be based on correcting a heritable immunologic condition. In this case, susceptible animals could be the targets for immunotherapy involving vaccines of immunomodulation. A final option would be genetic selection to remove susceptible animals from the herd. Producers with a high degree of risk for parasite-induced production losses, such as organic producers of producers in geographic areas with environmental conditions favorable to high rates of transmission would benefit the most from this strategy. In contrast, producers at low risk could take a more conservative approach and select against susceptibility when other factors were equal.

Animals↗

Efficacy of benzimidazole carbamate on an intestinal fluke co-infected with nematodes.

The efficacy of a single dose of benzimidazole, drugs commonly used for the treatment of Ascaris and hookworm, was evaluated against one of the tiny-sized intestinal flukes, Haplorchis sp in the endemic area where mixed infections of roundworms and flatworms existed. At day 7 after treatment, albendazole (400 mg) induced 42.5% cure rate, mebendazole (500 mg) a cure rate of 32.4%, on the other hand, praziquantel (40 mg/kg) gave 94.6% cure rate and the placebo at 15.9%. At the single dose, benzimidazole could not completely expelled the haplorchid; but could reduce one third to two fifth of the infection, similar to the drugs efficacy against Trichuris infection.

Albendazole↗

Polymyositis caused by a new genus of nematode.

We report two patients who presented with increasing malaise and myalgia, and had biopsy-proven polymyositis. Their conditions deteriorated after corticosteroid treatment, and repeat muscle biopsies showed adult and larval nematodes. Anthelminthic treatment was completely successful in both cases. The infecting nematode appears to belong to a new genus and is, to our knowledge, the first known muspiceoid nematode to infect humans. Its life cycle and the route of infection are unknown.

Adult↗

Faecal egg counts in Angora goats following natural or experimental challenge with nematode parasites: within-flock variability and repeatabilities.

Phenotypic variation in faecal worm-egg count (FEC) was studied in 100 Angora goats between 6 months and 2 years old as part of a wider investigation into the potential use of FEC as a selection trait in breeding goats for improved natural resistance to nematode infection. Shortly after weaning at 4-5 months old the goats were subjected to a procedure involving successive natural and experimental challenges, which was continued at 6-monthly intervals until the goats were 2 years old. Experimental challenges consisted of Trichostrongylus colubriformis infective larvae administered orally at a dose rate of 1000 larvae/kg liveweight. FECs resulting from both natural and experimental infections varied considerably between individual goats as well as between different sampling times. Mixed model analysis of variance of natural infections confirmed that a significant component of this variation was attributable to differences between individual goats (P < 0.01) as opposed to other potential sources of variability, indicating that although generally deemed to be more susceptible than sheep, Angora goats do nevertheless vary in their ability to resist nematode infection. Rank correlations between pairs of FECs derived from the same infection were relatively high for both natural and experimental infections (r = 0.63-0.70 and 0.54-0.79, respectively; all P < 0.01). However, those between mean FECs measured on successive 6-monthly sampling occasions were considerably lower and more variable (r = 0.01-0.41 and 0.19-0.62 for natural and experimental infections, respectively). Nevertheless, FECs resulting from natural infections showed moderate correlations with those from corresponding experimental infections (P < 0.01) on all except the first sampling occasion (r = 0.23-0.61). Correlations were marginally higher when FECs from experimental infections were adjusted for infecting dose size (r = 0.32-0.63). FECs resulting from experimental infections were significantly higher on the first sampling occasion (at 6 months) than on subsequent occasions (P < 0.01) suggesting that some degree of resistance to nematode establishment had developed in the goats by 12 months old. On the basis of unadjusted FECs there appeared to be no marked increase in resistance beyond that age, although analyses based on FECs adjusted for infecting dose size suggested an increase in acquired resistance up to 18 months of age. FECs following natural challenge in goats which had also received experimental infections were not significantly different from those exposed only to natural challenge, indicating that the drench-abbreviated experimental infections had had no immunizing effect. Does that had given birth and were lactating at 24 months had significantly higher FECs than 'dry' does in the flock (P < 0.01), suggesting that like sheep, goats exhibit a post-parturient relaxation of immunity. The use of FECs as potential indicators of resistance status in goats and as a criterion for the selection of breeding animals is discussed.

Animals↗

The promoter of the nematode resistance gene Hs1pro-1 activates a nematode-responsive and feeding site-specific gene expression in sugar beet (Beta vulgaris L.) and Arabidopsis thaliana.

The Hs1pro-1 gene confers resistance to the beet cyst nematode Heterodera schachtii in sugar beet (Beta vulgaris L.) on the basis of a gene-for-gene relationship. RNA-gel blot analysis revealed that the transcript of Hs1pro-1 was present in uninfected roots of resistant beet at low levels but increased by about fourfold one day after nematode infection. Treatments of plants with external stimuli including salicylic acid, jasmonic acid, gibberellic acid and abscisic acid as well as wounding or salt stress did not result in changes in the gene transcription, indicating de novo transcription of Hs1pro-1 upon nematode infection specifically. To study transcriptional regulation of Hs1pro-1 expression at the cellular level, a 3082 bp genomic fragment representing the Hs1pro-1 promoter, isolated from the YAC-DNA housing the Hs1pro-1 gene, was fused to the beta-glucuronidase reporter gene (1832prm1::GUS) and transformed into susceptible beet roots and Arabidopsis plants, respectively. Fluorometric and histochemical GUS assays on transgenic beet roots and Arabidopsis plants carrying the 1832prm1::GUS construct demonstrated that the Hs1pro-1 promoter is functional in both species and drives a nematode responsive and feeding site-specific GUS-expression. GUS activity was detected as early as at initiation of the nematode feeding sites and GUS staining was restricted to the nematode feeding sites. To delineate the regulatory domains of the Hs1pro-1 promoter, fusion genes with various 5' deletions of the Hs1pro-1 promoter and the GUS gene were constructed and analysed in transgenic beet roots as well. Cis elements responsible for feeding site-specific gene expression reside between -355 and +247 from the transcriptional initiation site of Hs1pro-1 whereas an enhancer region necessary for higher gene expression is located between -1199 and -705 of the promoter. The Hs1pro-1 promoter drives a nematode feeding site-specific GUS expression in both sugar beet and Arabidopsis suggesting a conserved mechanism of regulation of Hs1pro-1 expression in these two species.

Animals↗

Constitutive secretion of the granule chymase mouse mast cell protease-1 and the chemokine, CCL2, by mucosal mast cell homologues.

BACKGROUND: The mucosal mast cell (MMC) granule-specific beta-chymase, mouse mast cell protease-1 (mMCP-1), is released systemically into the bloodstream early in nematode infection before parasite-specific IgE responses develop and TGF-beta1 induces constitutive release of mMCP-1 by homologues of MMC in vitro. Intraepithelial MMC may also express the chemokine CCL2 (monocyte chemotactic protein-1) during nematode infection but the expression of this chemokine by MMC homologues has not been investigated. OBJECTIVE: To investigate the expression and to compare the mechanisms of constitutive release of the chymase, mMCP-1, and the chemokine, CCL2. METHODS: MMC homologues were generated by culturing bone marrow cells in the presence of TGF-beta1, IL-3, IL-9 and stem cell factor (SCF). The intracellular distribution of mMCP-1 and CCL2 was examined by confocal microscopy. The involvement of the Golgi complex and of protein synthesis in the constitutive release of mMCP-1 and CCL2 was investigated using the Golgi-disrupting agent brefeldin A and cycloheximide to block protein synthesis. Secreted analytes were quantified by ELISA. RESULTS: mMCP-1 colocalized with Golgi matrix protein 130 but was most abundant in the granules, whereas CCL2 was not found in the granules but appeared to be located uniquely in the Golgi complex. Extracellular release of mMCP-1 was significantly inhibited ( approximately 40%) by cycloheximide and by the Golgi-disrupting agent brefeldin A, indicating both continuous protein synthesis and transportation via the Golgi complex are required for optimal mMCP-1 secretion. A similar but more marked inhibitory effect with both compounds was demonstrated on the constitutive secretion of CCL2. CONCLUSION: The culture conditions that promote mMCP-1 expression and release by MMC homologues also promote the expression and release of CCL2. Constitutive release involves de novo protein synthesis and requires a functional Golgi complex, suggesting that similar mechanisms of extracellular secretion operate for both mediators.

Animals↗

Heligmosomoides polygyrus and Trypanosoma congolense infections in mice: a laboratory model for concurrent gastrointestinal nematode and trypanosome infections.

A murine model using Heligmosomoides polygyrus and Trypanosoma congolense has been developed for studying the effects of concurrent chronic gastrointestinal nematode and trypanosome infections. Female outbred mice were infected either with 500 infective larvae (L3) of H. polygyrus or with 10(4) bloodstream forms of T. congolense or both. In concurrent infections, animals were dosed with both parasites simultaneously or the trypanosomes were injected 5 or 10 days after the mice were infected with the nematode. The course of infection was monitored by routine parasitological and immunological techniques for 30 days after the H. polygyrus infection. Concurrently infected mice were severely compromised, except when T. congolense was superimposed on a 10-day-old (adult) H. polygyrus infection. In H. polygyrus-infected mice, simultaneous or subsequent infection with trypanosomes did not markedly influence worm establishment or fecundity, but the female worms were slightly stunted. Surviving mice displayed a markedly reduced antibody response to H. polygyrus antigens and a slightly reduced antibody response to T. congolense antigens.

Animals↗