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Preferential expression of a plant cystatin at nematode feeding sites confers resistance to Meloidogyne incognita and Globodera pallida.

The expression patterns of three promoters preferentially active in the roots of Arabidopsis thaliana have been investigated in transgenic potato plants in response to plant parasitic nematode infection. Promoter regions from the three genes, TUB-1, ARSK1 and RPL16A were linked to the GUS reporter gene and histochemical staining was used to localize expression in potato roots in response to infection with both the potato cyst nematode, Globodera pallida and the root-knot nematode, Meloidogyne incognita. All three promoters directed GUS expression chiefly in root tissue and were strongly up-regulated in the galls induced by feeding M. incognita. Less activity was associated with the syncytial feeding cells of the cyst nematode, although the ARSK1 promoter was highly active in the syncytia of G. pallida infecting soil grown plants. Transgenic potato lines that expressed the cystatin OcIDeltaD86 under the control of the three promoters were evaluated for resistance against Globodera sp. in a field trial and against M. incognita in containment. Resistance to Globodera of 70 +/- 4% was achieved with the best line using the ARSK1 promoter with no associated yield penalty. The highest level of partial resistance achieved against M. incognita was 67 +/- 9% using the TUB-1 promoter. In both cases this was comparable to the level of resistance achieved using the constitutive cauliflower mosaic virus 35S (CaMV35S) promoter. The results establish the potential for limiting transgene expression in crop plants whilst maintaining efficacy of the nematode defence.

Journal Article↗

Immune responses of sheep to the parasitic nematode Trichostrongylus colubriformis: infections in Thiry-Vella loops.

Some aspects of the local immune response to nematode infection were examined by introducing fourth-stage Trichostrongylus colubriformis larvae into Thiry-Vella loops in worm-free and T. colubriformis-resistant sheep. Approximately 15% of the larvae introduced into the worm-free sheep survived and matured into adult worms, whereas less than 1% survival in resistant sheep. The amount of IgG1, IgG2, IgM and albumin discharged from loops in worm-free but not resistant sheep all increased following infection with larvae. Secretions from resistant sheep contained approximately three times as much IgA as secretions from the worm-free group and, in addition, their plasma IgG1 level was significantly higher. Antibodies against T. columbriformis were found in both plasma and intestinal secretions, the highest titres being found in the resistant sheep. During worm expulsion from loops in the resistant sheep neither immunoglobulin levels nor antibody titres in the secretions increased significantly.

Animals↗

IFN inhibits inflammatory responses and protective immunity in mice infected with the nematode parasite, Nippostrongylus brasiliensis.

Mice infected with the gastrointestinal nematode parasite Nippostrongylus brasiliensis (Nb) develop responses associated with enhanced production of IL-4 (increased serum IgE levels and intestinal mucosal mastocytosis) and IL-5 (tissue and peripheral blood eosinophilia). The antagonistic effects of IFN on IL-4-mediated responses prompted an examination of the effects of IFN on the host response to Nb. Treatment with rIFN-alpha and rIFN-gamma induced a marked increase in parasite egg production (fecundity) in BALB/c mice infected with Nb and delayed intestinal expulsion of adult worms. Treatment with rIFN-alpha or rIFN-gamma also inhibited the rise in peripheral blood eosinophilia that follows inoculation with Nb, and the intensity of pulmonary perivascular tissue eosinophilia. However, Nb-induced increases in serum IgG levels and intestinal mastocytosis were only temporarily delayed by IFN. Induction of endogenous IFN production by injection of fixed Brucella abortus into mice infected with Nb also resulted in an increased worm fecundity and delayed adult worm expulsion. These effects were ablated when mice given Brucella abortus also received injections of neutralizing anti-IFN antibodies. Thus, IFN inhibit host protective immunity to Nb, perhaps by interfering with the production and effects of Th2 cytokines.

Animals↗

Pathology of pyogenic liver abscess in children.

Little is known about preexisting lesions in livers of children with pyogenic liver abscess (PLA). Study of these lesions may elucidate possible predisposing factors for the disease. In Vitória, state of Espirito Santo, Brazil, PLA in children is frequently associated with helminthic infections and eosinophilia. We hypothesize that nematode infection with larvae migrating through the liver is a predisposing factor for PLA, because the infection induces immunomodulation and likely trapping of bacteria in liver granulomas. In this report, we describe observations of 22 cases of PLA in children studied at autopsy (16 cases) or in surgical biopsies (6 cases), including 17 boys and 5 girls ranging in age from 1 to 13 years (mean 4.6 years, median 3.5 years). Multiple abscesses in both lobes were found in 13 cases and a single abscess was found in the right lobe in 10 cases. All cases showed histologically classical pyogenic inflammation without morphological evidence of amoebiasis. In six cases there were granulomas similar to those caused by larva migrans visceralis (from Toxocara or other nematodes) in liver tissue not affected by the abscess. Nematode antigens in central areas of necrosis of granuloma in all six cases and fragments of a larva, possibly of Toxocara, were found on samples immunohistochemically stained with polyclonal anti-Toxocara antibodies. There were numerous eosinophils in abscesses with Charcot-Leyden crystals. Eosinophils were found frequently in portal triads far from the abscess wall. In four cases, in which bile duct ascariasis was found, worms were noted in the bile ducts, and eggs were found in liver parenchyma surrounding the abscess in two cases. Foreign-body granulomas were found in one case in which penetrating trauma was the cause of abscess. In one case there was one histiocytic granuloma whose origin was not determined. The observation of six cases of granuloma similar to larva migrans visceralis (or produced by other nematode larva) in liver tissue not directly affected by the abscess supports the hypothesis that helminth infections with larva migrating through the liver are a predisposing factor for pyogenic hepatic abscess in children.

Adolescent↗

Nematode genera diversity in cattle: similarity of between-sire progenies.

Breeding cattle for resistance to nematode infection is mostly based on egg excretion. This, however, does not allow for generic identification of the nematodes involved. Unless we know whether the selected resistance is directed against one or several particular genera, a strong bias could be introduced in the selection programs. In order to estimate the likelihood of this potential bias we investigated nematode genera diversity in the progeny of four sires in 1992 and seven sires in 1994. Three of the four Aberdeen Angus sires used in 1992 were related while the seven sires in 1994 were unrelated. Diversity was assessed using at least ten individual faecal cultures for each progeny group during each of the two sampling periods (beginning and end of grazing period, April and September). It was estimated by the relative proportion of each genera (Ostertagia, Cooperia, Haemonchus, Trichostrongylus and Oesophagostomum) or by either the Shannon (genera diversity) or Pielou (genera evenness) index. The Shannon index was repeatable when measured at 2-week intervals within the same progeny group on ten random faecal samples. No significant difference was recorded between sire genera diversity over the two sampling periods. This indicated that hosts have a limited effect on the nematode genera diversity as assessed by faecal cultures, and that the selection of resistant hosts could probably be achieved using faecal egg counts.

Animals↗

Study on the elimination of Angiostrongylus costaricensis first stage larvae in the experimental infection of Swiss mice.

Abdominal angiostrongylosis is a nematode infection wild rodents. Human infection may result in severe abdominal disease and has been reported from several countries in the Americas. The domestic mouse, Mus musculus, has not been found with natural infection and, like other urban rodents, should not be considered a natural host for Angiostrongylus costaricensis. Quantification of parasitic forms released for transmission may better express the coevolutionary status in parasite-host relationship. With this objective, five groups of experimentally infected Swiss mice were followed for up to 155 days post-infection (PI) days and the quantification of first stage larvae (L1) output revealed: an irregular elimination of L1 and a huge variation in the patency period (1 to 114 days) and in the number of L1 eliminated daily by individual animals (1 to 6340 L1/g). Overall mortality was 72% (range: 28% to 100%) at seven weeks PI. In conclusion, abdominal angiostrongylosis in M. musculus presents high mortality and a very variable and irregular elimination of L1 in feces.

Angiostrongylus↗

Microarray analysis of selection lines from outbred populations to identify genes involved with nematode parasite resistance in sheep.

Gastrointestinal nematodes infect sheep grazing contaminated pastures. Traditionally, these have been controlled with anthelmintic drenching. The selection of animals resistant to nematodes is an alternative to complete reliance on drugs, but the genetic basis of host resistance is poorly understood. Using a 10,204 bovine cDNA microarray, we have examined differences in gene expression between genetically resistant and susceptible lambs previously field challenged with larval nematodes. Northern blot analysis for a selection of genes validated the data obtained from the microarrays. The results identified over one hundred genes that were differentially expressed based on conservative criteria. The microarray results were further analyzed to identify promoter motifs common to the differentially expressed genes. Motifs identified in upregulated gene promoters were primarily restricted to those promoters; however, motifs identified in downregulated gene promoters were also found in the promoters of upregulated genes but not in the promoters of genes whose expression was unaltered. Protein Annotators' Assistant was used for lexical analysis of the differentially expressed genes, and Gene Ontology was used to look for metabolic and cell signaling pathways associated with parasite resistance. Two pathways represented by genes differentially expressed in resistant animals were those involved with the development of an acquired immune response and those related to the structure of the intestine smooth muscle. Genes involved in these processes appear from our analysis to be key genetic determinants of parasite resistance.

Actins↗

Influence of plasma A esterase on anthelmintic action of haloxon in sheep.

Controlled trials were conducted to evaluate the anthelmintic action of haloxon in 2 phenotypes of lambs, 1 having an A esterase in plasma which rapidly hydrolyzes di-(2-chloroethyl)aryl phosphates and the other without this enzyme. A total of 116 lambs, 57 with and 59 without the plasma A esterase, 6 to 9 months old, harboring naturally acquired nematode infections were used in 3 trials. Haloxon was administered orally at 20, 25, and 35 mg/kg of body weight. Nematodes against which haloxon was evaluated in the abomasum were Ostertagia circumcincta and Trichostrongylus axei and in the small intestine were T vitrinus, T colubriformis, Nematodirus spathiger, and N filicollis. The anthelmintic efficiency of haloxon did not differ in the 2 phenotypes of sheep.

Animals↗

Exploiting the effect of dietary supplementation of small ruminants on resilience and resistance against gastrointestinal nematodes.

This paper focuses on targeted nutritional supplementation as a means to reduce the requirement for chemotherapeutic control of gastrointestinal nematode infection of small ruminants and considers the limitations to practical application. Supplementary feeding, particularly with additional dietary protein, can assist resilience to infection during times when metabolic resources are being directed towards dealing with the pathophysiological effects of infection and away from production of meat, milk and fibre. Substantial experimental evidence from studies of both sheep and goats supports this hypothesis particularly in relation to young lambs and kids after weaning and in ewes around parturition. In addition, nutritional supplementation frequently increases resistance to infection, as indicated by decreased faecal worm egg counts and worm burdens. As a result, supplementation has the potential to reduce the requirement for anthelmintic treatment. Practical application of this knowledge can, however, be quite complex in many small ruminant production systems. In general, strategic supplementation should target those times when nutrient requirements are greatest and provide those nutrients which are deficient whether protein, energy, minerals or trace elements. Complexity arises when we consider that nutrient requirements will differ between localities for different species and breed of host, at different stages of growth and reproduction, with differing seasonal availability of forage, with different species of nematodes and different levels of established infections and exposure to infective stages. As a starting point, the provision of nutrients to optimize rumen function and animal performance in the particular production system should assist in maintaining resilience to nematode infection. Provision of nutrients in excess of this requirement, if economically feasible, may yield further benefits in some situations and reduce the need for alternative control measures for gastrointestinal nematode parasites.

Animal Feed↗

The efficacy of three anthelmintic drugs given in a single dose.

OBJECTIVE: To compare the efficacy of a single dose mebendazole 500 mg from the original manufacturer and a formulation locally produced by State Pharmaceuticals Manufacturing Corporation of Sri Lanka and albendazole 400 mg in mass treatment of soil-transmitted nematode infections. DESIGN: Randomised trial. SETTING: Government owned plantations in low-country regions of Sri Lanka with poor sanitary facilities. SUBJECTS: 399 children 3 to 15 years of age infected with at least one type soil-transmitted nematode. INTERVENTION: Mebendazole 500 mg or albendazole 400 mg given as single dose treatment. MEASUREMENTS: Cure rates and egg reduction rates calculated from egg counts of pre- and post-treatment stool samples using the quantitative Kato-Katz technique. RESULTS: All three drugs were effective against Ascaris infection with cure rates above 95%. The efficacy against Trichuris trichuria was uniformly poor. For hookworm infection albendazole appeared to be more effective in a single dose with cure rates of 77.9% compared with 28.7% and 35.8% for the two mebendazole formulations; corresponding egg reduction rates were 95.4% compared with 72.0% and 74.5%. CONCLUSION: Albendazole is the drug of choice for mass deworming where hookworm disease is prominent. There was no statistically significant difference between the original and locally produced mebendazole.

Adolescent↗

Cells, cytokines and other molecules associated with rejection of gastrointestinal nematode parasites.

Infections with gastrointestinal nematodes represent a major problem for human health and animal health and production. The physical size of the parasite, its changing life-cycle stages and the relative inaccessibility to host cells pose unique challenges to the immune system, which has evolved specialized strategies for parasite control. This paper reviews the work performed in the authors' laboratories to identify components that are involved in the natural rejection response against ruminant gastrointestinal nematode parasites, in particular Haemonchus contortus in sheep. The results of these studies indicate that stage-specific antibodies act in concert with effector cells, in particular globular leukocytes (intraepithelial mast cells) and eosinophils, appropriately activated/primed by type 2 (T2) cytokines, to initiate different mechanisms of parasite expulsion and killing. In addition, other molecules, in particular carbohydrate binding galectins, may be involved in strengthening the final effector phase of the rejection response.

Animals↗

Effects of continuous ivermectin treatment from birth to puberty on growth and reproduction in dairy heifers.

The effect of continuous ivermectin treatment from birth to puberty on growth and reproductive performance was studied in Holstein heifer calves grown on pastures in comparison to naturally nematode-infected, untreated animals. Ivermectin effectively abated the presence of nematode eggs in feces. Eggs per gram (EPG) in parasitized animals increased rapidly from wk 12 to 18 of age and then decreased. Animals treated with ivermectin grew faster than untreated ones, and differences in body weight became significant at 6 wk of life, even before eggs appeared in the feces of either treatment group. Ivermectin-treated heifers reached puberty 3 wk earlier than infected ones as assessed with serum progesterone concentrations (ivermectin, 30.4 +/- .8 vs untreated, 33.7 +/- 1.3 wk of age). This delay was not directly related to body weight. In addition, pelvic area at 39 wk and at 15 mo of age was increased in treated heifers (8 and 11%, respectively) compared with parasitized animals. No differences in the wither heights were observed. We conclude that ivermectin treatment in dairy heifers may increase growth rate during development, advance the onset of ovarian function, and positively affect yearling pelvic area.

Animals↗

Quantitative trait loci associated with parasitic infection in Scottish blackface sheep.

This study aimed to identify quantitative trait loci associated with endoparasitic infection in Scottish Blackface sheep. Data were collected from 789 animals over a 3-year period. All of the animals were continually exposed to a mixed nematode infection by grazing. Faecal samples were collected in August, September and October each year at ca. 16, 20 and 24 weeks of age; Nematodirus spp. eggs were counted separately from the other species of nematodes. Blood samples were collected in October from which immunoglobulin A (IgA) activity was measured and DNA was extracted for genotyping. In total, 139 Microsatellite markers were genotyped across eight chromosomal regions (chromosomes 1, 2, 3, 5, 14, 18, 20 and 21) in the sires and progeny were genotyped for the markers that were polymorphic in their sire. Evidence was found for quantitative trait loci (QTL) on chromosomes 2, 3, 14 and 20. QTL associated with specific IgA activity were identified in chromosomes 3 and 20, in regions close to IFNG (chromosome 3) and the MHC (chromosome 20). QTL associated with Nematodirus FEC were identified on chromosomes 2, 3 and 14. Lastly, QTL associated with non-Nematodirus Strongyle FEC were identified on chromosomes 3 and 20. This study has shown that some aspects of host resistance to gastrointestinal parasites are under strong genetic control, therefore these QTL could be utilised in a marker-assisted selection scheme to increase host resistance to gastrointestinal parasites.

Animals↗

Response of cattle treated with a fenbendazole slow release bolus to challenge from nematodes the following season.

Nematode infection of cattle treated in their first year at pasture with the fenbendazole slow release bolus ('Bolus group') was compared during the second year with that of untreated cattle. Ostertagia was the most prevalent parasite associated with Cooperia. Except for the Dictyocaulus spp. which caused clinical signs of bronchitis in the 'Bolus' group, the infection during the second year resulted in a moderate response of the cattle whatever the group. Rises in both pepsinogen and gastrin levels were correlated with the number of Ostertagia L3 on herbage. Damages in the abomasal mucosa were more frequent and severe in the 'Bolus' group where more inflammatory signs were observed in spite of a smaller number of worms. Nevertheless, the differences in total weight gains were not significant thanks to a compensatory effect during the second part of the grazing season in the 'Bolus' group. Hypotheses related to a minimum threshold of infection during the first year necessary to develop high enough protection during the second year are discussed. The pathological effects of gastrointestinal nematodes seem to vary more according to the inflammatory response than to the number of worms.

Animal Feed↗

First case of Lagochilascaris minor infection in Bolivia.

Few cases of human infection with Lagochilascaris minor have been reported in the literature. They occurred in Central and South America and the Caribbean. We describe the first patient with this nematode infection in Bolivia.

Adolescent↗

A new nematode species Goezia leporini n. sp. (Anisakidae) from cultured freshwater fish Leporinus macrocephalus (Anostomidae) in Brazil.

This paper describes nematode infection in the cultured freshwater fish Leporinus macrocephalus (Osteichthyses: Anostomidae) collected at Batatais, São Paulo State, Brazil. Of a total of 32 examined fish, 21 (65%) were infected with Goezia leporini n. sp. (Nematoda: Anisakidae) with mean infection intensity of 4.1 parasites. The nematodes presented total length greater than G intermedia, G. holmesi, G. pelagia, G. minuta, G. kliksi, G. sinamora, G. nonipapillata, G. alii, G. moraveci, G. brasiliensis, and G. brevicaeca. The main difference was a great number of preanal papillae in males when compared to G. brasiliensis and G. brevicaeca. The present description also differs from that of G. brasiliensis with respect to spicule length and distance of vulva from the anterior extremity.

Animals↗

Villus epithelial injury induced by infection with the nematode Nippostrongylus brasiliensis is associated with upregulation of granzyme B.

Intestinal parasite infections induce thymus-dependent villus atrophy, but the effector mechanisms directly responsible for the development of villus atrophy are not thoroughly understood. In this study, we analyzed the expression of cytotoxic factors or ligands in athymic nude rnu/rnu rats and their littermate euthymic rnu/+ rats infected with the nematode Nippostrongylus brasiliensis. Morphometric analyses showed that partial villus atrophy developed 10 days after infection in euthymic but not in athymic rats, whereas crypt hyperplasia occurred in both types of animal. Reverse transcription-polymerase chain reaction analyses of the isolated jejunal epithelial fraction showed that the development of villus atrophy in euthymic rats was positively correlated with an increase of granzyme B transcript levels but not with Fas ligand or tumor necrosis factor-alpha expression. In addition, the number of granzyme B-immunoreactive cells was increased significantly in euthymic rat villus epithelium and the propria mucosa after infection. The CD8+ cell number did not change significantly. Collectively, these findings showed that significant increases in the number of cells that express the cytotoxic factor granzyme B occur in the nematode-infected small intestine of immunocompetent hosts. The type of cells that express granzyme B and their role in the progression of enteropathy remain to be elucidated.

Animals↗

Serum IgG reactivity with 41-, 31-, and 28-kDa larval proteins of Strongyloides stercoralis in individuals with strongyloidiasis.

Proteins from a deoxycholate-soluble extract of Strongyloides stercoralis infective larvae were separated by SDS-PAGE, blotted onto nitrocellulose paper, and reacted with sera from individuals with confirmed S. stercoralis infections (n = 100), suspected S. stercoralis infections in whom no larvae could be detected (n = 27), and other nematode infections (40 with Wuchereria bancrofti, 20 with Onchocerca volvulus, 20 with Necator americanus, and 20 with mixed Ascaris lumbricoides and Trichuris trichiura infections). Immunodominant proteins of approximately 41, 31, and 28 kDa were recognized by IgG in 91%, 88%, and 90%, respectively, of sera from those with confirmed strongyloidiasis; in 100%, 100%, and 93% of sera from those with suspected strongyloidiasis; and in 9%, 12%, and 14% of sera from those infected with other nematodes. IgG reactivity to each of these proteins was a more specific means of immunodiagnosis than the currently used indirect ELISA; the methods were equally sensitive.

Animals↗