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Trypanosome-induced modulation of responses to concurrent helminth infection.

Infections with African trypanosomes are known to suppress immune responses to vaccines and to gastrointestinal nematode infections in livestock. Experimental infections with Trypanosoma brucei (Tb) and the gastrointestinal nematode Nippostrongylus brasiliensis (Nb) in mice were used to identify possible mechanisms involved in interference with anti-worm responses and to examine the effects of host genotype on the extent of suppression seen. Concurrent infections with T. brucei resulted in a prolongation of worm survival and a dramatic increase in faecal egg output. Infection also resulted in a marked suppression of the proliferative response of mesenteric lymphocytes (MLNC) to in vitro mitogenic stimulation. When MLNC from concurrently infected mice were stimulated in vitro with the mitogen ConA they released more IFN-gamma and less IL-5 than cells from mice infected only with N. brasiliensis. These data are interpreted in terms of a trypanosome-mediated influence on the development of host-protective type-2 T helper cell responses against N. brasiliensis. The degree to which T. brucei altered the kinetics of the nematode infection was influenced by the particular mouse strain concerned.

Animals↗

Alternatives to anthelmintics for the control of nematodes in livestock.

Efficient and welfare-friendly livestock production demands the control of nematode infection. Current control measures rely upon anthelmintic treatment but are threatened by the widespread evolution of drug-resistance in parasite populations. Several methods have been advocated to control nematodes without relying on effective anthelmintics. These include grazing management, biological control, nutritional supplementation, vaccination, and genetic approaches. Each method has its advantages and disadvantages. There are several grazing management schemes that can reduce the severity of infection but they are insufficient on their own to control infection. Biological control includes the use of predatory fungi to control nematode populations and the use of pasture species that can reduce the intensity of infection. Fungi can control nematodes but the current requirement for daily feeding means that this approach will be most useful for animals that are handled daily. Feeding supplementary protein can control nematode infection. The method is simple but can be expensive and may not be cost-effective for some marginal enterprises. Genetic approaches include the use of resistant breeds and selective breeding. Some breeds will thrive in conditions that kill animals from other breeds but substitution of resistant breeds is not always feasible. Selective breeding is effective and inexpensive but requires a high level of expertise. The most appropriate method or set of methods to minimize the adverse consequences of nematode infection may vary among farms.

Animal Husbandry↗

Genetic resistance to parasitic infection.

The ability of animals to resist infections with parasites is genetically determined and therefore variable between individuals or breeds of a given host species. Such variation may involve innate (non-immunological) and acquired (immunologically mediated) resistance mechanisms, and is determined by both major histocompatibility complex (MHC)-linked and non-MHC genes. Resistance is inherited as a dominant trait, with heritability often exceeding 0.3. Genetic variation can be exploited to improve the capacity of domestic animals to resist parasitic infection. Methods to achieve this exploitation are discussed in relation to gastrointestinal nematode infections in sheep.

Animals↗

In vivo rearing of Thripinema nicklewoodi (Tylenchida: Allantonematidae) and prospects as a biological control agent of Frankliniella occidentalis (Thysanoptera: Thripidae).

Methods are described for the in vivo production of the nematode Thripinema nicklewoodi (Siddiqi), an obligate parasite and potential biological control agent of western flower thrips Frankliniella occidentalis (Pergande). Nematode infection is not lethal but causes sterilization of adult female hosts. Both fertilization and horizontal transmission of T. nicklewoodi is achieved in 1.5-ml microcentrifuge tubes (infection arenas), in the presence of 100% humidity, a temporary food source and preferably a damp substrate. Following exposure to infection arenas, F. occidentalis are reared on excised bean leaves Phaseolus vulgaris (L.) in polypropylene containers for 2 wk at 25 degrees C to allow the reproduction and development of a single generation of nematodes within infected hosts's abdominal cavity. To identify infected hosts after this incubation period, thrips are isolated in microcentrifuge tubes and monitored for free-living nematodes being released along with frass. Infected thrips are reintroduced back into infection arenas to inoculate further thrips to maintain the culture. We documented the output of the rearing procedure using a standard method and following simple manipulation of several individual parameters of the infection technique. The standard method was the most efficient, and resulted in an increased (output/input) ratio of infected thrips of approximately 2; i.e., the number of infected thrips approximately doubles each generation. Monitoring infected thrips revealed that nematodes were first released between 12-14 d postinfection and for an average of 7.9 d at 25 degrees C; highlighting the potential to reuse infective thrips between infection arenas. The possibility of using T. nicklewoodi as an inoculative agent against F. occidentalis infesting floricultural crops is discussed.

Animals↗

An Arabidopsis thaliana pectin acetylesterase gene is upregulated in nematode feeding sites induced by root-knot and cyst nematodes.

By using differential display, gene expression was investigated in Arabidopsis thaliana roots shortly after nematode infection, and a putative pectin acetylesterase (PAE) homolog (DiDi 9C-12) was found to be upregulated. PAEs catalyze the deacetylation of pectin, a major compound of primary cell walls. mRNA in situ hybridization experiments showed that the expression of DiDi 9C-12 was enhanced very early after infection in initiating giant-cells and in cells surrounding the nematodes. Later on, the level of DiDi 9C-12 mRNA was lower in giant-cells and transcripts were mainly found in parenchyma, endodermis, and pericycle cells of the root gall. Twenty days after infection, DiDi 9C-12 transcripts could no longer be detected. DiDi 9C-12 transcripts were also found in young syncytia and in the cells surrounding the expanding syncytium. Our results suggest that plant parasitic nematodes can modulate the rapid growth of the feeding cells and the expansion of the root gall by triggering the expression of DiDi 9C-12. PAEs, which probably act together with a range of other pectin-degrading enzymes, could be involved in softening and loosening the primary cell wall in nematode-infected plant roots.

Amino Acid Sequence↗

[Natural development of gastrointestinal helminthiasis in calves born during the dry season in Guaira, State of São Paulo, Brazil].

Proceeding with their observations on bovine nematode epidemiology in Guaíra, State of São Paulo, the authors carried a study on the natural development of nematode infections in crossbred calves born at the dry season. The fecal nematode egg counts per gram (e.p.g.) performed every fortnight were positive at the start, in October, when calves were two months old, and reached the peak when they were five months old; cultures of faeces revealed larvae of Cooperia, Haemoncus, Oesophagostomum and Trichostrongylus. From this time on, until 12.5 months old, when the calves were slaughtered, the variable e.p.g. decreased continually, the average being, in the last examinations, 40 e.p.g. Strongyloides eggs were present in the faeces when the calves were two months old but they disappeared by the 5th month of age. The post-mortem examination revealed the following species of nematodes: Haemoncus similis, Haemoncus contortus, Trichostrongylus axei, Bunostomum phlebotomum, Cooperia punctata, Cooperia pectinata, Oesophagostomum radiatum, Trichuris discolor and Dictyocaulus viviparus.

Animals↗

Heligmosomoides polygyrus: CD4+ but not CD8+ T cells regulate the IgE response and protective immunity in mice.

Oral inoculation of BALB/c mice with infective larvae of Heligmosomoides polygyrus resulted in chronic infection characterized by the release of parasite eggs in the feces for several months. The actual number of eggs per gram of feces was dependent on the dose of the inoculum. Serum IgE in infected mice peaked at a level of greater than 70 micrograms/ml during Weeks 3 through 6 following inoculation, and high levels of IgE (greater than 40 micrograms/ml) persisted for over 14 weeks. Protective immune responses resulted in reduced egg production and the development of markedly fewer adult worms in the small intestines following a challenge inoculation. The role of CD4+ and CD8+ T cells in these responses was examined by depletion in vivo of either T cell subpopulation with rat mAb specific for the appropriate determinants. Mice treated with anti-CD4 during a primary infection had increased EPG which was due primarily to an increase in worm fecundity (eggs produced per adult female). A challenge inoculation of mice that had been cleared of the primary infection with an anthelmintic drug induced a protective response that reduced development of new adult worms by 70-80% and their fecundity by greater than 90%. This protective response was abrogated by injection of mice with anti-CD4. Serum IgE diminished when adult worms were removed after anthelmintic treatment. A more precipitous drop in serum IgE followed successive treatments of mice with an anthelmintic and anti-CD4. In addition, the anamnestic serum IgE response to a challenge inoculation was reduced by over 80% in anti-CD4-treated mice. Anti-CD8 treatment had no appreciable effect on the immunological or parasitological parameters measured following a challenge inoculation with H. polygyrus. Thus, CD4+ T cells regulate host protective immunity, worm fecundity, and IgE levels in an H. polygyrus infection. This experimental system may be particularly suitable for analysis of chronic nematode infections of humans and livestock because of the responsiveness of the parasite in vivo to changes in host immune function.

Animals↗

Parasites and associated pathology observed in pinnipeds stranded along the Oregon coast.

Forty-two seals and sea lions found dead along the Oregon Coast were examined for parasites and associated pathology. Nematode infections of the lung and/or gastrointestinal tract were the primary cause of death in 5 of 42 animals examined. New distribution records were established for Pricetrema zalophi and Zalophotrema hepaticum. New host records include Z. hepaticum and Diphyllobothrium cordatum in the Steller's sea lion (Eumetopias jubatus); Nanophyetus salmincola in the California sea lion (Zalophus californianus); P. zalophi in the harbor seal (Phoca vitulina); and P. zalophi, Trigonocotyle sp. and Otostrongylus circumlitus in the northern elephant seal (Mirounga angustirostris).

Animals↗

The distribution of pepsinogen within the abomasa of cattle and sheep infected with Ostertagia spp. and sheep infected with Haemonchus contortus.

The effect of nematode infections on the production of pepsinogen by ruminants was investigated immunohistochemically and biochemically. Abomasal tissues were collected from parasite-naive cattle and sheep, from sheep infected with predominantly Ostertagia circumcincta, sheep infected experimentally with Haemonchus contortus and cattle infected with Ostertagia ostertagi. Pepsinogen was also assayed biochemically in homogenates of fundic mucosae from sheep infected with predominantly O. circumcincta. Infection with Ostertagia spp. parasites was associated mainly with nodular hyperplasia, resulting in increased numbers of cells that produce both pepsinogen and mucus. Measured biochemically, nodules contained more pepsinogen than adjacent more normal mucosa (p < 0.05), and this effect was largely attributable to the greater mass of nodules. Infection of sheep with H. contortus was associated with generalised hyperplasia, characterised by increased numbers of mucopeptic cells and in at least one animal with reductions in parietal cell numbers. At the same time, the zymogen granule content of chief cells was reduced. Similar changes were occasionally seen in sheep infected predominantly with O. circumcincta. Generalised hyperplasia is likely to be indicative of the presence of ambulatory parasitic stages as opposed to those confined to nodules. The potential for the enhanced production of pepsinogen by increased numbers of cells with a joint mucous cell and zymogenic cell phenotype may offset decreases in the numbers of chief cells or reductions in chief cell activity.

Abomasum↗

Efficacy of purified schistosoma japonicum egg antigens for ELISA serodiagnosis of human Schistosomiasis japonica: specificity and sensitivity.

At present, there is no consensus that purified schistosome egg antigens offer any advantage in the diagnosis of schistosomiasis by enzyme linked immunosorbent assay (ELISA). Previously, we demonstrated by multiple techniques that the major serologic antigens in Schistosoma japonicum soluble egg antigen (SEA) are glycoproteins, and that the glycoproteins with highest specificity and sensitivity are hydrophobes. We therefore tested these materials for their specificity, sensitivity and cost effectiveness in the ELISA. In this study we used five SEA fractions that varied in their purity and antigenicity. The order of immunologic specific activity in the ELISA, measured by titration of a standard sera pool, was: hydrophobic glycoproteins (highest), crude SEA glycoproteins, hydrophilic glycoproteins, crude SEA, and SEA proteins (lowest). Complexity (purity) of these materials were (in rank order), hydrophilic glycoproteins (purest), hydrophobic glycoproteins, crude glycoproteins, SEA proteins, and crude SEA (most complex). Epidemiologic sensitivity in the ELISA was tested on limited but well characterized populations. At high antigen coating concentration (0.5 microgram/well), the only antigen fraction with poor sensitivity was SEA proteins. There was little difference in epidemiologic sensitivity between the purer fractions with highest immunologic sensitivity (hydrophobic glycoproteins and crude SEA glycoproteins) and the crude SEA which possesses intermediate immunologic sensitivity. Differences in epidemiologic sensitivity were most pronounced when wells were coated at an antigen concentration (0.1 microgram/well) where crude SEA began to fail. Specificity for all preparations, assessed by reactivity with sera from patients with other trematode infections and with cestode and nematode infections, was excellent. The clinical sensitivity of the ELISA employing crude S. japonicum SEA is so high, and the specificity so good, that the increased immunologic sensitivity of partially purified antigens had little effect on epidemiologic sensitivity. This is not true for the S. mansoni ELISA where crude antigens had inferior sensitivity and specificity.

Antibodies↗

Impaired balance of interleukin-4 and interferon-gamma production in infections with Schistosoma mansoni and intestinal nematodes.

In chronic infection with Schistosoma mansoni, IL-4 and IFN-gamma are co-modulated in opposite directions. This was shown when testing sera and cell culture supernatants from 31 Brazilian patients with schistosomiasis before, and three months after treatment with praziquantel. Thorough examinations were undertaken to account for polyparasitism with intestinal nematode infections involving tissue migrating larval stages that may induce analogous changes. Controls free of S. mansoni included a group (n = 17) matching the schistosomiasis patients' parasitation by intestinal nematodes and a group (n = 16) free of helminths other than T. trichiura. Serum IL-4 was greater than 20 pg/ml in 81% of schistosomiasis patients but in only 35 and 25%, respectively, of controls with and without intestinal nematodes. IL-4 data correlated inversely with the mitogen-induced IFN-gamma synthesis. Generation of IL-4 in response to phorbol esters was related to the intensity of infection by schistosomes and intestinal nematodes. The parasitological status three months after therapy with either praziquantel or mebendazole revealed a dichotomy: whereas the ratio of IFN-gamma to IL-4 generated in vitro was identical in uninfected controls and in patients who cleared the parasites, failure to eliminate the parasites was associated with lower IFN-gamma/IL-4 ratios generated in vitro.

Female↗

Impact of vitamin E or selenium deficiency on nematode-induced alterations in murine intestinal function.

The effects of deficiencies in the antioxidant nutrients, vitamin E and selenium, on the host response to gastrointestinal nematode infection are unknown. The aim of the study was to determine the effect of antioxidant deficiencies on nematode-induced alterations in intestinal function in mice. BALB/c mice were fed control diets or diets deficient in selenium or vitamin E and the response to a secondary challenge inoculation with Heligmosomoides polygyrus was determined. Egg and worm counts were assessed to determine host resistance. Sections of jejunum were mounted in Ussing chambers to measure changes in permeability, absorption, and secretion, or suspended in organ baths to determine smooth muscle contraction. Both selenium and vitamin E deficient diets reduced resistance to helminth infection. Vitamin E, but not selenium, deficiency prevented nematode-induced decreases in glucose absorption and hyper-contractility of smooth muscle. Thus, vitamin E status is an important factor in the physiological response to intestinal nematode infection and may contribute to antioxidant-dependent protective mechanisms in the small intestine.

Animals↗

The effect of two levels of dietary protein on resistance and resilience of dairy goats experimentally infected with Trichostrongylus colubriformis: comparison between high and low producers.

Numerous studies have examined the interactions between protein nutrition and the response to nematode parasitism in sheep, but very few in goats. Compared with other ruminants, goats are less resistant to nematode infection. In addition, in dairy goats, high producing animals have been shown to be less resistant and less resilient to infection compared to low producing ones. The objective of the present study was to examine the consequences of protein supplementation on both resistance and resilience of dairy goats to nematode trickle infection, taking into account the initial level of milk production of the animals. During a 14-week period, 40 milking goats received a high protein (HP) diet supplying 130% of the protein requirements, and 38 goats were fed a intermediate protein (IP) diet (120% of the protein requirements). In addition, half of each group was given a weekly trickle infection with Trichostrongylus colubriformis larvae, the other part of the flock remained non-infected. Faecal egg counts (FEC), eosinophil counts and pathophysiological data (urea, albumin and inorganic phosphate concentrations in the serum) were measured twice a month. Milk production data (milk yield, protein and fat contents) were also recorded every 15 days. The results showed that FECs were lower (p < 0.05) and eosinophil counts higher (p < 0.05) in the animals receiving the HP diet suggesting that resistance was enhanced by protein supplementation. Meanwhile, milk parameters (related to resilience) were not affected by the level of protein in the diet when considering the whole groups. In contrast, in the high producing goats, the milk production and milk composition parameters were improved with the HP diet. To conclude, we have seen that the expression of both resistance and resilience did not appear when the coverage of the protein requirements was insufficient. Because the milk production is dependent on the protein supply, we suggest that there is a competition in the use of the protein between the development of resistance and the milk production.

Animal Nutritional Physiological Phenomena↗

Evaluation of Duddingtonia flagrans in reducing infective larvae of Haemonchus contortus in feces of sheep.

Consequences of nematode infections due to Haemonchus contortus are a serious constraint for the sheep industry worldwide. Development of anthelmintic resistance and increasing concern about the impact of anthelmintic use dictate the need of alternative control. Such an alternative is using the nematode trapping fungus Duddingtonia flagrans to reduce infective larvae levels on pasture. Two trials were conducted to determine the effect of D. flagrans in reducing infective larvae (predominantly H. contortus) in feces. The first trial determined the dose effect of D. flagrans in reducing infective larvae in feces. Eighteen ewes were dewormed to remove existing infections and randomly assigned to six treatment groups: 5 x 10(4), 1 x 10(5), 2.5 x 10(5), 5 x 10(5), 1 x 10(6) or no (control) spores of D. flagrans per kg of body weight mixed in their feed for 7 days. Fecal samples were collected daily from these and from infected donor ewes. Feces from individual-treated ewes were mixed with equal amounts of donor ewe feces, theoretically approximating oral dose spore concentrations of 2.5 x 10(4), 5 x 10(4), 1.25 x 10(5), 2.5 x 10(5), 5 x 10(5) and no spores, and were cultured. Across dosages and during the 7 days of fungus feeding, percent reduction of infective larvae ranged from 76.6 to 100.0%. The second trial determined the effect of D. flagrans at the dose of 10(5) spores per kg body weight on reducing infective larvae in feces from naturally infected lambs. Twenty lambs were randomly assigned to either treatment or control groups based on fecal egg count. Treatment lambs were fed spores mixed in feed for 7 days. Feces were collected daily and cultured. During the 7 days of fungus feeding, the percent reduction of infective larvae ranged from 82.8 to 99.7%. Results of these trials demonstrated that the nematode trapping fungus D. flagrans was highly effective in reducing infective larvae in sheep feces and should be considered as a biological control agent for integrated nematode control programs.

Administration, Oral↗

Treatment of a granuloma caused by Halicephalobus gingivalis in a horse.

A 12-year-old Arabian gelding with a granuloma caused by the nematode Halicephalobus gingivalis caudal and dorsal to the left orbit was examined. There was no evidence of dissemination of the nematode to the kidneys or the CNS, and the horse was treated with ivermectin (1.2 mg/kg [0.55 mg/lb] of body weight, p.o., every 2 weeks for 3 treatments). The granuloma was surgically debulked 2 days after the first dose of ivermectin. The granuloma resolved with no evidence of nematode infection after 18 months. Halicephalobus gingivalis is a ubiquitous saprophytic nematode that has been reported to infect humans and horses. The nematode may form granulomas in the integument or may disseminate to various organs with a tropism for the CNS and kidneys. Once clinical signs of CNS involvement develop, the disease is rapidly fatal.

Animals↗

Relationship between bronchopulmonary nematode larvae and relative abundances of Spanish ibex (Capra pyrenaica hispanica) from Castilla-La Mancha, Spain.

The excretion of bronchopulmonary nematode infective larvae was evaluated in 160 faecal samples of Spanish ibex (Capra pyrenaica hispanica) collected from 13 populations in Castilla-La Mancha, south-central Spain in September 2003. Intensities and prevalences were compared with pasture availability, abundances of wild and domestic ungulates at both levels, i.e. for populations and for faeces in a two-step procedure. Protostrongylid larvae showed similar infection rates (mean intensity: 1.56+/-0.12, n=94; mean prevalence: 25.62+/-6.86%, n=160) to Dictyocaulus spp. (mean intensity: 1.03+/-0.11, n=48; mean prevalence: 30.00+/-7.11%, n=160). At the population level, positive correlations were found between the prevalences of both bronchopulmonary taxa. The prevalence in both groups, but not intensity, also correlated positively with Spanish ibex abundance indexes both for the populations and individual faeces. These findings suggest that: (i) parasite spreading across Spanish ibex populations in Castilla-La Mancha could respond to host density-dependent processes; and (ii) these populations may have similar exposition and/or susceptibility to both bronchopulmonary taxa resulting in similar host-parasite patterns, despite their different life cycles. Bronchopulmonary outputs in the Spanish ibex from Castilla-La Mancha seems not to represent a health risk for this endemic wild ungulate but may be useful in any health surveillance scheme for the increasing populations of Spanish ibex.

Animals↗

Current research and future prospects in the development of vaccines against gastrointestinal nematodes in cattle.

Vaccination is being considered as the most feasible alternative for anthelmintic drugs to control gastrointestinal nematode infections in cattle. However, despite the identification of several candidate protective antigens, no vaccines against gastrointestinal nematode parasites are currently available. The main problems that hamper the development of nematode vaccines in ruminants are that vaccination with recombinant nematode proteins produced in bacterial or eukaryotic expression systems did not induce a protective immune response and no suitable antigen delivery system is available for presentation of protective worm antigens to the bovine mucosal immune system. The present review will focus on recent advances and remaining obstacles in vaccine development against gastrointestinal nematodes in cattle, in particular against the abomasal parasite Ostertagia ostertagi.

Animals↗