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Genetic and other constraints on resistance to infection with gastrointestinal nematodes.

Infections with gastrointestinal (GI) nematodes are among the most prevalent infections of man. Not only are they common and widespread in populations throughout the world, but they are frequently chronic and occur repeatedly throughout the lifetime of an individual. It is now well established that such parasites can be highly immunogenic and that their environment, the gastrointestinal tract, is well equipped to mount potent immune and inflammatory responses. The abundance and long-term survival of GI nematodes therefore present a paradox. This review takes the standpoint that man as a species has the capacity to produce effective and protective responses against GI infections, but that this capacity is subject to a number of powerful constraints, arising from parasite evasion, depression of response capacity and deficiency in response capacity. These constraints are discussed in the light of evidence drawn from experimental studies with animal models.

Age Factors↗

In vivo effects of Fagara leaves on sheep infected with gastrointestinal nematodes.

Infections of the gastrointestinal tract with parasitic nematodes, and particularly Haemonchus contortus, represent one of the main pathological constraints on the breeding of small ruminants in tropical countries. Owing to the constant development of anthelmintic resistance in worm populations, alternative control methods are being evaluated, including an assessment of the value of the traditional plants. The effect of the consumption of Fagara leaves on sheep infected with gastrointestinal nematodes was therefore measured both under controlled conditions and in a field survey. In sheep experimentally infected with 2500 third-stage larvae of H. contortus, a three-day administration of Fagara was associated with a decrease in egg excretion and a significant reduction in the fertility of female worms, without changes in the worm number. A reduction in egg output was also confirmed from the results obtained of a survey or naturally infected sheep performed in smallholder farms. This survey also indicated that the repeated consumption of small amounts of Fagara leaves was more effective than a single three-day administration. Results of both studies suggest that Fagara leaves might be an alternative option to chemotherapy to modulate the epidemiology of H. contortus in tropical countries.

Animal Feed↗

Osteopenia and reduced serum alkaline phosphatase activity in grazing lambs naturally infected with gastrointestinal nematodes.

The effect of gastrointestinal nematode infections on bone development was investigated in growing sheep on pasture. Forty-five weaned lambs from six groups in a two-factorial design incorporating stocking rate (SR; low, medium and high) and presence or absence of infection on pasture were sampled in the late grazing season. Worm counts were performed at slaughter, and the left metacarpal bones were excised for bone assessment. Faecal egg counts and worm burdens, primarily of Ostertagia circumcincta and Trichostrongylus vitrinus, were considerably higher in the high SR infected group ("I-High") than in comparable animals at low or medium SRs, whereas uninfected groups showed negligible egg excretion. Clinical biochemistry revealed significantly reduced serum concentrations of albumin, calcium and alkaline phosphatase in infected lambs. Nematode infections were associated with significant reductions in bone mineral density (30% at high SR), measured by dual energy X-ray absorptiometry, and in bone size (9%). Histomorphometry indicated thinning of the trabecular structure and reduced bone formation in the infected groups, particularly the I-High group. Bone mineral density, bone tissue volume and structural changes were strongly associated with log-transformed worm counts. The study showed that lambs suffering from moderate to heavy degrees of naturally acquired gastrointestinal nematode infection developed marked osteopenia after weaning, i.eduring the later part of the grazing season.

Abomasum↗

Dynamics of carbon dioxide release from insects infected with entomopathogenic nematodes.

The quality of an insect as a host to an entomopathogenic nematode infective juvenile depends in part on whether or not the insect is already infected and on the stage of that infection. Previous research has shown that nematode response to hosts can change after infection and that, for uninfected hosts, CO(2) can be an important cue used by infective stage juveniles during attraction. We hypothesized that CO(2) production from an insect changes after it is infected, and that these changes could influence nematode infection decisions. Changes in CO(2) released by two insect species (Galleria mellonella and Tenebrio molitor) after infection by one of four nematode species (Steinernema carpocapsae, Steinernema feltiae, Steinernema glaseri, or Steinernema riobrave) were measured. Measurements were taken every 2h from time of initial exposure to nematodes up to 224 h after infection. Dead (freeze-killed) and live uninfected insects were used as controls. Infected G. mellonella showed two distinct peaks of CO(2) production: one between 20 and 30 h and the other between 70 and 115 h after exposure to the nematodes. Peaks were up to two times higher than levels produced by uninfected insects. Infected T. molitor showed only one peak between 25 and 50h. We found differences in peak height and timing among nematode and insect species combinations. The influence of these changes in CO(2) production on IJ attraction and infection behavior remains to be determined.

Animals↗

Nematode parasitism in adult dairy cows in Belgium.

Over a period of 1 year, from November 1997 to October 1998, the abomasa, blood and faecal samples of 121 dairy cows in Belgium were collected and examined for nematode infections. Nematodes were present in the abomasa of 110 animals. Ostertagia was found in all 110, Trichostrongylus was seen in 65 and Haemonchus in 14 abomasa. Overall, 91% of all trichostrongyles recovered were Ostertagia. The geometric mean total number of Ostertagia was 2750, with an average of 74% inhibited early fourth stage larvae (EL4). Between November and February >90% of the Ostertagia worm burden were EL4 stages. The majority of the animals (56%) harboured a low Ostertagia burden (100-5000) and 15% had a high burden (>10,000). Sixty-four percent of the coprocultures were positive and the genera recovered were Ostertagia sp. (100%), Trichostrongylus sp. (42%), Oesophagostomum (32%), Haemonchus sp. (29%) and Cooperia sp. (16%). A seasonal pattern was evident for serum Ostertagia-specific antibodies and for serum pepsinogen concentration, with the highest levels during the summer, and low values during the winter. Dictyocaulus viviparus specific antibodies were detected in the serum of eight (7%) animals.

Animals↗

Effect of feeding urea-molasses blocks with incorporated fenbendazole on grazing dairy heifers naturally infected with gastrointestinal nematodes.

Between June 1999 and August 2000, the effects of feeding medicated urea-molasses supplement blocks on the growth of dairy heifers in a marginal area of central Kenya were assessed by comparing the live-weight gain of supplemented and unsupplemented heifers grazing the same pasture. Thirty-nine heifers with an average age of 9.6 months were initially treated orally with albendazole (10 mg/kg body weight) and assigned to 3 groups: group I was fed urea-molasses blocks with incorporated fenbendazole (MUMB), group II was fed urea-molasses blocks (UMB) and group III heifers (control) received no block supplementation (NBS). Body weights of the heifers and faecal egg counts (FECs) were measured monthly and larval cultures were made of positive faecal samples of each group. The mean cumulative live-weight responses of the MUMB and UMB groups were significantly greater than the NBS group (P < 0.05). However, at the end of the experimental period, the mean weight gain of the MUMB group did not differ from that of the UMB group (P > 0.05). The FECs were moderate to low in all groups and decreased progressively with increasing age of the animals; FECs for the urea-molasses-supplemented groups remained significantly lower than those of the NBS group throughout the experimental period (P < 0.05). Haemonchus and Trichostrongylus were the predominant nematode genera found in the heifers, but Cooperia, Bunostomum and Oesophagostomum were also present. These results indicate that feeding of urea-molasses blocks substantially reduced production losses attributable to nematode infection of young grazing cattle, and confirms previous observations that well-fed animals are better able to overcome the effects of helminth infections.

Administration, Oral↗

Increased expression of interleukin-5 (IL-5), IL-13, and tumor necrosis factor alpha genes in intestinal lymph cells of sheep selected for enhanced resistance to nematodes during infection with Trichostrongylus colubriformis.

Cytokine gene expression in cells migrating in afferent and efferent intestinal lymph was monitored for extended time periods in individual sheep experimentally infected with the nematode Trichostrongylus colubriformis. Animals from stable selection lines with increased levels of either genetic resistance (R) or susceptibility (S) to nematode infection were used. Genes for interleukin-5 (IL-5), IL-13, and tumor necrosis factor alpha (TNF-alpha), but not for IL-4, IL-10, or gamma interferon (IFN-gamma), were consistently expressed at higher levels in both afferent and efferent lymph cells of R sheep than in S sheep. However, only minor differences were observed in the surface phenotypes and antigenic and mitogenic responsiveness of cells in intestinal lymph between animals from the two selection lines. The IL-4 and IL-10 genes were expressed at higher levels in afferent lymph cells than in efferent lymph cells throughout the course of the nematode infection in animals of both genotypes, while the proinflammatory TNF-alpha gene was relatively highly expressed in both lymph types. These relationships notwithstanding, expression of the IL-10 and TNF-alpha genes declined significantly in afferent lymph cells but not in efferent lymph cells during infection. Collectively, the results showed that R-line sheep developed a strong polarization toward a Th2-type cytokine profile in immune cells migrating in lymph from sites where the immune response to nematodes was initiated, although the IFN-gamma gene was also expressed at moderate levels. Genes or alleles that predispose an animal to develop this type of response appear to have segregated with the R selection line and may contribute to the increased resistance of these animals.

Animals↗

IL-5 contributes to worm expulsion and muscle hypercontractility in a primary T. spiralis infection.

Enteric nematode infections lead to increased interleukin (IL)-5 expression, eosinophilic inflammation, and intestinal smooth muscle hypercontractility. Although eosinophils release inflammatory mediators that cause smooth muscle contraction, the role of IL-5 and eosinophils in enteric smooth muscle hypercontractility is unclear. IL-5-deficient mice and their wild-type controls were infected with the nematode Trichinella spiralis. Intestinal parasites and eosinophils were counted, and jejunal longitudinal muscle contractility was assessed. During infection, IL-5 gene expression increased significantly in wild-type mice and was accompanied by significant intestinal eosinophilia in wild-type but not IL-5-deficient mice. Although both strains developed increased muscle contractility during infection, contraction was significantly less in the IL-5-deficient mice at days 16 and 21 postinfection. In addition, parasite expulsion was transiently delayed at day 16 in IL-5-deficient mice. Thus, in the nematode-infected mouse, IL-5 appears essential for intestinal eosinophilia and contributes to, but is not essential for, the development of muscle hypercontractility. IL-5 also appears to play a minor role in expelling a primary T. spiralis infection from the gut.

Animals↗

Efficacy of abamectin against natural infections of gastrointestinal nematodes and lungworm of cattle with special emphasis on inhibited, early fourth stage larvae of Ostertagia ostertagi.

The anthelmintic efficacy of abamectin (avermectin B1) was evaluated against gastrointestinal nematodes, including Ostertagia ostertagi inhibited larvae and lungworm, in yearling crossbred beef heifers during late spring. The calves were grazed on contaminated pasture for 10 weeks and then held under conditions free of nematode infection for 3 weeks prior to allotment and treatment on 5 June. Thirteen calves were randomly assigned to two groups of six by restricted randomization on body weights; the extra lightest calf was assigned to the non-treated control group. Group 1 calves were treated with abamectin at 200 micrograms kg-1 body weight by s.c. injection and Group 2 calves were not treated; all were killed at 14 days after treatment. Ostertagia ostertagi was present in all controls; arithmetic mean numbers of adults, developing fourth stage larvae (L4) and inhibited EL4 were 7683, 605 and 36,102, respectively. Other nematode genera present in controls in sufficient numbers for the experiment were Haemonchus placei adults, Trichostrongylus axei adults, Cooperia spp. adults, Oesophagostomum radiatum adults, Bunostomum phlebotomum adults, Dictyocaulus viviparus adults and E5 (immature adults). Abamectin was highly effective (consistently greater than 99% efficacy and P less than 0.05) in removing all nematodes present in treated calves as represented in non-treated controls, including the primary target of Ostertagia ostertagi inhibited EL4. The lowest efficacy was 93.8%, against D. viviparus E5.

Animals↗

Nutrition and parasite interaction.

This overview focuses on the interaction between nutritional status and gastrointestinal nematode infection in ruminants and considers: (i) the influence of the parasite on host metabolism; and (ii) the effect of host nutrition on the establishment and survival of parasite populations, the development of the host-immune response and the pathophysiology of infection. Gastrointestinal nematodes reduce voluntary feed intake and efficiency of feed utilisation, a key feature being an increased endogenous loss of protein into the gastrointestinal tract. Overall there is movement of protein from productive processes into repair of the gastrointestinal tract, synthesis of plasma proteins and mucoprotein production. Although reduction in feed intake is a major factor contributing to the reduced performance of parasitised ruminants, the underlying mechanisms of the anorexia are poorly understood. Supplementation of the diet with additional protein does not appear to affect initial establishment of nematode infections but the pathophysiological consequences are generally more severe on lower planes of protein nutrition. The main effect of protein supplementation is to increase the rate of acquisition of immunity and increase resistance to reinfection and this has been associated with an enhanced cellular immune response in the gastrointestinal mucosa. The unresponsiveness of the young lamb can be improved by dietary protein supplementation. Recent trials have shown that growing sheep offered a free choice between a low and a high protein ration are able to modify their diet selection in order to alleviate the increase in protein requirements which result from gastrointestinal nematode infection. Studies on the influence of nutrition on the expression of genotype have shown that the benefits of a superior genotype are not lost on a low protein diet whereas a high protein diet can partially emeliorate the disadvantages of an inferior genotype. In addition to dietary protein both macro-minerals and trace elements can influence the host-parasite relationship.

Animal Feed↗

Altered expression of goblet cell- and mucin glycosylation-related genes in the intestinal epithelium during infection with the nematode Nippostrongylus brasiliensis in rat.

Intestinal nematode infection induces marked goblet cell hyperplasia and mucus secretion, but the mechanisms of regulation of the changes still remain to be elucidated. In the present study, epithelial cells were isolated from the rat small intestine at various times after Nippostrongylus brasiliensis infection, and the levels of expression of goblet cell- and mucin glycosylation-related genes were estimated by semi-quantitative reverse transcription (RT)-PCR. Among the genes investigated, mucin core peptide (MUC) 2, sialyltransferase (Siat) 4c and trefoil factor family (TFF) 3 were upregulated as early as 2-4 days post-infection, suggesting that they are associated with an early innate protective response. Seven days post-infection and thereafter, when the nematodes reached maturity, significant upregulation of MUC3, MUC4, resistin-like molecule beta (Relmbeta) and 3O-sulfotransferase (3ST)1 was observed, while 3ST2 expression levels increased after the majority of the worms were expelled from the intestine. Similar alterations of glycosylation-related gene expression were also observed in mast-cell-deficient Ws/Ws rats, suggesting that mast cells in the epithelium are not relevant to the upregulation of these genes. The present finding that the expression level of each goblet cell- or glycosylation-related gene was altered differently during the time course of infection indicates the progression of sequential qualitative changes in the mucus layer after infection.

Animals↗

Cytokine mRNA expression profiles in rats infected with the intestinal nematode Nippostrongylus brasiliensis.

Although the immune responses to intestinal nematode infection have been well studied and have been shown to be strongly driven by Th2-associated cytokines in mice, such information has been limited with respect to rats. We investigated changes in levels of the mRNAs encoding interleukin-2 (IL-2), IL-3, IL-4, IL-5, IL-10, and gamma interferon in the mesenteric lymph nodes of rats infected with Nippostrongylus brasiliensis by reverse transcription-PCR in comparison with immunoglobulin E (IgE)/IgG2a antibody, eosinophil, basophil, and mucosal mast cell responses. In the two rat strains used, Brown Norway and Fischer-344, which show different responses to allergens, serum IgE increased to much higher levels in the former than in the latter 2 weeks after infection. Intestinal mastocytosis was observed much earlier and more intensely in Brown Norway rats than in Fischer-344 rats, but the degrees of peripheral eosinophilia and basophilia did not differ between the two strains. In both strains, IL-3, IL-4, and IL-5 mRNA expression increased and peaked around 7 to 14 days after infection, while expression of IL-2, IL-10, and gamma interferon mRNAs did not change notably throughout the experimental period. The highest IL-4 mRNA expression was observed slightly earlier in Brown Norway than in Fischer-344 rats, but levels of IL-3 and IL-5 mRNAs peaked synchronously in both strains. The amounts of mRNAs encoding these three cytokines were always higher in Brown Norway than in Fischer-344 rats. It is suggested that in rats, Th2 or Th2-like cells are also induced after nematode infection, and IgE elevation is mainly related to increased IL-4 gene expression.

Animals↗

Inhibition of autoimmune type 1 diabetes by gastrointestinal helminth infection.

Gastrointestinal nematode infections are prevalent worldwide and are potent inducers of T helper 2 responses with the capacity to modulate the immune response to heterologous antigens. Parasitic helminth infection has even been shown to modulate the immune response associated with autoimmune diseases. Nonobese diabetic (NOD) mice provide a model for studying human autoimmune diabetes; as in humans, the development of diabetes in NOD mice has been linked to the loss of self-tolerance to beta cell autoantigens. Previous studies with the NOD mouse have shown that helminth and bacterial infection appears to inhibit type 1 diabetes by disrupting the pathways leading to the Th1-mediated destruction of insulin-producing beta cells. The aim of our study was to examine whether infection with the gastrointestinal helminths Trichinella spiralis or Heligmosomoides polygyrus could inhibit the development of autoimmune diabetes in NOD mice and to analyze the mechanisms involved in protection and the role of Th2 responses. Protection from diabetes was afforded by helminth infection, appeared to inhibit autoimmune diabetes by disrupting pathways leading to the destruction of beta cells, and was mediated by seemingly independent mechanisms depending on the parasite but which may be to be related to the capacity of the host to mount a Th2 response.

Animals↗

Cells containing IgE in the intestinal mucosa of mice infected with the nematode parasite Trichinella spiralis are predominantly of a mast cell lineage.

To determine whether IgE+ cells in the intestinal mucosa of nematode-infected mice were of a mast cell or a lymphocyte lineage, the intestinal mucosae of mast cell-deficient w/wv mice were examined for IgE+ cells after inoculation with Trichinella spiralis muscle-stage larvae. Immunofluorescence staining techniques were used to detect IgE associated with cells in the intestinal mucosa. Comparisons were made among four strains of mice, w/wv (mast cell-deficient), +/+ (normal congenic littermates of w/wv), BALB/c, and SJL, that were either uninfected controls or inoculated with T. spiralis. Tissue sections from the small intestine of T. spiralis-infected BALB/c, SJL, and +/+ mice were fixed in ethanol and were stained with an affinity-purified F(ab')2 rabbit anti-mouse IgE followed by FITC goat anti-rabbit IgG. Large numbers of cells in the intestinal mucosa exhibited bright fluorescence. When other sections of intestines from these mice were processed in Carnoy's fixative and were stained with alcian blue at low pH (a metachromatic stain for mast cells) or alcian blue followed by immunofluorescence staining for IgE, large numbers of mast cells were observed in the intestinal mucosa, and 70 to 90% stained positively for IgE. There was a considerable number of cells in the intestinal mucosa which were IgE+ but which did not stain with alcian blue. Few alcian blue-positive cells and no IgE+ staining cells were present in the intestinal mucosa of control, uninfected +/+, BALB/c, and SJL mice. To determine whether these IgE+ alcian blue-negative cells were of a lymphocyte or a mast cell lineage, the mast cell-deficient w/wv mouse strain was examined after infection with T. spiralis. In contrast to BALB/c, SJL, or +/+ mice, few cells in the intestinal mucosa of T. spiralis-infected w/wv mice stained with alcian blue or were positive for IgE. However, when the IgE response in the MLN of the w/wv mice was compared to the IgE response of BALB/c, SJL, and +/+ mice, numerous IgE+ cells, but no alcian blue-positive cells, were observed in the parenchyma of the MLN from all four strains of T. spiralis-infected mice. In addition, flow microfluorometric analysis of MLN cells stained for surface IgE in suspension showed a comparable proportion of IgE-bearing cells, which were mostly B lymphocytes, among all four strains of T. spiralis-infected mice.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Efficacy of homeopathic treatment against natural infection of sheep by gastrointestinal nematodes.

The efficacy of the homeopathic treatment with the Fator Vermes, administered according to the manufacturer's recommendations, was evaluated against gastrointestinal nematodes infections in sheep. The experiment was divided into two phases: in the first phase (January/06/2004 to April/30/2004), the animals of the treated (n=10) and control (n=10) groups were treated individually with conventional anthelmintics to avoid deaths. In the second phase (April/30/2004 to July/06/2004), the sheep from the group that received the Fator Vermes were treated as they had been in the previous phase, while the control group animals were treated with conventional anthelmintics at 14 day intervals. In the first phase of the experiment, there was no significant difference (P>0.05) between group means regarding egg counts in feces (EPG), weight gain, or packed cell volume (PCV). Meanwhile, in the second phase, the control group sheep had a significantly higher weight gain, higher PCV values, and lower EPG counts. Infective larvae of Haemonchus spp., Trichostrongylus spp., Cooperia spp., and Oesophagostomum spp. were identified in the fecal cultures. After six months of daily treatment with the Fator Vermes, it was not possible to substantiate the product's benefits in both sheep health and productivity or in the prophylaxis of gastrointestinal nematode infections.

Animals↗

Variation in infection levels with gastrointestinal nematodes in first-year grazing calves in The Netherlands.

ELISAs with crude adult Ostertagia and crude adult Cooperia antigens were used in a survey to monitor the levels of gastrointestinal nematode infections of 21 dairy calf herds. This survey was performed at the end of the grazing season of 1994 in The Netherlands. The mean extinction value of antibody levels of each herd was compared with those of four groups of reference sera of animals that had experienced moderate infection levels (one group), low infection levels (two groups) or no infection (permanently housed group). This was based on faecal examination, pasture larval counts, serum pepsinogen values and worm counts in tracer and main trial calves. The results of the ELISAs indicated a wide variation in infection levels. Most herds (90%) had infection levels equal to, or even lower than, those of the low infection-reference groups. Herds that had been moved every 2 or 3 weeks had lower infections than set stocked herds. The length of the grazing season showed a positive correlation with Ostertagia, but not Cooperia, antibody titres. There was no evident correlation between infection levels and anthelmintic treatment. The implications of these observations in relation to management and nematode control are discussed.

Animal Feed↗

Cow-calf herds in eastern Germany: status quo of some parasite species and a comparison of chemoprophylaxis and pasture management in the control of gastrointestinal nematodes.

Infections with gastrointestinal parasites (Eimeria spp., Cryptosporidium spp., Buxtonella sulcata, Fasciola hepatica, Moniezia spp. and trichostrongyles) and lungworms were monitored in five cow-calf herds in the north German lowlands. Estimated prevalences of infections with Eimeria spp. (predominantly Eimeria bovis) ranged between approximately 2 and 48%. The highest prevalences were found during late summer and autumn. On one farm Cryptosporidium spp. were detected in July and August (prevalence: 8.5 +/- 2.7% and 6.7 < 2.0%). The latter finding coincided with diarrhoea in many calves. Buxtonella sulcata was found during the entire study period in highly variable estimated prevalences ranging between zero and 73%, but without any obvious association with clinical disease. Fasciola hepatica was detected on four out of five farms at estimated prevalences of approximately 1-20%. Lungworm infections played a minor role in at least three of five study herds. The estimated prevalence of trichostrongyle infections rose from August until November whereas the intensity of infection did not change significantly. No difference in the intensity of infection could be detected between one farm on which infections with gastrointestinal nematodes were controlled only by moving the animals to an uninfected pasture in July, and three other herds on which strategic anthelmintic control was in place.

Animal Husbandry↗

Relationship between intermediate host taxon and infection by nematodes of the genus Rhabdochona.

We describe the intermediate and definitive hosts of the fish nematodes Rhabdochona coronacauda and R. denudata honshuensis and discuss the relationships between parasitism and the feeding habitats of their intermediate hosts. We found that the principal intermediate hosts of the two nematodes were filter-feeding mayflies of the genera Ephemera, Photamanthus and Isonychia. Ephemera strigata seemed to be the most important intermediate host of these nematodes. Adult R. coronacauda were found mainly in Hemibarbus longirostris and Rhinogobius flumineus, which are benthic fishes that feed on benthic aquatic insects, including E. strigata. For R. coronacauda, therefore, the feeding habits of the definitive hosts facilitate host alternation by this species. However, adult R. denudata honshuensis were found in cyprinids. In particular, Zacco temmincki was the principal natural definitive host in our study area. Since Z. temmincki is a swimming predator, E. strigata nymphs that burrow in the substrate are not the main prey of this species. This indicates that the transmission of R. denudata honshuensis hardly occurs from E. strigata nymph to Z. temmincki, suggesting another, unknown transmission route.

Animals↗