Gastrointestinal Nematode Parasites Research Articles

Predatory activity of Pochonia chlamydosporia fungus on Toxocara (syn. Neoascaris) vitulorum eggs

Toxocara (Neoascaris) vitulorum is a gastrointestinal nematode parasite of young ruminants, responsible for high mortality rates in parasitized cattle and buffalo calves. The objective of this work was to compare the predatory capacity under laboratory conditions of four fungal isolates of the nematophagous fungus Pochonia chlamydosporia (VC1, VC4, VC5 and VC12) on T. vitulorum eggs in 2% water-agar (2% WA). T. vitulorum eggs were plated on 2% WA Petri dishes which contained cultured fungal isolates and control plates without fungi. After 10 and 15 days one hundred eggs were removed and classified according to the following parameters: type 1, biochemical and physiological effect without morphological damage to the eggshell, type 2, lytic effect with morphological alteration of the eggshell and embryo and type 3, lytic effect with morphological alteration of eggshell and embryo in addition to hyphal penetration and internal egg colonization. The fungal isolates were effective in the destruction of T. vitulorum eggs presenting the type 3 effect at 10 and 15 days after contact with the fungus. No nematophagous fungi were observed in the control group during the experiment. There was no variation in the predatory capacity of the fungal isolates (P > 0.01) at the intervals of 10 and 15 days. These results indicate that P. chlamydosporia (VC1, VC4, VC5 and VC12) negatively influenced the development of T. vitulorum eggs and can be considered a potential candidate for the biological control of nematodes.

P-glycoprotein interfering agents potentiate ivermectin susceptibility in ivermectin sensitive and resistant isolates ofTeladorsagia circumcinctaandHaemonchus contortus

P-glycoprotein (P-gp) homologues, belonging to the ATP Binding Cassette (ABC) transporter family, are thought to play an important role in the resistance of gastro-intestinal nematode parasites against macrocyclic lactones. The aim of this study was to investigate the influence of various P-gp interfering compounds on the efficacy of ivermectin (IVM) in sensitive and resistant nematode isolates. The feeding of IVM resistant and sensitive Teladorsagia circumcincta and Haemonchus contortus first-stage larvae (L1) was assessed using a range of IVM concentrations (0.08-40 nm) with or without P-gp inhibitors: valspodar, verapamil, quercetin, ketoconazole and pluronic P85. The P-gp inhibitors were selected on the basis of their ability to interfere with P-gp transport activity in an epithelial cell line over-expressing murine P-gp. In the presence of P-gp interfering agents, the in vitro susceptibility to IVM of both sensitive and resistant isolates of T. circumcincta and H. contortus was increased. These results show that compounds interfering with P-gp transport activity could enhance IVM efficacy in sensitive isolates, and also restore IVM sensitivity in resistant nematodes. These results support the view that ABC transporters can play an important role in resistance to IVM, at least in the free-living stages of these economically important gastro-intestinal nematodes.

Ostertagia ostertagi antibodies in milk samples: Relationships with herd management and milk production parameters in two Mediterranean production systems of Spain

The present study analyzed Ostertagia ostertagi antibodies by indirect ELISA in milk samples in two cattle systems in Mediterranean Spain to indirectly monitor gastrointestinal nematode (GI) parasitism effects on production. Individual samples from 10 animals and the corresponding milk herd samples were collected from 133 herds in Girona (intensive management) and 123 herds in Minorca (extensive management). Both locations showed high and significant positive relationships between average optical density ratios (ODR) of individual animals and ODR in their milk tank. Although antibodies levels were low, there were significantly higher in Minorca. Negative correlations between ODR values and milk production were found in both systems. Importantly, in Minorca, average herd milk production was higher in the herds that treated their animals against GI nematodes compared to those that did not treat. The ELISA technique was valuable to indirectly assess differences in the level of GI nematode infection even in cattle production systems with low levels of infection.

Efficacy of sericea lespedeza hay as a natural dewormer in goats: Dose titration study

Gastrointestinal nematodes (GIN) parasitism is the greatest threat to economic sheep and goat production in the southern USA, and there is widespread prevalence of GIN resistance to broad-spectrum anthelmintics in this region. A natural alternative for controlling GIN in small ruminants is feeding hay of sericea lespedeza [SL, Lespedeza cuneata (Dum.-Cours., G. Don)], a perennial warm-season legume high in condensed tannins. To determine the level of SL needed to reduce GIN infection, a confinement study was completed with 32 Spanish/Boer/Kiko cross yearling bucks offered one of four diets with 75% hay and 25% concentrate (n=8, 2pens/treatment, 4 goats/pen). The hay portion of each diet consisted of a combination of ground SL (0%, 25%, 50%, and 75% of the diet) and bermudagrass [BG, Cynodon dactylon (L.) Pers.; 75%, 50%, 25%, and 0% of the diet]. The bucks were allowed to acquire a natural GIN infection on pasture prior to moving to the pens. After a 3-week adjustment period in the pens, the goats were stratified by fecal egg count (FEC) and packed cell volume (PCV), randomly assigned to treatments and pens, and then fed the treatment diets for six weeks. During the experimental period, fecal and blood samples were collected from individual animals weekly to determine FEC and PCV, respectively. Adult worms from abomasum and small intestines were collected for counting and identification of species at slaughter. Goats fed SL hay at 25%, 50%, and 75% of the diet had 45.3% (P=0.2048), 66.3% (P=0.0134), and 74.5% (P=0.0077) lower FEC than control animals (75% BG hay) after 21 days. The 50% and 75% SL goats had 84.6% (P=0.0625) and 91.9% (P=0.0340) lower FEC than controls by day 42. The 75% SL-fed goats tended to have higher (P=0.0624) PCV and had fewer (P=0.035) abomasal worms than control animals, while PCV and adult worm numbers of the 50% and 25% SL goats were not different from controls. The optimum level of SL hay in the diet for reducing worm numbers of small ruminants appears to be 75%, whereas 50–75% SL reduces FEC, which could lead to reduced larval infection on pasture.

Are Scottish Blackface ewes more resistant to nematode parasite infection than Greyface ewes?

The extent of periparturient relaxation of immunity (PPRI) to gastrointestinal nematode parasites is sensitive to metabolisable protein (MP) scarcity but also varies between breeds of sheep (Houdijk, 2008). For example, under ad libitum feeding, Scottish Blackface ewes had a lower extent of PPRI than Greyface ewes (Zaralis et al., 2008). Such between-breed variation in PPRI may not necessarily be associated with genetic resistance per se but could arise from a higher nutrient demand of the more productive Greyface ewes (Houdijk, 2008). This experiment was designed to test the hypothesis that if the extent of PPRI has only a nutritional basis, then the sensitivity of PPRI to MP scarcity will not differ between the breeds when MP feeding is adjusted for between-breed differences in MP demand.

The transcriptomes of the cattle parasitic nematode Ostertagia ostartagi

Ostertagia ostertagi is a gastrointestinal parasitic nematode that affects cattle and leads to a loss of production. In this study, we present the first large-scale genomic survey of O. ostertagi by the analysis of expressed transcripts from three stages of the parasite: third-stage larvae, fourth-stage larvae and adult worms. Using an in silico approach, 2284 genes were identified from over 7000 expressed sequence tags and abundant transcripts were analyzed and characterized by their functional profile. Of the 2284 genes, 66% had similarity to other known or predicted genes while the rest were novel and potentially represent genes specific to the species and/or stages. Furthermore, a subset of the novel proteins were structurally annotated and assigned putative function based on orthologs in Caenorhabditis elegans and corresponding RNA interference phenotypes. Hence, over 70% of the genes were annotated using protein sequences, domains and pathway databases. Differentially expressed transcripts from the two larval stages and their functional profiles were also studied leading to a more detailed understanding of the parasite's life-cycle. The identified transcripts are a valuable resource for genomic studies of O. ostertagi and can facilitate the design of control strategies and vaccine programs.

Growth and meat quality of kids of indigenous Greek goats ( Capra prisca) as influenced by dietary protein and gastrointestinal nematode challenge

The effect of dietary protein and gastrointestinal nematode (GIN) parasitism on growth and meat quality of growing kids was assessed using sixty (60) kids in three groups ( n = 20); A: control, B: regularly treated with ALBENDAZOLE ® and C: supplemented with dietary protein. The kids grazed in a pasture contaminated with L3 larvae of GIN. Growth and condition score were assessed at 21-day intervals. After 86 days all kids were slaughtered. Carcasses were assessed for conformation, fatness, ultimate pH and other meat quality characteristics. Parasitic challenge was assessed by means of faecal egg counts (FEC), pasture larvae and adult nematodes in the GI tract of kids at slaughter. Groups C and B had higher growth rates and body condition score and produced significantly heavier ( P < 0.05) carcasses with better ( P < 0.01) conformation and fatness when compared to those of group A. Total unsaturated and monounsaturated fatty acids were higher ( P < 0.05) in fat tissue of groups B and C. Group A had the highest FEC and group C had the lowest ( P < 0.05) FEC. The parasitic challenge of L3 on pasture reached its highest point at 42 days and there were significant ( P < 0.01) differences between the numbers of Teladorsagia spp., Trichostrongylus spp., Haemonchus contortus, Oesophagostomum spp. and Chabertia spp. found in the GI tract of kids between the three groups; group A had the highest numbers. Overall, the results showed that the increased protein content in the diet of growing kids grazing on a pasture contaminated with L3 nematode larvae resulted in the production of acceptable carcasses.

Ovicidal and larvicidal activity of crude extracts of Maesa lanceolata and Plectranthus punctatus against Haemonchus contortus

The widespread development of anthelmintic resistance and high cost of the conventional anthelmintic drugs, has limited the control of gastrointestinal nematode parasites of sheep and goats and hence led to evaluation of medicinal plants as an alternative source of anthelmintics. In the current study, in vitro ovicidal and larvicidal activity of the leaves and fruits of the aqueous and hydro-alcoholic extracts of Maesa lanceolata and aerial parts of Plectranthus punctatus were evaluated on the egg and larvae of Haemonchus contortus using egg hatch assay and larval development test. All extracts of plants tested have shown complete inhibition of egg hatching at or below 1 mg/ml. ED 50 for egg hatch inhibition ranged from 0.11 to 0.29 mg/ml, for both the aqueous and hydro-alcoholic extracts of Plectranthus punctatus and Maesa lanceolata. All extracts have shown dose dependent inhibition of larval development with variable results. The complete inhibition (100%) at the maximum concentration tested (50 mg/ml) was obtained only for hydro-alcoholic extract of the fruits of Maesa lanceolata and the lowest inhibition (50.33%) was recorded for the hydro-alcoholic extract of the leaves of the same plant. The overall findings of the present study has shown that Plectranthus punctatus and Maesa lanceolata contain possible anthelmintic compounds and further evaluation of different extracts and fractions of these plants should be carried out.

The mannose receptor binds Trichuris muris excretory/secretory proteins but is not essential for protective immunity.

Trichuris muris is a natural mouse model of the human gastrointestinal nematode parasite Trichuris trichiura and it is well established that a T helper type 2-dominated immune response is required for worm expulsion. Macrophages accumulate in the large intestine of mice during infection and these cells are known to express the mannose receptor (MR), which may act as a pattern recognition receptor. The data presented here show for the first time that T. muris excretory/secretory products (E/S) induce bone-marrow-derived macrophages (BMDM) to produce several cytokines and have MR-binding activity. Using alternatively activated BMDM from MR knockout mice it is shown that the production of interleukin-6 partially depends on the MR. Infection of MR knockout mice with T. muris reveals that this receptor is not necessary for the expulsion of the parasite because MR knockout mice expel parasites with the same kinetics as wild-type animals and have similar cytokine responses in the mesenteric lymph nodes. Furthermore, despite acting to reduce serum levels of proinflammatory mediators, absence of the MR does not lead to increased gut inflammation after T. muris infection when assessed by macrophage influx, goblet cell hyperplasia and crypt depth. This work suggests that, despite binding components of T. muris E/S, the MR is not critically involved in the generation of the immune response to this parasite.

Herbal dewormer fails to control gastrointestinal nematodes in goats

Gastrointestinal nematode (GIN) parasitism is the most important disease of small ruminants. Control is usually based on the use of chemical anthelmintics (dewormers); but these are prohibited from use in organic livestock, and the effectiveness of chemical anthelmintics in conventional operations is limited by high levels of anthelmintic resistance. Consequently, herbal dewormers are increasing in popularity as an alternative to chemical dewormers for GIN control. However, the effectiveness of herbal dewormers remains unproven. The objective of this study was to examine the effectiveness of a commercially available herbal dewormer to control GIN in goats. Lactating ( n = 16) and young ( n = 8) dairy goats grazed poor quality mixed grass pastures between March and July 2006 at the Heifer International Ranch in Perryville, AR. Goats were supplemented with grass hay and concentrate. Goats were untreated or administered herbal dewormer ( n = 12/treatment) according to manufacturer recommendations. FAMACHA scores (1 = red or healthy; 5 = severely anemic) were determined and fecal samples collected for fecal egg count (FEC) determination every 14 days between Days 0 (day of first herbal treatment) and 112. FAMACHA scores in the herbal treated group were greater than in the untreated control group ( P < 0.005), indicating a higher level of anemia. FEC were greater for herbal treated goats on Pasture A compared with B by Day 42, but similar among groups thereafter. FEC ( P < 0.03) and FAMACHA scores ( P < 0.001) were greater in lactating than in non-lactating goats. Herbal dewormer treatment yielded no measurable health benefits indicating that the herbal dewormer failed to control GIN in these goats.

Duddingtonia flagrans: controle biológico de nematodeos de bovinos a campo

O controle biológico é um método para diminuir uma população pela utilização de antagonista natural. No presente estudo, testou-se a eficácia do fungo nematófago Duddingtonia flagrans no controle de nematódeos parasitos gastrintestinais de bovinos criados à campo no município de Júlio de Castilhos. Foram utilizados 20 bezerros, distribuídos igualmente em duas áreas formadas por pastagem nativa. O grupo A foi tratado com o fungo D. flagrans, cultivado em sorgo, numa concentração de 1x10(6)clamidósporos kg-1 de peso animal, misturados em ração de manutenção, diariamente, durante oito meses. O grupo B serviu como controle e não recebeu fungo, apenas ração. Foram coletadas amostras para contagem de ovos por grama de fezes (OPG) semanalmente. Mensalmente, foram realizadas coproculturas para identificar as espécies de larvas de nematódeos, a pesagem dos animais e a coleta de pasto para contagem das larvas na pastagem. Dados de temperatura e índice pluviométrico foram registrados diariamente. O OPG foi reduzido no grupo tratado, em média 56,8,% nos últimos três meses de experimento, variando entre 40,4 e 67,1% no grupo tratado (P&lt;0,001). A coprocultura demonstrou que os principais nematódeos encontrados em ambos os grupos foram dos gêneros Cooperia e Haemonchus. A contagem de larvas na pastagem obteve um percentual de redução 77,1% no grupo tratado ao final do experimento (P&lt;0,01). Pôde-se concluir com este estudo que o papel do fungo Duddingtonia flagrans, é, sem dúvida, importante, principalmente, na diminuição do OPG e na redução significativa de larvas na pastagem. Portanto, este fungo nematófago é uma ferramenta biológica eficaz para ser empregado em um controle integrado de nematódeos de bovinos criados a campo.

Availability of gastro-intestinal nematode larvae to sheep following winter contamination of pasture with six nematode species on the Northern Tablelands of New South Wales

This experiment was designed to determine the infective consequences of winter pasture contamination by sheep infected with 6 species of trichostrongylid gastro-intestinal nematode (GIN) at Armidale in the New England region of NSW. Ten helminthologically inert pasture plots were contaminated using ‘donor’ sheep harbouring artificial infections of Haemonchus contortus, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Trichostrongylus axei, Nematodirus spathiger and Teladorsagia circumcincta. A single plot was contaminated in March to serve as a positive control and three plots in each of the winter months of June, July or August of 2007. Weekly assessments of 24 h faecal output and worm egg count of all donor sheep, combined with differentiation of larval cultures enabled calculation of total egg deposition per plot for each GIN species for each month of contamination. All plots were provided with supplemental spray irrigation sufficient to remove desiccation as a cause of non-development. The infective consequence of each monthly contamination was assessed by grazing each plot with two tracer sheep for the last 14 days of each month following contamination until a final grazing in November. Translation for each GIN species was expressed as the percentage of eggs deposited translating to worms counted in tracer sheep. The GIN parasites fell into two distinct groups differing greatly in translational success. Translation of egg to establishment in the tracer host was ineffective for H. contortus, T. axei and T. colubriformis but effective for T. vitrinus, T. circumcincta and N. spathiger. Overall mean total translation for each of these species throughout the experiment was respectively 0.00%, 0.00%, 0.00%, 0.34%, 0.45% and 0.48%. We conclude that winter contamination of pastures in this region with eggs of the latter 3 species is capable of impacting on the health of sheep subsequently grazing these pastures. Where H. contortus and T. colubriformis predominate, paddocks selected for spring lambing may be grazed safely over the winter months without contributing significantly to pasture contamination. These findings are particularly relevant to the preparation of clean spring lambing paddocks in this region using grazing management.

Grazing willow ( Salix spp.) fodder blocks for increased reproductive rates and internal parasite control in mated hoggets

A grazing experiment, conducted for 116 days from 19 January to 15 May in the late summer/autumn of 2006 at Massey University's Riverside dry-land Farm, near Masterton (New Zealand), compared effects of grazing willow ( Salix spp.) fodder blocks or perennial ryegrass ( Lolium perenne)/white clover ( Trifolium repens) pasture, during the pre-mating and mating periods, on gastrointestinal nematode parasite control and reproductive rates in 7-month old ewe hoggets. A rotational grazing system with 348 weaned Romney ewe hoggets was used, with 116 hoggets per treatment (control pasture with regular anthelmintic drenching, control pasture with trigger drenching and willow fodder blocks with trigger drenching). Hogget live weights (LW), dag scores (DS), faecal nematode egg counts (FEC) and reproductive rates at foetal ultrasound scanning, lambing, tail docking and weaning were measured. Total N concentration (35 g/kg dry matter; DM), organic matter digestibility (OMD; 0.68) and metabolisable energy (ME; 10 MJ/kg DM) were similar for fodder block pasture and control pasture; the selected tree fodder had lower concentrations of N (24 g/kg DM) and was higher in OMD (0.74) and ME (10.4 MJ/kg DM). Tree fodder contained higher concentrations of condensed tannins (CT; 22.9 g/kg versus 1.6 g/kg DM) than control pasture and herbage in willow fodder blocks. Grazing willow fodder blocks increased LW gain (approximately 97 g/day versus 86 g/day) and increased reproductive rate corrected to equal LW by approximately 17 lambs/100 hoggets mated, due to increases in both oestrus activity and conception rates (hoggets pregnant/100 mated). Calculated daily DM (1.41 kg) and ME (14 MJ) intakes were similar in all groups. Calculated daily CT intake (6.0 g versus 2.1 g) and CT intake/100 g CP intake (2.2 g versus 0.7 g) were higher for willow fodder block hoggets than for the control pasture groups, and this may have increased the flow of undegraded dietary protein to the small intestine leading to increases in reproductive rates of this group. Grazing willow fodder blocks failed to reduce the number of anthelmintic drenches (3) needed to maintain FEC below 1000 eggs/g wet faeces, but was successful in reducing DS compared to grazing conventional ryegrass/white clover pastures. Grazing willow fodder blocks can be beneficial in sustainable farming systems since tree fodder can sustain growth rates, increase reproductive rates and reduce dag formation in parasitized hoggets.

The effectors responsible for gastrointestinal nematode parasites, Trichinella spiralis, expulsion in rats

Helper T (Th2) cells type 2 have a central role in host protective responses to gastrointestinal nematode parasite, Trichinella spiralis infection, but the actual effector mechanisms involved in parasite expulsion are still uncertain. Recent evidences suggest that mast cell recruitment and activation may associate with parasite elimination from host intestines in mice. On the other hand, IgE production may induce defensive responses to primary infection with the helminth in rats. The differences of host effector mechanisms to the same experimental infections might disturb our understanding of the host protective mechanisms to gastrointestinal nematode parasite infection. In order to redefine these differences, we investigated in detail the relationship between intestinal immune responses and worm expulsion following T. spiralis infection among several rat strains including mutants. As a result, there were significant correlations of parasite expulsion with mast cell hyperplasia in addition to serum IgE level. Moreover, mast cell-deficient and dysfunction rats showed delayed worm elimination from their gut. Therefore, the present study suggests that mast cells should also be one of the prominent effector cells involved in T. spiralis parasite expulsion in rats as well as mice.

Evaluation of anthelmintic activity of Iris hookeriana against gastrointestinal nematodes of sheep

The objective of this study was to evaluate the anthelmintic efficacy of Iris hookeriana Linn. rhizome against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for an in vivo study. The in vitro study revealed anthelmintic effects of crude aqueous extracts and crude ethanolic extracts on live Trichuris ovis worms (P < or = 0.05) as evident from their paralysis and/or death at 8 h after exposure. The aqueous extracts of I. hookeriana resulted in a mean worm motility inhibition of 54.0%, while ethanolic extracts resulted in a mean worm motility inhibition of 84.6%. The mean mortality index of aqueous extracts was 0.55, while for ethanolic extracts it was 0.85. The lethal concentration 50 for aqueous extracts was 0.45 mg ml- 1 and for ethanolic extracts it was 0.15 mg ml- 1. The in vivo anthelmintic activity of aqueous and ethanolic extracts of I. hookeriana in sheep naturally infected with mixed species of gastrointestinal nematodes demonstrated a maximum (45.62%) egg count reduction in sheep treated with ethanolic extracts at 2 g kg- 1 body weight on day 10 after treatment, closely followed by ethanolic extracts at 1 g kg- 1 body weight on day 10 after treatment (43.54% egg count reduction). The aqueous extracts resulted in a maximum of 31.53% reduction in faecal egg counts on day 10 after treatment with 1 g kg- 1 body weight. Thus ethanolic extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of alcohol-soluble active ingredients in I. hookeriana. From the present study it can be suggested that I. hookeriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

The effects of feeding sericea lespedeza hay on growth rate of goats naturally infected with gastrointestinal nematodes1

Goat production is increasing in the United States due to high ethnic demand, but infection with gastrointestinal nematode (GIN) parasites is a major constraint to the industry. Increasing GIN resistance to chemical anthelmintics worldwide has led to the development of alternative control strategies, including use of forages containing condensed tannins (CT). An experiment was designed using infected and dewormed male kids (Kiko x Spanish, 6 mo old, 18.9 +/- 3.25 kg) fed diets containing 25% concentrate and either 75% sericea lespedeza [SL; Lespedeza cuneata (Dum-Cours.) G. Don], a high CT forage (87 to 181 g of CT/kg), or 75% bermudagrass [BG; Cynodon dactylon (L.) Pers.] hay (n = 10/treatment). The kids were weighed every 14 d, and fecal and blood samples were taken weekly for fecal egg counts and packed cell volume determination, respectively. Fecal cultures were processed every 14 d to determine CT effect on larval development. At slaughter, adult GIN were collected from the abomasum and small intestines for counting and speciation. Blood samples were also analyzed for plasma urea-N, and ruminal VFA and pH were determined. The infected SL-fed kids had consistently lower (P < 0.05) fecal egg counts than the infected BG goats throughout the trial and greater (P < 0.05) packed cell volume beginning by d 77. Average daily gain was greater (P < 0.001) in kids fed SL- than BG-based diets, regardless of infection status (104.3 +/- 5.0 and 75.5 +/- 4.8 g/d, respectively). Total VFA and acetate concentrations were greater (P < 0.001) in the BG- than in SL-fed goats, whereas propionate levels were unaffected by diet. Acetate:propionate ratio (P = 0.01) and plasma urea-N (P = 0.03) levels were greater in BG-fed goats, whereas rumen pH was greater (P < 0.001) in the SL-fed goats. Feeding SL hay can reduce GIN infection levels and increase performance of goats compared with BG hay.

Epidemiology of gastrointestinal nematode parasitism in a commercial sheep flock and its implications for control programmes

The epidemiology of nematode infections in a uk commercial crossbred sheep flock was studied from January 2004 to January 2005. The ewes were treated orally with moxidectin when they were...

Cyclotides: Natural, Circular Plant Peptides that Possess Significant Activity against Gastrointestinal Nematode Parasites of Sheep

The cyclotides are a novel family of backbone-cyclized cystine-knot containing peptides from plants that have been shown to possess insecticidal activity against Helicoverpa larvae, an important pest of corn and cotton. In the current study, we investigated the in vitro effects of the cyclotides on the viability of egg, larval, and adult life stages of two species of economically important gastrointestinal nematode parasites of livestock, Hemonchus contortus and Trichostrongylus colubriformis. The cyclotides showed significant activity in inhibiting development of nematode larvae and motility of adult worms. Activities were comparable to some currently used anthelmintic compounds in these in vitro assay systems. A series of alanine mutants of the prototypic cyclotide kalata B1 were assayed against larvae to determine regions of the peptide responsible for activity. It was observed that anthelmintic activity was dramatically reduced as a consequence of the mutation of a large number of residues that are found clustered on one surface. Activities toward larvae were equivalent in the naturally occurring L-isomer of kalata B1 and a synthetic all-D-isomer, indicating that there is no chiral requirement for anthelmintic activity. The clustering of important residues and the lack of chiral selectivity further support the proposed mode of action of the cyclotides, which involves a membrane-based interaction rather than an interaction at a specific receptor. The cyclotide-induced leakage of a fluorescent dye from vesicles used as a model membrane mimetic further confirms the membrane lytic ability of cyclotides. The relative potency of kalata B1 and kalata B2 in causing membrane leakage is consistent with the order of their anthelmintic activity. These results demonstrate that the cyclotides show potential for use in the control of gastrointestinal nematode parasites.

Th2 Cytokine-Induced Alterations in Intestinal Smooth Muscle Function Depend on Alternatively Activated Macrophages

Enteric nematode infection induces a strong type 2 T helper cell (Th2) cytokine response characterized by increased infiltration of various immune cells, including macrophages. The role of these immune cells in host defense against nematode infection remains poorly defined. The present study investigated the role of macrophages and the arginase pathway in nematode-induced changes in intestinal smooth muscle function and worm expulsion. Mice were infected with Nippostrongylus brasiliensis and treated with clodronate-containing liposome to deplete macrophages or given S-(2-boronoethyl)-I-cysteine in drinking water to inhibit arginase activity. Segments of intestinal smooth muscle were suspended in organ baths to determine responses to acetylcholine, 5-hydroxytryptamine, or nerve stimulation. The phenotype of macrophages was monitored by measuring mRNA expression of the specific molecular markers by real-time polymerase chain reaction or viewed by immunofluorescence staining. Infection increased the infiltration of macrophages and up-regulation alternatively activated macrophage markers by a mechanism dependent on interleukin-4 (IL-4) or interleukin-13 (IL-13) activation of signal transducer and activator of transcription 6. Elimination of alternatively activated macrophages blocked smooth muscle hypercontractility and the increased smooth muscle thickness, and impaired worm expulsion. In addition, specific inhibition of arginase activity interfered with smooth muscle contractility, but only partially affected the protective immunity of the host. These data show that the phenotype of macrophages is determined by the local immune environment and that alternatively activated macrophages play a major role in the effects of Th2 cytokines, IL-4 and IL-13, on intestinal smooth muscle function.

Trade‐offs and benefits: implications of promoting a strong immunity to gastrointestinal parasites in sheep

The search for sustainable control regimes for gastrointestinal nematode parasites in grazing livestock has resulted in strategies that include assisting sheep to promote a strong immune response. However, the immune response imposes a substantial nutritional cost to the animal which, when nutrient supply is limited, is likely to result in the diversion of nutrients away from productive function to immunological tissues. Therefore, this review examines the potential trade-offs between immune function and productivity that may arise in young grazing sheep. It is hoped that future manipulations intended to promote a strong immunity to gastrointestinal parasites in sheep may be able to minimize these trade-offs to ensure that the productivity of the host is not compromised.