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PubMed2004

Ectopic expression of a Haemonchus contortus GATA transcription factor in Caenorhabditis elegans reveals conserved function in spite of extensive sequence divergence.

Comparative analysis between Caenorhabditis elegans and other nematode species offers a powerful approach to study gene function. C. elegans also has great potential as a surrogate expression system to study the function of genes from parasitic nematode species where transgenic methodologies are unavailable. However there is little information on the extent to which the biology of C. elegans is conserved with other nematode species and very few parasitic nematode genes have yet been functionally expressed in C. elegans. We have identified and characterised a homologue of the C. elegans GATA transcription factor elt-2, a central regulator of endoderm development, from the parasitic nematode Haemonchus contortus. The H. contortus ELT-2 polypeptide is present in endoderm nuclei throughout embryonic and post-embryonic development, except for in the infective L3 stage, and our experiments reveal that the development of the H. contortus endodermal lineage is strikingly similar to that of C. elegans. Sequence conservation between the H. contortus and C. elegans ELT-2 polypeptides broadly reflects function since the major region of sequence identity corresponds to the DNA binding domain. However, the overall level of sequence identity is remarkably low with the only other major region of identity corresponding to an unusual zinc finger domain. In spite of this, ectopic expression of the H. contortus elt-2 gene in transgenic C. elegans is sufficient to activate a programme of endodermal differentiation demonstrating that function is highly conserved. This approach of ectopic expression using an inducible promoter provides an effective way in which to use C. elegans for the in vivo functional analysis of parasitic nematode genes.

Amino Acid Sequence↗

PubMed2004

Genomic organization of an avermectin receptor subunit from Haemonchus contortus and expression of its putative promoter region in Caenorhabditis elegans.

Avermectins and milbemycins are believed to exert their anthelmintic effects by binding to glutamate-gated chloride channels (GluCls). Two GluCl subunits have been localized in the pharynx in Caenorhabditis elegans, and the pharynx has been implicated as a major target for avermectins in C. elegans. However, in parasitic nematodes, the pharyngeal localization of the GluCl subunits needs to be determined. The HcGluCla gene encoding an alpha-type GluCl subunit has been cloned from Haemonchus contortus previously. To investigate the expression site of the HcGluCla gene we have isolated a 1439bp 5'-flanking region and determined the genomic organization of this gene. The HcGluCla gene is composed of 12 exons separated by 11 introns and spans approximately 7.3kb of genomic DNA. Analysis of the 1439bp 5'-flanking region of the HcGluCla gene revealed that it contained TATA, CCAAT boxes, and several other consensus transcriptional factor recognition sequences. The 1439bp 5'-flanking region and the first exon and intron and part of the second exon of the HcGluCla gene were fused to green fluorescence protein (GFP) reporter gene and microinjected into the gonads of C. elegans. After microinjection of the construct into C. elegans, four stable transformed lines were established and assayed for GFP expression. The transformed animals exhibited fluorescence in the two pairs of MC and M2 pharyngeal neurons, but no expression was detected in the muscle cells. Expression of HcGluCla in pharyngeal neurons suggests a mechanism for the effects of avermectins and milbemycins on pharyngeal function in parasitic nematodes.

3' Untranslated Regions↗

PubMed2004

mRNA sequences for Haemonchus contortus intestinal cathepsin B-like cysteine proteases display an extreme in abundance and diversity compared with other adult mammalian parasitic nematodes.

Cathepsin B-like cysteine protease (cbl) genes produce the most abundant mRNAs ( approximately 16%) detected in the adult female intestine of the parasitic nematode Haemonchus contortus. CBL enzymes appear to digest host proteins and are vaccine candidates for immune control of H. contortus and potentially other parasitic nematodes. Hence, it is important to quantify the extent of diversity of H. contortuscbl genes. Here, expressed sequence tags (ESTs) were used to assess both the size and diversity of the H. contortuscbl gene family. Contig analysis of 686 cbl ESTs from a USA isolate resolved 123 clusters. ESTs were grouped into discrete sets and analyzed using an additive model. Discovery of new cbl clusters increased with each set and reached a terminal rate of about 1 per 10 ESTs. The extreme diversity was unique to cbls relative to other genes investigated and was ascribed to specific cbl clades. Sixty percent of cbl clusters from a UK isolate were shared with those identified in the USA isolate, suggesting conservation of cbl gene repertoires across regions, although minor to moderate geographic variation cannot be excluded. Sequence comparisons also suggested high potential for antigenic diversity among CBL proteins, which is relevant to vaccine strategies. Compared to other parasitic nematodes of mammals, the extreme abundance and diversity of intestinal cbl transcripts appear to be relative specializations for H. contortus. Therefore, adaptations related to nutrient acquisition may vary markedly among these parasitic nematodes.

Amino Acid Sequence↗

PubMed2005

RNA interference in Haemonchus contortus: suppression of beta-tubulin gene expression in L3, L4 and adult worms in vitro.

We have used RNAi to target two beta-tubulin genes in the parasitic stages of Haemonchus contortus in vitro. Soaking exsheathed-L3, L4 and adult worms in medium containing dsRNA resulted in a significant decrease (greater than 1000-fold in some cases) in the expression of the specific beta-tubulin transcript, as measured by quantitative PCR. During the initial 24h exposure to the dsRNA, the gene suppression effect was quite specific to the targeted gene. Six days after initial exposure to dsRNA, treated L3 worms were less able to migrate through a filter mesh, indicating decreased motility, and showed less development to the L4 stage than control larvae. The gene suppression effect occurs in exsheathed L3 larvae despite the fact that this life stage does not have functioning mouthparts, indicating that uptake of dsRNA does not depend on its ingestion. Suppression occurred with dsRNA presented in 'naked' or liposome-encapsulated forms, indicating that a liposome formulation was not necessary for uptake to occur. Adult worms also showed significant gene suppression, however, they did not show any reduced motility compared to controls over a 3-day period. Adult worms treated with ivermectin to paralyse their pharynx, still showed significant gene suppression, again suggesting that uptake of dsRNA does not require ingestion. We have shown that soaking in dsRNA is an effective method for RNAi with the parasitic stages of H. contortus, and, hence, may offer significant potential as a tool for studying gene function in this parasite species.

Animals↗

PubMed2006

Caenorhabditis elegans ivermectin receptors regulate locomotor behaviour and are functional orthologues of Haemonchus contortus receptors.

The target site for the anthelmintic action of ivermectin is a family of nematode glutamate-gated chloride channel alpha subunits (GluClalpha) that bind the drug with high affinity and mediate its potent paralytic action. Whilst the action of ivermectin on the pharyngeal muscle of nematodes is relatively well understood, its effect on locomotor activity is less clear. Here we use RNAi and gene knockouts to show that four GluClalpha subunits are involved in regulating the pattern of locomotor activity in Caenorhabditis elegans. A Haemonchus contortus orthologue of these subunits, HcGluClalpha3, has been shown to be expressed in the motor nervous system and here we have shown that it is a functional, as well as a structural, orthologue by virtue of the observation that it can restore normal motor movement in the C. elegans GluClalpha mutant, avr-14(ad1032), when expressed under the control of the avr-14 promoter. This supports the contention that ivermectin exerts its paralytic action on parasitic nematodes through activation of GluCl channels in the motor nervous system. Furthermore, functional complementation in C. elegans provides a method to further the understanding of this important class of anthelmintic targets.

Animals↗

PubMed2006

Ivermectin selection on beta-tubulin: evidence in Onchocerca volvulus and Haemonchus contortus.

Ivermectin resistance is common in trichostrongylid nematodes of livestock, such as Haemonchus contortus. This anthelmintic is the only drug approved for mass administration to control onchocerciasis caused by the nematode parasite, Onchocerca volvulus. In parts of West Africa up to 18 rounds of ivermectin treatment have been administered to communities and there are reports of poor parasitological responses to treatment. Understanding ivermectin resistance and ivermectin selection is an important step to reduce selection pressure for resistance, and to develop molecular markers which can be used to monitor the development of resistance and its spread. Here we report evidence that ivermectin selection changes the frequency of beta-tubulin alleles in both the sheep parasite, H. contortus, and the human parasite, O. volvulus. In O. volvulus we have been able to look at the frequency of beta-tubulin alleles in O. volvulus obtained before any ivermectin was used in humans in Africa, and following its widespread use. In H. contortus, we have been able to look at the frequency of beta-tubulin alleles in a strain which has not seen any anthelmintic selection and in an ivermectin selected strain derived from the unselected strain. We have found ivermectin selects on beta-tubulin in both of these nematode species. In the case of O. volvulus, we had previously reported that ivermectin selects for specific single nucleotide polymorphisms in the O. volvulus beta-tubulin gene. This polymorphism results in three amino acid changes in the H3 helix of beta-tubulin, as well as deletions in an associated intron. We report a simple PCR assay to detect the amplicon length polymorphism, resulting from these intronic deletions, which can be used to monitor the frequency of the beta-tubulin allele selected for by ivermectin in O. volvulus.

Africa, Western↗

PubMed2006

RNAi in Haemonchus contortus: a potential method for target validation.

RNA interference (RNAi) is a method for the functional analysis of specific genes, and is particularly well developed in the free-living nematode Caenorhabditis elegans. There have been several attempts to apply this method to parasitic nematodes. In a recent study undertaken in Haemonchus contortus, Geldhof and colleagues concluded that, although a mechanism for RNAi existed, the methods developed for RNAi in C. elegans had variable efficacy in this parasitic nematode. The potential benefits of RNAi are clear; however, further studies are required to characterize the mechanism present in parasitic nematodes, and to improve culture systems for these nematodes to monitor the long-term effects of RNAi. Only then could RNAi become a reliable assay of gene function.

Animals↗

PubMed2005

Lack of effects of quebracho and sainfoin hay on incoming third-stage larvae of Haemonchus contortus in goats.

The effects of tannins on adult populations of Haemonchus contortus, Trichostrongylus colubriformis and Teladorsagia circumcincta in goats are characterised mainly by a decrease in egg excretion without any significant changes in worm number. In contrast, the impact of tannins on T. colubriformis or T. circumcincta third-stage larvae (L3) is associated with a significant reduction in worm establishment. The objective of the present study was to examine the effects of quebracho extract tannins on H. contortus L3. The consequences of consumption of sainfoin hay were also examined. Twenty-one naïve kids were divided into three experimental groups. Group Q received quebracho extract and group S received sainfoin hay from days D3 to D5. Group C remained as an infected control group. All kids received 1500 L3 H. contortus on D0, D1 and D2. On D18, post-infection, the kids were slaughtered and the worm populations compared in the different groups. Compared to the control values, the worm counts decreased, respectively, by 33% and 38% in groups Q and S but the differences were not significant. No differences were found in pathophysiological measurements between the three groups. The results confirm differences in tannin effect according to nematode species but not parasitic stage.

Anacardiaceae↗

PubMed2004

Vaccination against the nematode Haemonchus contortus with a thiol-binding fraction from the excretory/secretory products (ES).

Fractionated excretory/secretory products (ES) of adult Haemonchus contortus were evaluated as protective antigens. The proteins were successively eluted from a Thiol Sepharose column using 25 mM cysteine followed by 25 mM Dl-dithiothreitol (DTT). Sheep were vaccinated three times and challenged with 5000 third stage infective larvae (L3) of H. contortus. Highest level of protection was found in sheep vaccinated with the DTT-eluted fraction in which egg output and worm burden were reduced by 52 and 50%, respectively, compared to the adjuvant control group. There was a positive correlation between fecundity (number of eggs per female) and the cumulative EPG or worm burden. Serum and mucus antibody levels of ES-specific immunoglobulins increased after immunizations and after challenge for IgG, IgA and IgE. The harvesting of H. contortus from animals clustered per group revealed the presence of cysteine protease activity in the ES of all groups but in addition to that, metalloprotease activity was also detected in the groups vaccinated with the DTT-eluted fraction, total ES and adjuvant only, in contrast to previous batches of ES (completely inhibited by E64) obtained from non vaccinated animals.

Abomasum↗

PubMed2004

Protection studies in sheep using affinity-purified and recombinant cysteine proteinases of adult Haemonchus contortus.

Vaccination with a membrane-bound thiol Sepharose-binding fraction (TSBP) of adult Haemonchus contortus has been shown to confer significant levels of protection against homologous challenge in sheep. This fraction is greatly enriched for cysteine proteinase activity. Following fractionation of TSBP by anion-exchange chromatography on MonoQ, protection was found to partition with those fractions further enriched for cysteine proteinase activity. In this study, the cysteine proteinases of adult H. contortus TSBP were specifically purified by affinity chromatography using recombinant H. contortus cystatin, a potent cysteine proteinase inhibitor. Although only 1-1.5% of total TSBP bound to cystatin-Sepharose, this fraction contained 100% of the cysteine proteinase activity, as determined by gelatin substrate gel analysis. When used to immunise sheep, less than 3microg per dose of this cysteine proteinase fraction was found to confer a substantial and repeatable level of protection against homologous challenge infection, reducing faecal egg counts by 48 and 28% and worm burdens by 44 and 46% over two trials. Host serum immunoglobulin levels and abomasal mast cell and eosinophil numbers were evaluated, although no correlation with protection was observed. Three cathepsin B-like cysteine proteinases present in TSBP (hmcp1, 4 and 6) have been identified previously by cDNA library immunoscreening. The predicted mature forms of these three cysteine proteinases were expressed in bacteria as insoluble, GST-fusion proteins. Following solubilisation in urea/DTT, the protective capacity of a cocktail of recombinant proteins was evaluated in sheep. Although no reduction in faecal egg counts was observed, sheep vaccinated with recombinant cysteine proteinases showed a highly significant 38% reduction (P <0.01) in worm burdens.

Animals↗

PubMed2003

Ovicidal and larvicidal activity in vitro of Spigelia anthelmia Linn. extracts on Haemonchus contortus.

The rapid development of anthelmintic resistance, associated with the high cost of the available anthelmintic drugs, had limited the success of gastrointestinal nematodiasis control in sheep and goats and thus awakened interest in the study of medicinal plants as alternative sources of anthelmintics. Spigelia anthelmia extracts obtained with hexane, chloroform, ethyl acetate or methanol, were tested on Haemonchus contortus eggs and larvae via egg hatch and larval development tests. The extracts were evaluated at five concentrations: 3.1, 6.2, 12.5, 25.0 and 50.0 mg ml(-1). At 50.0 mg ml(-1), the ethyl acetate extract inhibited 100% of the egg hatching and 81.2% of the larval development. In a similar way the methanolic extract inhibited 97.4% of the egg hatching and 84.4% of larval development. These results suggest that utilization of S. anthelmia extracts may be useful in the control of sheep and goats gastrointestinal nematodes.

Animals↗

PubMed2004

High genetic correlation between resistance to Haemonchus contortus and to Trichostrongylus colubriformis in INRA 401 sheep.

To compare the protection acquired by infection with Haemonchus contortus or Trichostrongylus colubriformis, successive cross infections with the two species were conducted in two genetically similar groups of 150 ewe lambs from 30 sires of INRA 401 breed. At 6 and 7.5 months of age, each group received two doses of 10000 L3 separated by a treatment of one of the species, followed by a similar infection with the second species at 9 and 11.5 months. Faecal egg counts (FECs) were done 28 and 35 days after each dose. There were very high genetic correlation within and between species suggesting that the environmental conditions were very similar. The heritability of FEC of H. contortus ranged from 0.39 to 0.48 and the genetic correlations between FEC after the first and second infections with the same or different species were all near 1. Selection against this species could be done on the response to only one dose. For T. colubriformis, only the response to challenge infection had a similar heritability (0.47); the genetic correlations within and among species were also near one. These results will assist the selection of sheep resistant to nematode parasites.

Animals↗

PubMed2004

Effect of dexamethasone treatment on the immune response of Gulf Coast Native lambs to Haemonchus contortus infection.

Neonatal and weaner Gulf Coast Native (Native) lambs were studied to determine whether an immunological basis underlies their natural resistance to Haemonchus contortus infection. Neonatal Native lambs (n = 8) and weaner Native lambs (n = 15) were randomly assigned to a treatment or a control group. Lambs in the treatment group received dexamethasone by intramuscular injection three times a week for 10 weeks (neonatal) and 15 weeks (weaners). All lambs were monitored for fecal egg count (FEC), blood packed cell volume (PCV), and white blood cell differential counts on a weekly basis for the duration of the studies. Neonatal lambs were kept on pasture with their dams and weaner lambs were dewormed at weaning and kept in pens where they received trickle infections. Serum antibody titers to H. contortus whole worm antigen (WWA) were determined using ELISA. Lymphocyte proliferation assays on peripheral blood mononuclear cells (PBMC) were done to assess lymphocyte function. All lambs were vaccinated with killed Brucella abortus strain 19 to assess the effect of dexamethasone treatment on antibody response. All lambs were necropsied at the end of each study to recover the contents of the gastrointestinal tract for nematode enumeration and identification. The results showed that mean FEC and mean PCV of the treatment group was significantly higher and lower, respectively, than in the control group in both neonatal and weaner lambs from weeks 6 and 5, respectively. At necropsy, total nematode count was significantly higher in treatment groups than in the control groups. Serum antibody titers to H. contortus WWA were significantly lower in treated groups than in control groups. Treatment groups showed a consistent depression in lymphocyte percentage being significantly lower from week 6 in both neonatal and weaner lambs. No differences were found in the response of PBMC to mitogen stimulation between the groups. Lambs in the control groups showed strong positive brucellosis card tests and the treatment groups did not. Dexamethasone treatment resulted in depression of the immune response and loss of natural resistance of Native lambs to H. contortus infection. The results of these studies suggest that some aspects of the immune response may underlie the natural resistance of Native sheep to H. contortus infection.

Animals↗

PubMed2004

Variability in the resistance of the Nigerian West African Dwarf goat to abbreviated escalating trickle and challenge infections with Haemonchus contortus.

The West African Dwarf (WAD) goat is known to be relatively resistant to infection with Haemonchus contortus. An experiment was carried out to assess variability in resistance under escalating trickle and single pulse challenge infection protocols. Thirty two 7-8 month old goats were subjected to trickle infection beginning with a thrice weekly dose of 100 L3 and increasing to 500 L3 by week 5, the last dose being administered on D30. Infection was abbreviated with fenbendazole on D32 and the animals challenged with 4000 L3 (=404-741L3/kg body weight) on D46 and necropsied on D81. The goats exhibited marked individual variability in faecal egg counts (FEC) and worm burdens (Wb). For convenience, we arranged the goats into FEC classes 1, 2 and 3, corresponding to peak counts of <1000, 1000-2500 and >2500 epg and Wb classes 1, 2 and 3, with counts of <50, 50-100 and >100 worms, respectively, broadly reflecting relatively resistant, intermediate and poorly resistant response phenotypes. Wb classes broadly reflected the FEC phenotypes and although there were no significant differences between the Wb of the FEC classes at necropsy, significant differences in FEC were detected when analysis was carried out only on Wb classes. There was also a significant positive correlation between FEC class on each of the days of challenge (D67 to D80) and worm burden at necropsy. In general, the goats showed strong resistance/tolerance to challenge, with more than 80% having Wb not exceeding 5% of the challenge dose, and only in class three goats were weight loss and anaemia detected in some animals. PCV and weight changes also closely reflected the FEC and Wb classes, with a significant negative correlation between each and Wb. Infection level did not significantly affect the sizes of either male or female worms and no significant difference was detected between the levels of circulating eosinophils in any FEC or Wb class. Overall, the data are consistent with the existence of a range of response phenotypes, which reflect variations in resistance of WAD goats to H. contortus infection, the genetic basis of which will be explored in future experiments.

Animals↗

PubMed2004

Effect of copper oxide wire particles dosage and feed supplement level on Haemonchus contortus infection in lambs.

The objective of the experiment was to determine the optimal dose of copper oxide wire particles (COWPs) to reduce infection of Haemonchus contortus in male lambs. Five to six-month-old hair breed lambs were housed on concrete and fed 450 (L; n = 25) or 675 g (H; n = 25) corn/soybean meal supplement and bermudagrass hay. In July, lambs were inoculated with 10,000 L(3) larvae (97% H. contortus; Day 0). Lambs were administered 0, 2, 4, or 6 g COWP on Day 28. Concentrations of copper in the liver were determined. There were no effects of supplement level on concentrations of copper in the liver and a linear relationship existed between COWP treatment and concentrations of copper in liver (P < 0.001). Least squares means of the 0, 2, 4, 6 g COWP treatments were 62.2, 135.7, 161.1, and 208.4 ppm (P < 0.001). Between Days 0 and 28, PCV declined and by Day 42, PCV of all COWP-treated lambs was markedly higher than control lambs and remained higher (COWP x day, P < 0.05). By Day 21, PCV was greater in the H compared with the L group of lambs (P < 0.001). Within 14 days of COWP treatment FEC declined from more than 8000 eggs/g (epg) to less than 250 epg in all COWP-treated lambs (COWP x day, P < 0.001). The numbers of H. contortus in the abomasum were greatly reduced in all COWP-treated groups of lambs and remaining nematodes were predominantly males. FEC were greater in L versus H supplemented lambs and values decreased to a greater extent in H lambs when treated with COWP (supplement x COWP; P < 0.02). The 2 g COWP was effective in alleviating H. contortus infection and reducing number of egg-laying nematodes in the abomasum with the lowest concentration of copper in the liver of the COWP treatment groups. PCV values were more favorable for lambs fed the higher level of supplement, especially when FEC were greater than 8000 epg.

Abomasum↗

PubMed2004

Characterization of moxidectin resistant Trichostrongylus colubriformis and Haemonchus contortus.

The development of moxidectin resistance (MOX-R) in sheep parasitic gastrointestinal nematodes already carrying multiple resistances to other anthelmintic groups has made control of these strains very difficult. The anthelmintic resistance patterns of MOX-R strains of Trichostrongylus colubriformis and Haemonchus contortus were characterized to provide an insight into the remaining role of anthelmintics in the control of such strains. Homozygous MOX-R individuals of both genera were unaffected by moxidectin. For MOX-R heterozygotes a dose rate of 200 microg/kg abamectin (ABA) given orally removed 25% of H. contortus while 200 microg/kg MOX given orally achieved a 72% reduction. Doubling the dose rate of ABA improved the mean efficacy to 37%. Consequently, in H. contortus, the degree of dominance differs markedly between the two anthelmintics. A dose rate of 8 mg/kg levamisole and 185 mg/kg napthalophos achieved >95% reduction in worm count of the MOX-R homozygous H. contortus but only 85 and 7%, respectively against the MOX-R homozygous T. colubriformis.

Abomasum↗

PubMed2005

The modulatory influence of Trypanosoma brucei on challenge infection with Haemonchus contortus in Nigerian West African Dwarf goats segregated into weak and strong responders to the nematode.

Although Nigerian West African Dwarf (WAD) goats are relatively resistant to infection with Haemonchus contortus and are also trypanotolerant, natural outbreaks of both infections are known to occur. Despite their relative resistance to H. contortus WAD goats nevertheless show variability in response phenotype and it was of interest to examine the effect of this variability on the outcome of concurrent trypanosome infection. Trypanosoma brucei infections were established in goats that were initially classified as good or poor responders to H. contortus. Thirty-nine goats were exposed to an escalating infection with H. contortus, and on the basis of their mean faecal egg counts (FEC) were allocated to high FEC (poor responders, 18 goats with the highest FEC) or low FEC (good responders, 18 goats with the lowest FEC) classes. Nine uninfected naive control goats were included to provide reference baseline values. Retrospective analysis of parasitological and pathological parameters after allocation into high/low FEC classes showed that FECs differed significantly, in both classes packed cell volume (PCV) values fell relative to naive controls, neither class lost weight and both generated marked IgG responses. All goats received anthelmintic on day 61, half of each group was infected with 50 million trypanosomes and on day 67, excepting the controls, all goats were challenged with 3000 L3 of H. contortus. Trypanosome parasitaemia was generally low, and marginally, but not significantly, higher in the low compared with high FEC class, peaking 12-16 days after exposure in both groups and then falling to below microscopically detectable levels (although still detectable by sub-inoculation into mice) by week 3. At autopsy (days 109/110), worm burdens were significantly higher in the trypanosome-infected goats from the high FEC class, relative to all other groups. Trypanosome infected goats showed a tendency (although not significant) towards higher FEC and, irrespective of their FEC class, had lower PCV values although body weight did not vary significantly. All goats challenged with H. contortus had higher antibody levels than naive controls, but neither trypanosome infection nor FEC class affected the magnitude of responses. These results confirm that WAD goats comprise a range of response phenotypes to initial H. contortus infection and that trypanotolerance is a key trait of this breed. Although immunity to nematode infection develops even in poor responders, these animals harbour higher nematode burdens during concurrent infection with T. brucei.

Animals↗

PubMed2004

Relationship of abomasal histology and parasite-specific immunoglobulin A with the resistance to Haemonchus contortus infection in three breeds of sheep.

This study was carried out to evaluate the relationship of abomasal inflammatory cells and parasite-specific immunoglobulin A (IgA) in mucus, with the resistance to Haemonchus contortus infection in three breeds of sheep naturally infected with gastrointestinal nematodes. The breeds were the native Santa Ines sheep, and the European Suffolk and Ile de France breeds. Mast cells, eosinophils and globule leucocytes were enumerated in abomasal mucosa. Eosinophils within the sub-mucosa also were counted separately. Histamine concentration was estimated in abomasal tissue samples. Enzyme-linked immunosorbent assay was carried out in mucus samples to determine the level of IgA anti-H. contortus third and fifth instar. There were no significant differences among group means of these variables (P>0.05). The correlation coefficients between fecal egg counts (FEC)xmast cells (r=-0.490; P<0.05) and FECxeosinophils in sub-mucosa (r=-0.714; P<0.01) was significant in the Santa Ines sheep. In the Ile de France group, the correlation coefficients between globule leucocytesxFEC (r=-0.879; P<0.001) and histaminexworm burden (r=-0.833; P<0.01) were also significant. In the Santa Ines and Ile de France sheep, correlation coefficients between IgA anti-L3xworm burden and IgA anti-L3xFEC were negative. In general, inflammatory cells and IgA-parasite-specific in abomasum were inversely associated with H. contortus worm burden and FEC indicating that they may impair parasite development or fecundity in the three breeds of sheep. However, similar mean values of inflammatory cells and IgA were found in the resistant (Santa Ines) and in the susceptible (Suffolk and Ile de France) breeds of sheep. The enumeration of cells by histological assessment does not provide information on their functional activity, which may be different among breeds. Thus, the effect of breed on the functional activity of these and other inflammatory cells is an important area for further study.

Abomasum↗