Phylogenetic analysis of Trichostrongylus vitrinus isolates from southwest Iran.

BACKGROUND: Trichostrongylus is one of the most important zoonotic trichostrongylid nematodes, infecting mostly livestock. Data on its genetic characteristics are lacking in Iran. METHODS: We determined the phylogenetic relationships of Trichostrongylus species in three counties of Kohgiloyeh and Boyerahmad (K-B) province, southwest Iran. Small intestine and abomasum of 70 sheep and goats were investigated. RESULTS: A total of 35 isolates of Trichostrongylus worms were detected and all were genetically identified as Trichostrongylus vitrinus. Analysis of 321 bp of the internal transcribed spacer 2 (ITS2) of ribosomal DNA revealed 16 genotypes. All genotypes were single nucleotide polymorphisms, including some hypervariable points. All sequences were trimmed to 170 bp, compared with sequences on GenBank including short sequences from other endemic foci of Iran and other countries and all isolates were used to generate a maximum likelihood phylogenetic tree, which consisted of two clades A and B. Clade A included isolates from Iran, Russia, New Zealand, Australia and the UK; clade B only contained South African isolates. Most clade A isolates (north, southwest and west Iran, Russia, New Zealand, Australia and UK) were in a similar phylogenetic position. One subclade was detected in clade A (isolates from Southwest Iran, New Zealand and UK). CONCLUSIONS: We hypothesize that drug resistant T. vitrinus may account for its exclusive detection in our samples. The high similarity of genotypes from Iran, New Zealand and UK may be due to their close political relationships during the colonial era. More research is needed to understand better the phylogeny of T. vitrinus and its relationship with drug resistance and human transmission.

First Molecular Characterization of Trichostrongylus colubriformis Infection in Rural Patients from Chile.

PURPOSE: The aim was to characterize the infection by Trichostrongylus spp. in patients from Chile using a combination of molecular detection techniques and phylogenetic analysis relating the findings to clinical and epidemiological reports of the patients METHODS: Strongylid eggs were detected in seven patients by coproparasitological techniques. From each sample a fragment of the ITS-2 ribosomal gene was amplified by PCR, sequenced and analyzed by the Neighbor-Joining method. RESULTS: All the sequences and phylogenetic clusters corresponded to T. colubriformis. Two samples presented a single nucleotide polymorphism showing two possible haplotypes. Six patients presented gastrointestinal symptoms. All of them lived on farms and used sheep manure as fertilizer. CONCLUSION: T. colubriformis was the strongylid involved in the infections of these Chilean patients associated with the presence of livestock and agricultural practices that favor infection by this type of nematode.

Molecular detection of Trichostrongylus species through PCR followed by high resolution melt analysis of ITS-2 rDNA sequences.

Polymerase chain reaction followed by high resolution melting (PCR-HRM) analysis is a simple, rapid and accurate method for molecular detection of various nematode species. The objective of the present study was, for the first time, to develop a PCR-HRM assay for the detection of various animal Trichostrongylus spp. A pair of primers targeting the ITS-2 rDNA region of the Trichostrongylus spp. was designed for the development of the HRM assay. DNA samples were extracted from 30 adult worms of Trichostrongylus spp., the ITS-2-rDNA region was amplified using PCR, and the resultant products were sequenced and characterized. Afterwards, the PCR-HRM analysis was conducted to detect and discriminate Trichostrongylus spp. Molecular sequence analysis revealed that 24, 4, and 1 of the samples were T. colubriformis, T. vitrinus and T. capricola, respectively. Results from PCR-HRM indicated that complete agreement was relatively found between speciation by HRM analysis and DNA sequencing for the detection of Trichostrongylus species. The PCR-HRM analysis method developed in the present study is fast and low-cost; the method can be comparable with other molecular detection techniques, representing a reliable tool for the identification of various species within the Trichostrongylus genus.

Effects of Acacia mearnsii supplementation on nutrition, parasitological, blood parameters and methane emissions in Santa Inês sheep infected with Trichostrongylus colubriformis and Haemonchus contortus.

Gastrointestinal nematodes are a main problem for ruminant production, reducing animal performance and increasing environmental impact per unit of animal product generated. Tannin supplementation may lead to positive results regarding aspects such as parasitic infections and methane (CH4) emissions. Therefore, the objective of this experiment was to evaluate the effects of the condensed tannins (CT) extract made of powdered Acacia mearnsii bark (PAB) on nutrition, parasitic status and CH4 emissions in sheep artificially infected with Trichostrongylus colubriformis and Haemonchus contortus. Twenty 10-month old Santa Inês lambs (24.7 ± 3.14 kg of initial body weight) were used in a 50-day trial. Animals were divided in four treatment groups according to parasitic infection and PAB supplementation: two control groups without infections, one without PAB (C-) (n = 4) and one with PAB (C+) (n = 4); two infected groups, one without PAB (I-) (n = 6) and another receiving PAB (I+) (n = 6). Initially, animals were kept in individual pens where they were fed ad libitum chopped tifton 85 hay (Cynodon spp.) and 210 g/animal/day of concentrate. On the first day of experiment, animals of I- and I+ groups were artificially infected with infective larvae (L3) of T. colubriformis and H. contortus. Lambs were weighed fortnightly to calculate average daily body weight gain (ADG). Blood and faeces samples were also collected in the same moment of weighing for the evaluation of blood parameters and faecal egg count (FEC) respectively. After 40 days of experiment, measurements of CH4 emissions in small chamber system started and following that, apparent total tract digestibility (ATTD) assay was carried out in metabolic cages. In the end of experimental period (50 days), lambs were slaughtered and samples of abomasum and small intestine content were collected for worm count, identification, and eggs/female count. No significant (p > 0.05) treatment effects were verified for ADG, ATTD and worm count. Blood parameters were affected in both infected groups (p < 0.05) from the 28th experimental day onwards, when these animals started to show reduced red blood cells, haemoglobin and packed cell volume when compared to C- and C+. Decreased FEC was verified in I+ when compared to I- and also, H. contortus eggs/female worm for I+ was lower than for I- (p < 0.05). Both infected groups showed higher CH4 emissions than the control groups (p < 0.05). Results highlighted the anthelmintic potential of PAB and indicated methanogenic effect of parasitic nematode infections.

Diversity and prevalence of gastrointestinal parasites in two wild Galago species in Gabon.

In this study, we characterize the diversity and estimated infection levels of gastrointestinal parasites circulating in two galago species, Galago demidoff and G. thomasi in two sites situated in the Southeastern forests of Gabon. Our study reveals that eleven parasites including nine helminthes (Ascaris spp., Ankylostoma spp., Dicrocoelium spp., Gongylonema spp., Oesophagostomum spp., Lemuricola spp., Strongyloides spp. Trichostrongylus spp. and Trichuris spp.) and two protozoans (Balantidium spp. and Entamoeba spp.) may infect Galago spp. with high infection rates. The results show that: a very similar parasite spectrum is found in both host species; all the taxa identified were previously observed in other Primate species and/or Man. They also show that age, gender and forest type may influence infection rates and/or parasite diversity found in a particular host and/or geographic area.

Assessment of Real-Time Polymerase Chain Reaction for the Detection of Trichostrongylus spp. DNA from Human Fecal Samples.

Sporadic cases of Tricostrongylosis are reported in humans. Diagnosis of enteric Trichostrongylus relies primarily on coproscopic analysis but morphological identification is difficult because of similarity among nematode species. The method is time consuming and requires some expertise. To overcome these limitations, we developed a molecular approach by real-time polymerase chain reaction (PCR) to provide a rapid, specific, and sensitive tool to detect Trichostrongylus spp. in human feces. We designed primers and probe specific for Trichostrongylus rDNA region 5.8S and internal transcribed spacer 2. Three Italian family clusters were analyzed and DNA sequencing was performed to confirm real-time PCR results comparing with known GenBank sequence data. Sequence analysis showed ≥ 99% identity to Trichostrongylus colubriformis and Trichostrongylus axei. This study provides a molecular methodology suitable for fast and specific detection of Trichostrongylus in fecal specimens and to distinguish the zoonotic species.

Persistent challenge with Trichostrongylus colubriformis and Haemonchus contortus larvae does not affect growth of meat-breed lambs suppressively treated with anthelmintics when grazing.

This experiment tested the hypothesis that persistent challenge with anthelmintic susceptible Trichostrongylus colubriformis and Haemonchus contortus larvae would not affect growth of grazing, meat-breed lambs when suppressively treated with anthelmintics. The experiment was a 2×2 factorial design using 6-7 months old White Suffolk X Border Leicester/Merino (meat-breed) lambs which were either infected with 2000 T. colubriformis and 300 H. contortus L3/week (IF) or remained uninfected (UIF) for 9 weeks and were either treated (TX) with a combination of short and long-acting anthelmintics or remained untreated (UTX). Lambs grazed as one flock and were rotated between paddocks to avoid autoinfection from pasture. Lambs were humanely euthanised on day 63 and the abomasum and small intestine collected to determine total worm burdens and tissue antibody response specific to T. colubriformis. As expected, worm egg count (WEC) and worm burden were significantly higher in IF UTX lambs (p<0.001). WEC was dominated by H. contortus and peaked at 2,325 epg on day 63 but remained at zero for the other treatment groups for the duration of the experiment. Tissue antibody responses were evident in IF lambs (titres; 9982 vs 2767, p=0.012) but treatment had no effect (titres; 5912 vs 5349, p=0.829). Lambs grew an average of 2.6 kg during the experiment with no difference between IF TX and UIF TX groups (p=0.432). Elevated tissue antibody responses were not associated with differences in growth. Results from this experiment support the hypothesis that persistent larval challenge with anthelmintic susceptible H. contortus and T. colubriformis will not affect growth of grazing, meat-breed lambs when suppressively treated with effective anthelmintics. Therefore the use of sheep suppressively treated with effective anthelmintics appears to be a valid substitute for gastrointestinal nematode-free lambs in field experiments.

Simultaneous infection by four feline lungworm species and implications for the diagnosis.

Besides Aelurostrongylus abstrusus, other parasites belonging to the superfamily Metastrongyloidea, namely Oslerus rostratus, Troglostrongylus brevior and to the family Trichuridae, i.e. Eucoleus aerophilus (syn. Capillaria aerophila), have also been reported as agents of respiratory infection in domestic cats. A case of simultaneous infection by four feline lungworm species in Sardinia is herein described. An adult female cat (Felis silvestris catus), road-killed in the southeast part of Sardinia (municipality of Villacidro, province of Cagliari), Italy, was referred to the Laboratory of Parasitology of the Veterinary Teaching Hospital in Sassari. At necropsy, the lungs were examined and dissected under a stereomicroscope for the presence of parasites, and first-stage larvae (L1) of broncho-pulmonary nematodes were searched for in a faecal sample using the Baermann method. Parasites collected in the lungs were morphologically identified as A. abstrusus, E. aerophilus, and O. rostratus. In addition to the above species, L1s of Troglostrongylus spp. were detected at coproscopy but no adult specimen was found in the lungs. The morphological identification was confirmed by the molecular amplification and sequencing of cox1 mitochondrial gene, 18S and ITS2 ribosomal DNA. This finding stands as the first simultaneous infection by four feline lungworm species in the same animal, and as the first report of O. rostratus and E. aerophilus in Sardinia.

The distribution and anthelmintic resistance status of Trichostrongylus colubriformis, T. vitrinus and T. axei in lambs in New Zealand.

AIM: To determine the distribution of the three common Trichostrongylus spp. infecting sheep and their resistance status on farms throughout New Zealand, using PCR. METHODS: Cultures were prepared from faecal samples from 70 farms while conducting faecal egg count reduction tests (FECRT) in lambs between 2010 and 2012. Trichostrongylus-type infective stage larvae (L3) were recovered from cultures, derived from untreated control and albendazole-, levamisole- and ivermectin-treated groups of lambs on each of the farms involved, and these were identified to species using PCR analysis of the second internal transcribed spacer region of ribosomal DNA. The species composition of the larvae present in cultures from the untreated control groups was examined across all farms to assess any potential differences in geographical distribution. In addition, the species composition of larvae cultured from the untreated and anthelmintic-treated lamb groups were compared to determine which species exhibited resistance to each of the anthelmintics used in the FECRT. RESULTS: Of 67 farms with Trichostrongylus spp. present, 42 (63%) cultures from the untreated control groups contained all three Trichostrongylus spp. and no significant geographical patterns in their distribution were detected. Seven samples contained only one species. Irrespective of the anthelmintic efficacy levels, Trichostrongylus colubriformis dominated cultures prepared from lambs following treatment with albendazole (99.1 (95%CI = 97-100)% of larvae) or levamisole (81.6 (95%CI = 75.3-87.9)% of larvae), indicating the presence of widespread resistance in this species. In cultures prepared from levamisole-treated lambs, small numbers of T. axei larvae were also frequently present (5.4 (95% CI = 1.3-12.4)% of larvae). Resistance to ivermectin was not found in any of the three Trichostrongylus spp. after PCR identification. Although larvae were identified, based on length, as being Trichostrongylus spp., for 24 of the 48 samples cultured following treatment with ivermectin, 100% of the larvae present were identified as Teladorsagia circumcincta. CONCLUSIONS: As in previous surveys, all three Trichostrongylus spp. were common throughout New Zealand and no geographical patterns were detected in the current study. On all farms where resistance to albendazole and/or levamisole was indicated (i.e. efficacy <95%), the species identified as being resistant was T. colubriformis. Even where efficacies were >95%, T. colubriformis still tended to dominate in post-treatment cultures. While this could reflect a lower susceptibility of T. colubriformis to these anthelmintics, it seems more likely to indicate the presence of resistant genotypes in these populations. Similarly, T. axei also tended to be present after treatment with levamisole, which likely reflects a known lower susceptibility of this species to these anthelmintics.

Ovine nematodes in wild lagomorphs in Australia and first record of Trichostrongylus rugatus in free living lagomorphs.

Gastrointestinal parasites are a major problem for sheep graziers and anthelmintic resistance has been reported worldwide. Given that in experimental circumstances, sheep helminths are able to develop in rabbits and hares, and that hares are very mobile and sympatric with sheep in many regions of Australia, free-living lagomorphs were investigated regarding carriage of ovine nematode parasites under field conditions. We performed specific worm counts in the gastrointestinal tracts of 88 rabbits and 110 hares. We revealed that ruminant worms are common in hares (prevalence 79%) and that they are occasionally found in rabbits (9%). Statistical analyses showed that the ruminant worm Trichostrongylus colubriformis occurred frequently in hares whilst rabbits were commonly infected with lagomorph-specific Trichostrongylus retortaeformis. Detection of the ovine worm Trichostrongylus rugatus is reported for the first time in wild lagomorphs. The potential for cross-transmission between hares and sheep in the natural environment is much more prevalent than previously believed.

Molecular evidence of Trichostrongylus colubriformis and Trichostrongylus axei infections in humans from Thailand and Lao PDR.

Human trichostrongylosis has been reported in Thailand. Recent reports in Lao People's Democratic Republic concerning species identification urged us to investigate species distribution in Thailand. We report eight human cases in Thailand and Lao People's Democratic Republic that were found to be infected by Trichostrongylus colubriformis and T. axei identified and confirmed by molecular techniques. This evidence is the first molecular evidence of human T. colubriformis and T. axei infection in Thailand. Infection by these two species was apparently epidemic in these areas. It is necessary to proceed with more comprehensive veterinary and epidemiologic studies to enable the practical prevention and control of this parasitic zoonosis.

Morphological identification of parasitic nematode infective larvae of small ruminants and cattle: a practical lab guide.

In 2004, a new concept was introduced for simplifying identification of larvae of the common nematodes of cattle, sheep and goats that comprises estimates of the lengths of the sheath tail extensions of infective third-stage larvae (L3) of each genus and/or species to that of Trichostrongylus spp., instead of having to be dependent only on measurements in micrometre. For example, if the mean length of the sheath tail extension (the extension of the sheath caudad, beyond the caudal tip of the larva) of Trichostrongylus colubriformis and Trichostrongylus axei is assumed to be 'X', then that of Haemonchus contortus is 2.0-2.7 'X' - a difference that is not difficult to estimate. An additional new approach suggested now, particularly for L3 of species and/or genera difficult to differentiate (such as Chabertia ovina and Oesophagostomum columbianum), is to estimate the proportion of the larval sheath tail extension comprising a terminal thin, whip-like filament. For the experienced person, it is seldom necessary to measure more than one or two sheath tail extensions of L3 in a mixed culture, because the identity of most of the remaining L3 can thereafter be estimated in relation to those measured, without having to take further measurements. The aim of this article was to present the novel approach in the form of a working guide for routine use in the laboratory. To facilitate identification, figures and a separate organogram for each of small ruminants and cattle have been added to illustrate the distinguishing features of the common L3.

Changes in the body condition scores of Nigerian West African Dwarf sheep experimentally infected with mixed infections of Haemonchus contortus and Trichostrongylus colubriformis.

Weekly changes in body condition scores and body weights were determined in West African Dwarf sheep following trickle exposure to weekly escalating mixed infections with Haemonchus contortus and Trichostrongylus colubriformis starting with single doses of 500 infective larvae (L(3)) at wk1, 1000 L(3) at wk2, 2000 L(3) at wk3 and 4000 L(3) at wk4. Thirty-three animals were randomly assigned to two groups namely, Group 1 (n=27) and Group 2 (n=6). Group 1 animals were used for experimental infections and Group 2 animals served as uninfected controls. Body condition scores (BCS), body weights (BW), faecal egg counts (FEC) per gram of faeces and worm burden (Wb) were determined using standard procedures. Lambs in group 1 with epg ≤ 1000 were classified as strong responders (n=16), those with epg between 1000 and 10,000 as intermediate (n=5) and lambs with epg>10,000 on any sampling day were classified as weak responders (n=6). Based on this classification, the mean epg of lambs classified as strong responders was significantly lower (P<0.001) than the epg of lambs classified as weak responders. The body condition scores and body weights of the strong responder lambs were comparable with those of the uninfected control and they were significantly higher than those of the weak and intermediate responders from wk5 till the end of the experiment. There was an inverse and significant relationship between the average BCS of days 56 and 59 and the average FEC of days 56 and 59 (r(s)=-572; P=0.017; n=27) as well as with Wb (r(s)=-531; P=0.028; n=27). We concluded that changes in the body scores are good indicators of the intensity of gastrointestinal nematode infection in the Nigerian WAD sheep.

Anthelmintic efficacy of five tropical native Australian plants against Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats (Capra hircus).

The study of the anthelmintic properties of plants rich in plant secondary metabolites can provide ecologically sound methods for the treatment of parasites on grazing animals. The purpose of the present study was to evaluate the anthelmintic effect of five tropical native Australian plant species rich in plant secondary metabolites on adult Haemonchus contortus and Trichostrongylus colubriformis in experimentally infected goats. Thirty young, nematode-free goats were infected with 2500 H. contortus and 5000 T. colubriformis infective larvae thrice weekly for a week (day 1-7 of the experiment). On day 27 after first infection, the goats were allocated into six groups of five animals per group. From day 28 to day 35, fresh leaves from Acacia salicina, Acacia nilotica, Eucalyptus corymbia, Casuarina cunninghamiana and Eucalyptus drepanophylla were included in the goats diet. Five groups were offered leaves from one of these plant species and one group, the untreated control, received only the basal diet formulated with 20% Medicago sativa and 80% Avena sativa. Following plant material administration, the goats were monitored daily until day 40 and then slaughtered on day 41. Total faecal worm egg output, total production of larvae recovered from faecal cultures, total post-mortem worm burdens and the per capita fecundity of female worms were estimated. The toxicity of the plant species for the goats was measured by histopathological analyses of liver and kidney samples. Results showed that goats feeding on the plant material rich in plant secondary metabolites had significantly lower egg output compared to the control goats (P<0.05). A similar response was found for larval production in both H. contortus and T. colubriformis supporting that egg output was affected in both species. Although the total worm burdens were not affected by the plant material (P>0.05), the per capita fecundity was significantly reduced by E. corymbia, A. nilotica and A. salicina (P<0.05). No signs of toxicity were detected in the liver or kidney samples. It is concluded that goats can benefit from the short-term ingestion of plant secondary metabolites, which reduce the total faecal egg output and thus decrease the potential for re-infection from the pasture.

Immune response and performance of growing Santa Ines lambs to artificial Trichostrongylus colubriformis infections.

This study was carried out to evaluate the immune response and the impact of Trichostrongylus colubriformis infections on the performance of growing Santa Ines lambs. Thirty male lambs, 3-4 months of age, were maintained in individual pens and restrictively randomised by weight into 3 treatment groups: (1) infected group, artificially infected with 2500 T. colubriformis larvae, three times a week, for 13 weeks, and fed ad libitum; (2) Pair-Fed Group, non-infected and fed with the same amount of food consumed by the infected animal of the same class on the previous day; and (3) control group, non-infected and fed ad libitum. Refused feed was weighed daily to assess the food intake of each lamb. Animals were weighed weekly and blood and fecal samples were collected. At the end of the trial, all lambs were euthanized to determine worm burden and collect intestinal tissues and mucus samples for histological and immunological analysis. The infected group presented eosinophilia, increased number of inflammatory cells in the mucosa, in addition to an increased production of specific immunoglobulins against T. colubriformis, which partially prevented the establishment of infective larvae. As a consequence of parasitism, the infected lambs presented reduced serum albumin concentrations and demonstrated severe small intestine lesions, such as villous atrophy and epithelial erosion, which impaired the digestion and absorption of nutrients, causing a significant loss in performance. The infected group presented a 37% reduction in daily weight gain (107.26 ± 10.8 g/day), when compared with the control group (171.07 ± 7.15 g/day). The infected lambs also demonstrated the worst food conversion, i.e., each animal needed to consume on average 10.05 ± 0.52 kg of food to gain 1 kg live weight. The voluntary hay intake depression in infected animals was small, and statistical difference (P<0.01) was seen only on two occasions (ninth and 12th weeks), in comparison with the control group. In conclusion, Santa Ines lambs infected with T. colubriformis demonstrated a reduction in performance, caused by the damages produced by the adult nematodes in intestinal mucosa, and also by the immunopathological changes associated with the infection.

Short report: Human Trichostrongylus colubriformis infection in a rural village in Laos.

In Lahanam Village, Savannakhet Province, Laos, 125 of 253 villagers (49.4%) were found by fecal examination to harbor hookworm eggs. The eggs were heterogeneous in morphology and size, suggesting infections of mixed nematode species. To confirm the hookworm egg species, on a voluntary basis, 46 hookworm egg-positive participants were treated with albendazole, and post-treatment adult worms were collected from purged fecal samples. The common human hookworm was found in only 3 participants; 1 case of Necator americanus, and 2 cases of Ancylostoma duodenale. In contrast, adult Trichostrongylus worms were expelled from most participants (43 of 46, 93.5%). The Trichostrongylus species were confirmed by morphology and internal transcribed spacer 2 sequences; all worms were of the same species (T. colubriformis). In addition, some Trichostrongylus worms were obtained from a goat in the same village and identified as T. colubriformis. The results suggested that T. colubriformis was the main zoonotic species causing hookworm infections in the village.

Reciprocal effects between host phenotype and pathogens: new insights from an old problem.

Relationships between the host phenotype and pathogen infection are assumed to reflect either causes or consequences of the infection. In fact, these processes are likely to co-occur, even in the same phenotypic traits. For example, hosts with high ingestion rates have a higher growth rate but are also more infected by trophically transmitted pathogens that subsequently reduce the host growth rate. Here, we briefly review the empirical evidence suggesting reciprocal effects in host-pathogen interaction. We then provide a 'verbal' model that aims to predict how reciprocal effects can bias our interpretation of the relationship between host phenotype and pathogen infection. Finally, we outline technical avenues for explicitly considering reciprocal effects in the future and discuss their fundamental and applied implications.

Haemonchus contortus and Trichostrongylus colubriformis did not adapt to long-term exposure to sheep that were genetically resistant or susceptible to nematode infections.

We tested the hypothesis that Haemonchus contortus and Trichostrongylus colubriformis would adapt to long-term exposure to sheep that were either genetically resistant or susceptible to H. contortus. Sheep genotypes were from lines with 10 years prior selection for low (resistant, R) or high (susceptible, S) faecal worm egg count (WEC) following H. contortus infection. Long-term exposure of H. contortus and T.colubriformis to R or S genotypes was achieved using serial passage for up to 30 nematode generations. Thus, we generated four nematode strains; one strain of each species solely exposed to R sheep and one strain of each species solely exposed to S sheep. Considerable host genotype differences in mean WEC during serial passage confirmed adequate nematode selection pressure for both H. contortus (R 4900 eggs per gram (epg), S 19,900 epg) and T. colubriformis (R 5300 epg, S 13,500 epg). Adaptation of nematode strain to host genotype was tested using seven cross-classified tests for H. contortus, and two cross-classified and one outbred genotype test for T. colubriformis. In the cross-classified design, where each strain infects groups of R, S or randomly bred control sheep, parasite adaptation would be indicated by a significant host genotype by nematode strain interaction for traits indicating parasite reproductive success; specifically WEC and, for H. contortus strains, packed cell volume. We found no significant evidence of parasite adaptation to host genotype (P>0.05) for either the H. contortus or T. colubriformis strains. Therefore, we argue that nematodes will not adapt quickly to sheep bred for nematode resistance, where selection is based on low WEC, although selecting sheep using a subset of immune functions may increase adaptation risk. Our results support the hypothesis that nematode resistance is determined by many genes each with relatively small effect. In conclusion, selection of sheep for nematode resistance using WEC should be sustainable in the medium to long-term.

Cross-species utility of microsatellite markers in trichostrongyloid nematodes.

The development of microsatellite markers for parasitic nematodes has been hampered by technical difficulties in isolation and PCR amplification. We have investigated the potential for circumventing these problems using microsatellites from 3 trichostrongyloid species on a panel of 7 species. Ten of the 22 PCR primer pairs tested amplified in species other than the target species, usually in closely related species, and 2 new variable loci were discovered in the sheep parasite Trichostrongylus vitrinus. This study provides evidence that cross-species testing of microsatellite primers can be an effective alternative to isolation de novo.