A survey of gastrointestinal nematode species in red deer (Cervus elaphus) farms in New Zealand using PCR.

Gastrointestinal nematodes are recognised as an animal health issue for farmed red deer. The aim of this study was to explore the range of species infecting farmed deer herds and their farm-level prevalence in New Zealand. Faecal samples were collected from 12-24-month-old deer (n = 6-26; mean 19) on 59 farms located in the North (n = 25) and South (n = 34) Islands. Sub-samples of faeces were pooled by farm and cultured to recover third stage larvae. Twenty four larvae were randomly selected and identified to species using a multiplex PCR (total = 1217 larvae). At farm-level the most prevalent nematodes were Oesophagostomum venulosum 83% (n = 49) and the deer-specific nematodes in the subfamily Ostertagiinae (=Ostertagia-type) including, Spiculoptera asymmetrica 73% (n = 43), Ostertagia leptospicularis 47% (n = 28), Spiculoptera spiculoptera 47% (n = 28). The recently identified Trichostrongylus askivali was present on 32% (n = 19) of the farms and Oesophagostomum sikae on 17% (n = 10). In the analysis of the total number of larvae identified, the proportion was in similar order, 45% (n = 548) were O. venulosum, 14% (n = 173) S. asymmetrica, 10% (n = 124) S. spiculoptera, 9% (n = 114) O. leptospicularis, T. askivali, 3% (n = 40) and only 2% were O. sikae (n = 20). This study is the first to show the farm-level prevalence of nematode species in deer in New Zealand and the first to use PCR as a diagnostic tool. It provides data consistent with cross-infection from sheep/cattle to deer, and provided tentative insights into the proportions of the main GIN species across the deer population including O. sikae and T. askivali which have only recently been identified in New Zealand.

Efferent intestinal lymph protein responses in nematode-resistant, -resilient and -susceptible lambs under challenge with Trichostrongylus colubriformis.

The mechanisms underlying resistance to challenge by gastrointestinal nematode parasites in sheep are complex. Using DIGE, we profiled ovine lymph proteins in lambs with host resistance (R), resilience (Ri) or susceptibility (S) to a daily trickle challenge with the nematode Trichostrongylus colubriformis. Efferent intestinal lymph was collected prior to infection (day 1) and on days 5 and 10 post-infection. Eight proteins identified by LC-MS/MS, showed differences relating to host genotype. Of these, Serpin A3-3 and Serpin A3-7 have not been reported previously in the lymph proteome. Three acute phase proteins showed significant differences relating to interactions between breeding line and parasite challenge, including complement C3ß, C3α and haptoglobin (Hp) ß. In the R lambs C3α was significantly up regulated (P<0.05) on day 10, while in the Ri lambs Hp ß was significantly down regulated (P<0.05). In the S lambs, levels of C3ß were up regulated and levels of Hp ß down regulated (both P<0.05) on day 10. Hence we demonstrate that acute phase inflammation proteins contribute to differences in the innate immune response of sheep to challenge by T. colubriformis. The findings may lead to the development of new approaches to combat nematode infestations in sheep production systems. BIOLOGICAL SIGNIFICANCE: Breeding lines of sheep with resistance (R), resilience (Ri) or susceptibility (S) to nematode infections provide an experimental model to examine the biological mechanisms underlying the ability of some sheep to expel worms and remain healthy without the use of an anthelmintic. Using proteomics we identified differences in the expression of acute phase lymph proteins in the R, Ri and S lambs. The results will assist the development of alternative control strategies to manage nematode infections in livestock.

A multiplex PCR-based method to identify strongylid parasite larvae recovered from ovine faecal cultures and/or pasture samples.

A multiplex PCR-based method was developed to overcome the limitations of microscopic examination as a means of identifying individual infective larvae from the wide range of strongylid parasite species commonly encountered in sheep in mixed sheep-cattle grazing situations in New Zealand. The strategy employed targets unique species-specific sequence markers in the second internal transcribed spacer (ITS-2) region of ribosomal DNA of the nematodes and utilises individual larval lysates as reaction templates. The basic assay involves two sets of reactions designed to target the ten strongylid species most often encountered in ovine faecal cultures under New Zealand conditions (viz. Haemonchus contortus, Teladorsagia circumcincta, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirus spathiger, Chabertia ovina, and Oesophagostomum venulosum). Five species-specific primers, together with a pair of "generic" (conserved) primers, are used in each of the reactions. Two products are generally amplified, one by the generic primer pair regardless of species (providing a positive PCR control) and the other (whose size is indicative of the species present) by the appropriate species-specific primer in combination with one or other of the generic primers. If necessary, any larvae not identified by these reactions can subsequently be tested using primers designed specifically to detect those species less frequently encountered in ovine faecal cultures (viz. Ostertagia ostertagi, Ostertagia leptospicularis, Cooperia punctata, Nematodirus filicollis, and Bunostomum trigonocephalum). Results of assays undertaken on >5500 nematode larvae cultured from lambs on 16 different farms distributed throughout New Zealand indicated that positive identifications were initially obtained for 92.8% of them, while a further 4.4% of reactions gave a generic but no visible specific product and 2.8% gave no discernible PCR products (indicative of insufficient or poor quality DNA template). Of the reactions which yielded only generic products, 91% gave positive identifications in an assay re-run, resulting in a failure rate of just ∼ 0.4% for reactions containing amplifiable template. Although the method was developed primarily to provide a reliable way to identify individual strongylid larvae for downstream molecular applications, it potentially has a variety of other research and practical applications which are not readily achievable at present using other methods.

Managing anthelmintic resistance: is it feasible in New Zealand to delay the emergence of resistance to a new anthelmintic class?

The recent registration in New Zealand of the first new class of broad-spectrum anthelmintic, for use against nematode parasites of ruminants, in nearly three decades has raised the possibility that parasite management practices could be improved to minimise the emergence of resistance to the new drug. A review of knowledge pertaining to the selection of anthelmintic resistance in nematode parasites of sheep highlights a number of management practices which could be altered to achieve this. A number of previously common practices such as whole-flock treatment of adult ewes around lambing, and treatment of lambs as they are moved onto pastures with low parasite contamination have been clearly identified as high risk for selecting resistant parasites. Once high-risk practices have been identified steps can be taken to either eliminate their use or mitigate the associated risk. Much of the focus on the management of resistance around the world is on the retention of susceptible genotypes in refugia. While approaches to retaining unselected parasites are likely to vary around the world, empirical studies indicate that the practice is likely to be effective at slowing the development of resistance. The challenge for farmers and advisors will be to strike a balance between retaining sufficient susceptible parasites to usefully delay the development of resistance while not unduly compromising animal performance and farm profitability. The merits of combining different classes of anthelmintic in order to slow the development of resistance remains somewhat contentious in some countries. However, the attributes of oral anthelmintics are such that they seem likely to meet most, if not all, of the criteria for combinations to be highly effective at slowing the build-up of resistance in nematode parasites. It is evident that considerable progress has been made in understanding the factors involved in selecting anthelmintic-resistant nematodes since the last broad-spectrum anthelmintic class was released in the early 1980s. Therefore, it should be possible to manage a new class of anthelmintic in such a way as to significantly extend its effective life. The challenge is likely to be in convincing farmers of the merits of adopting such pro-active strategies.

Heritability of resistance to infestation with the body louse, Bovicola ovis, in Romney sheep bred for differences in resistance or resilience to gastro-intestinal nematode parasites.

The inheritance of resistance to louse infestation and the related allergic skin disease, cockle, was examined in Romney lambs. The lambs used in the study were the 2001- and 2004-born progeny of four experimental breeding lines ("Resistant", "Susceptible", "Resilient" and "Control") developed as part of a long-term study of the genetics of host resistance (maintenance of low faecal egg count (FEC) under nematode challenge) or resilience (maintenance of health and productivity under nematode challenge irrespective of FEC) to nematode parasites in sheep. Between 13 and 22 progeny (equally distributed between males and females, where possible) from each of five sires in each line were selected each year for this trial. All lambs (n=701) were examined for lice (Bovicola ovis) before artificial infestation; in 2001 the lambs were free of natural infestation, whilst in 2004 naturally acquired infestation was evident. In November 2001 and May 2002, approximately 60 B. ovis were transferred to each lamb, followed by monitoring at approximately 2-monthly intervals until August 2002. Similar procedures, but with fewer monitoring times, were repeated on the 2004 lambs. Overall, lambs in the Control line were significantly more susceptible to louse infestation and cockle compared with those in the other three lines (P<0.001). Least squares-means (SEM) of log-transformed louse score for the control, resistant, susceptible and resilient lines, respectively, were 2.178 (0.045), 1.499 (0.050), 1.618 (0.050) and 1.587 (0.044), and for cockle score were 1.36 (0.05), 0.76 (0.05), 0.95 (0.05) and 0.78 (0.05). From all progeny together, the heritability of log-transformed louse score was 0.22 (Standard Error (SE) 0.06) in autumn and 0.34 (SE 0.08) in winter, with a value of 0.44 (SE 0.09) when these data were combined. These estimates were similar to those obtained for resistance to gastro-intestinal nematodes in these breeding lines, using log-transformed FECs. Heritability estimates for cockle score in autumn, winter and when combined were 0.06 (SE 0.04), 0.45 (SE 0.09) and 0.40 (SE 0.09), respectively. The genetic correlations of mean log-transformed louse score with mean cockle score and levels of two different louse antigens in wool were, respectively, 0.97 (SE 0.04), 0.96 (SE 0.08) and 0.95 (SE 0.09). However, there was no significant genetic correlation between louse scores and FEC. These results suggest that selective breeding would be effective in reducing louse infestation and cockle in sheep, but that differences in louse burdens were not related to differences in nematode burdens as indicated by FECs.

Parastrongyloides trichosuri, a nematode parasite of mammals that is uniquely suited to genetic analysis.

Commonly studied nematode parasites have not proven amenable to simple genetic analyses and this has significantly reduced the available research options. We introduce here a nematode parasite of mammals, Parastrongyloides trichosuri, which has features uniquely suited for genetic analysis. This parasite has the capacity to undergo multiple reproductive cycles as a free-living worm and thereby amplify the numbers of its infective L3s in faeces. Culture conditions are presented that permit facile laboratory maintenance of this worm for >90 free-living life cycles (to date) without the need for re-entry into a permissive host. Even after long maintenance as a free-living worm, culture conditions can be manipulated to favour development of infective L3 worms, which remain able to successfully infect their marsupial hosts. The switch to infective L3 development is triggered by a secreted factor contained in culture medium conditioned by multiple generations of free-living worm culture. It is simple to perform single pair crosses with P. trichosuri to carry out Mendelian genetics in the laboratory and this has been done multiple times with sibling pairs to generate highly inbred lines. Lines of worms can readily be cryopreserved and recovered. Over 7000 expressed sequence tags have been produced from cDNAs at different life cycle stages and used to identify single nucleotide polymorphisms and microsatellites as genetic markers. Free-living worms live only a few days on average while the patency of parasitic infections can last for several months. Since we show this is not the result of re-infection, we conclude that parasitic worms have a lifespan capacity at least 20-30 times longer than their free-living counterparts. We discuss how it should be possible to exploit these unique features of P. trichosuri as a model for future studies that explore the genetic basis of longevity and parasitism.

Resistance to prophylactic treatment with macrocyclic lactone anthelmintics in Teladorsagia circumcincta.

Sixty-four Romney ewe lambs were allocated to 12 groups on the basis of liveweight. Four groups (n=5) were administered oral ivermectin (IVM), 4 (n=6) oral moxidectin (MOX) and the remaining 4 (n=5) controlled-release capsules containing IVM (IVM-CRCs). Nine and 10 days later, the groups within each treatment type were challenged with infective-stage larvae (L3) of 1 of 4 different isolates of Teladorsagia circumcincta (two doses each of 5000 L3). The first of these (S) was known to be anthelmintic-susceptible; the second (OR) was a multiple anthelmintic-resistant strain recovered from the field following therapeutic failure of both ivermectin and moxidectin and subsequently maintained in the laboratory without further anthelmintic selection; the third (R) was derived from OR but had been passaged for five generations indoors with each generation being screened with all three broad-spectrum anthelmintic classes; and the fourth (RxS) was an F1 cross between the R and S isolates. As anticipated, because of its limited residual activity, IVM had no significant effect on the establishment, 9 and 10 days post-treatment, of any of the parasite isolates. In contrast MOX, which has greater residual activity, was highly effective at preventing the establishment of the S isolate but showed no significant effect against the OR, R or RxS isolates. The IVM-CRC was also highly effective at preventing the establishment of the S isolate and furthermore it significantly reduced establishment of both the OR and RxS isolates, although it had no significant effect against the R isolate. The results suggest that with respect to the establishment of T. circumcincta L3s following anthelmintic treatment, macrocyclic lactone (ML) resistance is effectively a dominant trait in the presence of MOX, while it behaves as a partially dominant/recessive trait under treatment with IVM-CRCs. The potential implications of this finding in relation to selection for ML resistance in T. circumcincta are discussed.

Resistance to therapeutic treatment with macrocyclic lactone anthelmintics in Ostertagia circumcincta.

Eighty-eight lambs were allocated to one of four groups which were then dosed with 10,000 infective-stage larvae (L3) of one of four populations of Ostertagia circumcincta; the first (S) was an isolate known to be anthelmintic-susceptible; the second (OR) was a multiple anthelmintic-resistant isolate which had been recovered from the field following therapeutic failure of both ivermectin and moxidectin and subsequently maintained in the laboratory without further anthelmintic selection. The third (R) was derived from OR but had been passaged for five generations in the laboratory with each generation being screened with all three broad-spectrum drench families; the fourth (R x S) was an F1 cross between the S and R isolates. On patency, each of the four infection groups was sub-divided into five treatment groups, one of which received no anthelmintic while the others were administered either oral ivermectin (IVM-oral), controlled-release capsules containing ivermectin (IVM-CRCs), oral moxidectin (MOX-oral) or injectable MOX (MOX-inj). Neither formulation of IVM reduced FEC in the R, R x S and OR infected lambs compared to their untreated controls, but significant reductions were observed in all cases following MOX-oral or MOX-inj treatment. Similarly, neither IVM formulation significantly reduced the numbers of R or R x S worms compared to their untreated controls, although the numbers of OR worms were reduced in both cases (P<0.05). Direct comparisons of efficacy across the isolates, however, indicated that neither formulation was any more effective against R x S or OR worms than against the more highly selected R worms. In contrast, both MOX formulations significantly reduced worm numbers of all the resistant isolates compared to their respective untreated controls; furthermore, worm burdens of R x S were reduced significantly more than burdens of R (P<0.05). Reductions in OR burdens, which were intermediate between the two, did not differ significantly from either. The results are consistent with published work on Haemonchus contortus, which suggests that macrocyclic lactone (ML) resistance is expressed as a dominant trait under treatment with IVM. However, these data differ from the H. contortus studies in suggesting that ML resistance in O. circumcincta may effectively be rendered incompletely dominant or recessive by treatment with MOX.

Breeding sheep in New Zealand that are less reliant on anthelmintics to maintain health and productivity.

Breeding sheep that are less reliant on the use of anthelmintics to maintain acceptable health and productivity is one of several options that may assist farmers to manage the growing anthelmintic-resistance problem in New Zealand and meet consumer demands to minimise drug usage in livestock. Although it has been known for many years that genetic factors contribute to the ability of sheep to cope with roundworm challenge, attempts to selectively breed for such factors have occurred only recently. Two host traits, 'resistance' and 'resilience', are thought to be involved in limiting the deleterious effects of roundworms on the health and productivity of sheep, and the associated need for treatment with anthelmintics. Over the last decade, considerable advances have been made in our understanding of the feasibility and implications of breeding for these traits under practical conditions. The experimental breeding lines developed as part of this research are now proving to be a valuable resource for further work directed at identifying genes and/or genetic markers associated with host resistance and understanding the mechanisms of host immunity to nematode parasites in ruminants. This review provides an up-to-date summary of the results of research in this field, with particular reference to dual-purpose sheep in New Zealand, and describes how this information is being applied in practical breeding programmes.

Faecal egg counts in Angora goats following natural or experimental challenge with nematode parasites: within-flock variability and repeatabilities.

Phenotypic variation in faecal worm-egg count (FEC) was studied in 100 Angora goats between 6 months and 2 years old as part of a wider investigation into the potential use of FEC as a selection trait in breeding goats for improved natural resistance to nematode infection. Shortly after weaning at 4-5 months old the goats were subjected to a procedure involving successive natural and experimental challenges, which was continued at 6-monthly intervals until the goats were 2 years old. Experimental challenges consisted of Trichostrongylus colubriformis infective larvae administered orally at a dose rate of 1000 larvae/kg liveweight. FECs resulting from both natural and experimental infections varied considerably between individual goats as well as between different sampling times. Mixed model analysis of variance of natural infections confirmed that a significant component of this variation was attributable to differences between individual goats (P < 0.01) as opposed to other potential sources of variability, indicating that although generally deemed to be more susceptible than sheep, Angora goats do nevertheless vary in their ability to resist nematode infection. Rank correlations between pairs of FECs derived from the same infection were relatively high for both natural and experimental infections (r = 0.63-0.70 and 0.54-0.79, respectively; all P < 0.01). However, those between mean FECs measured on successive 6-monthly sampling occasions were considerably lower and more variable (r = 0.01-0.41 and 0.19-0.62 for natural and experimental infections, respectively). Nevertheless, FECs resulting from natural infections showed moderate correlations with those from corresponding experimental infections (P < 0.01) on all except the first sampling occasion (r = 0.23-0.61). Correlations were marginally higher when FECs from experimental infections were adjusted for infecting dose size (r = 0.32-0.63). FECs resulting from experimental infections were significantly higher on the first sampling occasion (at 6 months) than on subsequent occasions (P < 0.01) suggesting that some degree of resistance to nematode establishment had developed in the goats by 12 months old. On the basis of unadjusted FECs there appeared to be no marked increase in resistance beyond that age, although analyses based on FECs adjusted for infecting dose size suggested an increase in acquired resistance up to 18 months of age. FECs following natural challenge in goats which had also received experimental infections were not significantly different from those exposed only to natural challenge, indicating that the drench-abbreviated experimental infections had had no immunizing effect. Does that had given birth and were lactating at 24 months had significantly higher FECs than 'dry' does in the flock (P < 0.01), suggesting that like sheep, goats exhibit a post-parturient relaxation of immunity. The use of FECs as potential indicators of resistance status in goats and as a criterion for the selection of breeding animals is discussed.

Epidemiology of nematodosis in Romney lambs selectively bred for resistance or susceptibility to nematode infection.

Field trials were undertaken to compare nematode population dynamics, lamb productivity and levels of breech soiling in experimental flocks of Romney lambs selectively bred for increased resistance or susceptibility to nematode infection. In each year of the 2 year study, spring-born ewe lambs derived from Wallaceville Animal Research Centre's divergent nematode-resistant and nematode-susceptible breeding lines were grazed as separate flocks on matched farmlets from weaning (at 3 months old) until they were approximately 10-11 months old. Allocation of farmlets was reversed between Years 1 and 2 of the study to account for any possible paddock-related effects. Within each year both flocks were subjected to identical management conditions, including anthelmintic treatment (which was administered only when the overall mean faecal worm egg count measured across both genotypes reached 1500 eggs g-1). In both years, by mid-autumn (April) nematode larval infestation levels on pasture were approximately 5-6-fold greater on the farmlet grazed by susceptible (S) genotype lambs than on that grazed by their resistant (R) counterparts (Year 1: 2506 cf. 544 larvae kg-1 herbage; Year 2: 431 cf. 74 larvae kg-1 herbage). This led to 51-fold and 56-fold differences in faecal egg count between R and S lambs by late autumn (May) and winter (July) in Years 1 and 2, respectively. Although mean growth rates were similar in the R and S lambs over summer (while pasture infestation levels on the farmlets were still in the process of diverging), significantly higher growth rates occurred in the R than in the S lambs over autumn-winter in both years of the study (P < 0.01). In contrast, no significant differences in growth rate occurred in either year between male lambs derived from the nematode-resistant and nematode-susceptible breeding lines which were grazed together on another area of the Wallaceville farm from weaning until late autumn. Despite the substantially lower pasture infestation levels encountered by the R ewe lambs, they nevertheless temporarily suffered more breech soiling (dags) than their S counterparts (P < 0.01) in both years. Yearling fleece-weights of the R and S genotypes did not differ significantly in either year. Although the results of our study confirmed that there are potentially significant epidemiological benefits to be derived from breeding sheep for resistance to nematode infection, these benefits did not appear to be associated with large advantages in animal performance. Further work is needed to establish how these results should be interpreted with respect to anthelmintic drench requirements of genetically resistant animals.

Feasibility and implications of breeding sheep for resilience to nematode challenge.

There are potentially 2 different types of host response which could be targeted when seeking to breed sheep which require minimal frequency of anthelmintic treatment to maintain acceptable health and productivity in the face of nematode challenge. These are "resistance" (the ability to suppress the establishment and/or subsequent development of a parasite infection), and "resilience" (the ability to maintain relatively undepressed production while subjected to parasite challenge). Most studies have concentrated on resistance, and used faecal worm egg count (FEC) as the selection criterion. However, in Romney sheep in New Zealand, it has become apparent that "resistance to infection" does not equate to "disease resistance" in the strict sense. Although genetically low FEC Romney lambs have been shown to have significantly reduced burdens of the most important nematode species, they nevertheless appear to have no significant production advantages over their higher FEC counterparts when all are grazed together under the same larval challenge. Furthermore there is evidence of an unfavourable association between FEC and dagginess (soiling of the breech area) suggesting that some forms of host response to nematode challenge may result in diarrhoea. Our results suggest that the main benefits of selectively breeding for low FEC are likely to be derived indirectly as a result of reduced pasture contamination. This has led us to investigate resilience as a possible alternative breeding option to resistance. Over a period of 3 years approximately 14,000 progeny of 213 different rams were tested for resilience (assessed in terms of drench requirements using an individual "on demand" treatment procedure), as well as growth rate, dag-score, FEC and fleece-weight. Overall the heritabilities of various measures of resilience examined proved to be relatively low (ranging between 0.10 +/- 0.03 and 0.19 +/- 0.04), although estimates varied between flocks and years possibly as a result of differences in the severity of challenge. Nevertheless, results suggested that progeny-test selection for resilience would lead to lambs with higher growth rates and lower dags-scores when left undrenched on infective pasture. In contrast to some Australian results we found no positive association between resilience to mematode challenge and resistance to infection. If the potential benefits of both resistance and resilience are to be captured our results suggest that an index selection procedure will be needed.

Nematode burdens and immunological responses following natural challenge in Romney lambs selectively bred for low or high faecal worm egg count.

Breeding lines of Romney sheep, selected as lambs for consistently low or high faecal nematode egg count (FEC) following periods of natural challenge, have been maintained at Wallaceville for some years. In order to determine the extent to which FECs in low and high genotypes reflected their ability to resist the establishment of gastro-intestinal nematode burdens, we investigated the infection status and immune responses in 8- to 9-month-old progeny of selected rams from low and high FEC breeding lines following a period of grazing without anthelmintic treatment in autumn/early winter. In each of the 2 years of the study, outcross male progeny of the two lowest FEC (LFEC) (i.e. most 'resistant') and two highest FEC (HFEC) (i.e. most 'susceptible') rams from the divergent lines were slaughtered shortly after autumn/early winter FECs had been analysed. Post-mortem worm counts and examination of intestinal histology were then undertaken. Blood samples collected before slaughter in the second year of the study were assayed to measure serum levels of Trichostrongylus colubriformis-specific antibody and immunoglobulins (IgG1 and IgM), and numbers of circulating eosinophils. Overall, correlations between pre-slaughter FEC and total trichostrongyle burdens in the lambs proved to be very high (0.91 and 0.85, respectively, for the 2 years studied). In the first year, LFEC lambs, which were shedding only 28.6% as many strongyle eggs as their HFEC counterparts at slaughter, were found to harbour 37.6% as many adult trichostrongyle worms, while in the second year, LFEC lambs, which were shedding 16.1% as many strongyle eggs as their HFEC counterparts at slaughter, were found to harbour 33.5% as many adult trichostrongyle worms. Results, particularly in the second year, confirmed that significantly fewer worms of most of the important abomasal and small intestinal nematode species which infest lambs in New Zealand (i.e. Haemonchus contortus, Ostertagia circumcincta, Cooperia curticei, Nematodirus spathiger, T. colubriformis, and Trichostrongylus vitrinus) had established in the LFEC genotypes than in their HFEC counterparts. In addition, in utero egg counts of female intestinal Trichostrongylus spp. were significantly lower in LFEC lambs than in their HFEC counterparts, indicating a reduction in fecundity of those worms which did establish. There was also some evidence of an effect of host response on the developmental composition of burdens in the case of some worm species. In relation to host responses, numbers of globule leucocytes/mucosal mast cells in the intestinal mucosa were significantly higher (P < 0.01) in LFEC lambs than in HFEC lambs in both years of the study. Numbers of connective tissue type mast cells and eosinophils in the intestinal mucosa were also significantly higher in LFEC lambs but only in the second year of the study (P < 0.01 and P < 0.05, respectively). Numbers of circulating eosinophils did not differ significantly between the genotypes. T. colubriformis-specific antibodies, IgG1 and IgM to both L3 and adult worm antigens were all significantly higher (P < 0.01 or P < 0.05) in LFEC lambs than in HFEC lambs.

The immune responsiveness of Romney sheep selected for resistance or susceptibility to gastrointestinal nematodes: lymphocyte blastogenic activity, eosinophilia and total white blood cell counts.

Blastogenic activity, eosinophil and total white blood cell counts (TWBC) were examined over a period of 14 weeks in Romney lambs, genetically resistant or susceptible to gastrointestinal nematodes. The lambs were infected with 5000 infective Trichostrongylus colubriformis larvae twice weekly. Compared to preinfection levels, the blastogenic activity of unstimulated lymphocytes in lambs of both lines peaked at week 3, and was significantly higher in resistant than in susceptible lambs. These changes may have been due to in vivo polyclonal activation. Lymphocytes from susceptible sheep responded more strongly to Con A, PHA and PWM than cells from resistant sheep. Counts per minute (c.p.m) for Con A- and PHA-stimulated lymphocytes increased in both lines of sheep from week 2 to week 7 and then returned to initial levels. An increase in c.p.m. in PWM-stimulated cell cultures was observed from weeks 3 to 5 in both groups. The blastogenic activity for LPS-stimulated cultures was significantly higher for resistant than susceptible sheep at weeks 3 and 4. No significant correlations between the decline in faecal egg counts (FEC) and the blastogenic activity was observed. Eosinophil counts in peripheral blood began to increase one week earlier in resistant than in susceptible sheep. No significant correlation between FEC and eosinophil counts was observed in resistant lambs, whereas in susceptible lambs a significant correlation was found between FEC and eosinophil counts at some sampling times. TWBC in resistant lambs steadily increased with infections whereas susceptible lambs showed a decrease until week 5 and then steadily increased. There was no significant correlation between the decline in FEC and TWBC.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunization of sheep against parasitic nematodes leads to elevated levels of globule leukocytes in the small intestine lumen.

In sheep that had been given three immunizing infections with Trichostrongylus colubriformis and Ostertagia circumcincta infective (L3) larvae, drenched after the last infection and challenged with larvae of the same species, there was a significant increase in numbers of small intestine mucosal tissue globule leukocytes (TGLs) and lumenal globule leukocytes (LuGLs) compared with sheep that had only been drenched and challenged. There was a positive correlation between the numbers of LuGLs and TGLs in the small intestine but the ratio of these two cell types was lower in non-immunized than immunized sheep. In immunized sheep positive correlations were observed between LuGLs and levels of arylsulphatase and peroxidase in the intestinal mucus and between arylsulphatase and larval migration inhibition (LMI) activity in mucus. Lumen eosinophils correlated with blood eosinophils, serum antibody against T. colubriformis correlated with peroxidase in the mucus and blood eosinophils correlated with nematode specific IgM levels in the intestinal mucus. In the abomasum, TGLs were present but not LuGLs. Sheep repeatedly infected with T. axei also had significantly more LuGLs in the small intestine than control animals. Two sheep that had a surgically prepared isolated small intestinal loop, after oral infection with T. colubriformis had TGLs and LuGLs in the intact intestine, but not in the isolated loop. Significantly more LuGLs were produced in sheep by allowing repeated T. colubriformis L3 infections to develop to adult stages compared to sheep treated with the same number of larvae, but where the infections were terminated by drenching at various intervals.

An evaluation of an oral formulation of moxidectin against selected anthelmintic-resistant and -susceptible strains of nematodes in lambs.

The efficacy of an oral formulation of the newly developed parasiticide, moxidectin, was tested against benzimidazole-resistant Haemonchus contortus, Trichostrongylus colubriformis, and Nematodirus spathiger, levamisole-resistant Ostertagia circumcincta, and susceptible Cooperia curticei infections in weaned lambs. Thirty-two lambs were experimentally infected with mixed doses of the above strains of nematodes. They were allocated into four treatment groups by stratified randomisation using liveweights and faecal egg counts 28 days later. One group received moxidectin at 0.2 mg/kg liveweight, one group oxfendazole at 4.5 mg/kg liveweight, one group levamisole at 7.5 mg/kg liveweight and the last group remained untreated as the control. Worm burdens in the lambs at slaughter 10 days after oral treatment confirmed the resistance status of the nematode strains used, and showed that moxidectin had a greater than 99.9% efficacy (p<0.01) against all of them. No adverse effects due to treatment with moxidectin were observed in any of the animals.

Efficacy of a topical formulation of ivermectin against naturally acquired gastro-intestinal nematodes in weaner cattle.

The efficacy of a topical formulation of ivermectin against naturally acquired gastro-intestinal nematodes in weaner cattle was evaluated. At slaughter, 14-15 days after treatment, burdens of Ostertagia spp, Trichostrongylus axei and Oesophagostomum radiatum were significantly lower in the treated calves than in the untreated controls (p<0.01). Efficacies (based on geometric mean worm burdens of treated and control groups) were 99.6%, 95.1% and 100% respectively. The sizes of the Cooperia spp and Trichuris ovis burdens in the treated group did not differ significantly at the 5% level of confidence from those in the control group.

Economics of a dry-cow anthelmintic drenching programme for dairy cows in New Zealand. Part 1. Overall response in 47 herds.

Trials were conducted on 47 seasonal supply dairy farms (greater than 5500 cows) to assess the economics of a dry-cow anthelmintic drenching programme. The programme was administered during the autumn/winter of 1983 and consisted of two treatments, the first shortly after drying off (late April-early June) and the second shortly before calving (mid-July-late August). The effects of the programme on the body condition of cows over winter and their subsequent milk production and reproductive performance were assessed. Overall, the treatment did not result in a worthwhile improvement in cow condition over winter. However, there was a small, but significant overall increase in milk production (2.24 kg milkfat/cow/lactation = 51.5 l milk; P less than 0.01). Young cows (3 years old) did not respond significantly better than mature cows, but high quality cows (as assessed on the basis of the previous season's milk production) responded significantly better than those of poorer quality. The pre-calving condition of cows did not significantly influence the magnitude of their response. Calving data for the year following the trials indicated that conception rates and time of conception were unaffected by the drench programme. At prices prevailing at the time of the trials, the programme proved to be only marginally economic overall (approximately 1 kg milkfat/cow was required simply to cover the cost of the anthelmintic used). However, levels of response in the individual herds involved varied considerably.

Economics of a dry-cow anthelmintic drenching programme for dairy cows in New Zealand. Part 2. Influence of management factors and other herd characteristics on the level of response.

The results of a series of trials conducted on 47 seasonal supply dairy farms in the southern North Island of New Zealand, which indicated a statistically significant overall production response of 2.24 kg milkfat/cow/lactation following a 2-treatment dry-cow anthelmintic drenching programme (Bisset et al., 1987), were subjected to further analyses in an attempt to identify any management factors or herd characteristics which may have influenced the levels of production response to treatment in individual herds. The management factors examined included calf drenching practices, types of grazing system, mean pre-calving condition of the herds, and 'nutritional status' of the herds over the milking season. The influence of herd quality, herd size and geographical location were also examined. Considered separately, only calf drenching practice had a significant influence on the level of herd response to treatment. Increases in milkfat production/cow due to the treatment programme were significantly greater (P less than 0.05, t-test) on farms where calves had received a minimal number of drench treatments (less than or equal to 2) between weaning and the end of March (mid-autumn) (+5.32 kg/cow/lactation, P less than 0.01) (= 108.2 l milk), than on farms where calves had received regular 3-4 weekly treatments over the same period (+0.42 kg/cow/lactation, N.S.) While none of the other factors, considered in isolation, appeared to significantly influence the production response of herds, observations on possible interactive effects suggested that the grazing system employed was probably of importance in so far as it determined the level of exposure of cows to calf-contaminated pasture. Thus, the greatest mean response to the treatment programme occurred in herds where cows were overwintered on areas grazed during the milking season by calves which had received less than or equal to 2 drench treatments from weaning until the end of March (+5.95 kg milkfat/cow/lactation, P less than 0.01).