Comparative studies of experimental oral inoculation of pigs with Ascaris suum eggs or third stage larvae.

This experimental study was designed to compare the acquired resistance in pigs to Ascaris suum eggs following 4-weekly oral immunizations with either 200 A. suum infective eggs or 50 A. suum third stage larvae (L3). The two immunized groups (n = 7) together with an unimmunized control group (n = 7) of pigs were challenged orally with 50 infective A. suum eggs per kilogram bodyweight on day 19 after the last immunization. Seven days post-challenge the group immunized with eggs showed signs of resistance as evidenced by reduced lung larval counts compared with the challenge control group. Such significant resistance was not observed in the L3-immunized group. However, a markedly increased inflammatory liver reaction and white spot formation was demonstrated in the L3-immunized pigs after challenge compared with both control animals and egg-immunized pigs. On the day of challenge only the egg-immunized pigs mounted an anti-Ascaris antibody response both in serum and in lung lavage fluid. Ascaris-antigen induced increased histamine release from peripheral leucocytes following both immunization and challenge could only be demonstrated in the egg-immunized pigs. On day 7 post-challenge local IgA-anti-Ascaris antibodies were further demonstrated in bile of the egg-immunized group and in the small intestine of both immunized groups. In conclusion, oral A. suum egg immunization of pigs induced a significant reduction in lung larval counts upon challenge. In contrast, oral L3 immunization seemed to prime the pigs as observed by the presence of stunted lung larval growth and increased liver reaction post-challenge with A. suum eggs.

An epidemiological investigation of congenital Schistosoma japonicum transmission in Hubei Province, PR China.

A cohort study was conducted in Hubei Province, China, following serious flooding of the Yangtze River in the autumn of 1998 to investigate the possibility of congenital transmission of Schistosoma japonicum in humans. The cohort investigated was comprised of 205 women and their 208 infants born between 1 September and 30 December 1998. Blood and fecal samples from all the women and their infants were collected and examined for S. japonicum infection. Positive specific antibody titers were found in 14 (6.8%) of the mothers, but no fecal egg excretion was observed. All infants had negative specific antibody titers and no S. japonicum eggs were found in their feces. Hence, the present study coud not confirm congenital S. japonicum transmission in humans. Further studies are highly wanted to study the impact of prenatal exposure of S. japonicum on the offspring.

Postnatal challenge infections of congenitally Schistosoma japonicum-infected piglets.

The aim of the present study was to assess the effect of a congenital Schistosoma japonicum infection on the establishment, fecundity, and pathogenicity of a postnatal challenge infection. Five prenatally S. japonicum-infected piglets received a challenge infection (prenatal + challenge group), 5 prenatally infected piglets were followed without challenge (prenatal group), and 10 piglets, born by unexposed sows, served as challenge controls (challenge control group). Challenge infections were given 8 wk after the piglets were born (14 wk after the primary infection of the sows), and the study lasted another 11 wk. Variables included worm burden, tissue egg count, and liver pathology. Worm establishment and tissue egg count were comparable in the prenatal + challenge group and in the challenge control group, both exceeding at a statistically significant level those in the prenatal group. No difference in worm fecundity (eggs/female worms/g tissue) was seen between the 3 groups. Liver pathology (i.e., portal and septal fibrosis) was more severe in the challenge control group compared to the other groups. A congenital S. japonicum infection in piglets thus affected neither establishment nor fecundity of a postnatal challenge infection. In spite of this, the challenge infection gave rise to much less liver pathology than the similarly sized challenge control infection.

Comparison of the vaccine efficacy of gamma-irradiated Schistosoma japonicum cercariae with the defined antigen Sj62(IrV-5) in pigs.

Development of a vaccine against Schistosoma japonicum which can protect both man and the domestic animal zoonotic reservoirs of infection would be an invaluable tool in attempts to control this infection in those areas in which conventional control methods have failed to break transmission. The pig is a natural host of S. japonicum and because of its anatomical and immunological similarities to humans, it is a potentially valuable host for studies on S. japonicum in particular and schistosomes in general. Radiation-attenuated cercariae are highly effective in inducing immunity in experimental schistosomosis and there are promising reports of partial protection against schistosomes with recombinant-derived individual antigens. In the present study we have set out to establish a protocol for inducing protection with gamma-irradiated cercariae in pigs and to assess the protective capacity of recombinant and naked DNA formulations of Sj62, a 62kDa region of S. japonicum myosin. The corresponding S. mansoni version or Sj62, recombinant IrV-5, has previously been implicated in irradiated vaccine immunity in S. mansoni infections and has been shown to induce high levels of immunity in a variety of hosts. Groups of pigs were immunised three times at 2-week intervals with 2000 cercariae irradiated at 20krad, with Sj62 as a recombinant (rSj62) incorporated in Freund's adjuvant, a micellar preparation, or as a naked DNA construct. Vaccination with irradiated cercariae did not induce significant anti-Sj62 antibody but following intramuscular challenge with 2000 cercariae, the vaccinated pigs showed >95% resistance as assessed by reduced faecal egg output, worm tissue egg burdens and also reduced septal fibrosis. Immunisation with each of the Sj62 formulations induced significant anti-Sj62 antibody responses, the highest titre (>12,800) being with the Freund's preparation but none of the Sj62-immunised groups showed significant resistance to challenge. The data suggest that Sj62 shows little promise as a vaccine candidate for schistosomosis.

Comparison of worm control strategies in grazing sheep in Denmark.

Control of nematode parasites with reduced reliance on the use of anthelmintics was studied in 16 ewes with suckling twin lambs on contaminated pasture in Denmark. Ewes and lambs were treated with albendazole at turn-out 3 May. Ewes were removed from the groups on 26 July, and lambs were slaughtered on 11 October. The animals were allocated to 4 groups of 8 lambs and their 4 ewes. Group TS was treated with albendazole at weeks 3, 6 and 8 after turnout and set-stocked; group TM was similarly treated but moved to clean pasture in conjunction with the last drenching; group US was untreated and set-stocked, and group UM was left untreated but moved to clean pasture week 8 after turn-out. Supplementary feed was offered in June and August due to scarcity of pasture. Strategic treatments of ewes and lambs weeks 3, 6 and 8 after turn-out, with or without a move to clean pasture, were highly effective in controlling nematode infections for most of the season. This was reflected in better weight gains and carcass characteristics in the treated compared to untreated lambs, resulting in an average increase in the value of the product by 36%. The effect of moving without treatment (UM) on faecal egg counts was limited but peak pasture infectivity was reduced to less than 10% compared to the set-stocked group and weight gains of lambs were significantly better despite poor feed availability in late season. The study showed that under set-stocked conditions repeated anthelmintic treatments of both ewes and lambs in early season may ensure sufficient nematode control whereas moving animals to clean pasture without dosing was less efficient. The latter may, however, still be a viable option in organic and other production systems where routine use of anthelmintics is banned, particularly if weaning and moving are combined or a second move is performed.

The epidemiology of gastrointestinal nematodes of dairy cattle in central Kenya.

The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bimonthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March-June and October-December) and lowest during the dry seasons (July-September and January-February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.

Cytokine mRNA expression in pigs infected with Schistosoma japonicum.

The pig is a natural host of Schistosoma japonicum and a useful animal model of human disease. In the present study mRNA levels of Th1 (IFN-gamma) and Th2 (IL-10 and IL-4) cytokines were assessed by RT-PCR within tissues from infected pigs. Twelve Danish crossbred pigs were infected by intramuscular injection or orally with 1000 cercariae. Six other pigs served as non-infected controls. Liver and intestinal tissues were collected 10 weeks post-infection, and analysed for their relative levels of cytokine mRNA. Infected pigs developed a Th2 response as characterized by the increased level of mRNA encoding for IL-4 and IL-10 in their large intestine (caecum and colon). In contrast, levels of IFN-gamma did not differ between control and infected animals although variation between animals was observed. When comparing the immune response of orally and intramuscularly infected animals, we found that orally infected pigs produced higher IL-4 and IL-10 levels in their caecum and colon respectively. This stronger Th2 response correlated with a previously reported delay in maturation of infection following oral infection. The cytokine expression levels in tissue samples taken from lesion sites and in nearby areas, without obvious lesions, were then compared. Subsequent to an oral infection, the Th2 type cytokine production was higher in the lesion sites of the liver. In conclusion, this study is the first demonstration of IL-4 and IL-10 cytokine response in pig tissues during S. japonicum infection.

Schistosoma japonicum: interactions of successive infections in pigs and mice using polymerase chain reaction-based identification of individual worms.

The study reported here investigated acquired resistance of mice and pigs to challenge-infections with Schistosoma japonicum. Two morphologically indistinguishable isolates of the parasite (from the Anhui and Zhejiang provinces of China), which could be typed by polymerase chain reaction-linked restriction fragment length polymorphism analysis (PCR-RFLP), were used for the infections. In two parallel infection studies, 60 female outbred NMRI mice and 29 Danish Landrace/Yorkshire/Duroc crossbred pigs were used. Two of the groups received a primary infection with either the Anhui or the Zhejiang isolate, respectively. The remaining groups received a primary infection with the Zhejiang isolate and challenge-infections with the Anhui isolate at either week 2, 3, 4 or 6 post primary infection. The results of the study indicated that both mice and pigs are partially resistant to challenge-infection from week 4 post primary infection. Resistance appeared to decrease in pigs 6 weeks after primary infection, while it remained effective in mice. These results suggest that the mechanism responsible for acquired resistance in mice and pigs may not be the same and support the theory that worm burdens in pigs receiving repeated infection are in a balance between acquisition and loss of worms.

Schistosoma japonicum infection in the pig as a model for human schistosomiasis japonica.

Valuable information on human schistosomiasis japonica has been provided using primates and experimental rodent hosts. However, major drawbacks such as high costs and ethical concerns for the primate models and large biological deviations for the rodent models have led to the search for more appropriate models. Recent data on the pig indicate that this natural host for Schistosoma japonicum might be a realistic alternative. As only very few research groups have investigated the S. japonicum/pig model, the present review mainly deals with the experimental methods and the major host/parasite findings obtained from the authors own research group. With emphasis on a critical evaluation of the work, the results are compared to the scarce information existing on human schistosomiasis japonica. Like in humans, S. japonicum establishes mainly in the large intestinal veins, with high faecal egg counts during the acute phase of infection, which varies greatly within and between days. Concomitant resistance is another shared feature, but studies in pigs have indicated that the phenomenon is more complex than generally thought. Clinical signs as eosinophilia and diarrhoea with mucus and blood in the acute phase of infection and hepatomegaly, increased portal diameter, periportal fibrosis and ascites in chronic infections are common findings in both humans and pigs. Low protein diet aggravates the disease in pigs by increasing the establishment rates, the faecal egg excretion and the morbidity. A 100% cure rate is achieved when treating S. japonicum infected pigs with praziquantel at 40 mg/kg, and 4 weeks post treatment pigs remain resistant to reinfection. Lastly, human congenital S. japonicum infection has been confirmed in pigs but the implications of such infections for the pathogenesis of schistosomiasis japonica remain to be investigated.

Congenital transmission of Schistosoma japonicum in the rabbit.

Fourteen pregnant rabbits were each infected with 300 cercariae of Schistosoma japonicum and divided into two groups. Group M (n = 8) was infected during mid-gestation (the organogenetic stage) and group L (n = 6) was infected during late-gestation (the post-organogenetic stage). Mother rabbits and rabbit kittens were killed 45-60 days after infection and perfused in order to obtain worm counts. Furthermore, faecal egg counts and tissue egg counts from livers were obtained from the mother rabbits as well as the rabbit kittens. All mother rabbits became infected harbouring 207.6 +/- 20.2 and 220.0 +/- 27.5 adult worms in group M and L, respectively. In groups M and L, 13.5% and 46.7% of the kittens were infected, respectively. In 12 of 14 litters at least one kitten was infected. The infected kittens harboured between one and three adult S. japonicum. The livers of the kittens infected with a worm pair displaced lesions as a result of egg deposition. The results, therefore, show that congenital transmission of S. japonicumcan occur in rabbits. The close anatomical resemblance between the rabbit and human placenta may be indicative of the presence of congenital transmission of S. japonicum infection in humans.

The prevalence and intensity of helminth and coccidial infections in dairy cattle in central Kenya.

A survey of gastrointestinal parasite infections of young (< 6 months old), immature (6-12 months old) and adult (> 12 months old) dairy cattle on 16 farms in Kiambu District, Kenya was conducted during a dry season (September 1991 to January 1992) and during a wet season (March to July 1992). The survey was based on monthly coproparasitological examination of cohorts and worm counts in tracer calves. The effects of age, sex, farm and season on the prevalence and intensity of helminth and coccidial infections were determined. Faecal egg and oocyst counts revealed that the overall prevalences were: strongyles (including trichostrongyles) (85.5%), liver flukes (Fasciola gigantica) (34.0%), coccidia (30.9%) and tapeworms (9.6%). Eight species of the protozoan Eimeria were identified, the most prevalent species being E. bovis and E. zuernii. The most prevalent nematode genera were Haemonchus, Cooperia, Oesophagostomum and Trichostrongylus. Season, farm and age of the animals had a significant (p<0.05) influence on the intensity of infection with strongyles, liver flukes and coccidia, whereas the sex of the animals had no significant (p>0.05) effect on the prevalence or intensity of infections. A higher intensity of infection with strongyles and coccidia was found in the wet season than in the dry season (p<0.05). The age-specific intensity was in the following order: for strongyles, immature animals of 6-12 months of age had the highest egg counts, followed by young calves and adults. Calves had significantly (p<0.05) higher oocyst counts than immatures or adults. Liver fluke egg counts did not differ significant (p>0.05) between immatures and adult cattle.

An electrophoretic comparison of Schistosoma japonicum (Trematoda) from different provinces in the People's Republic of China suggests the existence of cryptic species.

Schistosoma japonicum from the People's Republic of China is considered to represent a single species comprising either 1 or 4 'strains'. We conducted an allozyme electrophoretic study to examine the extent of genetic variation in S. japonicum from mainland China. The allelic profiles of S. japonicum from 7 provinces were established at 16 enzyme loci. S. japonicum from Sichuan had 3-5 (19-31%) fixed differences compared with those from Zhejiang, Anhui, Jiangxi, Hunan, Hubei and Yunnan, suggesting that S. japonicum in mainland China represents a species complex. In addition, genetic markers were also established for different laboratory-maintained populations of S. japonicum which has significant implications for studying the biology of these organisms in human and animal hosts, and for the control and surveillance of human schistosomiasis in China.

PCR-based identification of individuals of Schistosoma japonicum representing different subpopulations using a genetic marker in mitochondrial DNA.

A mitochondrial NADH dehydrogenase I gene fragment (NDI) was sequenced for three laboratory maintained isolates of Schistosoma japonicum. Comparison of sequences representing the isolates (originally obtained from the Anhui and Zhejiang provinces of the People's Republic of China, and from the Philippines) revealed inter-isolate sequence variations of 0.2-0.6% and no intra-isolate variation was found. The sequences also indicated that while the amplification products of the Zhejiang and Philippine isolates contained a recognition site for the endonuclease RsaI, there was no such site in the Anhui isolate. This was tested by digesting amplification products from a number of individual worms with RsaI. Then an infection experiment was designed to test the value of this genetic marker for studies of the population biology of S. japonicum in the final host. For this, the two Chinese isolates were used. Three groups of mice (A-C) were exposed firstly to a primary infection and then challenge-infected at weeks 4 and 7 of the experiment. In group A the first infection was done with the Anhui isolate, and the two others with the Zhejiang isolate, thereby providing a specific, detectable cohort. In groups B and C the Anhui isolate was used for the second and third infection. All mice were perfused 5 weeks after the last challenge infection, and the NDI was subsequently amplified from DNA of the perfused worms and digested with RsaI. The digestion revealed that while infection groups A and B contained mixed populations of the Anhui and Zhejiang isolates, only Zhejiang worms were present in group C. We concluded that the absence/presence of the RsaI site in the NDI provides a useful marker for the delineation of cohorts of S. japonicum.

Congenital transmission of Schistosoma japonicum in pigs.

Congenital transmission of Schistosoma japonicum in pigs was investigated by experimentally infecting sows at four weeks gestation (n = 3), 10 weeks gestation (n = 3), or a few weeks prior to insemination (n = 2). None of the piglets born to sows infected prior to insemination or in early pregnancy were found to be infected. However, all of the piglets (n = 26) born to sows infected at 10 weeks gestation were found to harbor schistosomes with S. japonicum eggs recovered from both their feces and livers. The findings show that congenital S. japonicum infection of pigs can occur if sows are infected during mid-to-late pregnancy and may have important implications not only for pigs but also for other mammalian hosts of schistosomes, including humans.

Scanning for nucleotide variations in mitochondrial DNA fragments of Schistosoma japonicum by single-strand conformation polymorphism.

In this study, we employed a mutation scanning approach for the direct visual display of genetic variability in mitochondrial DNA (mtDNA) fragments within and among populations of Schistosoma japonicum from the People's Republic of China. Fragments of the NADH dehydrogenase 1 gene (ND1) and the cytochrome c oxidase subunit I (COI) were individually amplified from parasite DNA by polymerase chain reaction (PCR), denatured and subjected to single-strand conformation polymorphism (SSCP) analysis. Using ND1 and COI fragments, individuals representing different genotypes could be readily identified based on characteristic and reproducible SSCP profiles. The results demonstrated the usefulness of SSCP for the direct visual display of low-level sequence variation in mtDNA of S. japonicum prior to DNA sequence analysis. This approach has important implications for studying the genetic structure and biology of S. japonicum populations, and for analysing the inheritance of mitochondrial DNA.

Elucidation of Schistosoma japonicum population dynamics in pigs using PCR-based identification of individuals representing distinct cohorts.

The study reported here investigated the interactions of successive infections and acquired resistance of pigs to challenge infections of Schistosoma japonicum. Two morphologically indistinguishable geographical isolates from China (from Anhui and Zhejiang provinces) were used for the infections. The worms of the two isolates were distinguishable by PCR-linked restriction fragment length polymorphism analysis of the nicotinamide adenine dinucleotide phosphate dehydrogenase I gene of the mitochondrial genome. Thirty-two pigs divided into seven groups were used in the experiment. Two groups received a single infection by either the Anhui or the Zhejiang isolate. In Challenge Groups 1, 4, 6, 8 and 12, a primary infection of the Zhejiang isolate was followed by a challenge infection with the Anhui isolate at week 1, 4, 6, 8 or 12 after the primary infection. In this way it was possible to determine whether worms recovered by perfusion originated from the primary or the challenge infection. Only the challenge infection at week 1 resulted in a higher worm burden when compared with a single primary infection with the Zhejiang isolate. The results showed that challenge worms were able to establish, and that the proportion of worms originating from challenge infection increased at the later challenge infections, however without an increase in the total number of worms. In addition, mixed pairs of the two isolates were found in all challenge-infected groups. The results indicate that pigs are able to mount a partial resistance against re-infection with S. japonicum by 4 weeks after a primary infection, but that worms of the challenge infections eventually replace the primary infection. The finding of mixed pairs of the two isolates indicates that worms of S. japonicum are either polygamous or able to wait in solitude for up to 12 weeks for a partner.

Schistosoma japonicum in the pig: the influence of age on the host/parasite relationship.

The current study sought to elucidate a possible association between age and susceptibility to a primary infection with Schistosoma japonicum in pigs. Sixteen Landrace/Yorkshire crossbred specific pathogen-free pigs in three different age groups (group A-C), aged approximately 7, 24 and 37 weeks at the beginning of the experiment, were infected by intramuscular injections of 1,000, 1,500 or 2,400 cercariae, respectively. Fecal egg counts were obtained twice weekly from six to eight weeks post infection (wpi), and the pigs were killed 11 wpi. The number of worms collected were counted and sexed subsequent to perfusion. Tissue egg counts were estimated on samples from the liver. The worm recoveries for group A, B and C were 3.2%, 8.1% and 3.8%, respectively. No differences were observed between the male/female ratios of the three groups. The fecundity parameters, ie, fecal egg counts per mature female and liver egg counts per mature female, showed no significant differences between the three age groups. The results did not indicate any difference in susceptibility between the different age-groups of pigs to a primary infection with S. japonicum.

Schistosoma japonicum infection and serum and tissue concentrations of retinol and zinc in pigs.

The effects of Schistosoma japonicum infection on the concentrations of zinc in serum, liver, spleen and muscle and on the concentrations of retinol in serum and liver were studied in 48 pigs. Twenty-four of the pigs were each infected by intramuscular inoculation with 2000 cercariae of S. japonicum in medium and the rest were similarly inoculated with parasite-free medium, as controls. On each of weeks 4, 11, 17 and 24 post-inoculation (PI), 12 pigs (six of which were infected) were killed. Tissue samples were collected at necropsy. Blood samples were taken prior to infection and at necropsy from all pigs, and bi-weekly from the pigs killed 24 weeks post-infection. In an analysis of variance in which serum retinol was the dependent variable, the interaction infection x time was found to be significant (P = 0.009). The main reason for this significance was that the concentration of retinol in the sera collected from infected pigs at necropsy at 11 weeks PI was significantly lower than in the control pigs killed at the same time (P = 0.02). Although, overall, infection led to higher zinc concentrations in the liver (P = 0.04) and spleen tissue (P = 0.01), it had no apparent effect on liver retinol, muscle zinc or serum zinc. However, among the pigs which were tested bi-weekly, serum zinc was consistently lower in the infected pigs than in the controls (P = 0.01). The transient declines seen in the concentrations of retinol and zinc in sera from the infected pigs were not accompanied by similar changes in the tissue concentrations, and may reflect an acute-phase response to infection. Schistosoma japonicum infection in pigs is considered a useful model of S. japonicum infection (and probably also of S. mansoni infection) in humans. Similar effects, if they occur in the human infections, may lead to misclassification of vitamin-A and zinc status in endemic populations if this status is based on serum retinol and serum zinc.

False-positive Trichuris suis egg counts in pigs in relation to coprophagia.

Sixteen non-infected control pigs housed together with 16 pigs infected with Trichuris suis, excreted T. suis eggs in their faeces (range 20-4, 960 eggs per gram faeces (EPG)). When the control pigs were moved to clean pens, their egg counts dropped to zero EPG within five days. A significant correlation was found between T. suis egg counts of infected and control pigs penned together (r = 0.89, P < 0.001). These results suggest that false-positive faecal egg counts may be the result of coprophagia.

An abattoir survey of gastrointestinal nematode infections in cattle in the central highlands of Kenya.

The gastrointestinal tracts of 672 crossbred cattle were obtained from various abattoirs in Kiambu District, Kenya from August 1992 to July 1993, and examined for the presence of gastrointestinal nematodes. Eight nematode species were found in 583 (86.8%) of the animals. The nematodes were, in order of prevalence: Haemonchus placei (67.0%), Cooperia pectinata (53.0%), Cooperia punctata (41.7%), Oesophagostomum radiatum (38.4%), Trichostronglyus axei (24.3%), Nematodirus helvetianus (19.6%), Trichuris globulosa (9.7%) and Strongyloides papillosus (3.6%). The intensity of the nematode infection was moderate; the mean burden being less than 7000 worms. H. placei accounted, on average, for 52.3% of the total burden. The total burden was least during the dry seasons and increased gradually during the rainy seasons. Adult H. placei persisted in the host throughout the year and there was no indication of hypobiosis. The heaviest gastrointestinal worm burdens were detected in 1.5- to 3-year-old animals. These findings are discussed with regard to their relevance for strategic control of gastrointestinal nematodes in cattle.