Resistance to experimental infection with Fasciola hepatica in exotic and domestic breeds of sheep

Significant breed differences were detected in fecal egg counts and fluke counts following experimental infection of five breeds of sheep with metacercariae of F. hepatica. Two experiments compared the responses of Barbados Blackbelly, St. Croix, Florida Native, Finn X Rambouillet and Targhee sheep to primary and challenge infections (Expt. 1) and to multiple infections (Expt. 2). Barbados Blackbelly sheep were more susceptible to infection than the other breeds in both experiments. St. Croix and Florida Native sheep were the most resistant breeds in Expt. 1, while Targhee and Florida Native were the most resistant in Expt. 2. There was no evidence of acquired immunity in any breed in either experiment.     

Resistance of nonlambing exotic and domestic ewes to naturally acquired gastrointestinal nematodes

No breed differences in Haemonchus contortus burdens were found when nonlacfating exotic (St. Croix and Barbados Blackbelly), domestic (Finn-Dorset × Rambouillet) and $\text{1}\text{2}\text{exotic}\text{-}\text{1}\text{2}$ domestic ewes grazed fall pasture, but all ewes had significantly (p < 0.05) lower worm burdens at necropsy than tracer lambs grazing the same pasture. Florida Native ewes grazing the same pasture showed significantly (p < 0.05) less packed cell volume (PCV) decrease and significantly (p < 0.05) lower fecal egg counts than the other ewe breeds but were not available for necropsy. Tracer lambs had a significantly (p < 0.05) greater PCV decrease and significantly (p < 0.05) higher fecal egg counts than ewes.The results of this study suggest that breed differences in the periparturient rise in fecal egg counts of exotic and domestic ewes observed in an earlier study may have been caused by breed differences in the periparturient relaxation of immunity rather than breed differences in the ability to acquire immunity to worms.     

A comparison of the periparturient rise in fecal egg counts of exotic and domestic ewes

Ewes of three exotic breeds (Florida Native, Barbados Blackbelly and St. Croix) showed no periparturient rise in fecal egg counts (PPR) when housed from late fall through lambing and weaning. Domestic breed ewes (Rambouillet and Finn-Dorset × Rambouillet) showed a pronounced PPR 6–7 weeks post-lambing while St. Croix × domestic ewes showed an intermediate PPR under similar conditions. There was no PPR in non-lambing ewes.In a second experiment, breed differences in the PPR were not as pronounced when ewes were allowed to graze a contaminated pasture after lambing. Florida Native, St. Croix and $\text{3}\text{4}$ St. Croix × $\text{1}\text{4}$ domestic showed lower PPRs than the remaining breeds (Barbados Blackbelly, F₁ hybrids of exotic and domestic breeds and domestic ewes), but the magnitude of the PPR was generally small in all breeds. No relationship was observed between hemoglobin genotype and PPR in either experiment.It was concluded that a low or absent PPR is an important manifestation of breed resistance in exotic ewes and that this may be useful as a marker for breeding parasite-resistant sheep.     

The periparturient rise in fecal egg counts in three strains of Florida native ewes and its value in predicting resistance of lambs to Haemonchus contortus

The periparturient rise in fecal egg counts in three strains of Florida Native ewes and its value in predicting resistance of lambs to Haemonchus contortus. International Journal for Parasitology16: 185–189. Significant (P < 0.05) differences in the magnitude of the periparturient rise in fecal egg counts (PPR) occurred in ewes of three strains of Florida Native sheep. Non-lambing ewes (n = 12) and lactating University strain ewes (n = 31), a strain selected over a 26-year period for parasite resistance by survival at pasture without anthelmintic treatment, had lower fecal egg counts than ewes of either Maxcy strain (n= 17), a commercial strain treated regularly with anthelmintics, or Backlinie strain (n = 16), a strain of Florida Native sheep that had some crossbreeding to Hampshire and Suffolk in their genotype and were regularly treated with anthelmintics. Whereas both Maxcy and Backlinie ewes showed a pronounced PPR, none occurred in non-lambing ewes (n=12) and the slight rise in fecal egg counts observed in University ewes was not significantly different from that of the non-lambing ewes. Backlinie ewes having twin lambs (n = 5) had a higher PPR than Backlinie ewes with single lambs (n = 11) or Maxcy ewes with single lambs (n = 15), but all three groups had higher PPRs than University ewes with single lambs (n = 27). No differences occurred in fecal egg counts of non-lambing ewes regardless of strain. Six weeks after weaning fecal egg counts of all ewes were uniformly low regardless of strain or prior lactation status.Ram lambs born to these ewes showed no significant strain differences in Haemonchus contortus burdens after experimental infection, deworming with levamisole and reinfection, although University lambs (n= 13) had lowest mean worm burdens at necropsy (2437) followed by Maxcy lambs (n= 12, mean =3106) and Backlinie lambs (n= 13, mean =3670). There was no significant correlation between worm burdens in these lambs and the magnitude of the PPR in their dams. Eight to 16 weeks after turn out to a common parasite-infested pasture ewe lambs (n = 13) born to University ewes had lower fecal egg counts than similar ewe lambs (n=8) born to Backlinie ewes. Prior to this time differences were not significant and egg counts were generally low. The magnitude of the PPR in their dams was not significantly correlated with fecal egg counts or PCVs of individual ewe lambs.     

Parasitological and immunological responses of genetically resistant Merino sheep on pastures contaminated with parasitic nematodes.

One hundred and twenty lambs were grazed continuously from weaning until 9 months of age on 12 plots contaminated with larvae of three nematode species (Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia circumcincta). The lambs were sired by either a genetically resistant ram or susceptible rams (determined by the response of previous progeny to artificial H. contortus infection). Half the resistant and half the susceptible lambs were given strategic anthelmintic treatment and the remainder remained untreated. Faecal egg counts and blood packed cell volume were measured frequently in all animals. One and 5 months after weaning, two lambs from each plot were slaughtered, and worm burdens and larval establishment rates of the three species of nematode were estimated. At the second slaughter, leukotriene levels and larval migration inhibitory (LMI) activity were measured in mucus collected from the small intestine. The dominant species in all faecal samples and the gastrointestinal tract was T. colubriformis. Lambs of the resistant genotype had lower faecal worm egg counts, lower worm burdens and higher levels of resistance to larval establishment. There were no differences in larval migration inhibition (LMI) activity, but resistant lambs had higher levels of the leukotriene LTC4/D4/E4. Further, the resistant genotype, identified on responsiveness to artificial infections with H. contortus, was more resistant to infections of three important species acquired naturally from contaminated pastures. All these genetic differences were maintained while the lambs were subject to strategic anthelmintic treatment.     

Merino ewes bred for parasite resistance reduce larval contamination onto pasture during the peri-parturient period

The peri-parturient period is crucial for controlling worms as the acquired immunity of ewes is disrupted, resulting in an increase in faecal worm egg counts. Two hypotheses were tested in this experiment - that ewes bred for worm resistance would have lower faecal worm egg counts than unselected control ewes, during late pregnancy and lactation, under similar but separate grazing areas; and also that numbers of infective nematode larvae would be lower on pastures grazed by resistant ewes than pastures grazed by unselected control ewes. Faecal samples were collected from resistant and unselected ewes in late pregnancy and early lactation, during the winter rainfall season, and analysed for numbers of Trichostrongylus colubriformis and Teladorsagia circumcincta. Pasture samples were taken 1 week before and 7 weeks after lambing started and analysed for infective larvae. In all sheep, worm egg counts rose 2 weeks prior to lambing and continued into lactation. Worm egg counts were significantly lower in the resistant ewes from 1 week before lambing to 2 weeks after lambing. There were no differences in egg counts between single- and twin-bearing ewes in the resistant line. However, twin-bearing control ewes had significantly higher egg counts than single-bearing control ewes. Following lactation, plots grazed by resistant ewes had substantially less contamination with T. colubriformis larvae, but there were no differences in numbers of T. circumcincta larvae. Our results demonstrate that sheep bred for worm resistance has lower worm burdens during the peri-parturient phase and that lambs born to resistant ewes face a lower larval challenge during their introduction to grazing. In our environment, selection for low worm egg counts has produced sheep highly resistant to T. colubriformis, but has had less impact on resistance towards T. circumcincta.     

In vivo and in vitro egg production by Nematospiroides dubius during primary and challenge infections in resistant and susceptible strains of mice

Experiments were conducted to examine adult worm burdens, fecal egg output, and in vitro fecundity of Nematospiroides dubius in resistant LAF1 and susceptible CBA mice 12, 15, 18, and 21 days following primary and challenge infections. A strong correlation was obtained on the number of eggs produced by worms cultured in vitro and the egg production as assessed by fecal egg count. Worm counts, fecal egg counts, and in vitro fecundity were similar on all days studied following a primary infection in both mouse strains. However, after challenge infection, LAF1 mice showed lower worm burdens, fecal egg output, and in vitro egg production when compared to CBA mice. Although the egg production of surviving female worms from immune LAF1 mice was decreased, it never fell below a threshold of 100 eggs/day. The reduced fecundity may be a manifestation of a general anti-worm response rather than responses directed specifically at worm reproduction.     

Is the establishment rate and fecundity of Haemonchus contortus related to body or abomasal measurements in sheep?

The relationship among parasitological parameters, abomasal size and body size measurements was investigated in lambs following an experimental infection with Haemonchus contortus. In total, 100 lambs from five different genotypes (German Merino (GM), Texel × GM, Suffolk × GM, German Blackhead Mutton × GM and Ile de France × GM) were experimentally infected with 5000 infective third stage larvae of H. contortus at the time of weaning at 12 weeks of age. Four and six weeks after infection, individual faecal samples were collected for estimation of faecal egg counts (FECs). Furthermore, wither height, shoulder width, heart girth, loin girth and body length were taken at 18 weeks of life. Lambs were slaughtered and necropsied 7 weeks post-infection, and worm counts, abomasal volume and surface area were determined. Positive correlations were found between different body size parameters, body weight and abomasal sizes. FEC and worm counts were not significantly correlated either with body size parameters or with abomasal size. The mean worm burden was higher in GM than in crossbred lambs. There was no significant difference in abomasal size between GM and crossbred lambs. The results suggest that the variations between animals in worm burden following an experimental infection with H. contortus (worm resistance) are not influenced by body size parameters or abomasal sizes. Therefore, other factors, including genetic-based differences in resistance, must cause these findings between and within breeds.     

Variation in resistance to haemonchosis: selection of female sheep resistant to Haemonchus contortus.

Seventy female lambs (6-7 months old) which were exposed to natural infections of Haemonchus contortus were designated as responders or non-responders on the basis of 10 weekly cumulative faecal egg counts. Selected responder and non-responder lambs were treated with ivermectin, housed separately and 6 weeks post-housing, seven lambs from each group were given a trickle infection of Haemonchus contortus at 1000 L3 daily for 5 days per week up to 2 weeks and examined weekly for 10 weeks after first infection. Analysis of data revealed significantly lower mean faecal egg counts and non-significantly less weight loss in responder than non-responder lambs. Mean values of haemoglobin, packed cell volume, total serum protein and peripheral eosinophil counts were significantly higher in responders than non-responders. In contrast, serum pepsinogen concentration was significantly less in responders than in non-responders. At 10 weeks post-infection, there were fewer pathological lesions and significantly lower worm burdens in responders than in non-responders. These results demonstrate a distinct resistance in responders to Haemonchus contortus infection.     

Schistosoma japonicum: day to day variation in excretion and hatchability of parasite eggs in the domestic pig, Suis suis

The aim of the present study was to describe the course of an Schistosoma japonicum infection in individual pigs over time, with special regards to fecal egg counts and egg hatchability, emphasizing the extent of variation during and between days. Five specific pathogen free Danish Landrace/Yorkshire/Duroc crossbred male pigs were each given 3500 cercariae intramuscularly. From day 36 to 62 post infection, fecal samples were collected from each pig, morning and evening. Pigs were perfused 62 days post infection. Actual fecal egg counts, miracidial counts, and worm burdens were determined. The trend of fecal egg excretion was adequately described with a third order polynomial and logarithmic link function. Miracidial counts were related directly to morning fecal egg counts through a simple linear function. The study revealed a fairly similar overall pattern of egg excretion for all pigs, showing an increase in egg excretion until week 8, followed by a marked reduction to almost zero. In general, large fluctuations around this trend were seen for all pigs, as revealed in variations in egg counts within the same day as well as between days. However, as revealed by the estimated model lines, these fluctuations are due to random variation in egg density in stools rather than being caused by biologically determined cycles. An exact time for both first occurrence of eggs in feces (days 35-38 post infection) and for peak egg counts (days 44-48 post infection) was predicted for each individual pig. Furthermore, the model revealed that miracidial counts can be related directly to the expected fecal egg counts of the same day through a parameter, which represents hatchability. This hatchability parameter was found to be independent of time post infection for each pig, but large individual differences were seen in hatchability between the pigs.     

Population dynamics of Trichostrongylus colubriformis and Ostertagia circumcincta in single and concurrent infections.

Twenty-one-week-old worm-free pen-reared lambs were infected weekly with either 10,000 T. colubriformis larvae, 5000 O. circumcincta larvae, or with both species (15,000 larvae per week). Larval establishment and total worm burdens were estimated after 4, 7, 10 and 13 weeks of infection. Faecal egg counts and lamb bodyweights were measured weekly, and numbers of eosinophils in blood were estimated before infection and at weeks 5, 8 and 14. For both species of worms, the dynamics of infection (establishment, worm burdens, egg counts) were not affected by concurrent or pre-existing infection with the other species. Infection with T. colubriformis alone did not protect against O. circumcincta, but infection with O. circumcincta alone provided slight protection against the T. colubriformis larvae. Blood eosinophils increased between 5 and 8 weeks of infection and were similar for the three infections. This corresponded to the reduction in establishment for both species.     

Responses of hair and wool sheep to a single fixed dose of infective larvae of Haemonchus contortus

Body weights, fecal egg counts (FEC), and packed cell volumes (PCV) of hair sheep and wool sheep crossbred lambs were compared over 8 weeks following administration of a single dose of approximately 10,000 third-stage larvae of Haemonchus contortus. Hair sheep lambs (n=17) were reciprocal crosses between mainland USA populations of Barbados Blackbelly (BB) and Virgin Islands White (VIW) sheep. Wool sheep lambs (n=64) were from a crossbred composite of 50% Dorset, 25% Rambouillet, and 25% Finnish Landrace breeding. Lambs of both breed types continued to grow during the period of infection. Mean weights were higher for wool lambs (39.7±0.8 kg) than for hair lambs (28.2±1.5 kg). FEC increased to week 5 in both breed groups and remained elevated in wool lambs through week 7 but declined sharply in hair lambs after week 5. Mean FEC for weeks 4 through 8 were 4011±361 eggs per gram of feces (epg) in wool lambs, and 1135±196 epg in hair lambs. PCV declined through week 7 in wool lambs but stabilized and then increased after week 4 in hair lambs. Mean PCV in weeks 4 through 8 were 22.4±0.3% in wool lambs and 24.3±0.5% in hair lambs. These results suggest that Caribbean hair breeds may be able to contribute significantly to development of parasite-resistant sheep populations.     

A mechanistic model of developing immunity to Teladorsagia circumcincta infection in lambs

Acquired immunity influences the severity of parasitic disease, but modelling the effects of acquired immunity in helminth infections has proved challenging. This may be due to a lack of suitable immunological data, or to the perceived complexity of modelling the immune response. We have developed a model of T. circumcincta infection in domestic sheep that incorporates the effects of acquired immunity on parasite establishment and fecundity. A large data set from commercially managed populations of Scottish Blackface sheep was used, which included relationships between IgA activity and worm length, and between worm length and fecundity. Use was also made of a recently published meta-analysis of parasite establishment rates. This realistic but simple model of nematode infection emulates observed patterns of faecal egg counts. The end-of-season faecal egg counts are remarkably robust to perturbations in the majority of the parameters, possibly because of priming of the immune system early in the season, reducing parasite establishment and growth and, therefore, faecal egg counts. Lowering the amount of early infection leads to higher end-of-season egg counts. The periparturient rise in egg counts in ewes appears to have an important role in supplying infection for the priming of the immune response. This feedback in the immune priming suggests that nematode infections may be difficult to eliminate.     

The curvilinear relationship between worm length and fecundity of Teladorsagia circumcincta.

The variation among sheep in fecundity of Teladorsagia (Ostertagia) circumcincta was estimated by dividing the faecal egg count by the worm number following deliberate infection of mature Scottish Blackface lambs. Fecundity was skewed and ranged from 0 to 350 eggs per worm per day. Most animals had relatively low worm fecundities, but a small number of individuals had relatively high worm fecundities. However, as fecundity is a ratio of two imprecise estimates, extreme values may be statistical artefacts. Following both deliberate and natural infection, differences in worm fecundity were associated with differences in adult female worm length. In both infections, fecundity varied with worm length to the power 0.4. This relationship should assist the measurement of fecundity in studies of host immunity, in epidemiological modelling and in estimating the influence of density-dependent relationships.     

Worm burden and immunological responses in Corriedale and Crioula Lanada sheep following natural infection with Haemonchus contortus

The effects of natural infection with Haemonchus contortus were compared in 15 Corriedale lambs and 18 Crioula Lanada lambs kept in the grazing conditions after weaning. The following parameters were assessed weekly for 9 consecutive weeks: number of eggs per gram of feces (EPG), packed cell volume (PCV), total serum proteins (TSP) and albumin (ALB). At 10 weeks, the animals were slaughtered and autopsied to evaluate to the worm burden and eosinophil, mast cell and globule leukocyte counts in histological sections of the abomasum. EPG counts and worm burden were significantly lower in Crioula lambs, which also showed higher levels of PCV, TSP and ALB, as well as higher eosinophil and globule leukocyte counts. Mast cell counts were similar for the two breeds. These results indicate that, under intensive pasture conditions, the Crioula Lanada breed shows a better response to natural infection with H. contortus, suggesting greater resistance to parasitism.     

Genome-wide scan reveals important additive and non-additive genetic effects associated with resistance to Haemonchus contortus in Florida Native sheep

Florida Native sheep is among the sheep breeds best adapted to humid and hot climatic conditions such as those of Florida, USA, and have shown a superior ability to regulate nematode burdens. This is one of the oldest sheep breeds in North America and is an endangered species. To ensure genetic diversity and long-term survival of the breed, protection of the current genetic stock is critical and conservation efforts are required to promote its breeding and production. The objective of the present study was to investigate the importance of additive and non-additive genetic effects on resistance to natural Haemonchus contortus infections in Florida Native sheep using a whole genome scan. A total of 200 sheep were evaluated in the present study. Phenotypic records included faecal egg count (FEC, eggs/gram), FAMACHA® score, packed cell volume (PCV, %), body condition score and average daily gain (ADG, kg). Sheep were genotyped using the GGP Ovine 50K SNP chip and 45.2 k single nucleotide polymorphism (SNP) markers spanning the entire genome were available for quality control procedures. Mixed models were used to analyse the response variables and included the identity by state matrix to control for population structure. Bonferroni correction was used to control for multiple testing and a second arbitrary threshold (0.1 × 10^?3) was used. Fifteen SNPs with additive and non-additive genetic effects and located in Ovis aries chromosome OAR1, 2, 3, 6, 8, 10, 11, 12, 13 and 21 were associated with FEC, FAMACHA® score, PCV and ADG. These SNPs could be potential genetic markers for resistance to natural H. contortus exposure in Florida Native sheep.     

Field studies on inhibition of Haemonchus contortus.

The epidemiology of Haemonchus contortus was studied in north east England in lambs reared under field conditions with infected ewes. The parasite population was monitored by pasture larval counts, faecal egg counts and lamb postmortems. From 25 June to 11 November, eight lambs were slaughtered each month, together with eight worm-free tracer lambs. Postmortem worm counts showed that on 25 June, small numbers of adult H. contortus were present but no inhibited larvae. However, subsequently the number of worms recovered rose rapidly to 1000-5000 and the percentage inhibition increased to 57% in July, 75% in August and virtually 100% in September, October and November. On each sampling occasion no significant differences in worm count or percentage inhibition were recorded between experimental and tracer lambs. These results suggest that neither autumn climatic effects nor host immunity were responsible for inhibition in this strain of H. contortus.     

Prenatal Schistosoma japonicum infection in piglets: effect of repeated exposure of the dams on treatment efficacy and susceptibility to challenge infections

This study elucidated the fate of prenatal infections in piglets born by dams repeatedly infected before and during pregnancy with Schistosoma japonicum. Independent variables included repeated infections of the dams and treatment or challenge infection (or both) of the prenatally exposed piglets. Dependant variables were worm counts, fecal and tissue egg counts, weight gain, and gross pathological observations. Fifteen female piglets (the dams) were included, of which 6 received repeated infections with S. japonicum during 6 mo. All dams were inseminated and 10 wk pregnant; 12 of the dams were infected with S. japonicum, of which 6 had been repeatedly infected. Three dams remained uninfected. Eight weeks after delivery, the prenatally exposed piglets (the offspring) were grouped, and 6 of the 12 groups were treated with praziquantel. Four weeks after treatment, 5 groups of piglets were infected with S. japonicum. Groups of piglets were killed either 12 or 22 wk after delivery. Repeated infections of the dam did not prevent establishment of a congenital infection in the pig fetuses. Piglets born with a congenital infection were not resistant to a S. japonicum challenge infection given 12 wk after birth. Neither did praziquantel effectively cure the piglets nor did treatment of the prenatally infected piglets prevent establishment of a challenge infection given 4 wk after treatment. Results of the present study indicate that prenatal exposure, independently of the dam's infection status, may change the host response to challenge infections and treatment after birth.     

Population dynamics of Haemonchus contortus and Trichostrongylus spp. in sheep

Twenty-five lambs grazing a parasite-contaminated pasture were divided into four groups. One group of seven lambs were left untreated and three groups of six lambs each were treated with levamisole at 2 week, 3 week or 1 month intervals respectively. Body weights, fecal egg counts and packed cell volumes were determined every 2 weeks for 8 weeks. Lambs were then necropsied on the day they would have received another treatment, thus the maximum accumulation of parasites at each treatment frequency was measured. Treatment at 2 and 3 week intervals markedly suppressed fecal egg counts, but egg counts of sheep treated every month rose between treatments and untreated animals continually shed eggs. Treatment at all three frequencies provided only slight protection from blood loss, but it lessened weight loss. There was little difference between treatment groups in burdens of adult H. contortus, but burdens of T. axei and T. colubriformis and arrested larvae of H. contortus increased with decreasing frequency of treatment. The results support the findings of other workers that Trichostrongylus spp. have a long adult life span and continue to accumulate with time, whereas H. contortus may have a short adult life span and a rapid population turnover. In the face of constant reinfection, repeated anthelmintic treatment gives more protection from the effects of the long-lived parasites than from those with a rapid population turnover.     

The oral route as a potential way of transmission of Schistosoma bovis in goats.

The infectivity of Schistosoma bovis cercariae administered orally was evaluated in Sahelian goats. Compared to the percutaneous route, a single massive oral dose resulted in a worm burden and in fecal egg excretion reduced by one-half. Surprisingly, tissue egg counts were increased by more than 4-fold. Fecundity of individual female schistosomes was, therefore, markedly increased. When infective doses were administered weekly for 20 wk, both worm and egg burdens were doubled without modification of the individual worm pair fecundity. Repeated oral infections seem to have induced an acquired tolerance toward parasite antigens. These results confirm the epidemiologic relevance of the oral route in a host species inclined to become infected through drinking water rather than percutaneous exposures.