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.     

Mechanisms underlying resistance to nematode infection

Lambs show considerable genetic variation in faecal egg count following natural, predominantly Ostertagia circumcinta infection. This genetic variation is acquired and not innate. Worm length is positively associated with worm fecundity. The genetic variation in faecal egg count is a consequence of genetic variation in worm length and hence worm fecundity, and not of genetic variation in worm burdens. In contrast to lambs, mature sheep may be able to regulate both fecundity and worm numbers. In lambs, three factors account for the majority of the variation in worm length: the strength of the local IgA response against fourth-stage larvae, the specificity of this response against four molecules in particular, and the density-dependent influence of worm number.     

Rapid improvement of immunity to Teladorsagia circumcincta is achieved through a reduction in the demand for protein in lactating ewes

Protein supplementation can improve the resistance to parasites of periparturient ewes, as indicated by reduced nematode egg excretion and worm burdens. However, the rate at which this improvement can occur is largely unknown. We investigated the rate of improvement by assessing temporal changes in faecal egg counts after we experimentally reduced nutrient demand. Three groups of nine pregnant ewes each were trickle infected with Teladorsagia circumcincta from day(-70) to day(16) (parturition is day0). Two groups of twin-rearing ewes were fed at 0.8 (L22) or 1.2 (H22) times their assumed metabolizable protein requirements, and a third group was fed the same daily food allowances as L22 ewes, but one of their lambs was removed on day10 (L21). Ewes were slaughtered on day21 to assess worm burdens, in vitro larval establishment on abomasal explants, and mucosal inflammatory cells. Faecal egg counts of L22 ewes were higher than H22 ewes throughout lactation. After the removal of one lamb, faecal egg counts of L21 ewes decreased within 5 days to levels similar to H22 ewes. Relative to L22 ewes, L21 and H22 ewes had lower worm burdens, parasite per capita fecundity and in vitro establishment rates of both T. circumcincta and Haemonchus contortus. Mucosal mast cell and eosinophil counts were similar for all ewes, but H22 ewes had higher globule leukocyte counts than L22 and L21 ewes. The data suggest that a reduction in protein demand can rapidly improve periparturient immunity to T. circumcincta. This may be associated with increased parasite expulsion, reduced fecundity and non-parasite specific reduction of in vitro larval establishment.     

Immunity acquired by sheep from an experimental infection with haemonchus contortus

Adams D.B. and Beh K.J. 1981. Immunity acquired by sheep from an experimental infection with Haemonchus contortus. International Journal for Parasitology11: 381–386. A primary infection of sheep with a single dose of Haemonchus contortus larvae was traced by faecal egg counts until it had substantially declined after 55 weeks. These primed sheep were then given a sequence of two reinfections with the parasite. Comparison of faecal egg counts in primed sheep and in two separate groups of previously worm-free sheep showed that primary infection conferred significant immunity. This, however, was not sufficiently protective to prevent the development of further anaemia and faecal egg counts indicative of clinical haemonchosis. It is suggested that an adaptation in the host-parasite relationship which promotes the longevity of primary infection with H. contortus may also moderate the induction of acquired immunity.The titre of haemagglutinating antibody specific for H. contortus rose in serum during the course of primary infection, but the two reinfections did not stimulate a rise in titre. Titres of haemagglutinating antibody before reinfection did not correlate with subsequent faecal egg counts.     

Response to re-infection with Brachylaima cribbi in immunocompetent and immunodeficient mice

The course of infection in C57BL/6J mice re-infected with Brachylaima cribbi was assessed by comparing faecal egg excretion of re-infected mice with age- and sex-matched mice receiving a primary infection only. For both male and female mice there was a significant reduction in the mean number of eggs per gram of faeces at the peak of infection 4 weeks after the challenge infection compared with mice receiving a primary infection only. There was no significant difference in the duration of the infection. This experiment was repeated using age-matched male mice but on this occasion all mice were killed and dissected 4 weeks after the challenge infection and mean eggs per gram of faeces, worm burden and fecundity determined. There was no significant difference in the worm burdens of the re-infected mice compared with age-matched animals receiving a primary infection only. However, there were significant differences in the mean faecal eggs per gram and worm fecundity with the challenge infection group having lower egg counts and reduced fecundity. An enzyme-linked immunosorbent assay using whole worm antigens was developed and used to determine mouse anti-B. cribbi serum antibody levels during the course of infection. Anti-B. cribbi serum antibody absorbance ratios increased six- to sevenfold by 4 weeks after a primary infection beyond which a constant level was maintained. The course of challenge infection in non-obese diabetic severe combined immunodeficient mice showed no significant differences in egg excretion, worm burden or fecundity when primary and challenge infections were compared. These results indicate that the immune response invoked by a previous B. cribbi infection in immunocompetent mice affects fecundity but does not affect the establishment or duration of infection.     

Population dynamics of Trichostrongylus colubriformis in sheep: model to predict the worm population over time as a function of infection rate and host age

The developing immunity of sheep to Trichostrongylus colubriformis infections was described by a mathematical function. The rate of adult establishment was assumed to be a measure of the host's acquired immunity to this parasite. Prediction of establishment from infection rate and host age was used to estimate worm burden, worm rejection and arrested development.     

Time of onset and target of immune reactions in sheep with acquired immunity against Haemonchus contortus

Adams D. B. 1982. Time of onset and target of immune reactions in sheep with acquired immunity against Haemonchus contortus. International Journal for Parasitology12: 439–443. Nonimmune sheep and sheep rendered immune by infection with the abomasal nematode, Haemonchus contortus, were infected with the parasite and treated at various times with the glucocorticosteroid, dexamethasone. The results show that this immunosuppressant drug abolished acquired but not innate immunity to H. contortus and that acquired responses were not important in restraining the fecundity of adult worms during primary infection. By treating immune sheep with dexamethasone during reinfection, it was shown that the responses acting against the establishment of infection commence later than the fourth day after larval administration and are complete by the seventh day. H. contortus more advanced in development than the fourth-larval stage were relatively insusceptible to this manifestation of acquired immunity.     

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.     

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.     

Trichinella spiralis: antifecundity and antinewborn larvae immunity in swine

The development of antifecundity and antinewborn larvae immunity in swine infected with Trichinella spiralis was investigated. In primary infections, adult female worm fecundity dropped sharply after 3 weeks, although adults could be recovered from the small intestine for at least 7 weeks after infection. In challenge infections of pigs infected previously, adult female worm fecundity was depressed up to 51% and the adults were expelled within 3 weeks. Since immune pigs are almost completely resistant to the secondary establishment of muscle larvae, this suggested the existence of immune effector mechanisms also acting on the newborn larvae. This was supported by observations, using an indirect fluorescent antibody assay, that pig antibody bound to the surface of the newborn larvae. Passive transfer of immune pig serum resulted in a large reduction in muscle larvae burden in both infected pig and rat recipients. Adult female worm fecundity in such immune serum recipients was reduced only by 20% and worm survival in the intestine was unaffected. These results indicate that immunity to the newborn larvae, in addition to antifecundity effects, are responsible for the high levels of acquired resistance to T. spiralis in swine.     

Energy deficits suppress both systemic and gut immunity during infection.

Protein-energy malnutrition and gastrointestinal nematode infections widely coexist in developing countries. Evidence is provided demonstrating the profound impact of dietary energy deficiency on immune function. Energy-restricted (ER) mice infected with a gastrointestinal nematode showed impaired lymphocyte proliferation and reduced production of Th2 cytokines and lower levels of IgE, parasite-specific IgG1, and eosinophils, which led to higher worm burdens and fecundity. We conclude that mild ER, without concurrent protein malnutrition, can modulate protective immunity from (a) activation early during a primary infection to (b) the expression of acquired immunity during reinfection in both systemic and gut-associated lymphoid tissues.     

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.     

Observations on the epidemiology and interactions between myxomatosis, coccidiosis and helminth parasites in a wild rabbit population in Scotland

A study of the epidemiology of myxomatosis and the protozoan liver parasite, Eimeria stiedae, in a population of wild rabbits in Scotland from 1977 to 2010 is reported. Rabbits were collected on a monthly basis resulting in a total of 5,337 animals examined for the infections. The investigation showed that within any 1 year over the 34 years of the investigation the percentage of rabbits with myxomatosis varied between 0 and 24 %, while for E. stiedae infections fluctuated between 3 and 42 %. There were strong seasonal trends in the prevalence of myxomatosis with over 16 % being infected in September and October, and for E. stiedae, peaks of over 40 % of livers infected were recorded in July. From 2007 to 2010, faeces were also examined for coccidia oocysts and parasitic nematode eggs. Rabbits infected with the myxoma virus had mean oocyst counts of 73,665 per gram faeces, while rabbits not infected with the myxoma virus had counts of 31,952 oocysts per gram. Comparable figures for nematode faecal egg counts were 911 per gram in myxomatosis-infected animals and 427 per gram in uninfected animals. The elevated nematode faecal egg counts in rabbits with myxomatosis reflects increased worm burdens already reported, but the elevated counts of coccidial oocysts have not previously been reported. This would suggest that myxomatosis could compromise rabbit immunity to nematodes and coccidia.     

Faecal egg counts provide a reliable measure of Trichostrongylus tenuis intensities in free-living red grouse Lagopus lagopus scoticus

The reliability of different egg counting methods for estimating the intensity of Trichostrongylus tenuis infections in red grouse, Lagopus lagopus scoticus, was investigated in the autumn, when grouse may harbour high parasite intensities. Possible limitations to the use of these methods were also examined. Faecal egg counts were found to accurately estimate T. tenuis worm intensities, at least up to an observed maximum of c. 8000 worms. Two egg counting methods (smear and McMaster) gave consistent results, although the exact relationship with worm intensity differed according to the method used. Faecal egg counts significantly decreased with increasing length of sample storage time, but egg counts were reliable for estimating worm intensity for three weeks. The concentration of eggs in the caecum was also found to reliably estimate worm intensity. However, egg counts from frozen gut samples cannot be used to estimate worm intensities. These results conclude that, despite some limitations, faecal and caecum egg counts provide useful and reliable ways of measuring T. tenuis intensities in red grouse.     

Identification and characterisation of an aspartyl protease inhibitor homologue as a major allergen of Trichostrongylus colubriformis

Allergens were identified from the gastrointestinal nematode of sheep, Trichostrongylus colubriformis, by probing Western blots of infective larvae (third stage) somatic antigen with IgE purified from the serum of sheep grazed on worm contaminated pasture. A 31 kDa allergen was frequently recognised by sera from immune sheep, particularly those deriving from a line that has been genetically selected over 23 years for parasite resistance. Using a proteomic approach, the 31 kDa allergen was identified as an aspartyl protease inhibitor homologue. The entire coding sequence of T. colubriformis aspartyl protease inhibitor (Tco-api-1) was obtained and the mature protein expressed in Escherichia coli. Anti-Tco-API-1 antibodies revealed that a commonly observed 21 kDa T. colubriformis allergen species is a truncated form of Tco-API-1. Specific IgE responses to T. colubriformis aspartyl protease inhibitor were significantly correlated with the degree of resistance to nematode infection as measured by faecal egg count in sheep. Surprisingly, IgE responses to Tco-API-1 were not correlated with breech soiling (dag score), which is thought to be caused, in part, by allergic hypersensitivity to worms. Therefore, a specific IgE response to this allergen may be a suitable marker for identifying lambs at an early age that will develop strong immunity to gastrointestinal nematodes.     

Population dynamics of Trichostrongylus colubriformis in sheep: mathematical model of worm fecundity

A mathematical model was constructed to predict the egg production of Trichostrongylus colubriformis worms as a function of worm age and host experience of infection. The model set egg production at zero until the worm was 14 days old, when a linear increase to maximum egg production levels occurred over 7 days. It was assumed that egg production remained at maximum levels until a threshold total worm burden was exceeded, when an exponential decline in egg production occurred. The rate of decline was assumed independent of worm age or worm burden. The estimated parameters (maximum egg production, threshold, lag and rate of decline) were optimized by fitting values predicted from the model to faecal egg counts observed in continuously infected sheep, giving R2 = 0.80. The model was validated against faecal egg counts obtained in two other continuous infection experiments, one performed at the same laboratory and the other in Britain.     

Genetic resistance to helminths. The influence of breed and haemoglobin type on the response of sheep to re-infection with Haemonchus contortus.

The influence of genetic factors on acquired resistance to Haemonchus contortus infections in sheep was investigated. Animals whose primary infections were terminated with an anthelmintic failed to develop any immunity against subsequent challenge as judged by worm numbers. Nevertheless, all were better able to retard the development and reduce the fecundity and haematophagic activities of their parasite populations than animals undergoing primary infections. High levels of resistance, as judged by all these parameters, were observed in most animals when the challenge larvae were superimposed on existing worm populations. The patterns of worm establishment and disease indicated that genetic factors operated in determining resistance, since fewer worms became established and less severe clinical and pathophysiological changes were observed in Scottish Blackface than in Finn Dorset sheep with the same haemoglobin type. Similar advantages were displayed by animals with haemoglobin AA and to a lesser extent those with haemoglobin AB over haemoglobin BB types. The importance of breed was further indicated by the occurrence of 'self-cure' in the majority of the Scottish Blackfaces but in only one Finn Dorset. There was no evidence that this reaction was associated with haemoglobin type.     

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.     

Polyclonal and specific antibodies mediate protective immunity against enteric helminth infection

Anti-helminth immunity involves CD4+ T cells, yet the precise effector mechanisms responsible for parasite killing or expulsion remain elusive. We now report an essential role for antibodies in mediating immunity against the enteric helminth Heligmosomoides polygyrus (Hp), a natural murine parasite that establishes chronic infection. Polyclonal IgG antibodies, present in naive mice and produced following Hp infection, functioned to limit egg production by adult parasites. Comparatively, affinity-matured parasite-specific IgG and IgA antibodies that developed only after multiple infections were required to prevent adult worm development. These data reveal complementary roles for polyclonal and affinity-matured parasite-specific antibodies in preventing enteric helminth infection by limiting parasite fecundity and providing immune protection against reinfection, respectively. We propose that parasite-induced polyclonal antibodies play a dual role, whereby the parasite is allowed to establish chronicity, while parasite load and spread are limited, likely reflecting the long coevolution of helminth parasites with their hosts.     

Acquired immune heterogeneity and its sources in human helminth infection

Similarities in the immunobiology of different parasitic worm infections indicate that co-evolution of humans and helminths has shaped a common anti-helminth immune response. However, recent in vitro and immuno-epidemiological studies highlight fundamental differences and plasticity within host-helminth interactions. The 'trade-off' between immunity and immunopathology inherent in host immune responses occurs on a background of genetic polymorphism, variable exposure patterns and infection history. For the parasite, variation in life-cycle and antigen expression can influence the effector responses directed against them. This is particularly apparent when comparing gastrointestinal and tissue-dwelling helminths. Furthermore, insights into the impact of anti-helminthic treatment and co-infection on acquired immunity suggest that immune heterogeneity arises not from hosts and parasites in isolation, but also from the environment in which immune responses develop. Large-scale differences observed in the epidemiology of human helminthiases are a product of complex host-parasite-environment interactions which, given potential for exposure to parasite antigens in utero, can arise even before a parasite interacts with its human host. This review summarizes key differences identified in human acquired immune responses to nematode and trematode infections of public health importance and explores the factors contributing to these variations.