Echinococcus granulosus in sheep: transfer from ewe to lamb of 'Arc 5' antibodies and oncosphere-killing activity, but not protection.

Fourteen ewes were orally dosed with 2000 E. granulosus eggs at 2 weeks of age, were mated at 19 months, and produced lambs when the cysts were 2 years old. One week after parturition, all 14 ewes had 'Arc 5' antibodies in their serum, as did 11/14 of their lambs. Fourteen uninfected ewes were immunized three times before parturition with E. granulosus eggs injected intramuscularly. Cysts grew at the first, or first and second site, but not the third, indicating that the ewes were immune prior to parturition. Most sera from these ewes and their lambs, and from 14 control ewes and their lambs, produced precipitin arcs with hydatid cyst fluid, but no 'Arc 5'. All lambs were challenged with 500 eggs 2 weeks after birth. At necropsy, cyst numbers within groups ranged from 3 to > 200, but there was no significant difference between the three groups of lambs. The immunized ewes did not pass a protection to their lambs that was effective when the lambs were challenged. 'Arc 5' antibodies were induced by prolonged infection with cysts, and were not seen in the sera of ewes immunized with eggs, although the eggs developed into cysts at the injection sites. 'Arc 5' antibodies did not protect lambs against infection, and were not correlated with protection in ewes. Subcutaneous injection of oncospheres into four ewes from each group at the time of lamb challenge showed that the immunized ewes were immune to this method of challenge, but the infected and control ewes were not.(ABSTRACT TRUNCATED AT 250 WORDS)     

The use of a water-in-oil adjuvanted vaccine in an attempt to immunize lambs against Taenia ovis cysts in the presence of maternal antibody

An attempt was made to actively immunize lambs during the period when they had maternally-acquired antibody, so that there would be no time when lambs were susceptible to Taenia ovis infection. A slow-release water-in-oil adjuvanted vaccine containing T. ovis oncosphere products was used. Half the ewes were vaccinated prior to parturition and lambs were vaccinated at approx. 5 weeks of age. At necropsy, after challenge infection, vaccinated lambs had a mean of 39 cysts, compared to 131 in unvaccinated control lambs (p < 0.05), but there was no effect on development or survival of the established cysts. The presence of passively-acquired anti-oncosphere antibody at the time of vaccination did appear to influence the degree of antibody response, and the level of protection induced. However, there was no correlation between levels of antibody at vaccination and levels of protection induced, because of the considerable individual variation between lambs. In non-vaccinated control lambs, the serological response to challenge infection was also highly variable.Some vaccinated lambs developed a strong primary serological response and were mostly (5/7) fully protected. Lesser degrees of primary response in other lambs did not fully protect. All vaccinated lambs gave a rapid secondary response after challenge, but it was too late to be protective. The immunization procedure, although effective in some lambs and partially effective in most, cannot be recommended.     

Active immunization of ewes against ovine placental lactogen increases birth weight of lambs and milk production with no adverse effect on conception rate

In two experiments, 16 Booroola-Assaf and 35 Assaf ewe-lambs were actively immunized at 5 months of age against recombinant ovine placental lactogen (oPL). At 9 months of age, the ewe-lambs were mated for the first time and then introduced into a frequent mating-system. Anti-oPL antibody titers, reproductive performance, maternal serum levels of oPL during pregnancy, lamb birth weight and milk production of the ewes were followed in the immunized ewes and in their non-immunized control counterparts. All the immunized ewes developed anti-oPL antibodies, which interfered with oPL bioactivity in an in vitro cell proliferation assay. Conception rates did not differ (P>0.05) between immunized and non-immunized ewes. Abundant antibody-bound non-active oPL detected in sera of immunized ewes by western blotting indicated enhanced oPL production by the placenta following immunization. An increase (P<0.02) in serum oPL bioactivity, but not immunoreactivity, was observed in the immunized ewes in late gestation relative to control ewes. The average litter size was 1.83 and 1.32 lambs born per ewe lambing in the first and second experiments, respectively. Average birth weights of lambs born to the immunized ewes were higher (P<0.01) than for lambs born to control ewes by 10, 17 and 39% for those born as singles, twins and triplets, respectively. Immunized ewes produced 19 and 33% more milk (P<0.02) than the control ewes in the first 3.5 months of the first and second lactations, respectively. These findings do not suggest a role for oPL in maternal recognition of pregnancy, but they strongly suggest important roles for oPL in fetal growth and mammogenesis. Immunization of ewes against oPL may thus represent a novel practical technique for enhancing birth weights of lambs born to prolific sheep, as well as milk production by both dairy and mutton ewes.     

Evaluation of progesterone, estrogens and placental lactogen in pregnant ewes following experimental infection with an abortive agent chlamydia psittaci "Var-Ovis"

The plasma levels of progesterone, free and conjugated estrone and of placental lactogen have been determined in three groups of pregnant ewes inoculated with an abortive agent (Chlamydia psittacci var. ovis) on day 66 of gestation. In the control group, ewes were immunized against this abortive agent, and lambed after a normal duration of pregnancy; the blood levels of the three studied hormones showed normal patterns. In the two other groups, the ewes which were not immunized, had a shortened gestation and gave birth in most cases to dead lambs. In some animals, plasma levels of estrone followed an apparently normal evolution; in others the amplitude of the prepartum rise was lower than in the control group. Moreover, plasma levels of progesterone and of placental lactogen were lower in the animals which aborted than in the controls. This effect on the secretion of these two placental hormones probably reflects the action of the abortive agent on the placenta.     

Antibody responses of experimentally infected lambs to antigens collected during in vitro maintenance of the adult, metacestode or oncosphere stages of Taenia hydatigena and Taenia ovis with further observations on anti-oncospheral antibodies

The antigenicity and specificity of crude antigens collected during the in vitro maintenance of Taenia hydatigena and T. ovis, excretory/secretory (ES) antigens, were assessed in a peroxidase microenzyme-linked immunosorbent assay (ELISA), using sera from lambs given experimental monospecific infections with T. hydatigena, T. ovis, Echinococcus granulosus or Fasciola hepatica. ES antigens of larval cysts of T. ovis and T. hydatigena were less reactive than those of adult or oncosphere stages. Strong interspecific cros-reactions occurred between all antigen preparations, and these antigens offered no better specificity than crude somatic extracts. IgG1 was the major immunoglobulin detected in sera from lambs experimentally infected with T. ovis or T. hydatigena using antigens prepared from sonicated oncospheres. Discrete peaks of anti-oncospheral antibodies were detected following initial and challenge infections with eggs (whether the homologous or heterologous species), when sera were assayed with a PBS sonicate or an ES antigen from oncospheres. However, when oncospheres solubilised with sodium deoxycholate were used, the antibody response was prolonged and resembled that reported previously when somatic extracts of adult and metacestode stages were used as antigen. The results showed that oncospheres share antigens in common with other life-cycle stages, but also support the notion that they may possess some unique stage-specific antigenic determinants.     

Studies on the abomasal pathology of immunized and non-immunized sheep infected with Haemonchus contortus

Five adult ewes and five lambs were repeatedly immunized with weekly doses of 10,000 irradiated Haemonchus contortus L3 before challenge with 100,000 and 10,000 normal larvae respectively. Two groups of non-immunized ewes and lambs were similarly challenged and one animal from each of the four groups killed on days 3, 5, 7, 10 and 24 post-challenge. The cellular changes in the abomasal mucosa were less marked in the non-immunized groups than in the immunized animals and appeared later in the lambs than in the ewes. Thus, in the immunized ewes increases in the numbers of mast cells and eosinophils were evident within five days of challenge whereas similar changes appeared later in the immunized lambs. Also marked lymphoid aggregates at the base of the mucosa and in the submucosa were detected only in the immunized ewes. However, both immunized and non-immunized ewes showed rises in the numbers of IgA plasma cells after challenge which were not evident in either group of lambs.     

Vaccination against Taenia solium cysticercosis in pigs using native and recombinant oncosphere antigens.

Pigs were immunised with antigens derived from Taenia solium oncospheres or with a pool of three recombinant antigens from Taenia ovis, and subsequently challenged with T. solium eggs. The native oncosphere antigens induced 83% protection against viable, and 89% protection against the total number of cysticerci established following the challenge infection. Immunisation with the recombinant T. ovis antigens induced 93% protection against the establishment of viable cysticerci, and 74% protection against the total number of cysticerci. These results, and those achieved elsewhere with Taenia saginata and T. ovis, support the possibility of developing a practical vaccine to assist in the control of transmission of T. solium through pigs.     

The establishment rate of Ostertagia circumcincta and Trichostrongylus colubriformis in lactating Romney ewes

The ability of lactating Romney ewes to resist establishment of ingested infective-stage larvae (L3) of Ostertagia circumcincta and Trichostrongylus colubriformis was measured in the field. Three groups of seven single-lamb-bearing ewes were selected on the basis of uniformity of lambing date from a large flock held on pasture. Either 2, 4 or 6 weeks after parturition, groups of ewes were dosed with 24000 L3 of known oxfendazole-resistant parasite strains; 12000 of each species. Ten to 14 days later the ewes, along with their lambs, were transferred from the field to indoor pens. Twenty-five days after the challenge dose the ewes were drenched with oxfendazole to remove any field-derived infection and 3 days later slaughtered for worm counts. Mean establishment of the resistant parasites was low at all times, with the highest rate recorded being 6.1% for O. circumcincta 2 weeks after parturition. Establishment of O. circumcincta 4 and 6 weeks after parturition, and of T. colubriformis at all times, never exceeded 2%. By comparison, mean establishment in lambs held indoors and parasite free for 13 weeks prior to infection, was 24.9% and 47.1% for O. circumcincta and T. colubriformis, respectively. These results indicate that the lactating ewes were exhibiting a substantial ability to prevent establishment of ingested larvae. The results of this and other similar studies suggest that the dynamics of parasitism in lactating Coopworth and Romney ewes in New Zealand is substantially different to that in Merino ewes in Australia, and that these differences influence optimal strategies for the management of anthelmintic resistance in the two countries.     

The effect of selection of both sire and dam on the response of F1 generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae

Windon R. G. and Dineen J. K. (1981). The effect of selection of both sire and dam on the response of F₁ generation lambs to vaccination with irradiated Trichostrongylus colubriformis larvae. International Journal for Parasitology11: 11–18. Rams and ewes, tested for responsiveness to vaccination with irradiated T. colubriformis larvae at an early age, were mated on the basis of responder × responder and non-responder × non-responder. Progeny were vaccinated at 8 and 12 weeks of age with 20,000 irradiated larvae, treated with anthelmintic at 16 weeks and challenged with 20,000 normal larvae at 17 weeks. Faecal egg counts of progeny from responder matings were significantly lower than progeny from non-responders, and within each mating type, ewe lambs had markedly lower egg counts than ram lambs. The level of circulating complement-fixing antibodies to T. colubriformis larval extract were inversely related to egg counts. Thus, ewe progeny from responder matings had the highest serum antibody levels, non-responder ram progeny had the lowest levels and responder rams and non-responder ewes had similar intermediate levels. In vitro responses of cells stimulated with T. colubriformis L₃ antigen were greater in progeny from responder matings, whereas responses to bacterial lipopolysaccharide were higher in progeny from non-responder matings. The results confirm that the response to vaccination at an early age is genetically determined, and show that the response of progeny is most vigorously expressed when both sires and dams have been selected.     

Protection from murine cysticercosis by immunization with a parasite cysteine protease

Central nervous system infection by Taenia solium cysts causes neurocysticercosis, a common neurological infection in the Third World. We have previously isolated cysteine proteases from Taenia crassiceps and T. solium. In this study, we immunized BALB/c mice with the purified T. solium cysteine protease and challenged them with Taenia crassiceps. Immunized mice had a 72% reduction in parasite burden compared to mice that received no immunization. Immunized mice developed antigen specific lymphocyte proliferation. These data support further studies of the T. solium cysteine protease as a vaccine candidate.     

Serum and skin surface antibodies and their associations with sheep biting lice, Bovicola ovis, on experimentally infested sheep

The sheep biting louse ( Bovicola ovis ) feeds superficially on the skin of sheep but appears to stimulate an immune response. In this study we examined the association between louse infestation and serum and skin surface antibodies. Louse numbers were monitored on experimentally infested Polypay and Columbia ewes for two years and on their lambs in the second year. Serum and skin wash samples were tested for antibodies to soluble extracts of B. ovis , Stomoxys calcitrans and Musca autumnali s by enzyme linked immunosorbent assay (ELISA). In addition, the effects of skin wash extracts on B. ovis were examined in vitro .
The titre of anti- B. ovis antibodies in the serum did not differ significantly between infested and naive ewes. However, there was an increase in serum antibody titre which coincided with periods of high louse density in ewes with high louse counts. Infested lambs had higher serum antibody levels than naive lambs. Substantial cross reactivity was evident among extracts of the different insects.
Densities of lice on the ewes during population decline were negatively related to the titre of skin surface antibodies. Skin washings collected from sheep during B. ovis population decline reduced the number of louse progeny when incorporated into louse diet. These results indicate that B. ovis stimulates an immune response in sheep and suggest that compounds on the skin surface may play a role in the regulation of louse populations.     

In vitro oncosphere-killing assays to determine immunity to the larvae of Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium

Taeniid cestodes infect humans and livestock, causing considerable morbidity and mortality, as well as economic loss. Substantial progress has been made toward the production of recombinant vaccines against cysticercosis in livestock animals. Further development of these vaccines would be aided if a reliable in vitro test were available to measure host-protective immune responses in vaccinated animals. Here, we describe in vitro oncosphere-killing assays for the quantification of host-protective serum antibodies against Taenia pisiformis, Taenia ovis, Taenia saginata, and Taenia solium in rabbits, sheep, cattle, and pigs, respectively. Activated oncospheres of T. pisiformis, T. ovis, T. saginata, and T. solium were incubated in vitro in culture medium, test serum, and a source of complement, and oncosphere killing was assessed after 10 days of culture. In vitro oncosphere killing reflected the presence of specific antibody, and the oncosphere-killing assay typically indicated immunity to the homologous parasite that had been determined in vivo. This study describes the first reliable oncosphere-killing assays for T. pisiformis, T. ovis, T. saginata, and T. solium. These assays will be used for further research into the optimization of recombinant vaccines against cysticercosis.     

Protection of Merino lambs against Haemonchus contortus by trickle infection of neonates

In two trials, lambs were trickle infected with 400 or 1000 infective larvae of Haemonchus contortus (HcL(3)), twice weekly for 4 weeks from the day of birth. Following anthelminthic treatment at 6 weeks (Trial 1) or 7 weeks of age (Trial 2), lambs were challenged 1 week later with a trickle infection totalling 5000 (Trial 1) or 10000 HcL(3) (Trial 2). In both cases, significant protection (P<0.05) ranging from 42 to 79% was achieved against egg and worm counts. Serum antibody responses as well as abomasal lymph node cell proliferation and production of interferon-gamma or interleukin (IL)-5 did not differ significantly between immunised and control lambs. The results are consistent with earlier findings that neonatal lambs can generate protective immunity against Trichostrongylus colubriformis, but the underlying mechanism(s) remain to be determined.     

Studies of stage-specific immunity against Trichostrongylus colubriformis in sheep: immunization with adult parasites.

Merino sheep immunized by the adoptive transfer of adult T. colubriformis for 8 weeks were significantly protected against a challenge infection of 20,000 larvae. Two additional groups of sheep received a primary infection of 9000 adult worms which were allowed to persist for 14 weeks before one group was drenched. When both groups were challenged 10 days later with 30,000 larvae, serial necropsies of these and naive sheep revealed that worm rejection did not occur until 7-10 days after challenge. By comparison with the rapid rejection of larval challenges from sheep immunized with normal primary infections, the results suggest that the antigens which elicited rejection in these experiments are stage-specific and were only present or synthesized in sufficient quantities when parasites had developed for 1 week.     

Iodine nutrition in ewes. 2. Effects of low to high iodine intake by ewes on the I content of biological fluids and plasma immunoglobulins G in newborn lambs

1. In two experiments samples of blood and organs were taken from newborn and young lambs born to five groups of ewes that received different dietary iodine (I) levels. Dietary I contents (mg/kg dry matter [DM]) in pregnancy and lactation, respectively, were: Experiment 1--0.36 and 0.26 for group C, 2.01 and 1.94 for group D; Experiment 2--0.13 and 0.12 for group A, 0.22 and 0.20 for group B, 10.77 and 8.88 for group E. 2. I intake of ewes had no effect on birthweight, body weight gain, or mortality between birth and wk 2 of life. No difference between the groups was recorded in the weights of brain, heart, lungs, and liver of lambs killed at birth. However, the weight of both the thyroid lobes in lambs from group E was lower than that of the other groups. The I content of the thyroid lobes of lambs from group A was 30-40% lower than that of the other groups. 3. The plasma inorganic iodine (PII) of lambs from birth to d42 of life was affected by ewe I intake, except for groups A and B, during the first 16 h of life. The PII of lambs increased with the I intake via milk in each group. The ratio of PII of lambs at birth/PII of ewes at d7 prepartum was 3.77, 2.96, 1.68, 1.39 and 8.62, respectively, for groups A, B, C, D and E. 4. The high I intake by group E ewes induced a higher plasma T4 concentration in lambs at birth. The decrease in plasma immunoglobulins G (IgG) recorded for the lambs in this group might be explained by this increase in plasma concentration.     

Congenital abnormalities in newborn lambs following Akabane virus infection in pregnant ewes

To clarify the pathogenicity of Akabane virus for ovine embryos, pregnant ewes were inoculated intravenously with the virus. As a result, all of them were affected with viremia and showed an increase in neutralizing antibody 2 weeks after inoculation. The virus was recovered from many organs of embryos which were inoculated with it at 29--45 days of pregnancy and sacrificed 9--30 days later. In particular, some of these embryos which were sacrificed 15 days after inoculation were found suffering from systemic infection. A large quantity of virus was recovered from the organs all over the body of them. No virus, however was recovered from any organ of embryos which were inoculated with the virus at 81 days of pregnancy and sacrificed 30 days later. Abnormal changes were observed in neonatal lambs born from ewes inoculated with the virus at 30--50 days of pregnancy. They were especially severe when the virus was inoculated at 30 days of pregnancy. They consisted of ankylosis of the limbs, scoliosis, hydranencephaly, porencephaly, stillbirth with dwarfism, and death after birth with dwarfism and weakness. Nothing abnormal was found in any neonatal lambs born from ewes inoculated with the virus at 91--101 days of pregnancy. When embryos exceeded 64 days of intra-uterine life more than 29 days after virus inoculation, it was possible to detect immunoglobulin, IgM or IgG or both, and antibody from the serum. Attempts failed to detect either immunoglobulin from embryos less than 59 days of intrauterine life. No IgA was detected from the serum of any embryo. In almost all the neonatal lambs born from ewes inoculated with the virus at 28--101 days of pregnancy, neutralizing antibody was detected from the serum at the time of birth.     

Effect of immunization on reproducvive performance, embryo quality and progesterone in Rasa Aragonesa ewes actively immunized against androstenedione or passively immunized against testosterone

A total of 217 Rasa Aragonesa ewes were used to test two immunization treatments: 1.Active immunization against androstenedione: ewes immunized in previous matings (androstenedione, reimmunized; AR groups, n=58) or not (first immunization; AF groups n=64) were boosted either 2 or 4 wk before mating. 2.Passive immunization against testosterone: antisera were injected either at sponge withdrawal (zero time; T0 group, n=21) or 1 wk previously (Tl group, n=22). We used 52 ewes as controls (C group). Half of each group was used either to record reproductive performance or to embryo viability assessment. Prolificacy was significantly increased in ewes which reached a moderate antibody level, independently of the treatment. Fertility was lower in AR ewes that attained a high antibody titre (P<0.01). The percentage of viable embryos recovered was lower in AF ewes (P<0.01), and in ewes whose testosterone antibody titre was high (P<0.05), compared to C group. It was proven that similar or lower antibody levels were more harmful for ewes from AF and Tl than for ewes from AR or T0 groups. The proportion of nonfertilized recovered ova was not significant. Progesterone levels were notably increased in AR ewes (P<0.001) independently of ovulation rate and were positively correlated to antibody titre at mating (P<0.01) but these events were not observed in T ewes. These findings indicate that after androgen immunoneutralization, only those ewes having antibody titres within a limited range at mating had improved reproductive performance. Further research is needed in order to understand the role that progesterone plays in immunized ewes.     

Gastro-intestinal nematode parasite populations in ewes and lambs and the origin and time course of infective larval availability in pastures

Changes in the numbers of infective larvae available on pasture and the development of helminth infection in spring-born lambs were studied in an environment with a mild winter and where pastures are grazed throughout the year. Prior contamination of pastures in autumn and early winter was compared with egg deposition by lambing ewes as sources of infection. Larvae derived from eggs deposited by lambing ewes did not appear on pasture until 8 weeks after the beginning of lambing but were present in increasing numbers for the last 7 weeks before weaning at 11–15 weeks of age. Lambs exposed to prior contamination became infected earlier than those exposed only to ewe contamination, but lamb egg output made no contribution to larval numbers on pasture before weaning. Lamb growth rates to weaning did not differ between groups exposed either to prior contamination alone, to ewe egg output alone or to both, but all groups carried appreciable infections at weaning. After weaning, lambs remained on the same pastures, and those exposed to ewe contamination suffered severe trichostrongylosis 5 weeks after weaning. Lambs exposed only to over-wintered contamination also developed severe trichostrongylosis 9 weeks after weaning. The results are compared with those of similar recent studies in Britain and their implications for control of infection are discussed.