Passive transfer of colostral immunoglobulins from ewe to lamb and its influence on neonatal lamb mortality

The transfer of immunoglobulin G (IgG) and immunoglobulin M (IgM) from ewe to lamb was quantitated to determine the occurrence of failure in passive transfer. Concentrations of IgG and IgM in ewe serum did not correlate with those in the colostrum. Colostrum from all ewes contained abundant amounts of immunoglobulins when compared with serum values, with IgG being selectively concentrated over IgM. Absorption through the intestinal tract of the lamb appeared to be a nonselective process, lacking predilection for IgG and IgM. All lambs tested 24 hours after birth absorbed colostral immunoglobulins to some extent; however, 13 (14%) of 91 clinically normal lambs demonstrated some failure of passive transfer. In contrast, failure of passive transfer was found in 27 (46%) of 59 lambs dying of natural causes between 24 hours and 5 weeks of age. Evidence presented emphasizes the importance of absorption of adequate amounts of immunoglobulins to enable the newborn lamb to survive the first few weeks of life.     

Sheep immunoglobulins and their transmission to the neonatal lamb

Extract

In the past decade great strides have been made in clarifying the types of immunoglobulins found in various species of domestic animals. This review summarizes the information presently* * This review cites material published up to the end of 1976. available on sheep immunoglobulins — their classes, quantity and distribution in serum and colostrum. the review also highlights recent studies of fetal immunological competence, presucking lamb serum immunoglobulins, the process by which colostral immunoglobulins are passively transferred to the neonate, and the effects both of colostrum administration and deprivation on the newborn lamb.     

Development of a new monoclonal antibody to ovine chimeric IgE and its detection of systemic and local IgE antibody responses to the intestinal nematode Trichostrongylus colubriformis

The J558L cell line, previously transfected with the ovine Cepsilon gene, was induced to secrete a chimeric IgE protein composed of the ovine heavy chain and a mouse light chain with MW of approximately 80 and 26 kDa, respectively. After purification, the chimeric protein was used to immunise BALB-c mice and monoclonal antibodies (mAbs) were generated. The mAb 2F1, which had greatest anti-IgE activity in preliminary screens, was chosen for further characterisation and an examination of systemic and local IgE responses to the intestinal nematode, Trichostrongylus colubriformis. The chimeric IgE protein was not recognised in enzyme linked immunosorbent assay (ELISA) by mAbs raised against ovine IgG1, IgG2, IgA or IgM. However, 2F1 was highly specific to the chimeric IgE protein, and did not cross-react with ovine IgG1, IgG2 or IgA. Western blot analysis also showed that 2F1 and secretory IgA (sIgA) did not cross-react, and that 2F1 and the anti-IgA mAb identified different MW bands from colostrum (approximately 200 and 400 kDa, respectively). 2F1 bound to mucosal mast cells (MMC) isolated from the intestines of lambs infected with T. colubriformis, but cultured bone marrow-derived mast cells (BMMC) required prior incubation with the chimeric IgE protein for this binding to occur. Distinctive staining of plasma cells and putative mast cells were observed using 2F1 on immunohistological sections of mesenteric lymph node and jejunum.ELISA incorporating 2F1 was able to detect >0.4 ng chimeric protein. Total IgE in ovine colostrum and intestinal homogenates was quantified using a capture ELISA, with known amounts of chimeric protein used to produce a standard curve. Colostrum from outbred Merino ewes had 0.55-11.05 ng ml(-1) total IgE, and their lambs, at necropsy after infection with a total of 18,000 T. colubriformis infective larvae over a 9-week period, had 45-620 ng g(-1) total IgE in intestinal tissue. Compared to genetically susceptible lambs, antigen-specific levels of IgE were significantly higher in genetically resistant lambs after infection with 4500 T. colubriformis infective larvae (TcL3) per week for 9 weeks (161.4 versus 44.8 geometric mean titres; P=0.043). In western blots, distinctive bands (19-21 and 27 kDa) from T. colubriformis larval antigen were differentially recognised by IgE, as identified by 2F1, in intestinal homogenates from genetically resistant animals.These results have demonstrated the value of 2F1 for quantification of IgE responses in samples derived from ovine fluids and tissues using ELISA, western blots and immunohistology. In this respect, it recognises native ovine IgE and does not require pre-treatment of the sample with denaturing agents or ammonium sulphate.     

Measurement in vitro of a larval migration inhibitory factor in gastrointestinal mucus of sheep made resistant to the roundworm Trichostrongylus colubriformis

Lambs were challenged by dosing with 2500 T. colubriformis larvae per day for 34 weeks, rested for 4 weeks and then re-challenged with the infective larvae for a further 10 weeks. A technique for the measurement of inhibition of larval migration from agar gels was used to investigate the antiparasitic activity of mucus, obtained from the small intestine and abomasum of the lambs, at the end of the re-challenge period. Measurements were also made on ileal digesta samples collected at different times during the development of the initial resistance, the rest period and the re-challenge period, and on faeces collected during the re-challenge infection. The mucus from the small intestine and abomasum paralysed and inhibited larval migration from agar gels significantly more (P less than 0.01 and P less than 0.05, respectively) than corresponding mucus from parasite-free control animals. This inhibitory activity was also detected (P less than 0.01) in the ileal digesta of infected animals from Week 6 of primary dosing. The magnitude of the inhibition in the ileal digesta increased with time during the infection period and was again detected during re-infection (P less than 0.01). It was not detectable in resistant sheep towards the end of the rest period. Slight inhibitory activity was detected in faeces after 2 weeks of re-infection. These observations suggest that substances secreted into the lumen of the small intestine of infected animals are responsible for resistance against ingested larvae.     

Subclinical infection with the nematode Trichostrongylus colubriformis increases gastrointestinal tract leucine metabolism and reduces availability of leucine for other tissues

Gastrointestinal (GI) tract leucine metabolism was measured in 6- to 9-mo-old lambs subjected to trickle infection with Trichostrongylus colubriformis larvae and in separate animals that were not infected. Animals prepared with a jejunal catheter and with indwelling catheters into the aorta and the portal- (PDV) and mesenteric- (MDV) drained viscera were infused simultaneously with [1-13C] and [5,5,5-2H3] leucine to determine GI tract sequestration of leucine from arterial and luminal amino acid pools by tracer and tracee arteriovenous concentration differences. Leucine oxidative losses and net fluxes were also determined across the GI tract. Infection had no detectable effect on whole-body leucine flux, but it increased total GI tract leucine sequestration by 24% (P<.05) and GI tract oxidative losses of leucine by 22 to 41% (P<.01). Net PDV fluxes of leucine were decreased by 20 to 32% during the infection. The infection did not alter either the proportion of precursor leucine used by GI tract metabolism that was derived from the arterial leucine pool (.84 to .88) or the proportional sequestration of digesta-derived leucine during "first pass" absorptive metabolism (.12 to .18). These findings help to elucidate the metabolic basis for the reduced growth rates and nitrogen retention observed when animals are subjected to subclinical nematode infection.     

The maternal to fetal transfer of immunoglobulins associated with placental lesions in sheep.

In this study we evaluated maternofetal transmission of immunoglobulins in ewes under conditions of altered placental morphology. Intravenous injection of human red blood cells was used to induce immunoglobulins in pregnant ewes. The hemagglutination test was used to detect antibody in maternal serum, fetal and placental fluids. Placental injury was induced by intravenous inoculation of Escherichia coli endotoxin or spores of Aspergillus fumigatus into pregnant ewes at days 99 or 100 of gestation respectively. Placental infarction, thrombosis of maternal placental vessels and variable neutrophil infiltrate characterized lesions produced by A. fumigatus. Endotoxin treated ewes developed marked placental edema, congestion, hemorrhage and focal loss of uterine epithelium. Human red blood cell agglutinating antibody was not detected in placental or fetal fluids obtained from ewes with either of the above placental lesions. Placentitis of undetermined etiology was observed in seven ewes. Two ewes had received A. fumigatus, two had received endotoxin and three were untreated ewes. Histological examination of their placentas revealed trophoblastic and endometrial epithelial necrosis and necrotizing vasculitis of the chorioallantois. Human red blood cell agglutinating antibody was detected only in the fetal and placental fluids of the seven ewes with these placental lesions. The nature of these lesions would have produced a functional confluence of the maternal and fetal circulations. Antibody transfer from dam to fetus was observed only in association with placental lesions which produced this confluence of circulations. The character of the placental lesions, rather than the mere presence of placental lesions apparently determined the transfer of immunoglobulins.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of four tropical tanniniferous plant extracts on the inhibition of larval migration and the exsheathment process of Trichostrongylus colubriformis infective stage

The anthelmintic (AH) effect of Acacia pennatula, Leucaena leucocephala, Lisyloma latisiliquum and Piscidia piscipula was evaluated in the infective larvae (L(3)) of Trichostrongylus colubriformis. Different concentrations of lyophilized extracts were tested using the larval migration inhibition (LMI) test. An inhibitor of tannins (the polyvinyl polypyrrolidone [PVPP]) was used to verify whether these compounds were responsible for the AH effects. Then, the effect of extracts on larval exsheathment was examined by observing the exsheathment process at 10-min intervals for 70 min. The LMI test showed a dose-dependant AH effect for A. pennatula, L. leucocephala and L. latisiliquum (P<0.01), but not for P. piscipula. The restoration of L(3) migration to values similar to those of controls after the addition of PVPP, indicates that tannins are involved in AH effects. Trichostrongylus colubriformis exsheathment was partially or totally blocked by the four plants extracts. Tropical tanniniferous plants evaluated in the current study may have potential as AH for the control of T. colubriformis if in vivo investigations indicate useful effects.     

The influence of Ostertagia circumcincta and Trichostrongylus colubriformis infections on the pharmacokinetics of febantel in lambs

Plasma concentrations of febantel and its major metabolites fenbendazole, oxfendazole and oxfendazole sulphone were determined after oral administration of 7.5 mg/kg febantel in lambs before and 28 days after infection with 50 000 L₃ larvae of Ostertagia circumcincta or Trichostrongylus colubriformis. The febantel concentrations were always very low and only in a few samples higher than the detection limit. The mean decrease in AUC for the three metabolites for the infected sheep in comparison to the parasite naive sheep was 13.9%± 4.1% (mean± SEM) and 23.7%± 5.3% in the 0. circumcincta infected and the T. colubriformis infected lambs respectively. This reduction was only significant for the T. colubriformis infected group.
In order to determine a more complete pharmacokinetic profile, febantel was injected intravenously at a dose of 2.5 mg/kg in a further study.     

In vitro activity of Peltophorum africanum Sond. (Fabaceae) extracts on the egg hatching and larval development of the parasitic nematode Trichostrongylus colubriformis

Trichostrongylus colubriformis is an important cause of parasitic gastroenteritis in ruminants, where it causes protracted diarrhoea, rapid loss of weight, loss of production and death. The in vitro efficacy of extracts of Peltophorum africanum was determined against this parasitic nematode. Eggs and larvae of T. colubriformis were incubated at 23 degrees C in the extracts of the leaf, bark and root of P. africanum at concentrations of 0.008-25 mg ml-1 for 2 and 5 days, respectively. Thiabendazole and water were used as positive and negative controls, respectively. Inhibition of egg hatching and larval development increased significantly (P<0.05) with increasing concentrations of the extracts. Concentrations of 0.2-1.0 mg ml-1 of the extracts of leaf, stem bark, and root bark of P. africanum completely inhibited the hatching of eggs and development of larvae. No eggs and larvae of T. colubriformis could be observed in wells incubated with all the three extracts at concentrations of 5 and 25 mg ml-1. The in vitro model results support the traditional use of P. africanum against nematode parasites. Further research is required to isolate and structurally identify the active anthelmintic compounds, and to improve methods of plant extraction of the effective anthelmintic components that will be readily adaptable for use by rural communities against helminthosis.     

Occurrence of IgE in foals: evidence for transfer of maternal IgE by the colostrum and late onset of endogenous IgE production in the horse

IgE is the key antibody involved in type I allergies. Allergen mediated crosslinking of IgE bound to high affinity Fc-receptors on mast cells and basophils stimulates cellular degranulation and release of inflammatory mediators and cytokines. In this report, we demonstrate that IgE antibodies can be transferred from the mother to offspring in horses via the colostrum. We found a clear correlation between the IgE concentration in colostrum and the total IgE concentration in foal sera on day 2 after birth (r_sp = 0.83). Maternal IgE was detected in foal sera by ELISA and on peripheral blood leukocytes of foals by flow cytometry. Both serum and cell membrane-bound IgE were undetectable in newborn foals before colostrum uptake and peaked on days 2–5 after birth. Cell-bound IgE became undetectable at 2 months after birth. Serum IgE disappeared from the circulation within the first 3–4 months of age. These kinetics suggest that the IgE antibodies which are detectable in foals during the first 4 months after birth are of maternal origin only. The endogenous IgE production was found to begin at 9–11 months of age, when IgE could be detected on peripheral blood leukocytes and in foal sera again. After 18 months of life, the total IgE concentrations in foal sera were comparable to those detected in their dams. The late onset of endogenous IgE production offers an explanation for observations that IgE mediated allergies are generally not observed in horses before puberty. The roles of the passively transferred maternal IgE in newborn foals are not yet known, but could be manifold, ranging from passive immunity and induction of immunoregulatory functions to determinative influences of maternal IgE on the antibody repertoire in the offspring.     

The effect of diet fed to lambs on subsequent development of Trichostrongylus colubriformis larvae in vitro and on pasture

Contrasting herbage diets were fed to lambs to evaluate their effect on subsequent development of Trichostrongylus colubriformis larvae in faeces and on pasture. The diets had either no condensed tannin (CT), lucerne (Medicago sativa cv. Otaio), white clover (Trifolium repens cv. Tahora), or had moderate to high concentrations of CT, sulla (Hedysarum coronarium cv. Grassland Aokau), Lotus corniculatus (cv. Grasslands Goldie), L. pedunculatus (cv. Grassland Maku), Dorycnium pentophyllum, and Dorycnium rectum. Trials were carried out in summer (warm) and in autumn (cool and moist). In summer, egg viability was evaluated in vitro with egg hatch and larval development assays. In both seasons faeces were placed on pasture to compare recovery of eggs and larvae from faeces and larvae from herbage on the high and low fertility farmlets on the AgResearch Ballantrae Hill Country Research Station. D. rectum and D. pentophyllum diets decreased (P<0.01) egg hatching and larval development in laboratory assays relative to other diets. In summer, the number of larvae recovered from faeces placed on pasture was far greater (P<0.001) if the lambs had been fed lucerne than any other diet, whereas recovery was always lowest from faeces of sheep fed D. rectum and D. pentophyllum. Although dietary differences were lower in autumn than in summer, larval recoveries were lower (P<0.05) from faeces of lambs fed D. rectum and L. corniculatus than from white clover, lucerne and sulla diets. This study indicates that the diet of the host can have a significant impact on egg hatching and the subsequent development of T. colubriformis larvae in the laboratory and in the field. In particular, D. rectum consistently reduced T. colubriformis development. Effects measured in vitro generally under-estimated effects measured under field conditions.     

Assessing the agreement between Ostertagia ostertagi ELISA tests performed using the crude adult antigen and the adult and larval stage 4 excretory/secretory antigens

The objective of this study was to determine the agreement between ELISA tests conducted using three O. ostertagia antigens: crude adult worm, larval stage 4 (L4) excretory/secretory (ES) and adult ES. This study was carried out on 289 Holstein cows from five herds in Prince Edward Island and one herd in Nova Scotia. Composite milk samples of these cows were collected (between May and September 2002) from the respective provincial laboratories and sent to the Atlantic Veterinary College where each sample was tested for antibodies to O. Ostertagi using an indirect microtitre ELISA test. Results were expressed as optical density ratio (ODR) values. Each milk sample was tested with three ELISA tests, with each test using a different O. ostertagi antigen. There was a slight rise in ODR values of both adult antigens, between May and August, with higher values obtained using the adult ES antigen. L4 ES ODR values were generally higher than those for both adult antigens during the study period, except for May. There was a more dramatic rise in L4 ES ODR values between May and August. Rises in ODR in May and end of July coincided with periods of mass maturation of L4 to adult worms. The results of the study showed that the concordance correlation coefficient (CCC) between tests performed using both ES and the crude antigens were low (crude adult versus adult ES=0.31, crude adult versus L4 ES=0.30). The highest CCC was observed between tests done using both ES antigens (CCC=0.56). Generally, the study results suggest that the antibody response (detectable by the ELISA) is mainly directed against ES antigens (especially L4) than the crude adult worm antigen.     

The effect of daily challenge with Trichostrongylus colubriformis larvae on the nutrition and performance of immunologically-resistant sheep

Immunological resistance to Trichostrongylus colubriformis was developed in sheep by challenging them with 2500 larvae per day for 34 weeks. They were then rested for 24 weeks before being re-challenged with the same dose rate of the same larvae for 10 weeks. Nutritional, haematological and parasitological parameters were measured during the first 8 weeks of re-challenge infection. There were no faecal worm eggs excreted during the re-challenge infection. There was a small but significant increase in plasma-N leakage from Days 4 to 9 of dosing but this then declined to initial levels. There was no other detectable nutritional disturbance associated with re-infection. Re-challenge caused a rapid development of eosinophilia which peaked during Week 6 before declining to levels of the control animals by Week 8. Sheep resistant to T. colubriformis appear to be able to re-activate their immunological mechanisms when re-challenged with the parasite with little associated nutritional penalty.     

Trichostrongylus colubriformis: in vitro culture of parasitic stages and their use for the evaluation of anthelmintics

Approximately 50 per cent of fourth stage larvae of Trichostrongylus colubriformis taken from the gerbil, Meriones unguiculatus, on day 8 after infection, moulted to the young adult stage when cultured in a complex medium over a seven day period in vitro. Larvae at the late fourth stage of development were highly susceptible to certain benzimidazole, prebenzimidazole, imidazothiazole, pyrimidine, quaternary ammonium, organophosphorus and macrocyclic lactone anthelmintics when any of these were included at very low concentrations in the culture medium. However, few anthelmintics lacking activity against T colubriformis in vivo affected these larvae. An assay employing these larvae in vitro should offer a means for assessing the intrinsic activity of new compounds against T colubriformis in the absence of any complicating host pharmacokinetic factors, and could also be adapted for use as a high capacity preliminary screen. Thus it should now be possible to employ a target parasite at the earliest stages of a lead discovery programme obviating the need to use less relevant free-living nematodes or ones that are natural parasites of rodents.     

The influence of reproductive physiology and nutrient supply on the periparturient relaxation of immunity to the gastrointestinal nematode Trichostrongylus colubriformis in Merino ewes

A pen experiment was conducted to investigate the interaction of early-weaning and nutrient supply on the periparturient relaxation of immunity to the gastrointestinal nematode (GIN) Trichostrongylus colubriformis in Merino ewes. Mixed-age pregnant and non-pregnant (dry) ewes were infected with 8,000 T. colubriformis L(3)/week, and fed either a high or low quality diet. Following parturition, lambs were either removed from their mothers at 2 days of age or allowed to continue suckling. Systemic immunity began to wane during late pregnancy with circulating eosinophils and plasma total antibody (Ab) levels declining from day -37 (relative to the midpoint of lambing) and day -24, respectively. Pregnant ewes fed the low quality diet exhibited an increasing faecal worm egg count (WEC) from day -24 and had higher intestinal worm burdens on day 13, whereas ewes fed the high quality diet had a delayed transient rise in WEC of lower magnitude. Dry and early-weaned ewes remained highly resistant to T. colubriformis at all times. In the post-lambing/lactation period, ewes fed the high quality diet had higher levels of local total Ab and numbers of goblet cells (GC) in the small intestine on days 13 and 41. Lactating/suckled ewes had a lower anti-parasite local immune response as indicated by reduced titres of total Ab, IgG(1), IgM and IgA and lower numbers of mucosal mast cells (MMC), globule leukocytes (GL) and GC in small intestinal tissue compared to their dry and early-weaned counterparts. Early-weaning resulted in rapid recovery of blood eosinophils and total Ab. On day 13 post-lambing, titres of total Ab, IgG(1), IgM, IgA and IgE, and numbers of MMC and GL were greater than those measured in dry and suckled ewes. When fed the high quality diet, ewes had a higher dry matter (DM) intake, maternal weight, fat score, greater fat depth and eye muscle depth, birthed heavier lambs that had higher growth rates, and produced more milk. The physiological status of pregnancy resulted in a higher DM intake but lower measures of fat depth and eye muscle depth, and suckling led to an increase in DM intake but a reduction in body weight and fat score through mobilisation of fat and muscle reserves. Despite the marked effect of diet quality on production traits, some inconsistencies were observed between body composition and apparent parasite resistance, measured by WEC and worm counts, suggesting that the nutritional influence was not necessarily always mediated through changes in body composition. Although reproductive status affected blood leptin levels, diet had no effect within suckled ewes and therefore it was concluded that leptin has no causative role in maintaining the periparturient relaxation of immunity to T. colubriformis.     

A comparison of the infectivity of cryopreserved versus unfrozen infective larvae of Haemonchus contortus, Trichostrongylus colubriformis and Trichostrongylus axei. Results of the Onderstepoort Veterinary Institute and collaborators from 1977 to the present

The infectivity for sheep of cryopreserved infective larvae (L3) of various strains of Haemonchus contortus, Trichostrongylus colubriformis and Trichostrongylus axei is compared using previously published results of trials conducted at the Onderstepoort Veterinary Institute laboratories, and of collaborators. The means and ranges of development were similar for both frozen and unfrozen larvae of two of the three worm species reviewed. A mean of 33.4% (range, 12.7-63.0%) of cryopreserved H. contortus L3 developed, compared to a mean of 43.7% (range 2.4-78.7%) of unfrozen L3 of this worm species. The corresponding values for T. colubriformis were 33.0% (range 10.3-62.7%), and 33.5% (range 8.3-52.2%), respectively. In the case of T. axei, the development of the cryopreserved L3 (tested in only three trials) was markedly lower than that of unfrozen L3 in the single trial in which the latter was evaluated. It is concluded that development of cryopreserved L3 is probably similar to that of unfrozen L3 and that, for several reasons, maintaining nematode larvae in the frozen state in liquid nitrogen is a much superior method to that of one which entails cycling worm strains continually in their final hosts.