The response of lambs to vaccination at weaning with irradiated Trichostrongylus colubriformis larvae: segregation into 'responders' and 'non-responders'

Dineen J. K., Gregg P. and Lascelles A. K. 1978. The response of lambs to vaccination at weaning with irradiated Trichostrongylus colubriformis larvae: segregation into ‘responders’ and ‘non-responders’. International Journal for Parasitology8: 59–63. Groups of colostrum fed and colostrum deprived lambs were vaccinated with irradiated Trichostrongylus colubriformis larvae at weaning (3 months) and challenged with normal infective larvae. As there was no significant difference between the worm counts of these groups after challenge, it was concluded that the failure of lambs, generally, to respond to vaccination as vigorously as mature sheep, was not due to ‘feed-back inhibition’ of the immune response by the action of maternal antibody.However the results showed that the lambs segregated into ‘responders’ and ‘non-responders’. The mean worm count of responders was 1560 whereas for non-responders this was 24,340. A responder was defined as an animal whose worm count was less than the lower limit of the 99 % confidence interval of unvaccinated controls. These results suggest that genetically-determined factors play an important role in the responsiveness of lambs to vaccination.Whereas counts of globule leucocytes in duodenal tissues were negatively correlated with worm counts (i.e. positively correlated with resistance), counts of both eosinophils and neutrophils were positively correlated with worm counts. These findings suggest that either globule leucocytes are involved in the resistance mechanism, or they are by-products of cellular events involved in resistance. On the other hand it is unlikely that either eosinophils or neutrophils are involved in the mechanism of resistance.     

The relationship between ovine lymphocyte antigens and faecal egg count of sheep selected for responsiveness to vaccination against Trichostrongylus colubriformis

The purpose of this work was to determine if ovine lymphocyte antigen (OLA) type was associated with level of resistance and whether it could be used as a predictive test for identifying responsiveness of animals to vaccination against the parasite T. colubriformis. Antisera to ovine class I lymphocyte antigens were used to measure the frequency of three OLA types in merino sheep bred as either high or low responders to vaccination and challenge with T. colubriformis. A particular combination of lymphocyte antigens (SY la + lb) appeared to be associated with low faecal egg counts (resistant), and high faecal egg counts (susceptible) appeared to be associated with other OLA types, such as SY2. When high and low responder data were pooled, 56 ewes with SY la + lb, had a mean count of 149 eggs per gram (e.p.g) of faeces and 69 ewes with SY2 had 560 e.p.g. For 28 rams with SY la + lb, the mean count was 317 e.p.g. and 51 rams with SY2 had 1143 e.p.g. Overall these results were statistically highly significant. Separate analysis of each line showed that high responders, with SY la + lb had a mean count slightly below those with all other combinations within the same line, but this was not statistically significant. The SY la + lb type was not present in low responder males but in 11 low responder females this type was associated with significantly increased egg count. It was concluded that the case has yet to be proven that SY la + lb is a predictive marker for parasite resistance and not merely a foundation effect resulting from the selection and mating of particular sires.     

Vaccination against the nematode Trichostrongylus colubriformis. II. Attempts to protect guinea-pigs with worm acetylcholinesterase.

Soluble material extracted from T. colubriformis fourth stage larvae was fractionated by membrane ultrafiltration, gel filtration and ion-exchange chromatography. The acetylcholinesterase (AChE) peak obtained by gel filtration protected guinea-pigs against infection but it was contaminated by worm allergen. However, there was no relationship between the AChE content of fractions obtained by membrane ultrafiltration and ion-exchange chromatography and their ability to stimulate protective immunity. Purified T. colubriformis AChE, prepared by ion-exchange chromatography and free from demonstrable allergen did not stimulate protective immunity whereas another fraction, containing less than one-thousandth the amount of AChE, was effective in doing so.     

The response of immune sheep to challenge with Trichostrongylus colubriformis: enteric plasma loss and secretion of biogenic amines

Sheep were challenged with a single large dose of larvae after vaccination with irradiated Trichostrongylus colubriformis larvae. Worm counts performed on vaccinated sheep 6 or 7 days after challenge (DAC) showed that they were solidly immune and only retarded L3 larvae were recovered at this time. Enteric plasma loss (EPL) in vaccinated animals increased immediately after challenge to peak 4–6 DAC and then decreased to pre-challenge levels. In contrast, a substantial rise in EPL did not occur in unvaccinated sheep until 10 DAC. Secretion of histamine into the duodenum of immunized sheep increased significantly from 2 to 7 DAC with the highest value at 6 DAC which corresponded with a lower duodenal tissue histamine level at this time. Histamine and 5HT secretion into the small intestine of previously uninfected sheep gradually increased during 12 weeks of a trickle infection of 3000 normal T. colubriformis larvae per week. The results indicate that rejection of incoming larvae by immune sheep is accompanied by an intestinal inflammatory response involving secretion of biogenic amines and a concurrent plasma loss.     

Histamine and leukotriene concentrations in duodenal tissue and mucus of lambs selected for high and low responsiveness to vaccination and challenge with Trichostrongylus colubriformis

Lambs selectively bred for high responsiveness or low responsiveness to vaccination with irradiated Trichostrongylus colubriformis were vaccinated and challenged. Duodenal tissue histamine concentrations in both high and low responder lambs were lower at 3 days than at 28 days after challenge. At 3 days after challenge, histamine concentrations were higher in both male and female high responder lambs than in low responder lambs whereas at 28 days concentrations were increased only in high responder females. At 3 days after challenge, histamine concentrations were generally lower in mucus than in tissues, but levels were again higher in mucus from high responder groups. In duodenal tissue at 3 days after challenge, leukotriene C₄ and B₄ concentrations were similar in high and low responder animals. At the same time, concentrations of both leukotrienes were higher in mucus than in tissues, with high responder female lambs having the highest concentration. It is suggested that increased levels of histamine and leukotrienes in mucus and tissue are associated with larval rejection or exclusion.     

Attempts to probe the antigens and protective immunogens of Trichostrongylus colubriformis in immunoblots with sera from infected and hyperimmune sheep and high- and low-responder guinea pigs

Comparison of antisera from sheep during primary infection and following vaccination and challenge with Trichostrongylus colubriformis, with antisera obtained following primary infection of high- and low-responder guinea pigs, failed to reveal different antigenic patterns in proteins separated from fourth stage larval extracts by two-dimensional electrophoresis and probed by the immunoblot technique.Generally, serum IgG reacted specifically with worm antigens of mol. wt greater than 94,000, whereas protection against challenge infection was elicited most effectively in the guinea pig by fractions in the 67,000–94,000 range.Most distinct separations of larval proteins by SDS-polyacrylamide gel electrophoresis were obtained by extraction of live larvae and the extracts used within 2–3 days.     

The response of sheep to parenteral vaccination and immunizing infections against the abomasal nematode, Haemonchus contortus

Adams D. B., Beh K. J. and Davies H. I. 1982. The response of sheep to parenteral vaccination and immunizing infections against the abomasal nematode, Haemonchus contortus. International Journal for Parasitology12: 445–449. Subcutaneous injection of relatively large amounts of unfractionated homogenates of adults plus infective larvae of Haemonchus contortus emulsified in complete Freund's adjuvant produced a degree of protective immunity when challenge infection was given eight weeks after the first or only dose of vaccine. In an attempt to improve acquired immunity, parenteral vaccination was either followed or preceded by a short immunizing infection with H. contortus, which was terminated by anthelmintic before patency. This treatment aimed at stimulating general responsiveness to worm antigens and invoking mucosal immunity in the abomasum. Disparate results were obtained; immunizing infections either increased immunity or made sheep more susceptible to challenge infection. In this latter situation, the unresponsiveness associated with primary infection with H. contortus may have been increased.     

Vaccination against the nematode Trichostrongylus colubriformis. I. Vaccination of guinea-pigs with worm homogenates and soluble products released during in vitro maintenance

Vaccination of guineapigs with homogenates of fourth larval stage and adult Trichostrongylus colubriformis and soluble products released by fourth stage larvae during in vitro maintenance, induced a high level of protection against challenge infection with the parasite.     

Attempts to vaccinate ewes and their lambs against natural infection with Haemonchus contortus in a tropical environment

A vaccine containing integral membrane glycoproteins from the intestine of Haemonchus contortus was evaluated in three groups of grazing sheep each containing 13 ewes and their 16 lambs naturally infected with gastrointestinal nematodes. Two groups were vaccinated with either 5 or 50 μg of the antigen per immunisation, while the third, the control group, received adjuvant alone. The sheep were immunised six times at 3 week intervals, partly because the vaccine antigens are hidden and thus no immunological boost would be delivered by subsequent infection and partly because the level of Haemonchus spp. challenge was expected to be high. The vaccinated ewes, first immunised approximately 1 month before lambing, showed a circulating antibody response but no signs of reduced anaemia or Haemonchus spp. egg counts, compared with control ewes. Several ewes with severe haemonchosis in all three groups had to be given precautionary treatment with anthelmintic drugs. In contrast, vaccinating their lambs with either 5 or 50 μg of the antigen per immunisation resulted in 10 fold higher antibody titres. In the case of the lower antigen dose this was associated with significantly less anaemia, 72% reduction in the overall number of Haemonchus spp. eggs produced and significantly fewer worms compared with control lambs. It is hypothesised that the heavily pregnant or lactating ewes did not have sufficient physiological reserves to mount a protective response following vaccination in the tropical weather and high challenge conditions that prevailed. Nevertheless, the vaccine could afford useful protection for lambs against H. contortus.     

The effect of vaccinating infection during pregnancy and dietary protein supply on the peri-parturient immune response of sheep to infection with Teladorsagia circumcincta and Trichostrongylus colubriformis larvae

It is well established that dietary protein supply can influence the peri-parturient breakdown of immunity to nematode parasites but there is no information on the importance of exposure to nematode larvae during pregnancy for this response. We investigated this by exposing housed pregnant sheep, scanned as carrying two lambs, to a vaccinating infection with a trickle mixed infection of Teladorsagia circumcincta and Trichostrongylus colubriformis larvae (L3) or to no infection during weeks − 9 to − 4 relative to parturition. At the beginning of week − 3 all sheep were treated with anthelmintic to remove any vaccinating worm burden and from week − 2 to week +6 received a trickle challenge infection with the same nematodes. Within each vaccinating treatment there were two nutritional treatments (no. = 20 per subgroup) designed to provide 1.5 or 1.0 and 1.3 or 0.8 of metabolisable protein (MP) requirement during pregnancy and lactation, respectively. Five ewes were necropsied during weeks +1 and +3 to measure worm burdens and mucosal inflammatory cells and the remainder maintained until week +6. Serum levels of total, IgA and IgE antibodies against L3 antigen of each nematode were measured.Scanning errors and lamb losses resulted in some ewes carrying and/or rearing only one lamb. Numbers of lambs reared was therefore introduced as a treatment effect. Vaccinating infection delayed the peri-parturient rise in faecal egg count (FEC) by an average of 2 weeks but its effect on FEC during the first 6 weeks of lactation was smaller and less persistent than that of dietary MP supply and single- v. twin-suckling.Populations of both nematodes were lower in association with high MP supply, vaccination and single suckling. These changes were associated with increases in numbers of mucosal mast cells (MMC) as a result of both increased MP supply and vaccination. Evidence for a more rapid return of host ability to limit populations of the abdominal nematode T. circumcincta than of the intestinal nematode T. colubriformis was associated with fewer eosinophils and more globule leucocytes (GL) in abomasal than in intestinal tissue.None of the serum antibody isotypes was affected by dietary protein supply. Total and IgA antibodies were maintained by a current larval (vaccinating) intake. IgA titres, however, increased progressively during pregnancy, especially in twin-bearing ewes. IgE titres appeared to be sensitive primarily to the reproductive cycle itself, peaking around parturition.This work supports the conclusion that availability of MP supply influences the recruitment and activity of cells of the immune armoury of the gastro-intestinal tract to nematode parasites. The precise outcome may differ with site and/or nematode species.     

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.     

Association between the prion protein genotype and animal performance traits in a large multibreed sheep population

Genetic susceptibility to scrapie, a fatal disease of sheep and goats, is modulated by polymorphisms in the prion protein (PrP). Neither the frequency of the PrP genotypes nor their association with animal performance has been investigated in a large multibreed Irish sheep population. Scrapie genotypes were available on 16 416 animals; the breeds represented included purebred Belclare (733), Charollais (333), Suffolk (739), Texel (1 857), Vendeen (191), and crossbreds (12 563). Performance data on lambing, lamb and ewe performance as well as health traits were available. The association between alternative approaches of describing the PrP genotype (i.e. 15 individually called PrP genotypes, five genotype classes representing susceptibility to scrapie, or number of ARR haplotypes) and animal performance were quantified using animal linear mixed models. All 15 of the possible scrapie genotypes were detected, although the frequency differed by breed. The frequency of the five PrP haplotypes in the entire population were 0.70 (ARR), 0.15 (ARQ), 0.11 (ARH), 0.02 (AHQ) and 0.01 (VRQ); the most susceptible haplotype (VRQ) was only detected in purebred Texels and crossbreds. No association was detected between the PrP genotype of either the animal or dam and any of the lambing traits (i.e. lambing difficulty score, perinatal mortality and birth weight). With the exception of ultrasound muscle depth, no association between the PrP genotype and any of the lamb performance traits (i.e. lamb BW and carcass) was observed. Lambs carrying the category four PrP genotype (i.e. ARR/VRQ) had 1.20 (SE = 0.45) mm, 1.38 (SE = 0.12) mm, 1.47 (S = 0.25) mm shallower ultrasound muscle depth relative to lambs of the less susceptible scrapie categories of 1, 2, 3, respectively (P < 0.05). Nonetheless, no association between PrP genotype and lamb carcass conformation, the ultimate end goal of producers, was detected. Ewe litter size, body condition score or lameness did not differ by PrP genotype of the ewe (P > 0.05). For ewe mature BW, ARH/VRQ ewes differed from most other ewe PrP genotypes and were, on average, 3.79 (SE = 1.66) kg heavier than ARR/ARR genotype ewes. Lamb dag score differed by dam PrP genotype (P < 0.05), although the differences were small. Results from this study show that scrapie is segregating within the Irish sheep population, but the PrP genotype was not associated with most traits investigated and, where associations were detected, the biological significance was minimal. This suggests minimal impact of selection on PrP genotype on performance, at least for the traits investigated in the present study.     

The association between frequency of thiabendazole treatment and the development of resistance in field isolates of Ostertagia spp. of sheep

Martin P. J., Anderson N., Lwin T., Nelson G. and Morgan T. E. 1984. The association between frequency of thiabendazole treatment and the development of resistance in field isolates of Ostertagia spp. of sheep. International Journal for Parasitology14: 177–181. Isolates of Ostertagia spp. were obtained from grazing sheep 3,4 and 5 years after nil, planned (five per year) and regular (3-weekly) treatments with thiabendazole (TBZ). Levels of resistance to TBZ were measured by an in vitro egg hatch assay and a controlled anthelmintic efficiency assay. Isolates from planned treatment groups showed an increase in the level of resistance; the lethal concentrations of TBZ to 50% of eggs (LC₅₀s) were 3, 3 and 6 times the LC₅₀s of isolates from nil treatment groups for years 3, 4 and 5 respectively. The LC₅₀s of isolates from regular treatment groups were 14 times higher than those from nil treatment groups in each year. To assess the potential for an increase in level of resistance, additional egg assays were done 14 days after treatment with 44 mg kg^t?1 of TBZ on sheep infected with the planned group isolates for each year. This treatment raised the LC₅₀S for years 3, 4 and 5 respectively by 3, 2 and 1–5 times the LC₅₀s of the same isolates which had not been exposed to additional TBZ treatment. The controlled anthelmintic efficiency assay using 44 mg kg^t?1 of TBZ produced a significant reduction in the number of adult and immature worms from the nil isolate but failed significantly to reduce the number of worms from the planned and regular isolates. A three component analysis resolved the nonlinear trends of the log dose-probit plots in egg hatch assays for isolates from planned treatment groups into subpopulations of susceptible, hybrid and resistant individuals each with different levels of resistance. The proportions of these subpopulations changed in accordance with the level of resistance observed in each year.     

An association between the acute phase response and patterns of antigen induced T cell proliferation in juvenile idiopathic arthritis

The aim of this research was to determine whether all memory T cells have the same propensity to migrate to the joint in patients with juvenile idiopathic arthritis. Paired synovial fluid and peripheral blood mononuclear cell proliferative responses to a panel of antigens were measured and the results correlated with a detailed set of laboratory and clinical data from 39 patients with juvenile idiopathic arthritis. Two distinct patterns of proliferative response were found in the majority of patients: a diverse pattern, in which synovial fluid responses were greater than peripheral blood responses for all antigens tested; and a restricted pattern, in which peripheral blood responses to some antigens were more vigorous than those in the synovial fluid compartment. The diverse pattern was generally found in patients with a high acute phase response, whereas patients without elevated acute phase proteins were more likely to demonstrate a restricted pattern. We propose that an association between the synovial fluid T cell repertoire and the acute phase response suggests that proinflammatory cytokines may influence recruitment of memory T cells to an inflammatory site, independent of their antigen specificity. Additionally, increased responses to enteric bacteria and the presence of αEβ7 T cells in synovial fluid may reflect accumulation of gut associated T cells in the synovial compartment, even in the absence of an elevated acute phase response. This is the first report of an association between the acute phase response and the T cell population recruited to an inflammatory site.     

Genome-wide association study dissects the genetic control of plant height and branch number in response to low-phosphorus stress in Brassica napus

Background and AimsOilseed rape (Brassica napus) is one of the most important oil crops worldwide. Phosphorus (P) deficiency severely decreases the plant height and branch number of B. napus. However, the genetic bases controlling plant height and branch number in B. napus under P deficiency remain largely unknown. This study aims to mine candidate genes for plant height and branch number by genome-wide association study (GWAS) and determine low-P-tolerance haplotypes.MethodsAn association panel of B. napus was grown in the field with a low P supply (P, 0 kg ha⁻¹) and a sufficient P supply (P, 40 kg ha⁻¹) across 2 years and plant height and branch number were investigated. More than five million single-nucleotide polymorphisms (SNPs) were used to conduct GWAS of plant height and branch number at two contrasting P supplies.Key ResultsA total of 2127 SNPs were strongly associated (P < 6·25 × 10⁻⁰⁷) with plant height and branch number at two P supplies. There was significant correlation between phenotypic variation and the number of favourable alleles of associated loci on chromosomes A10 (chrA10_821671) and C08 (chrC08_27999846), which will contribute to breeding improvement by aggregating these SNPs. BnaA10g09290D and BnaC08g26640D were identified to be associated with chrA10_821671 and chrC08_27999846, respectively. Candidate gene association analysis and haplotype analysis showed that the inbred lines carrying ATT at BnaA10g09290Hap1 and AAT at BnaC08g26640Hap1 had greater plant height than lines carrying other haplotype alleles at low P supply.ConclusionOur results demonstrate the power of GWAS in identifying genes of interest in B. napus and provided insights into the genetic basis of plant height and branch number at low P supply in B. napus. Candidate genes and favourable haplotypes may facilitate marker-based breeding efforts aimed at improving P use efficiency in B. napus.     

Think globally,act locally: the role of local demographics and vaccination coverage in the dynamic response of measles infection to control

The global reduction of the burden of morbidity and mortality owing to measles has been a major triumph of public health. However, the continued persistence of measles infection probably not only reflects local variation in progress towards vaccination target goals, but may also reflect local variation in dynamic processes of transmission, susceptible replenishment through births and stochastic local extinction. Dynamic models predict that vaccination should increase the mean age of infection and increase inter-annual variability in incidence. Through a comparative approach, we assess national-level patterns in the mean age of infection and measles persistence. We find that while the classic predictions do hold in general, the impact of vaccination on the age distribution of cases and stochastic fadeout are mediated by local birth rate. Thus, broad-scale vaccine coverage goals are unlikely to have the same impact on the interruption of measles transmission in all demographic settings. Indeed, these results suggest that the achievement of further measles reduction or elimination goals is likely to require programmatic and vaccine coverage goals that are tailored to local demographic conditions.     

Inhibitory effect of ethylenediaminetetraacetate (EDTA) on the porcine lymphocyte response to mitogens and reversal of the effect with metal ions

The effect of ethylenediaminetetraacetate (EDTA) on the mitogen response of porcine lymphocytes and the role of metal ions in reversal of the inhibitory effect of EDTA were determined. Porcine lymphocyte responses to mitogens were totally suppressed when serum used to supplement Ca²⁺, Mg²⁺-free minimum essential medium (MEM) was dialyzed against saline or saline with 0.2 or 0.60 mM EDTA, but the responses were only partially reduced when the same serum was added to RPMI-1640 medium. The inhibition observed in MEM could be reversed by adding 1×10⁻³ M Ca²⁺ and 1×10⁻³ M Mg²⁺ to the dialyzed serum. Serum treated directly with 0.60 mM EDTA completely suppressed blastogenesis in lymphocyte cultures maintained in RPMI-1640 or Ca²⁺, Mg²⁺ free MEM. The inhibitory effect of EDTA-treated serum could be completely reversed by adding Zn²⁺ or a combination of Zn²⁺ with other cationic ions, or partially reversed by adding Ni²⁺ or Fe³⁺. Zn²⁺ was the most effective ion, in that it was the only ion that, when alone added to the serum, could completely restore lymphocyte responses to phytohemagglutinin (PHA) or pokeweed mitogen (PWM).     

HDL3 reduces the association and modulates the metabolism of oxidized LDL by osteoblastic cells: a protection against cell death

Oxidized low density lipoproteins (OxLDL) are known to promote atherosclerosis, but it is only recently that OxLDL have been associated with alterations of the functions of bone-forming osteoblasts and osteoporosis. Although high density lipoproteins (HDL) are recognized for their anti-atherogenic action, there is less information about their ability to protect against osteoporosis. Therefore, we investigated the capacity of HDL3 to prevent the cell death induced by OxLDL in human osteoblastic cells. Simultaneous exposure of the cells to HDL3 and OxLDL abolished the reduction of cell viability monitored by MTT activity measurement and the induction of apoptosis determined by annexin V staining indicating that HDL3 prevent the apoptosis of osteoblasts induced by OxLDL. This protection correlated with the displacement by HDL3 of OxLDL association to osteoblasts, signifying that OxLDL binding and/or internalization are/is necessary for their cytotoxic effects. We also found that exposition of osteoblastic cells to HDL3 prior to incubation with OxLDL reduced cell death and preserved the lysosomal integrity. This protection was correlated with an increase of SR-BI expression, a modification of OxLDL metabolism with less global uptake of OxLDL and greater selective uptake of cholesterol from OxLDL. These results strongly suggest that, as for atherosclerosis, HDL may exert beneficial actions on bone metabolism.