Injection site lesions of footrot vaccines in sheep

Three trials were conducted to assess the size and frequency of lesions induced by an oil-based and an alum-precipitated aqueous B. nodosus commercial footrot vaccine at two injection sites (neck and brisket) with or without prior skin swabbing with methanol. Animals used were previously unvaccinated and clean and dry at time of vaccination. Trial 1 contained 57 2-tooth Booroola Merino x Coopworth ewes, trial 2 contained 60 mixed-age Corriedale ewes, and trial 3 contained 39 rams of mixed age and sex. Firm nodules or plaques with indurated edges and with a maximum dimension of 1cm or more appeared soon after vaccination and were still very common (50-100% of sheep vaccinated) 5-6 weeks after injection. A variable proportion exhibited a liquefied central area of necrosis and later discharged necrotic material. The oil-based vaccine produced significantly larger lesions than the alum-precipitated aqueous vaccine. Lesion size at the brisket site was significantly larger than the neck site in ewes but not in rams. Skin preparation with methanol had no effect on lesion frequency or size.     

Duration of resistance to experimental footrot infection in Romney and Merino sheep vaccinated with Bacteroides nodosus oil adjuvant vaccine

Groups of 10-12 Romney and Merino wethers were challenged simultaneously with homologous experimental footrot infection after having received the second of 2 doses of Bacteroides nodosus (strain 198) vaccine 0, 4, 8, 12, 16 and 20 weeks previously. Inoculations were carried out 28 days apart and unvaccinated sheep of both breeds were challenged as controls. Most Romneys that had been vaccinated up to 16 weeks prior to challenge were resistant to footrot whereas 8 of 10 controls were susceptible. This resistance was lost by about 20 weeks after vaccination. By contrast, protection against challenge in vaccinated Merinos lasted only about 4-5 weeks, although residual benefits of vaccination were apparent after longer intervals from the reduced number and severity of foot lesions among vaccinates compared with controls. Agglutinin titres, which did not differ markedly after similar intervals between Romneys and Merinos, reached maximum levels between 4 and 8 weeks after the second vaccine dose and subsequently declined. Although peak titres were generally recorded at the time of maximum protection in Merinos, the relationship between agglutinin levels and resistance in the Romneys was ill-defined.     

Protection of sheep against experimental footrot by vaccination with pili purified from Bacteroides nodosus

Merino sheep vaccinated with either whole Bacteroides nodosus organisms, a crude surface antigen preparation or highly purified pili (>99% homogeneity) in oil adjuvant, developed significant resistance to artificial footrot infection when compared with unvaccinated control sheep inoculated with saline-in-oil emulsion (Freund;s incomplete adjuvant) alone. The pili-vaccinated sheep generally had higher K-agglutinating antibody titres than sheep vaccinated with whole B. nodosus. These results confirmed the role of B. nodosus pilus protein both as a protective antigen and the K-agglutinogen. Vaccines prepared with Freund;s incomplete adjuvant containing either purified pili, crude pili or B. nodosus whole cells did not produce significantly different injection-site reactions.     

The effect of time between doses on serological response to a recombinant multivalent pilus vaccine against footrot in sheep

SUMMARY The response of sheep to a recombinant multivalent footrot vaccine containing pilus antigens was examined after the administration of two doses of vaccine at Intervals ranging from 2 to 52 weeks. Agglutinating antibody titres were measured 3 weeks after the second vaccination and showed that lengthening of the interdose interval results in higher agglutinin titres. The capability of sheep to mount an increasingly strong immune response as the interval between doses is increased provides an opportunity to maximise the usefulness of vaccination by administering the first dose well before an expected footrot transmission period. This advantage of increasing the interdose interval has not been reported for traditional, whole-cell footrot vaccines, and use of the new pilus vaccine in this manner may improve prospects for disease control. Furthermore, sheep given a third dose either 6 or 12 months after their initial two-dose vaccination program achieved significantly higher titres than those elicited after the second dose, suggesting the likelihood of further improvement in disease control in successive seasons.     

Vaccination against lumpy jaw and measurement of antibody response in wallabies (Macropus eugenii)

Successful protection against lumpy jaw disease in a colony of captive wallabies (Macropus eugenii) was induced by vaccination with a commercial ovine footrot vaccine. No mortalities attributable to lumpy jaw were observed in 69 vaccinated animals while six of 42 unvaccinated control wallabies died of the disease. Vaccinated animals exhibited significant increases in antibody titres to Bacteroides nodosus after the first and second doses of vaccine. Titres were measured by an enzyme-linked immunosorbent assay.     

Colostral transfer of Bacteroides nodosus antibodies in sheep.

Ovine contagious foot rot may cause lameness in sheep, resulting in decreased wool growth and low weight gain. Affected neonatal lambs are difficult to treat, and treatment is labor intensive; thus, a method of prevention is warranted. Vaccination of ewes with a multivalent vaccine in an oil adjuvant induced development of antibody to the somatic O antigen of Bacteroides nodosus, and this antibody was detected in serum of newborn lambs after consumption of colostrum from the vaccinated ewes. Antibody titers were determined in 48 unvaccinated ewe/lamb pairs, and in 50 once-vaccinated and 78 twice-vaccinated pairs. Serum and colostrum O-agglutinin titers to B nodosus were determined by a microtitration agglutination test. Lambs from vaccinated ewes had significantly (P less than 0.05) higher O-agglutinin titers than those from unvaccinated ewes, and double vaccination of ewes resulted in the highest potentially protective titers (greater than 1:2,400) in ewes and lambs.     

Impact of footrot vaccination and antibiotic therapy on footrot and contagious ovine digital dermatitis

Footrot and contagious ovine digital dermatitis (CODD) are common causes of foot disease of sheep in the UK. The study reported here is a split flock randomised treatment trial undertaken on a group of 748 fattening lambs on a UK sheep farm affected by CODD and footrot. The sheep were randomly assigned to one of two treatment protocols. In protocol A, all sheep were given two doses of footrot vaccine (Footvax, MSD), plus targeted antibiotic therapy (long-acting amoxicillin, Betamox LA, Norbrook Pharmaceuticals) to sheep with foot lesions likely to be associated with a bacterial infection. In protocol B, the sheep only received targeted antibiotic therapy. Sheep were re-examined and foot lesions recorded five and nine weeks later. New infection rates in the footrot vaccinated group were lower compared with the vaccinated group for both CODD (18.2 per cent compared with 26.4 per cent, P=0.014) and footrot (12.55 per cent compared with 27.5 per cent, P<0.001). Recovery rates were unaffected for CODD (80.46 per cent compared with 70.97 per cent, P=0.14) but higher for footrot (92.09 per cent compared with 81.54 per cent, P=0.005) in sheep which received the vaccine. On this farm, a footrot vaccine efficacy of 62 per cent was identified against footrot and 32 per cent against CODD infection. An association between a sheep having footrot at visit 1 and subsequently acquiring CODD was identified (odds ratio [OR] 3.83, 95 per cent CI 2.61 to 5.62, P<0.001). These results suggest a role for infection with Dichelobacter nodosus in the aetiopathogenesis of CODD on this farm.     

Effectiveness of vaccination in controlling ovine footrot

Efficacy of reducing footrot by use of a commercial vaccine was evaluated over 2 yr with 223 ewes from five crossbred genotypes. Ewes were assigned randomly within genotype, age and initial footrot status to a 2 x 3 factorial array of vaccination in the 1st yr and fall vaccination, winter vaccination or control in the 2nd yr. The flock contained a single Bacteroides nodosus serotype (XV) known to be highly cross-reactive with one of the vaccine serotypes (British serotype H). Vaccination reduced footrot incidence by 61% (P less than .01) in the 1st yr and by 45% (P less than .01) in the 2nd yr. Vaccination of previously uninfected ewes did not significantly reduce the rate of new infection, but vaccination of infected ewes reduced re-infection by 92% (P less than .01). Vaccination in the 1st yr failed to produce any carryover or additive protection in the 2nd yr. Genotypes differed in both footrot incidence (P less than .05) and footrot reduction following vaccination (P less than .05), but the effect was inconsistent between years. Vaccination increased serum antibody titers in both years (P less than .01). Among vaccinates in the 1st yr, infected ewes had lower titer levels than uninfected ewes (P less than .01), but no correlation between titer level and incidence was observed in the 2nd yr.     

Differences between breeds of sheep in their responses to Bacteroides nodosus vaccines

In a field experiment on irrigated pasture, sheep of several breeds were vaccinated twice, subcutaneously, in the upper neck, with Bacteroides nodosus vaccine containing either depiliated cells (DC vaccine), or whole, piliated cells (WC vaccine) and the responses were measured over the following 14 weeks. DC vaccine was as effective as WC vaccine in protecting against the development of foot-rot under conditions of moderate challenge, although the WC vaccine induced significantly higher pilus agglutinating antibody titres. Foot-rot developed in significantly more vaccinated Merinos (Peppin and Saxon strains) than in Romney Marsh, Dorset Horn or Border Leicester breeds. Agglutinating antibody titres after WC vaccination were significantly lower in the Peppin Merino than in the other sheep for the first 6 weeks, while after DC vaccination the titres remained elevated longer in the Border Leicester and Saxon Merino and were significantly higher from 6 weeks onwards. Reactions at the inoculation sites were generally larger in the British breeds than in the Merinos and among the former the reactions were largest, most numerous and most frequently discharged their contents in the Dorset Horn. Bodyweight gains in all vaccinated sheep were initially reduced, compared with controls, but the differences were no longer significant after the eighth week.     

The incidence of caseous lymphadenitis in Alberta sheep and assessment of impact by vaccination with commercial and experimental vaccines.

In Alberta, caseous lymphadenitis (CLA) is one of the leading causes of lamb and mutton carcass condemnation. In this study, serologic results confirmed a high (50-94%) incidence of exposure to Corynebacterium pseudotuberculosis, the causative agent of CLA, in mature, unvaccinated sheep in southern Alberta. To assess the efficacy and impact of vaccination with 2 commercial (Glanvac-6 and Case-Vac) and 1 experimental (WC+ MDP-GDP) CLA vaccines, a series of 3 field trials in 3249 ewes and lambs was conducted in affected flocks from 1992-1996. Efficacy was assessed from the serological response to vaccination, prevalence and size of injection site reactions by treatment, and the incidence of CLA abscesses. Overall, agglutinating antibody titres to C. pseudotuberculosis in lambs vaccinated with WC+MDP-GDP and Case-Vac remained significantly elevated above nonvaccinated control lambs for the 12 mo period after the initial vaccination. Lambs vaccinated with the WC/MDP-GDP maintained higher titres (P < 0.06) than those vaccinated with Case-Vac for the period from 6 to 12 mo after vaccination. Agglutinating antibody titres for lambs vaccinated with Glanvac did not differ from those of controls at any point during the 12 mo period after vaccination. The number of injection site reactions was elevated in lambs vaccinated with Glanvac as compared to those vaccinated with WC+MDP-GDP but the size of injection site reactions did not significantly differ. Sheep vaccinated with WC+ MDP-GDP also had a reduced incidence of putative CLA abscesses, although confirmation of the presence of C. pseudotuberculosis was only successful in a small number of instances.     

Studies on the antigenic structure of Bacteroides nodosus.

The antigenic mosaics of three Bacteroides nodosus isolates (198, 199 and 127) were studied to elucidate the nature of the protective immunogen. In vaccinated sheep the three isolates induced high homologous serum agglutinin titres but it was also apparent that 198 and 199 shared a major surface antigen not present on 217. This major cross-reacting antigen was not detected with rabbit antisera. The fimbriae, consisting predominantly of protein, induced high homologous titres in rabbits and represented the type-specific antigen. Lipopolysaccharide (LPS) from each of the isolates induced low agglutinin titres and high 2-mercaptoethanol-sensitive indirect haemagglutinating antibody titres. The heat-stable LPS contained at least two common carbohydrate O antigen determinants but no type-specific O antigens were detected.     

Field trials with a killed, nine-strain, oil adjuvanted Bacteroides nodosus footrot vaccine in sheep

Field trials were set up on five properties to determine the cure and protection rates of a killed, nine-strain, oil adjuvanted Bacteriodes nodosus footrot vaccine, under natural conditions. Foot infections were scored at the time of the first vaccination and at approximately four weeks and ten weeks later. Cure rates were 34-47% for vaccinated sheep, and 18-47% for untreated sheep. Using the Australian Licensing Authorities calculation, the cure rates for the vaccine ranged from -40% to 44%. A lack of suitable conditions for spread of footrot meant that protection rates could not be determined.     

Broomfield Corriedales: a strain of sheep selectively bred for resistance to footrot

Broomfield Corriedales originate from a 15-year programme of selective breeding from sheep that evaded footrot when deliberately subjected to field challenge. To determine whether this policy may have resulted in improved resistance to footrot, trials were set up in which representative animals from the Broomfield blood-line were challenged with footrot by either experimental or natural infection, together with Corriedale sheep of similar age and sex but from flocks in which the disease is not known to occur. In another approach, the occurrence of footrot was compared in the offspring of Broomfield and of non-Broomfield sires mated with randomly selected ewes in a commercial Corriedale flock in which the condition was prevalent. Finally, the effects of natural challenge in Broomfield lambs were compared with those in a selection line of Corriedale lambs of which both parents had a history of chronic footrot disease. Footrot affected significantly fewer sheep of the Broomfield blood-line and to a markedly lesser degree than those chosen for comparison under identical challenge conditions. Where present, foot infections in Broomfield animals also tended to be less invasive and less persistent. Reduced footrot prevalence was also demonstrated in three birth cohorts of first-cross offspring of Broomfield sires relative to the offspring of other sires, though differences were not significant at all inspections. These findings, considered as a supplement to the historical development of the Broomfield flock, offer encouraging prospects for breeding programmes to enhance the resistance of sheep to footrot disease.     

Comparison of alum-absorbed or non-alum-absorbed oil emulsion vaccines containing either pilate or non-pilate Bacteroides nodosus cells in inducing and maintaining resistance of sheep to experimental foot rot

Highly pilate (P) or non-pilate (NP) cells of Bacteroides nodosus were compounded into oil emulsion (O) either with or without prior absorption onto alum (A). The abilities of these four preparations (referred to as PAO, NPAO, PO and NPO vaccines) to stimulate antibody production and to protect sheep from foot rot were compared. Two injections of PAO vaccine protected sheep against homologous challenge 12 weeks after the second dose by PO, NPO and NPAO vaccines were less effective. Sheep were protected against homologous challenge for 14 weeks after a single dose of PAO vaccine and for 22 weeks after three doses; an ameliorative effect was still evident 40 weeks after the third dose. Protection against challenge with two heterologous strains was demonstrated at six weeks after three doses of vaccine. A numerical system of scoring the lesions also confirmed that foot rot in vaccinated sheep challenged outside the 'protective' period of the vaccine was somewhat less severe than in controls. PAO vaccine induced much higher and more persistent titres of agglutinins than the other vaccines tested. There was a relationship between agglutinin titres and resistance to homologous challenge.     

布鲁氏菌弱毒疫苗粘膜免疫及检测方法的研究

为研究布鲁氏菌弱毒疫苗粘膜免疫及其检测方法,本实验采用粘膜点眼途径对健康母羊接种布鲁氏杆菌猪2号疫苗(S2)、牛19号疫苗(A19)和羊强毒株(M16),筛选布鲁氏杆菌病鉴别检测方法。将12月龄～14月龄母羊60只随机分为3组,以常规疫苗推荐剂量进行半量粘膜点眼接种。采集血液、淋巴、脏器进行布鲁氏菌病血清学检测和细菌学分离以及PCR检测。结果表明:布鲁氏菌弱毒疫苗抗体水平持续6个月,其中血清学的试管凝集试验、半胱氨酸凝集试验与补体结合试验的阳性符合率达到100%。细菌分离期为6个月,乳腺、乳腺淋巴、髂淋巴分离率较高;而强毒株M16的抗体水平和细菌分离持续12个月以上。结果显示以常规血清学和细菌学检测方法在点眼免疫布鲁氏菌S2、A19苗6个月后可以进行野毒感染和疫苗免疫畜的鉴别诊断。     

Eradication of virulent footrot from sheep and goats in an endemic area of Nepal and an evaluation of specific vaccination

Programmes based on the identification and treatment of cases and the culling of animals refractory to treatment had failed to eradicate virulent footrot from two districts in the western region of Nepal. From 1993 to 1996 vaccination against two endemic virulent strains of Dichelobacter nodosus was tested for its potential to contribute to the eradication of footrot from the region. Only sheep and goats which had been free of signs of footrot at three inspections at monthly intervals before their annual migration to alpine pastures were eligible for inclusion. From November 1992, the treatment of cases identified during inspections included the injection of specific vaccine. Successfully treated cases migrated with their flocks but were excluded from the vaccine trial. Non-responding cases were culled. Forty combined flocks of sheep and goats (approximately 9500 animals) were used initially to compare three vaccination regimens. Eleven flocks (sheep and goats) were treated with two doses of specific vaccine (group A), nine (sheep and goats) were treated with commercial vaccine followed by specific vaccine (group B) and 10 (sheep and goats) were treated with two doses of commercial vaccine (group C) in March to April 1993 before the annual migration; 10 flocks (sheep and goats) remained unvaccinated (group D). Only sheep and goats free of signs of footrot were allowed to migrate. Nevertheless, virulent footrot recurred in many flocks three months later. However, its prevalence was significantly lower in group A than in the other three groups combined. Groups A, B and C then received the specific vaccine before their migrations in 1994 to 1996; group D remained unvaccinated. The annual programme of inspection and identification and treatment of cases continued for seven years, but the vaccinations ceased after four years. There was no recurrence of virulent footrot after November 1993. After the first season the virulent strains of D nodosus used in the specific vaccine could no longer be isolated, although antigenically distinct, benign strains of the organism persisted in cases of benign footrot.     

Diagnosis of footrot in goats: application of ELISA tests for response to antigens of Dichelobacter nodosus

Goats are an important natural host for footrot and are infected with Dichelobacter nodosus that have virulence characteristics similar to those of sheep strains. However, the humoral response of goats to D. nodosus antigens and the possibility of a serological diagnosis of footrot in goats have not been studied. With the aim of evaluating a diagnostic ELISA test, we investigated the primary immune response of goats to experimental and natural infection, the memory response in recovered animals, and the transfer and persistence of colostral antibodies in kids. Footrot stimulated the goat's immune system and, as in sheep, under-running lesions were the primary stimulus for production of anti-D. nodosus antibodies. The immune response could be detected in ELISA using either fimbrial or outer membrane protein (KSCN) antigens of D. nodosus. Antibody titres resulting from infection declined quickly after recovery and reached pre-infection levels within 3-4 months. Previously affected animals, however, mounted a memory response when injected with purified D. nodosus antigens. Antibody levels attained after anamnestic challenge were correlated with the maximum levels attained during infection, and were therefore indicative of the infection status. Anti-D. nodosus antibodies were also transferred to kids via colostrum, but these antibodies did not persist and therefore were unlikely to interfere with the diagnostic ELISA after 3 months of age. Though these ELISA tests were highly specific, their sensitivity was rather low. Therefore, they are only suitable for a herd diagnosis of footrot in goats and are dependent on the development of advanced under-running infections in a proportion of affected goats.     

Quantitation by ELISA of pili and sheep antibodies to the pili of Bacteroides nodosus

ELISA systems have been developed to quantitate the isotypic antibody response of sheep naturally infected with B. nodosus isolate 198 or injected with pili from isolate 198 in oil emulsion vaccines. The predominant humoral antibody detected following vaccination was IgG1, with substantially lower amounts of IgG2 and IgM. The antibody response was relatively specific for the pilus antigen from isolate 198. Although weak cross reactivities were detected with antiserums to some other isolates, ELISA IgG antibody titres in excess of 200 offer a tentative identification of the isolates of B. nodosus involved in natural outbreaks of footrot. A related ELISA was also developed to quantitate the amount of pili in cell suspensions and crude preparations of pili used in vaccines.     

The use of an autogenous Dichelobacter nodosus vaccine to eliminate clinical signs of virulent footrot in a sheep flock in Bhutan

An outbreak of virulent footrot was investigated in a flock of 605 Merino cross-bred sheep in Bhutan. Conventional control methods in the preceding eight years had reduced its prevalence from 36-79% in different components of the flock to about 15% overall. Only one serogroup (B) of Dichelobacter nodosus was identified among 40 isolates cultured from affected sheep. A vaccine prepared from this strain was used in a pilot trial to compare the response of 14 treated and 14 untreated sheep. All affected, vaccinated animals in this trial healed quickly and were protected against re-infection while additional cases developed among untreated sheep during a period favourable for the spread of footrot. The serogroup B vaccine was administered to the whole flock for two successive years. No other footrot treatment was given during these or subsequent years. The whole flock was examined three times, foot by foot, for two years and twice yearly for another two years. When vaccination began there were 88 affected sheep in the flock, an affected sheep being defined as an animal with a foot-score of 2 or greater in one or more feet. There were neither affected sheep in the flock 30 days after the first dose of vaccine nor were any identified in later inspections. Virulent footrot, originating from the farm under investigation, persisted in neighbouring village flocks during this period. It was concluded that whole flock specific D. nodosus vaccination made a major contribution to the elimination of all clinical signs of footrot from the flock of 605 sheep where the condition had previously persisted for 10 years.     

Treatment of ovine virulent footrot with zinc sulphate/sodium lauryl sulphate footbathing

Trials were conducted on 2 commercial sheep flocks in the Gippsland region of Victoria to determine the efficacy of treating ovine virulent footrot by footbathing in aqueous zinc sulphate solution (20% w/v). The effects of foot paring, parenteral penicillin, vaccination and addition of sodium lauryl sulphate (SLS) to the footbaths were assessed. Trial 1 comprised 297 sheep with an initial prevalence of footrot of 33% and most lesions were severe and chronic. Treatment of sheep with unpared feet by zinc sulphate footbathing for 1h did not result in a significant reduction in footrot prevalence (n = 120, cure rate 33%) whereas a significant (P less than 0.01) response was obtained by footbathing for 1h with zinc sulphate/SLS (n = 120, cure rate 55%). Trial 2 comprised 1,042 sheep with a pretreatment footrot prevalence of 71% and predominantly severe lesions. In this flock all treated sheep were footbathed in zinc sulphate/SLS for 1h on 2 occasions, 5 days apart and the effects of additional surgical and parenteral treatments were assessed. Foot paring had a significant detrimental effect on cure rate (P less than 0.01). The administration of procaine penicillin at the time of the first footbathing with zinc sulphate/SLS made no significant improvement to the rate of cure. Footrot vaccine given 8 and 2 weeks prior to footbathing did not cure significantly more sheep than footbathing alone, but the results were significantly better than from foot paring plus footbathing, and from combined foot paring, footbathing and parenteral penincillin treatment (P less than 0.01). The cure rate was 84% for sheep that were only footbathed, 72% for those foot pared and footbathed, 72% for those foot pared, footbathed and given penicillin, and 88% for those vaccinated and footbathed.