Transmission of virulent footrot between sheep and goats

OBJECTIVE: To determine the infectivity of ovine and caprine strains of Dichelobacter nodosus for both sheep and goats. DESIGN: Pen experiments in which 20 sheep and 19 goats were challenged directly with the two strains, and transmission experiments on pasture, using donors infected by experimental challenge. RESULTS: Sheep and goat strains of D nodosus infected both animal species in experimental challenges. Animals so infected transmitted footrot to both sheep and goats on pasture plots. A significantly smaller proportion of goats than sheep was infected when challenged with either strain. The interval between exposure and development of footrot in goats was longer than in sheep when recipient animals were exposed to infected donors on pasture. The disease was less invasive in goats than in sheep. CONCLUSIONS: With the strains of D nodosus used there was no evidence of host specificity. Direct transmission of footrot can occur between sheep and goats in the same environment. There is a need to include goats in ovine footrot eradication programs and vice versa.     

Effect of climatic region on the clinical expression of footrot of lesser clinical severity (intermediate footrot) in sheep

OBJECTIVE: To determine if the clinical classification of intermediate footrot (IFR) is changed to virulent footrot (VFR) by a transfer of the infected flock to a region where climatic conditions are more favourable for the transmission of the disease. DESIGN: Clinical examination of two groups of Merino wethers infected with IFR; one group of 309 in a region considered less favourable for footrot and another group of 343 at a second site considered more favourable. PROCEDURES: After characterising the form of footrot at the first site, infection was established at the second site by mixing 142 wethers from the first site with 201 unrelated wethers considered to be free of IFR and VFR. Observations of clinical characteristics were made over a 16 month period during which an outbreak of footrot occurred. Clinical assessments were made by inspecting every foot of every sheep at regular intervals and allocating a footscore. Evidence that the same clonal lines of D. nodosus were responsible for the footrot at both sites was provided by serotyping of isolates and using omp gene RFLP as a molecular epidemiological tool. RESULTS: The disease at the first site was classified as IFR because 7% of the sheep developed a maximum footscore (MFS) of 4, the most severe category, despite relatively low rates of transmission. When the outbreak occurred at the second site, which was more suitable for footrot transmission, the maximum proportion of the flock that developed a MFS of 4 was 3.6%, confirming the initial classification of IFR. CONCLUSIONS: When a flock infected with IFR was moved to a region where climatic conditions were more favourable for footrot transmission, the clinical classification of the disease remained the same in both the original flock and in sheep exposed to the infection for the first time.     

Severity and persistence of footrot in Merino sheep experimentally infected with a protease thermostable strain of Dichelobacter nodosus at five sites

Objective To test the hypothesis that ovine footrot associated with a thermostable protease strain of Dichelobacter nodosus undergoes self cure or is sustained as an annually recurring disease, depending on the environment.
Design and procedure Forty Merino sheep from a single blood line were infected with a protease thermostable strain of D nodosus a t each of five sites in Western Australia. Footrot lesions and microscopic evidence of D nodosus were recorded every fortnight for 2.5 years, supplemented by laboratory culture. Rainfall, soil and air temperature, pasture quantity and composition and soil types were also recorded. Flocks that apparently self cured were relocated to a more favourable site for footrot in the final spring season.
Results The maximum prevalence of feet with clinical footrot lesions was 80.6, 1.3, 14.4, 3.8 and 88.1% at the five sites. Severe footrot occurred for three consecutive spring seasons at one site that had clay loam soil and at least 3500 kg/ha total pasture dry matter annually. However, the infection was asymptomatic for up to 10 weeks between outbreaks. D nodosus was isolated from flocks for 2.5 years at only two sites, although there was microscopic evidence of the organism at other sites in the final year. A thermolabile variant (strain U6) of D nodosus was isolated from the two sites where footrot persisted.
Conclusion Depending on time and location, ovine footrot induced by a protease thermostable strain of D nodosus either self cured or persisted as annual outbreaks interspersed with periods of asymptomatic infection.     

A longitudinal study of the risks for introduction of severe footrot into sheep flocks in the south west of Norway

In 2008, ovine footrot was detected in Norway for the first time since 1948. By December 2012 it had spread to 99 flocks, all in the county of Rogaland in the south west of Norway, and 42% of which were located in the municipality of Rennesøy in Rogaland. The aim of this study was to investigate risk factors for contracting severe footrot in flocks of sheep. A flock was considered positive for severe footrot based on positive virulence test or by clinical signs in addition to a positive PCR test.     

Observations on the indirect transmission of virulent ovine footrot in sheep yards and its spread in sheep on unimproved pasture

SUMMARY: Virulent ovine footrot was transmitted accidentally to a group of 23 adult Merino sheep (flock B) after holding for 1 hour in sheep yards, which earlier the same day had contained another flock (flock A) with < 1% prevalence of sheep with footrot lesions. Sheep in flock B were rendered susceptible to virulent footrot by grazing 600 mm high unimproved pasture dominated by paspalum ( Paspalum dilatatum ) and kangaroo grass ( Themeda australis ) during warm, humid and wet weather. In addition to moisture, interdigital abrasions caused by the pasture might have predisposed the interdigital skin to infection with Dichelobacter nodosus .     

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.     

An anamnestic serological test for ovine footrot

Following recovery from ovine footrot, a proportion of sheep in a flock may carry the causative organism and spread it to other sheep if environmental conditions are favourable. Footrot affected sheep have elevated levels of serum antibody against Bacteroides nodosus, but these levels decline rapidly after clinical recovery. When challenged by subcutaneous injection with 470 micrograms of protein extracted from the cell membrane of B. nodosus, without adjuvant, sheep that had recovered clinically from virulent footrot produced a marked increase in specific serum antibody within 7 d, while antibody levels in footrot-free sheep injected with the same antigen, and in saline injected controls, did not increase over a period of 25 d. Artificial stimulation and serological detection of immune memory may be useful in footrot eradication programs by identifying sheep that have had clinical footrot infection. This procedure may be applicable to other diseases where antibody responses are inconsistent or transient.     

Field evaluation of ‘Defeat’ a commercial preparation based on copper 8-hydroxyquinolate,for the control of ovine footrot

Extract

Sir: — In early 1984, a new footbathing chemical named ‘Defeat’ (F.I.L. Industries Ltd, P.O. Box 4114, Mt Maunganui) became commercially available in New Zealand for controlling footrot in sheep. Described as both a footrot treatment and preventative, and having a rapid effect against footrot bacteria in sheep, the product is sold for use either as a footbathing preparation, or in conjuction with a portable foam pad that can be saturated with the solution and placed in yards or raceways for sheep to walk over. It is distributed in various pack sizes as a liquid water-soluble concentrate to be diluted 1:100 for footbath and foam pad applications, or 1:50 for direct application to the pared feet of sheep affected with footrot. No recommendations appear on the labels of the packs on the frequency or duration of footbathing treatments required to achieve statisfactory results. A claim that ‘Defeat’ kills the bacteria which cause footrot within 10 seconds of contact features in a promotional brochure issued with the product, this evidently being a reference to results of unspecified in vitro tests     

Experimental evaluation of a commercial footrot vaccine against native Canadian strains of Dichelobacter nodosus.

Two serotypes of the anaerobic bacterium Dichelobacter nodosus were used to experimentally infect young sheep resulting in infectious pododermatitis or footrot characteristic of the natural disease in sheep. The specific serotypes of D. nodosus were reisolated from the feet and identified using immunofluorescent microscopy of hoof scrapings. Prior immunization of sheep with a commercially available bacterin containing whole cell preparations of ten strains of D. nodosus resulted in serum IgG reactive to a serotype of D. nodosus common to the vaccine. Immunization also produced serum IgG reactive to a serotype of D. nodosus not incorporated in the vaccine. A less severe infection occurred in the immunized sheep than in the controls regardless of the serotype of bacteria used to infect them. Clinical lameness and lesion severity were milder in sheep infected with the serotype of D. nodosus common to the vaccine. Western blot analysis of sera from convalescent sheep showed cross-reactive antibodies to nonfimbrial cell surface proteins, as well as bacterial lipopolysaccharide. Such cross-reactivity may explain the partial protection seen in animals infected with a serotype distinctive from the ones in the vaccine. Despite the historical emphasis of fimbrial immunogens in ovine footrot this study using a new model of experimental ovine footrot suggests other surface antigens may also be important in protective immunity.     

Failure to eradicate ovine footrot associated with Dichelobacter nodosus strain A198 by repeated daily footbathing in zinc sulphate with surfactant

OBJECTIVE: To investigate the effect of repeated daily footbathing in zinc sulphate on virulent ovine footrot associated with S1, U1 and U5 zymogram types of Dichelobacter nodosus, including the highly virulent S1 strain A198. DESIGN: A field trial with experimentally infected sheep. PROCEDURE: At week 0, 50 sheep were infected with D. nodosus strains A198 (S1), C305 (U1), BC3993 (U5) and BC3995 (U5). At weeks 1 and 47, respectively, 169 and 235 uninfected sheep were added. At week 60, sheep were allocated to control and treatment groups each containing 220 sheep. Every 2 or 4 weeks to week 113, feet were inspected, and lesions were scored and sampled. Treatment sheep were footbathed in 15 to 18% (w/v) zinc sulphate with surfactant for 5 consecutive days (10 min per day) during week 61. During week 110, the footbathing protocol was applied again, this time to all surviving treatment and control sheep. RESULTS: BC3993 and A198 were isolated from 57% (162 of 285) and 20% (58 of 285), respectively, of new lesions sampled between weeks 3 and 13, and 21% (57 of 271) and 50% (136 of 271) between weeks 49 and 59. Percentages of new lesions associated with C305 and BC3995 remained constant. During the initial 17 weeks after footbathing at week 61, 90% (75 of 83) and 19% (95 of 490) of lesions in treatment and control sheep, respectively, were score 4 or 5, and 94% (47 of 50) and 38% (33 of 87) of those were associated with A198. CONCLUSIONS: Repeated daily footbathing did not eradicate virulent ovine footrot because strain A198 produced deep, covert lesions that facilitated the survival of D. nodosus.     

Effectiveness of small workshops for improving farmers knowledge about ovine footrot

OBJECTIVE: To determine sheep farmers' attitudes to and beliefs about ovine footrot, and to improve their knowledge about the diagnosis, control and eradication of this disease. METHOD: Eighteen workshops, involving 291 farmers, were conducted across Victoria in the spring of 1996. The workshops were designed as small-group discussions with a maximum attendance of 20 farmers to encourage active participation. All participants completed questionnaires before each workshop and 12 to 40 weeks after the last workshop. RESULTS: Before the workshops the farmers had a poor understanding of the principles of diagnosis, control and eradication of footrot. For example, only 50% knew the footrot organism survived in soil for less than 7 days, over two-thirds did not know the reason for paring sheep's feet during an eradication program, and only 31% realised cattle were a potential source of footrot infection for sheep. After the workshops, understanding about footrot was significantly improved; 87% said Dichelobacter nodosus survived in soil less than 7 days, 71% knew the reason for paring sheep's feet and 64% realised that cattle were a potential source of footrot infection. As well as improved knowledge, change of attitude among farmers is fundamentally important if virulent footrot is to be successfully controlled and eradicated. The workshops successfully initiated this process; 40% of farmers thought the workshops changed their attitudes to footrot, while 37% said they gained an increased understanding of other people's opinions about the disease. CONCLUSION: Farmers' poor understanding of ovine footrot is a constraint to the programs aimed at controlling this disease. Small group workshops may be an effective way to influence farmers' attitudes and beliefs, and could facilitate the effectiveness of regulatory disease control programs.     

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.     

Determination of prevalence,serological diversity,and virulence of Dichelobacter nodosus in ovine footrot with identification of its predominant serotype as a potential vaccine candidate in J&K,India

The aim of this study was to determine the prevalence, serological diversity, and virulence of Dichelobacter nodosus in footrot lesions of sheep and identification of its predominant serotype as a potential vaccine candidate. The overall prevalence of footrot in sheep was 16.19%, and ranged from 13.69 to 19.71%, respectively. A total of 759 flocks with 22,698 sheep were investigated for footrot and 2374 clinical samples were collected from naturally infected sheep exhibiting footrot lesions. Of the 2374 samples collected, 1446 (60.90%) were positive for D. nodosus by polymerase chain reaction (PCR). These positive samples when subjected to serogroup-specific multiplex PCR, 1337 (92.46%) samples carried serogroup B, 247 (17.08%) possessed serogroup E, 86 (5.94%) serogroup I, and one (0.069%) serogroup G of D. nodosus. While mixed infection of serogroups B and E was detected in 127 (8.78%), B and I in 46 (3.18%) and B, E, and I in 26 (1.79%) samples, respectively. The serogroup B of D. nodosus was the predominant (92.47%) serogroup affecting sheep population with footrot followed by serogroup E (19.91%) and serogroup I (4.57%), respectively. Virulent status of D. nodosus strains were confirmed by presence of virulence-specific integrase A (intA) gene and the production of thermostable proteases. The intA gene was detected in 709 (72.79%) samples while gelatin gel test carried out on 246 representative isolates all positive for intA gene produced thermostable proteases, confirming their virulence nature. The PCR-restriction fragment length polymorphism (PCR-RFLP) of whole fimA gene of serogroup B revealed the predominance of serotype B5 (82.97%) of serogroup B. This information suggests that serotype B5 is the predominant serotype of D. nodosus associated with severe footrot lesions in sheep in Jammu & Kashmir (J&K), India. Hence, this serotype can be a potential vaccine candidate for the effective control and treatment of ovine footrot.

     

Phagocytosis of Bacteroides nodosus by ovine peripheral blood leucocytes

Phagocytosis of Bacteroides nodosus by ovine peripheral blood leucocytes (PBL) was examined after organisms had been opsonized in sera from normal sheep, or from animals immune to, or infected with ovine footrot. Ingestion of bacteria, as assessed microscopically or by counting isotopically-labelled organisms spectrometrically was effected in suspensions by polymorphonuclear leucocytes (PMN). Opsonization of bacteria in immune serum, particularly its IgG₂ isotype, enhanced the rate of phagocytosis by PMN compared with that promoted by normal serum or medium alone. Whereas IgG₂ from immune serum also increased the rate of ingestion of B. nodosus by adherent PMN, IgM and IgG₁ from immune serum also initiated phagocytosis of bacteria by adherent ovine monocytes. Leucocytes from normal, immune or infected sheep of different breeds ingested B. nodosus with equal facility.     

Nodular hyperplasia of the ovine spleen

A field trial was carried out to compare 3 topical treatments for ovine footrot. They were preliminary hoof paring with weekly footbathing either in 10% formalin, or in 10% zinc sulphate + 0.2% anionic surfactant (‘Teepol’), or twice-weekly footbathing in the same zinc sulphate-surfactant mixture without hoof paring. Control sheep were not treated.

Within 21 days, the number and severity of active lesions of footscald and footrot among treated sheep were substantially reduced by comparison with the controls; no significant differences were ohserved between treatments. Treatment effects were also comparable against mild footrot infection of the sole or heel region, hut in the more advanced lesions the healing response to paring and zinc footbathing was superior to that from paring and formalin footbathing (P<0.01). Twice-weekly foothathing in zinc-surfactant solution was effective in preventing new footrot infection. No chronic toxic effects attributable to zinc suluhate at a concentration of 10% were demonstrable in the hoof integument either clinically or histologically.     

The effect of footrot on body weight and wool growth of sheep

Body weight and traits associated with production of wool were measured over a 2-year period between 1985 and 1987 in south-western New South Wales in a flock of Merino wethers experimentally infected with footrot. The disease was allowed to spread freely amongst 150 of the flock but kept at very low prevalence in the remaining 50 by preventive footbathing during transmission periods. Severe, underrunning footrot had a significant adverse effect on body weight, for each year of the trial. Body weight was most severely reduced at times of the year when footrot was spreading among animals and lesions were severe. The mean body weight of the infected group at the end of the 2 years of observation was 7.3 kg (11.6%) below that of the control group. Footrot also depressed wool growth, with the mean clean fleece weight of the infected group being 0.4 kg (8%) lighter than that of the controls at each of the 2 annual shearings. There were no consistent differences between the groups for the other wool characteristics measured.     

Serum antibody responses in sheep after natural infection with Bacteroides nodosus

Soluble outer membrane protein of Bacteroides nodosus extracted with potassium thiocyanate (KSCN) was employed as antigen in an enzyme linked immunosorbent assay (ELISA) to detect serum antibody in sheep naturally infected with a heterologous serogroup. Serum antibody responses in 55 sheep were monitored for 2 years and maximum levels were directly related to the severity of clinical foot lesions. Serum antibody levels rose 2 weeks after foot lesions developed and declined within several months of resolution of lesions. After the first footrot transmission period, antibody levels persisted significantly (P less than 0.001) longer in sheep that did not become affected in the next transmission period compared with sheep in which footrot recurred. Antibody response did not appear to result in resolution of foot lesions. ELISA using KSCN antigen gave similar results to whole cell ELISA where cells prepared from an homologous serogroup were used as antigen. Both these assays were more sensitive than ELISA in which heterologous whole cell antigen was used. Proteins extracted from the outer membrane of B. nodosus, which are known to be immunogenic in natural infection and common to different serogroups of B. nodosus, appear to be useful antigens for serological investigations of ovine footrot.     

Comparison of footbathing treatments for ovine footrot using formalin or zinc sulphate

A field trial was carried out to compare 3 topical treatments for ovine footrot. They were preliminary hoof paring with weekly footbathing either in 10% formalin, or in 10% zinc sulphate + 0.2% anionic surfactant ('Teepol'), or twice-weekly footbathing in the same zinc sulphate-surfactant mixture without hoof paring. Control sheep were not treated. Within 21 days, the number and severity of active lesions of footscald and footrot among treated sheep were substantially reduced by comparison with the controls; no significant differences were observed between treatments. Treatment effects were also comparable against mild footrot infection of the sole or heel region, hut in the more advanced lesions the healing response to paring and zinc footbathing was superior to that from paring and formalin footbathing (P<0.01). Twice-weekly footbathing in zinc-surfactant solution was effective in preventing new footrot infection. No chronic toxic effects attributable to zinc sulphate at a concentration of 10% were demonstrable in the hoof integument either clinically or histologically.     

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.     

A DEGRADING PROTEINASE TEST TO DISTINGUISH BENIGN AND VIRULENT OVINE ISOLATES OF BACTEROIDES NODOSUS

Bacteroides nodosus was recovered from naturally occurring cases of virulent ovine footrot (VFR) and benign footrot (BFR) using an artificial culture medium which incorporated trypticase, arginine, serine and 5% agar. A degrading proteinase (DP) test was developed which measured the proteinase activity of broth cultures of B. nodosus for a period of time after organism death to assess the stability of the enzyme. The spectrophotometric measurement of the release of dye from a hide powder--azure conjugate by the action of proteinase provided an objective analysis of enzyme activity. The DP test differentiated VFR and BFR isolates and promises to be a useful laboratory method for the diagnosis of benign and virulent footrot in sheep.