Risk factors associated with the infection of sheep with Dichelobacter nodosus.

Ovine footrot is a highly contagious foot disease caused by the gram-negative bacterium Dichelobacter nodosus (D. nodosus). In a recent report, we showed a prevalence of 42.9% D. nodosus positive swabs across Germany. In this follow-up study, we used real-time PCR results for D. nodosus and footrot scores of 9297 sheep from 208 flocks and collated these data with survey data on herd and animal characteristics and herd management. The aims of the present study were to investigate herd and animal factors associated with D. nodosus infection and footrot scores in individual sheep. Multivariable analyses with generalized mixed models showed that month of recording, breed, herdbook membership, use of antibiotics, and footbaths in the past 3-10 years, signs of footrot in the past 12 months and flock environment of the sheep, modelled as a random farm effect within region, were significant risk factors. Among the 21 different breeds, Romney had the lowest risk of D. nodosus infection, while Swifter had the highest risk and German Merino and German White Heath were the next breeds at highest risk of D. nodosus infection. The variance between farms in the prevalence of D. nodosus was large and accounted for 84% of the total variance in the mixed model analysis. We conclude that specific and as yet unknown effects influencing D. nodosus infections in flocks, as well as breed and weather, are the most important effects on D. nodosus infection in sheep, pointing towards the need to establish adequate infection control at farm level

Serological Diversity of Dichelobacter nodosus in German Sheep Flocks.

Footrot is one of the major causes of lameness in sheep and leads to decreased animal welfare and high economic losses. The causative agent is the Gram-negative anaerobic bacterium Dichelobacter nodosus. The prevalence of D. nodosus in 207 sheep flocks across Germany was 42.9%. Based on the sequence variation in the type IV fimbrial gene fimA, D. nodosus can be subdivided into ten serogroups (A-I and M). There are commercially available vaccines covering nine serogroups, but the efficacy is low compared to bivalent vaccines. The aim of this study was to investigate the diversity of serogroups in Germany at the flock and animal levels. In total, we detected at least one serogroup in 819 samples out of 969 D. nodosus-positive samples from 83 flocks using serogroup-specific singleplex PCR for the serogroups A-I. Serogroup A was most prevalent at the animal level, followed by serogroups B, H and C. At the flock level, serogroups A and B had the highest prevalence, each with 64%, but only 40% of flocks had both. The average number of serogroups per animal was 1.42 (range one to five) and, per flock, 3.10 (range one to six). The serogrouping showed within-flock specific clusters but were widely distributed, with 50 different combinations across the flocks. The factors associated with the number of serogroups per animal and single serogroups were the load of D. nodosus, footrot score, sheep breed and flock. Our results indicate that efficient vaccination programs would benefit from tailor-made flock-specific vaccines and regular monitoring of circulating serotypes in the flock to be able to adjust vaccine formulations for nationwide progressive control of footrot in Germany

Field Study on the Prevalence of Ovine Footrot, Contagious Ovine Digital Dermatitis, and Their Associated Bacterial Species in Swedish Sheep Flocks

Ovine footrot and contagious ovine digital dermatitis (CODD) cause lameness in sheep, affecting welfare and economics. Previous Swedish studies focused on individual slaughter lambs, leaving flock-wide prevalence less explored. This study examined the prevalence of footrot and CODD in Swedish sheep flocks, focusing on adult sheep. From 99 flocks, 297 swabs were analysed using real-time PCR for Dichelobacter nodosus, Fusobacterium necrophorum, and Treponema spp. Sampled feet were photographed and assessed using scoring systems for footrot and CODD. Results indicated footrot prevalences (footrot score >= 2) of 0.7% and 2.0% at the individual and flock levels, respectively, whereas there were no signs of CODD. The individual footrot prevalence was lower than that from a 2009 study but aligned with a 2020 study, both conducted on slaughter lambs. Dichelobacter nodosus, F. necrophorum, and Treponema spp. were found in 5.7%, 1.3%, and 65.0% of sheep, and in 9.1%, 3.0%, and 82.8% of flocks, respectively. Compared to the 2020 study, there was a notable decrease in F. necrophorum and Treponema spp., while D. nodosus was consistent. In conclusion, the findings show a low prevalence of footrot, CODD, D. nodosus, and F. necrophorum in Swedish sheep flocks. Continuous surveillance and owner education are important to maintain this favourable status

First report on outbreaks of contagious ovine digital dermatitis in Sweden

Background Contagious ovine digital dermatitis (CODD) is considered widespread in the United Kingdom but was only recently reported in mainland Europe, as one outbreak in Germany. The disease can cause severe lameness in sheep and, if left untreated, can lead to total avulsion of the hoof capsule. CODD is considered to have multifactorial and polymicrobial aetiology, in which Treponema medium/Treponema vincentii phylogroup, Treponema phagedenis phylogroup and Treponema pedis are believed to play a significant role. Footrot and CODD have a close connection and footrot is considered an important risk factor for CODD. Case Lameness, mainly in lambs aged 1.5 months, was reported on a farm in Sweden in spring 2018. The animals showed no signs of footrot and the causative agent, Dichelobacter nodosus, was not found. CODD was suspected but not confirmed, and the clinical signs subsided when the animals were turned out to pasture. In February 2019, young lambs and ewes were lame again and this time CODD was diagnosed. After treatment, the whole flock was slaughtered later in 2019 due to CODD. In autumn 2020, CODD was diagnosed on another Swedish farm, this time as part of a mixed infection with D. nodosus. The animals were treated with footbaths in zinc sulphate 10% by the farmer, but lameness recurred soon afterwards. The animals were treated, but ultimately the whole flock was slaughtered. No connection was found between the two farms. Conclusion The first two outbreaks of CODD in Sweden have been diagnosed and are described in this case report. If it spreads, CODD could have a negative impact on the Swedish sheep industry in terms of animal welfare, production and antibiotic use

Aspects of the aetiopathogenesis and diagnosis of ovine footrot

Footrot is prevalent in most sheep-producing countries. Two forms are generally recognised: virulent and benign. Virulent footrot has major economic and animal welfare impacts. Footrot has a complex aetiology; the clinical manifestations of the disease result from interactions between the essential causative agent, Dichelobacter nodosus, and the bacterial community of the foot. The severity of these manifestations varies according to environmental conditions and host susceptibility, thus clinical diagnosis of virulent footrot can be challenging, and laboratory tests are used to assist diagnosis. A comparative evaluation of phenotypic and genotypic virulence tests was undertaken, including the elastase test and a qPCR test targeting the aprV2 protease gene, which is thought to be a key virulence marker. The qPCR had a low diagnostic specificity, with aprV2-positive D. nodosus strains detected flocks with clinically benign footrot. As such, aprV2 was deemed an unreliable virulence marker. In contrast, expression of elastase was closely associated with virulence. D. nodosus strains are divisible into ten immunologically distinct serogroups (A to I, M). A culture-independent testing procedure was developed that enhanced the speed and accuracy of serogrouping, and a cPCR test targeting serogroup M was developed. Such a test was previously unavailable. The foot microbiomes of healthy and footrot-affected Merino sheep were characterised and compared using next-generation sequencing and analysis of the bacterial 16S rRNA gene. Fifteen bacterial genera were found to be preferentially abundant on the feet of footrot-affected Merino sheep, several of which were not previously known to contribute to the disease process. A pasture-based experimental model was also developed that provides a more accurate representation of the conditions in which footrot is naturally transmitted and expressed

The prevalence, virulence, and serogroups of Dichelobacter nodosus and prevalence of Fusobacterium necrophorum in footrot lesions of sheep and cattle in Morocco

Background and Aim: Footrot is a contagious disease of ruminants leading to severe economic losses. This study aimed to estimate the prevalence, virulence, and serogroups of Dichelobacter nodosus and the prevalence of Fusobacterium necrophorum in footrot lesions of sheep and cattle. Materials and Methods: A total of 106 pathogenic lesion samples were taken from 74 sheep and 32 cattle exhibiting typical footrot lesions and were analyzed for the presence of D. nodosus and F. necrophorum by real-time polymerase chain reaction (PCR). Both virulence and serogroup were estimated for D. nodosus positive samples. Results: Among the 106 samples, 89 were positive by PCR for F. necrophorum, D. nodosus, or both. Dichelobacter nodosus was detected at a rate of 78.3% versus 28.3% for F. necrophorum. Virulent D. nodosus strains were detected in 67.5% of positive samples, with a higher rate in sheep (73.4%) than in cattle (47.4%). Benign D. nodosus strains were detected in 57.8% of samples, with a lower prevalence rate in sheep (50%) than in cattle (84.2%). The positive samples of D. nodosus revealed the presence of three dominant serogroups (D, H, I) and three minor serogroups (G, C, A) by serogroup-specific multiplex PCR. Conclusion: The findings provided information on the prevalence of D. nodosus and F. necrophorum strains in footrot lesions of sheep and cattle in some regions of Morocco, which will be useful for developing an effective autovaccine for the prevention of this disease in cattle and sheep in these regions

First cases of contagious ovine digital dermatitis in Germany

Contagious ovine digital dermatitis (CODD) is a significant disease of the ovine foot characterised by severe lameness and progressive separation of the hoof horn capsule from the underlying tissue. Similar to bovine digital dermatitis (BDD), pathogenic members of the genus Treponema including the Treponema medium phylogroup, Treponema phagedenis phylogroup and Treponema pedis are frequently found together in CODD lesions. To date, CODD was only described in Ireland and the United Kingdom. In northern Germany, cases of an unusually severe lameness presented in a sheep flock that had been affected by footrot for several years. These cases were non-responsive to conventional footrot therapies, with some sheep exhibiting substantial lesions of the claw horn that resulted in horn detachment. Lesion swab samples were collected from both clinically affected and asymptomatic animals. In all clinically affected sheep, CODD-associated Treponema phylogroups were detected by polymerase chain reaction. This is the first report of CODD in Germany and mainland Europe, indicating a wider geographic spread than previously considered. In cases of severe lameness attributed to claw lesions in sheep that fail to respond to footrot treatment, CODD should be considered irrespective of geographic location

The in situ analysis of the microbial community associated with footrot of sheep

Footrot (FR) is a highly infectious and debilitating disease of sheep, which has a significant economic impact on the sheep farming industry, in the UK and worldwide and causes significant suffering of sheep. Despite some recent advances, FR remains a scientifically challenging disease to understand. To help improve our understanding of disease pathogenesis, two culture-independent techniques were developed to examine the microbial succession events between the causative agent, Dichelobacter nodosus and an accessory agent, Fusobacterium necrophorum, the latter also postulated to be involved in disease initiation. The two populations were monitored in relation to disease initiation and progression during a longitudinal study and disease presentation in tissue biopsies (in situ). Finally, the distribution of these two species of bacteria in the environment was examined to highlight possible sources of infection. The work in this thesis has demonstrated that FR is a disease where expression is related to D. nodosus load present in the ovine interdigital space. D. nodosus (rpoD) load increased from that on a healthy foot to one presenting with interdigital dermatitis (ID) and feet with a higher D. nodosus (rpoD) load were more likely to go on to develop FR one week later. FISH analysis of the D. nodosus population present within the epidermis also revealed similar findings; D. nodosus cell counts increased during stages of ID, but the organism was less frequently detected in biopsies from feet with FR. Suggesting that ID might be the most infectious stage of the disease process. A fact that needs to be highlighted to farmers to encourage treatment at this stage of disease. In contrast, F. necrophorum (rpoB) load did not correlate with ID presentation or prior to the development of FR, but increased the week of FR onset. FISH analysis also revealed that F. necrophorum cell counts were higher in feet with FR than those with ID. It is possible therefore that F. necrophorum may thrive in the altered environment of a foot presenting with FR, possibly contributing to disease persistence and severity. Finally, both pathogens were detected in a range of environmental samples from a farm with endemic FR, highlighting possible sources of infection and material, which once contaminated with D. nodosus and F. necrophorum may contribute to the spread of FR. This study has provided an improved understanding of the microbial population dynamics involved in the development of ID and FR in sheep, which may have implications for control and treatment practices not only in the UK, but world-wide

Higher whole-blood selenium is associated with improved immune responses in footrot-affected sheep

We reported previously that sheep affected with footrot (FR) have lower whole-blood selenium (WB-Se) concentrations and that parenteral Se-supplementation in conjunction with routine control practices accelerates recovery from FR. The purpose of this follow-up study was to investigate the mechanisms by which Se facilitates recovery from FR. Sheep affected with FR (n = 38) were injected monthly for 15 months with either 5 mg Se (FR-Se) or saline (FR-Sal), whereas 19 healthy sheep received no treatment. Adaptive immune function was evaluated after 3 months of Se supplementation by immunizing all sheep with a novel protein, keyhole limpet hemocyanin (KLH). The antibody titer and delayed-type hypersensitivity (DTH) skin test to KLH were used to assess humoral immunity and cell-mediated immunity, respectively. Innate immunity was evaluated after 3 months of Se supplementation by measuring intradermal responses to histamine 30 min after injection compared to KLH and saline, and after 15 months of Se supplementation by isolating neutrophils and measuring their bacterial killing ability and relative abundance of mRNA for genes associated with neutrophil migration. Compared to healthy sheep, immune responses to a novel protein were suppressed in FR-affected sheep with smaller decreases in FR-affected sheep that received Se or had WB-Se concentrations above 250 ng/mL at the time of the immune assays. Neutrophil function was suppressed in FR-affected sheep, but was not changed by Se supplementation or WB-Se status. Sheep FR is associated with depressed immune responses to a novel protein, which may be partly restored by improving WB-Se status (> 250 ng/mL)