Comparison of clinical cure rates from footrot and contagious ovine digital dermatitis using zinc sulphate foot bathing and topical oxytetracycline: A randomised trial.

BACKGROUND: This study reports the clinical cure rates of topical oxytetracycline and 10% zinc sulphate foot bathing for treatment of interdigital dermatitis (ID), footrot (FR) and contagious ovine digital dermatitis (CODD) in lambs. METHODS: The study was a randomised controlled trial of 75 lambs. Group A (n = 38) was foot bathed daily for 5 days in 10% zinc sulphate for 15 minutes and group B was treated with daily topical oxytetracycline for 5 days. On days 0, 7, 14, 28 and 42, lambs were scored for locomotion and foot lesions were recorded. RESULTS: The initial cure rates for ID were 96.20% and 97.00%; for FR, 100% and 95%; and for CODD, 90.09% and 83.33% for zinc sulphate and oxytetracycline, respectively. By day 42, these had changed to 53.16% and 61% for ID; 47.82% and 70% for FR; and 100% and 83.33% for CODD. There were no significant differences in cure rates between the treatments for most time points. LIMITATIONS: The sample size was small, and further studies in larger cohorts and different classes of sheep are required before the findings can be translated into recommendations for clinical practice. CONCLUSION: Both treatments achieved cure rates that are comparable to reported cure rates using systemic antibiotics and could be an effective alternative.

NF-κB signaling pathway mechanism in cow intertoe skin inflammation caused by Fusobacterium necrophorum.

Background: Fusobacterium necrophorum is the main pathogen inducing bovine foot rot. The infected site is often accompanied by a strong inflammatory response, but the specific inflammatory regulatory mechanism remains unclear. Aim: A cow skin explants model was established to elucidate the mechanism of F. necrophorum bacillus causing foot rot in cows, and to provide reference for future clinical practice. Methods: Cow intertoe skin explants were cultured in vitro, and F. necrophorum bacteria solution and nuclear factor-κB (NF-κB) inhibitor BAY 1-7082 were added to establish an in vitro infection model. Hematoxylin and eosin staining, terminal - deoxynucleotidyl transferase mediated nick end labeling, and immunohistochemistry were used to detect the pathological changes of the skin explants infected with F. necrophorum, the degree of tissue cell apoptosis, and the expression of the apoptosis-related protein Caspase-3, respectively. RT-qPCR, Western blot, and ELISA were used to detect the activation of the NF-κB pathway and inflammatory cytokines by F. necrophorum. Results: The intertoe skin structure of cows infected with F. necrophorum changed with different degrees of inflammation, and the degree of tissue cell apoptosis was significantly increased (P < 0.01). In addition, infection with F. necrophorum significantly increased the phosphorylation level of IκBα protein and up-regulated the expression level of NF-κB p65. The high expression and transcriptional activity of NF-κB p65 significantly increased the expression and concentration of the inflammatory cytokines TNF-α, IL-1ß, and IL-8, thus inducing the occurrence of an inflammatory response. However, inhibition of NF-κB p65 activity significantly decreased the expression of inflammatory factors in the intertoe skin of cows infected with F. necrophorum. Conclusion: F. necrophorum activates NF-κB signaling pathway by increasing the expression of TNF-α, IL-1ß, IL-8 and other inflammatory factors, leading to foot rot in dairy cows.

Volatile-Based Diagnosis for Pathogenic Wood-Rot Fungus Fulvifomes siamensis by Electronic Nose (E-Nose) and Solid-Phase Microextraction/Gas Chromatography/Mass Spectrometry.

Wood rot fungus Fulvifomes siamensis infects multiple urban tree species commonly planted in Singapore. A commercial e-nose (Cyranose 320) was used to differentiate some plant and fungi volatiles. The e-nose distinctly clustered the volatiles at 0.25 ppm, and this sensitivity was further increased to 0.05 ppm with the use of nitrogen gas to purge the system and set up the baseline. Nitrogen gas baseline resulted in a higher magnitude of sensor responses and a higher number of responsive sensors. The specificity of the e-nose for F. siamensis was demonstrated by distinctive clustering of its pure culture, fruiting bodies collected from different tree species, and in diseased tissues infected by F. siamensis with a 15-min incubation time. This good specificity was supported by the unique volatile profiles revealed by SPME GC-MS analysis, which also identified the signature volatile for F. siamensis-1,2,4,5-tetrachloro-3,6-dimethoxybenzene. In field conditions, the e-nose successfully identified F. siamensis fruiting bodies on different tree species. The findings of concentration-based clustering and host-tree-specific volatile profiles for fruiting bodies provide further insights into the complexity of volatile-based diagnosis that should be taken into consideration for future studies.

[First case description of contagious ovine digital dermatitis in Switzerland]. / Erstbeschreibung der Krankheit «Kontagiöse ovine digitale Dermatitis¼ in der Schweiz.

INTRODUCTION: Contagious ovine digital dermatitis (CODD) is an emerging infectious foot disease in sheep. To date, CODD has been described in Great Britain, Ireland, Sweden and Germany and now in Switzerland for the first time. Unlike foot rot, the CODD lesions do not spread from the interdigital space, but usually begin at the dorsal/abaxial coronary band. The changes can spread to the hoof wall and the sole and finally can lead to exungulation, similar to foot rot. Treponema spp. are often found in CODD lesions analogous to digital dermatitis (Mortellaro's disease) in cattle. Involvement of Dichelobacter nodosus (D. nodosus) is considered a risk factor, but the presence of the bacterium is not mandatory. In February 2022, ulcerative lesions in the dorso-axial coronary band area were noticed on both claws of the left forelimb in an ewe. Histology of the biopsy showed hyperkeratosis and erosion with exocytosis and crust formation. Treponema spp. PCR and fluorescence in situ hybridization (FISH) were positive for Treponema phylotype 1 (PT1). In addition, D. nodosus and Porphyromonas levii could be detected in the biopsy using PCR. A single local application of chlortetracycline spray led to clinical healing within two weeks, no recurrence was seen within the following two months. Three control sheep, which were kept together with the diseased sheep, did not show any clinical signs of CODD. Treponema spp could not be found in interdigital and coronary band biopsies by PCR or FISH. This is the first description of CODD in Switzerland and aims to sensitize veterinarians to CODD as a differential diagnosis for foot diseases in sheep.

INTRODUCTION: La dermatite digitale contagieuse ovine (contagious ovine digital dermatitis; CODD) est une maladie infectieuse des onglons des moutons d'importance croissante. À ce jour, la CODD a été décrite en Grande-Bretagne, Irlande, Suède et Allemagne, et maintenant pour la première fois également en Suisse. Au contraire du piétain, les lésions de CODD ne s'étendent pas à partir de l'espace interdigité, mais elles commencent en général au bord coronaire dorsal/abaxial. De là, les lésions peuvent s'étendre à la corne de la paroi et à la sole, ce qui peut finalement conduire à une perte complète de la boite cornée de l'onglon, comme en cas de piétain. En analogie à la dermatite digitale (maladie de Mortellaro) chez les bovins, des tréponèmes sont souvent mis en évidence dans les lésions de CODD. La présence de Dichelobacter nodosus (D. nodosus) est considérée comme un facteur de risque, mais elle n'est pas indispensable au développement de la CODD. Des lésions ulcératives dans la région du bord coronaire dorso-axial des deux onglons antérieurs d'une brebis ont été remarqués en février 2022. L'examen histologique de la biopsie de la lésion de CODD a montré une hyperkératose ainsi que des érosions avec de l'exocytose et la formation de croûtes. Aussi bien la PCR pour les Treponema spp. que l'hybridisation in-situ à fluorescence (FISH) étaient positives pour Treponema Phylotype 1 (PT1). De plus, D. nodosus et Porphyromonas levii ont été mis en évidence dans la biopsie. Une application locale unique de spray à la tétracycline après le prélèvement de la biopsie a conduit à une guérison clinique en deux semaines, et aucune récidive n'a été observée dans le deux mois suivants. Trois moutons de boucherie qui étaient détenus avec la brebis malade mais ne présentaient pas de lésions de CODD ont servi de contrôles négatifs. Des Treponema spp. n'ont été mis en évidence chez ces animaux, ni dans des biopsies du bord coronaire ni dans celles de l'espace interdigité. Cette étude représente la première description de la CODD en Suisse et est destinée à sensibiliser la profession vétérinaire à la CODD comme diagnostic différentel en cas de maladies des onglons chez les moutons.

Immunological response of lame sheep to clinical interdigital dermatitis and footrot: Procalcitonin, acute phase proteins, and pro-inflammatory cytokines.

In many countries, sheep lameness is a cause of economic concern and a contributing factor to a declining economy. This study aimed to investigate changes in procalcitonin (PCT), acute phase proteins (APPs), and cytokines (CYTs) in response to interdigital dermatitis and footrot in sheep under field conditions, to emphasize their role in the disease pathogenesis, diagnosis, as well as monitoring treatment response. Fifty-three sheep with foot diseases (26 clinical cases with interdigital dermatitis and 27 clinical cases with footrot) and 20 clinically healthy naemi sheep were used in this study. Real time PCR for detection of Fusobacterium necrophorum (F. necrophorum) and Dichelobacter nodosus (D. nodosus) revealed that, all samples collected from lame sheep (N = 53) were positive for D. nodosus (100 %), whereas F. necrophorum was detected in 19 out of 53 samples (35.84 %). The virulent D. nodosus was detected in 48 lameness cases where non-virulent D. nodosus were identified in 5 cases (in concurrent with F. necrophorum). The mean serum levels of PCT, C-reactive protein (CRP), serum amyloid A (SAA), haptoglobin (HP), fibrinogen (Fg) and CYTs (IL1-ß, IL-1α, IFN-γ, IL-6 and TNF-α) in sheep with clinical interdigital dermatitis and footrot were remarkably higher than those detected in control healthy sheep. The serum levels of PCT, CRP, SAA, HP, Fg, and CYTs markers in lame sheep pre- and post-treatment were measured. A substantial decline was detected in serum levels of tested biomarkers of lame sheep after 14 days of treatment. The ROC curves were created. The AUC was assessed to evaluate the accuracy of each variable in distinguishing diseased and healthy sheep. Based on the ROC curves and AUCs; PCT, CRP, SAA, HP, and CYTs were highly diagnostic and predictive for the treatment response of sheep with clinical interdigital dermatitis and footrot. Moreover, all tested biomarkers had a noteworthy role in disease immuno-pathogenesis. Nevertheless, PCT and CRP are better than other tested APPs and CYTs as diagnostic markers for interdigital dermatitis and footrot. However, PCT only has the ability to differentiate sheep with different lameness score.

Eficácia in sílico do florfenicol no tratamento de pododermatite infecciosa por Fusobacterium necrophorum em ovelhas / In sílico efficacy of florfenicol in the treatment of Fusobacterium necrophorum infectious pododermatitis in sheep

Objetivou-se comparar o efeito in silico do florfenicol nas doses de 20 e 30 mg/Kg em ovinos pelas vias intravenosa (IV) e intramuscular (IM), usando a modelagem PK/PD. Realizou-se uma simulação de Monte Carlo com base nos dados de concentração plasmática de um estudo publicado anteriormente. Calculou-se a área sob a curva (ASC) e as taxas de eficácia do florfenicol para os efeitos bacteriostático, bactericida e de erradicação bacteriológica. A dose de 20 mg/Kg IV demonstrou efeitos de erradicação de 100, 93 e 0% para CIM de 0,5, 1 e acima, respectivamente. O efeito bacteriostático foi de 99 e 90% para CIM de 4 e 2 µg/ml, enquanto o bactericida foi de 14% para CIM de 2 µg/ml. A dose de 30 mg/Kg IV apresentou 100% de erradicação para CIM de 1 µg/mL e 100% de efeito bactericida para CIM de 2 µg/mL. Há 100% de efeito bacteriostático em CIM de 4 µg/ml. As doses de 20 e 30 mg/Kg IM mostraram 100% de erradicação para CIM até 1 µg/mL e 0% para CIM maiores. O efeito bacteriostático foi mantido em 100% para uma CIM de 4 µg/mL em ambas as doses. Este estudo mostra o efeito de erradicação bacteriológica do florfenicol nas doses de 20 e 30 mg/Kg, IV e IM. Recomenda-se que seja feito um estudo de eficácia in vivo com a dose de 30mg/Kg IM em ovinos infectados por F. necrophorum com MIC superior a 2 µg/mL.

We aimed to compare the in silico effect of florfenicol at doses of 20 and 30 mg/Kg in sheep by intravenous (IV) and intramuscular (IM) routes, using PK/PD modeling. We performed a Monte Carlo simulation based on plasma concentration data from a previously published study. We calculated the area under the curve (AUC) and the efficacy rates of florfenicol to bacteriostatic, bactericidal, and bacteriological eradication effects. The dose of 20 mg/Kg IV demonstrated 100, 93, and 0% eradication effects for MICs of 0.5, 1, and above, respectively. The bacteriostatic effect was 99 and 90% for MIC of 4 and 2 µg/ml, while the bactericide was 14% for MIC of 2 µg/ml. The 30 mg/Kg IV dose showed 100% eradication for MIC of 1 µg/mL and 100% bactericidal effect for MIC of 2 µg/mL. There is a 100% of bacteriostatic effect at MIC of 4 µg/ml. Doses of 20 and 30 mg/Kg IM showed 100% eradication for MIC up to 1 µg/mL and 0% for MIC above. The bacteriostatic effect was maintained at 100% for a MIC of 4 µg/mL at both doses. This study shows the bacteriological eradication effect of florfenicol at doses of 20 and 30 mg/Kg, IV, and IM. Therefore, we recommend an in vivo efficacy study with a dose of 30mg/Kg IM in sheep infected with F. necrophorum with MIC greater than two µg/mL.

Prevalence of bacterial species associated with ovine footrot and contagious ovine digital dermatitis in Swedish slaughter lambs.

BACKGROUND: Ovine footrot and contagious ovine digital dermatitis (CODD) are contagious mixed bacterial infections with major impacts on animal health and production. In Sweden, ovine footrot and CODD were first detected in 2004 and 2019, respectively. In 2009, a voluntary control programme for footrot was established, and a prevalence study in slaughter lambs was conducted, however, the distribution of footrot and CODD-associated bacteria is still unknown. This study examined the prevalence of Dichelobacter nodosus, Fusobacterium necrophorum and Treponema spp., as well as the current prevalence of footrot and CODD, in Swedish slaughter lambs. RESULTS: A total of 2048 feet, from 512 slaughter lambs, were collected from eight slaughterhouses throughout Sweden in autumn 2020. All feet were visually examined for lesions of footrot and CODD and sampled for subsequent real-time polymerase chain reaction (PCR) analysis. Nine lambs (1.8%) had at least one foot affected with footrot (footrot score ≥ 2). A CODD grade 1 lesion was detected in a single lamb (0.2%). The prevalence of D. nodosus, F. necrophorum and Treponema spp. was 6.1%, 7.6% and 90.6%, respectively. The D. nodosus detected were benign strains. CONCLUSIONS: The prevalence of footrot in Swedish slaughter lambs has been significantly reduced, from 5.8 to 1.8%, during the past 11 years. This indicates that preventive measures, such as the national control programme and elimination of footrot from affected flocks, have been effective. A single lamb (0.2%) was found with a CODD lesion (grade 1). In Sweden, benign rather than virulent strains of D. nodosus seem to be the most common. Neither D. nodosus nor F. necrophorum were widespread among Swedish slaughter lambs, but both were more likely to be found in lambs with footrot. Treponema spp. was very commonly found in lambs with and without footrot, but there is a lack of information on the individual Treponema spp. present in Swedish slaughter lambs and their potential pathogenicity.

Interaction of 43K OMP of Fusobacterium necrophorum with fibronectin mediates adhesion to bovine epithelial cells.

Fusobacterium necrophorum, a Gram-negative anaerobe, is an important bovine pathogen that causes hepatic abscesses, foot rot, mastitis and endometritis. We have previously shown that the 43 kDa outer membrane protein (43 K OMP) of F. necrophorum is a porin protein that plays an important role in bacterial infections; however, the molecular mechanisms by which this protein mediates adhesion remain unclear. In this study, we investigated the role of 43 K OMP in F. necrophorum adhesion to bovine epithelial cells using 43 K OMP-deficient mutants, and identified the protein that interacts with 43 K OMP by immunoprecipitation-mass spectrometry. Our results indicated that the native 43 K OMP and recombinant 43 K OMP could bind to the cell membrane of MAC-T or bovine endometrial epithelial cells (BEECs). When F. necrophorum was preincubated with antibodies against the recombinant 43 K OMP or bovine epithelial cells were preincubated with 43 K OMP, the adhesion of F. necrophorum to MAC-T or BEECs decreased significantly (P<0.01). We successfully constructed a 43 K OMP-deficient strain (A25Δ43 K OMP) and bacterial attachment to MAC-T or BEECs was significantly higher with the F. necrophorum A25 strain than with mutant strain A25Δ43 K OMP (P<0.01). The deficiency of 43 K OMP reduced the binding of F. necrophorum to bovine epithelial cells by 90.5 %-94.9 %. Among the 39 potential differential proteins, fibronectin, collagen and myosin were selected as the target proteins, and direct interaction between 43 K OMP of F. necrophorum and fibronectin was demonstrated. Taken together, these results suggest that 43 K OMP plays a key role in adhesion of F. necrophorum to bovine epithelial cells through its interaction with fibronectin. These findings provide a theoretical basis for the pathogenic mechanism of F. necrophorum.

Ovine footrot in Southern Portugal: Detection of Dichelobacter nodosus and Fusobacterium necrophorum in sheep with different lesion scores.

The Mediterranean climate region of Alentejo in the Southern of Portugal is an important sheep production centre but little is known about the presence and characteristics of Dichelobacter nodosus in association with Fusobacterium necrophorum in the different footrot lesion scores. DNA from 261 interdigital biopsy samples, taken from 14 footrot affected flocks and from three non-affected flocks, were analysed for the presence of D. nodosus and F. necrophorum by real-time PCR. Both virulence and serogroup were determined for 132 and 53 D. nodosus positive biopsy samples, respectively. The co-infection with both bacteria was the commonest epidemiological finding associated with a greater disease severity. There was a statistically significant association (p = 0.002) between footrot-affected flocks and the presence of D. nodosus. Most D. nodosus positive samples were virulent (96.2 %) and belonged to serogroup B (90 %).

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.

Genomic heterogeneity of Dichelobacter nodosus within and between UK sheep flocks and between age groups within a flock.

BACKGROUND: Footrot and interdigital dermatitis are endemic infectious diseases in all sheep farming regions, impairing welfare and production. The development of efficacious vaccines against the primary causative pathogen has been hampered by the extensive antigenic diversity of Dichelobacter nodosus. Understanding the heterogeneity of the pathogen within and between flocks is essential if the feasibility of bespoke vaccine production is to be assessed for use in the U.K. RESULTS: In this study 56 ewe and lamb isolates from 9 flocks were compared by D. nodosus serogroup and Multi Locus Sequence Type which provides significantly enhanced discriminatory power for molecular epidemiology. Serogroup heterogeneity between flocks ranged from two to five unique serogroups per flock. Three flocks contained isolates of two serogroups, two flocks contained isolates of three serogroups and one flock included isolates of five serogroups. Analysis of 25 isolates from one flock with high prevalence of lameness, identified that serogroup and sequence type was significantly correlated with age. Significantly higher proportion of lambs were infected with serogroup B (principally ST85) as opposed to serogroup H (principally ST86), which predominated amongst adult sheep. CONCLUSIONS: Genomic heterogeneity of the pathogen was significantly lower within flock compared to heterogenicity observed between flocks. Furthermore, this study indicates that within a flock, the host-pathogen dynamics and susceptibility to particular D. nodosus strains may be age dependent.

Development and assessment of management practices in a flock-specific lameness control plan: A stepped-wedge trial on 44 English sheep flocks.

Lameness in sheep has economic and welfare implications, including loss of ewe body condition, lower lambing percentages, and poor lamb growth rates. It costs the UK sheep industry around £80 million per year. The majority of lameness is caused by the infectious diseases footrot and contagious ovine digital dermatitis, with white line separation, white line abscesses, and toe granulomas also reported by farmers. Most sheep farmers in the UK have other enterprises and care for their flock part-time. A lameness control plan (LCP) consisting of 37 management practices that covered all aspects of control of lameness was developed for part-time sheep farmers. Testing of the LCP was done using a stepped-wedge trial design with 7 visits to 44 flocks in England over 18 months. Flocks had 100-500 breeding ewes. Locomotion scoring was carried out at every visit, and farmers were interviewed every 6 months to record management practices. Clinical significance of changes in prevalence of lameness within each flock was analysed using the reliable change index (RCI). Management practices associated with prevalence of lameness were assessed using a multi-level multivariable over-dispersed Poisson model. The geometric mean (GM) prevalence of lameness at the start of the trial was 7.3% (95% confidence interval(CI) = 6.3-8.3%). Flocks with a clinically significant lameness reduction had a GM prevalence of lameness of 4.6% (95% CI = 4.1-5.2%), while flocks with a clinically significant increase in lameness had a GM prevalence of 10.5% (95% CI = 9.4-11.6%). Always separating lame sheep at treatment (relative risk (RR) = 0.60, 95% CI = 0.43-0.84) and culling sheep lame ≥2 occasions in a year (RR = 0.75, 95% CI = 0.61-0.92) were associated with a significant reduction in lameness. Compared with not footbathing at all, footbathing sheep when there were outbreaks of interdigital dermatitis reduced lameness (RR = 0.85, 95% CI = 0.75-0.96), however, 'routine' footbathing was not associated with a reduction in lameness. Housing during the previous lambing period was associated with a higher prevalence of lameness (RR = 1.23, 95% CI = 1.04-1.46). Vaccination in flocks with period prevalence ≥10% did not significantly reduce their risk of lameness compared to lower prevalence flocks. In conclusion, separating lame sheep at treatment, culling sheep lame ≥2 occasions per year, and only using a footbath to treat outbreaks of interdigital dermatitis are flock managements that contribute to improved control of lameness in flocks with part-time farmers.

A Novel 3D Skin Explant Model to Study Anaerobic Bacterial Infection.

Skin infection studies are often limited by financial and ethical constraints, and alternatives, such as monolayer cell culture, do not reflect many cellular processes limiting their application. For a more functional replacement, 3D skin culture models offer many advantages such as the maintenance of the tissue structure and the cell types present in the host environment. A 3D skin culture model can be set up using tissues acquired from surgical procedures or post slaughter, making it a cost effective and attractive alternative to animal experimentation. The majority of 3D culture models have been established for aerobic pathogens, but currently there are no models for anaerobic skin infections. Footrot is an anaerobic bacterial infection which affects the ovine interdigital skin causing a substantial animal welfare and financial impact worldwide. Dichelobacter nodosus is a Gram-negative anaerobic bacterium and the causative agent of footrot. The mechanism of infection and host immune response to D. nodosus is poorly understood. Here we present a novel 3D skin ex vivo model to study anaerobic bacterial infections using ovine skin explants infected with D. nodosus. Our results demonstrate that D. nodosus can invade the skin explant, and that altered expression of key inflammatory markers could be quantified in the culture media. The viability of explants was assessed by tissue integrity (histopathological features) and cell death (DNA fragmentation) over 76 h showing the model was stable for 28 h. D. nodosus was quantified in all infected skin explants by qPCR and the bacterium was visualized invading the epidermis by Fluorescent in situ Hybridization. Measurement of pro-inflammatory cytokines/chemokines in the culture media revealed that the explants released IL1ß in response to bacteria. In contrast, levels of CXCL8 production were no different to mock-infected explants. The 3D skin model realistically simulates the interdigital skin and has demonstrated that D. nodosus invades the skin and triggered an early cellular inflammatory response to this bacterium. This novel model is the first of its kind for investigating an anaerobic bacterial infection.

Inter-observer agreement for clinical examinations of foot lesions of sheep.

In sheep, the diagnosis of foot lesions is routinely based on physical examination of the hoof. Correct diagnosis is important for the effective treatment, prevention and control of both infectious and non-infectious causes of lameness. Therefore, the aim of this study was to evaluate the level of inter-observer agreement for clinical examination of ovine foot lesions. Eight observers of varying experience, training and occupation performed foot examinations on a total of 1158 sheep from 38 farms across North England and Wales. On each farm, a group of two to four observers independently examined a sample of 24 to 30 sheep to diagnose the presence or absence of specific foot lesions including white line lesions (WL), contagious ovine digital dermatitis (CODD), footrot (FR), inter-digital dermatitis (ID) and toe granuloma (TG). The inter-observer agreement of foot lesion assessments was examined using Fleiss kappa (κ), and Cohen's κ examined the paired agreement between the test standard observer (TSO) and each observer. Scoring differences with the TSO were examined as the percentage of scoring errors and assessed for evidence of systematic scoring bias. With the exception of WL (maximum error rate 33.3%), few scoring differences with the TSO occurred (maximum error rate 3.3%). This suggests that observers can achieve good levels of reliability when diagnosing most of the commonly observed foot conditions associated with lameness in sheep.

Whole-flock, metaphylactic tilmicosin failed to eliminate contagious ovine digital dermatitis and footrot in sheep: a cluster randomised trial.

The aim of this study was to evaluate the clinical success of whole-flock systemic tilmicosin and enhanced biosecurity in eliminating active contagious ovine digital dermatitis (CODD) from sheep flocks. Thirty flocks in the UK were randomly allocated to receive either treatment as usual (as per the farmer's normal routine) or whole-flock treatment with tilmicosin, together with isolation and extended treatment of clinically affected individuals and isolation and treatment of purchased sheep during the study period. All flocks were visited once at onset of the trial to examine all sheep. One year later, all sheep were re-examined to determine the presence/absence of clinical lesions. The primary outcome was the clinical elimination of CODD from flocks. Secondary outcomes were reduction in prevalence of CODD, clinical elimination of footrot and reduction in prevalence of footrot. The analysis included 11 control flocks and 13 intervention flocks, with initially 3460 and 4686 sheep, respectively. For CODD: at follow-up, in the intervention group, 6/13 (46 per cent) flocks had a prevalence of zero compared with 1/11 (9 per cent) in the control group (P=0.12). For footrot: at follow-up, no flocks had a prevalence of zero. Therefore, the intervention is not recommended for the elimination of CODD or footrot in the UK.