Evaluation of selected risk factors for different stages of digital dermatitis in Dutch dairy cows.

Digital dermatitis (DD) is a painful infectious disease in dairy cattle that causes ulcerative lesions of the skin just above the coronary band, mainly of the hind legs. Estimates for DD prevalence at cow level in the Netherlands range from 20% to 25%. In this study, risk factors for the various stages of DD were identified and quantified. The hind legs of 6766 cows on 88 farms were scored by trained interns, using the M-scoring system (M0-M4.1). Farms in this study were a convenience sample, based on the prevalence of DD as recorded at the latest herd trim, geographical location and willingness of the farmers to participate. A survey with questions about cow environment and herd management was conducted by the intern at the day of scoring. The data were collected between August 2017 and January 2018. DD was found on 38.6% of the scored legs; 49.8% of the cows had DD on at least one leg and M4 was the most frequent stage (20.9%). Not removing manure on a regular basis resulted in lower odds for M2, M4 and M4.1 compared to cleaning by automatic scrapers ten times a day or more (odds ratio [OR]= 0.16, 0.49 and 0.18, respectively). The odds for M2 and M4 lesions were higher in cows aged 3-5 years than in first-calved cows (OR> 1.5 and > 1.7, respectively). Rubber flooring in the passageways resulted in lower odds for both M1 and M2 (OR, 0.06 and 0.32, respectively). Prophylactic use of footbaths treatment with an alternative active compound resulted in significant higher odds for M4 lesions than formalin and a combination of formalin and copper sulphate (OR= 1.69 and 2.04 respectively). The odds for an M4.1 lesion were lower in cows from smaller herds (n = 50-100) compared to large herds (n >100; OR= 0.67).

Efficacy of quaternary ammonium salt-based disinfectant or chelated copper-zinc footbath solution in the treatment of digital dermatitis on one research dairy farm in Taiwan.

BACKGROUND: Footbaths can be used to manage digital dermatitis (DD), a common cause of lameness in dairy cattle. Copper sulfate and chelated copper-zinc (CZS) solutions in footbaths are potentially harmful to the environment. HYPOTHESIS/OBJECTIVES: To determine if a quaternary ammonium salt-based disinfectant (QASD) footbath is as effective as a chelated CZS solution in controlling DD in dairy cows. ANIMALS: Fifty-one lactating Holstein cows were randomly assigned to one of two treatment groups, with DD status based on the M-stage scoring system and locomotion score balanced between treatment groups. MATERIALS AND METHODS: The groups were treated with a 1% QASD or a 2.5% chelated CZS. Footbaths were performed once per week for 15 weeks. Logistic regression was used to analyse clinical improvement. All cows received appropriate medical treatment for DD and other hoof diseases. RESULTS: Clinical improvement rates were 67% in the QASD group and 38% in the CZS group (p = 0.03). Logistic regression analysis showed that the odds (95% confidence interval) for clinical improvement rate in the CZS group were 0.30 (0.095-0.948) times that of the QASD group (p = 0.04). The M0 score in the QASD and CZS groups increased significantly (p < 0.05) at the end of the 15 week study period. In the QASD group, the proportion of M2, M3 and M4 scores were significantly decreased (p < 0.05). CONCLUSION AND CLINICAL RELEVANCE: Over a 15 week period, QASD for footbathing was associated with a lower prevalence of active DD lesions than when using CZS.

Transmission and lesion progression of treponeme-associated hoof disease in captive elk (Cervus canadensis).

Treponeme-associated hoof disease (TAHD) is a debilitating disease of free-ranging elk (Cervus canadensis) in the northwestern U.S. While treponemes are associated with lesions, the etiology and transmissibility between elk are unknown. Our objective was to determine whether the disease can be environmentally transmitted to captive elk. Four individually housed treatment elk and 2 control elk were challenged with soil mixed with inoculum prepared from free-ranging elk hooves from TAHD-positive elk or autoclaved hooves from normal elk, respectively. The inoculum for each group was applied to the interdigital space and added to pre-existing soil in each pen. Eight challenges were conducted at 1-4-week intervals and lesion development was assessed during a 138-day challenge period that was followed by a 170-day monitoring period to document lesion progression. All treatment elk, but no control elk, developed gross and histologic lesions consistent with TAHD. Treponema phylotypes similar to those in bovine digital dermatitis in cattle were detected using 16S rRNA gene amplicon sequencing from lesions in all treatment elk, but no control elk, during the challenge period. Lesions progressed from ulcerations in the interdigital space to extensive ulceration and underrunning of the hoof capsule by 35 and 173 days following the initial inoculation, respectively. Lameness in treatment elk was correlated with lesion development (R = 0.702, p≤0.001), and activity of infected elk was reduced during the challenge (p≤0.001) and monitoring periods (p = 0.004). Body condition was significantly lower in treatment than control elk 168 days following the initial inoculation (p = 0.05) and at each individual elk's study endpoint (p = 0.006). Three of 4 treatment elk were euthanized when they reached humane endpoints, and one elk recovered. These results provide direct evidence that TAHD is a transmissible infectious disease in elk. As such, actions that reduce transmission risk can support disease management and prevention.

Broad-spectrum infrared thermography for detection of M2 digital dermatitis lesions on hind feet of standing dairy cattle.

Low-effort, reliable diagnostics of digital dermatitis (DD) are needed, especially for lesions warranting treatment, regardless of milking system or hygienic condition of the feet. The primary aim of this study was to test the association of infrared thermography (IRT) from unwashed hind feet with painful M2 lesions under farm conditions, with lesion detection as ultimate goal. Secondary objectives were to determine the association between IRT from washed feet and M2 lesions, and between IRT from unwashed and washed feet and the presence of any DD lesion. A total of 641 hind feet were given an M-score and IRT images of the plantar pastern were captured. Multivariable logistic regression analyses were done with DD status as dependent variable and maximum infrared temperature (IRTmax), lower leg cleanliness score and locomotion score as independent variables, and farm as fixed effect. To further our understanding of IRTmax within DD status, we divided IRTmax into two groups over the median value of IRTmax in the datasets of unwashed and washed feet, respectively, and repeated the multivariable logistic regression analyses. Higher IRTmax from unwashed hind feet were associated with M2 lesions or DD lesions, in comparison with feet without an M2 lesion or without DD, adjusted odds ratio 1.6 (95% CI 1.2-2.2) and 1.1 (95% CI 1.1-1.2), respectively. Washing of the feet resulted in similar associations. Dichotomization of IRTmax substantially enlarged the 95% CI for the association with feet with M2 lesions indicating that the association becomes less reliable. This makes it unlikely that IRTmax alone can be used for automated detection of feet with an M2 lesion. However, IRTmax can have a role in identifying feet at-risk for compromised foot health that need further examination, and could therefore function as a tool aiding in the automated monitoring of foot health on dairy herds.

Continuous activation of the IL-17F driven inflammatory pathway in acute and chronic digital dermatitis lesions in dairy cattle.

Objectives of the present study were to get a deeper insight into the course of the inflammatory pathways of digital dermatitis lesions in dairy cattle by investigating the gene expression patterns throughout the different clinical stages (M0 to M4.1) of the disease. Normal skin samples (M0) were used as a reference for comparing the gene expression levels in the other M-stages through RNA Seq-technology. Principal component analysis revealed a distinct gene expression pattern associated with digital dermatitis lesions in comparison to healthy skin with a further clustering of the acute M1, M2 and M4.1 stages versus the chronic M3 and M4 stages. The majority of the up-and downregulated genes in the acute and chronic stages can be placed into a common 'core' set of genes involved in inflammation, such as A2ML1, PI3, CCL11 and elafin-like protein, whereas the most downregulated genes included keratins and anti-inflammatory molecules such as SCGB1D and MGC151921. Pathway analysis indicated the activation of the pro-inflammatory IL-17 signaling pathway in all the M stages through the upregulation of IL-17F. These results indicate that digital dermatitis is associated with an excessive inflammatory immune response concomitant with a disrupted skin barrier and impaired wound repair mechanism. Importantly, despite their macroscopically healed appearance, a significant inflammatory response (Padj < 0.05) was still measurable in the M3 and M4 lesions, potentially explaining the frequent re-activation of such lesions.

Foot health and prevalence of Dichelobacter nodosus in 11 ungulate species at Berne Animal Park.

INTRODUCTION: Dichelobacter nodosus (D. nodosus) is the etiological agent of ovine footrot affecting mainly sheep worldwide, but also free-ranging wild ungulates such as Alpine ibex (Capra ibex ibex) and mufflon (Ovis orientalis orientalis). A nationwide ovine footrot eradication program is planned for the years to come, based on polymerase chain reaction (PCR)-testing of interdigital swab samples and regular footbathing. In this cross-sectional study, we clinically evaluated the foot health and analysed presence of D. nodosus in 11 different even-toed ungulate species (mainly European species) during a 13 months (2018-2019) period in Berne Animal Park. The foot lesions were scored for any clinical signs of pathologies as described in cattle and simultaneously for clinical signs of footrot as described for sheep, using a scale from 0 to 5 (while 0 describes clinically healthy feet and 5 loss of the horn capsule). From a total of 53 animals, 4-feet swab samples were taken from the interdigital cleft and subjected to real-time PCR assays to detect D. nodosus at animal level. Foot lesions were detected in five different species. In 3/5 muskoxen (Ovibos moschatus wardi), 7/12 Cretan wild goats (Capra hircus cretica) and 2/3 dwarf goats (Capra hircus aegagrus), they mainly consisted of white line disease, whereas in 9/10 European bison, dermatitis of the interdigital cleft was diagnosed. 1/3 alpaca was diagnosed with chorioptic mange of the heel area. None of the examined animals showed clinical signs of footrot (score 0), and neither benign (aprB2-positive) nor virulent (aprV2-positive) D. nodosus were detected in any of the samples. This study provides additional information to facilitate an efficient ovine footrot control program in Switzerland and suggests that captive wild even-toed ungulates do not pose a risk to the planned footrot control program.

INTRODUCTION: Dichelobacter nodosus (D. nodosus) est l'agent étiologique du piétin chez les ruminants, qui affecte principalement les moutons dans le monde, mais aussi les ongulés sauvages en liberté tels que les bouquetins (Capra ibex ibex) et les mouflons (Ovis orientalis orientalis). Un programme d'éradication du piétin ovin à l'échelle Nationale, basé sur des tests PCR (réaction de polymérisation en chaîne) d'écouvillons de l'espace interdigité et des pédiluves réguliers, est prévu dans les années à venir en Suisse. Dans cette étude transversale, nous avons évalué cliniquement la santé des onglons et recherché la présence de D. nodosus chez 11 espèces différentes d'animaux biongulés (principalement des espèces européennes) pendant une période de 13 mois (2018­2019) au Parc animalier de Berne. Les lésions des onglons ont été notées pour tout signe clinique de pathologie et de présence de piétin, comme cela est décrit chez les bovins et les moutons et en utilisant une échelle de 0 à 5 (où 0 décrit des pieds cliniquement sains et 5 la perte de la boîte cornée). Des écouvillons ont été prélevés dans l'espace interdigité des 4 pieds sur un total de 53 animaux et soumis à des tests PCR en temps réel pour détecter D. nodosus. Des lésions aux onglons ont été détectées chez cinq espèces différentes. Chez 3 boeufs musqués (Ovibos moschatus wardi) sur 5, 7 chèvres sauvages crétoises (Capra hircus cretica) sur 12 et 2 chèvres naines (Capra hircus aegagrus) sur 3, il s'agissait principalement de lésions de la ligne blanche, alors que dans 9 bisons sur 10, le diagnostic était une dermatite interdigitale. Un alpaga sur 3 a été diagnostiqué avec une gale chorioptique de la zone du paturon. Aucun des animaux examinés n'a montré de signes cliniques de piétin (score 0) et aucun D. nodosus bénin (aprB2-positif) ou virulent (aprV2-positif ) n'a été mis en évidence dans les échantillons. Cette étude fournit des informations supplémentaires pour améliorer l'efficacité du programme d'éradication du piétin ovin et suggère que les biongulés sauvages détenus dans les parcs zoologiques ne présentent pas de risque pour le programme d'éradication prévu en Suisse.

The role of vanilloid receptor type 1 (TRPV1) in hyperalgesia related to bovine digital dermatitis.

Bovine digital dermatitis is a contagious and chronic disease affecting the digits of dairy cattle worldwide. Tissue degradation may alter ionic channels and further activate vanilloid channels, more specifically the vanilloid receptor type 1 (TRPV1) that can generate and modulate hyperalgesia in cows affected with bovine digital dermatitis. The aim of this pilot study was to identify and quantify TRPV1 channels in dairy cows presenting with different stages of bovine digital dermatitis and compare these data according to the disease evolution and degree of hyperalgesia described in previous studies. Biopsies were taken from 15 lactating Holstein cows (23 lesions), and immunochemistry was performed to identify the number of TRPV1 fibers in the 4 M-stages of digital dermatitis and the control group. This pilot study had 5 experimental groups, M1 (5 samples), M2 (5 samples), M3 (4 samples), M4 (4 samples), and the control group (5 samples), with inclusion criteria was the presence of a bovine digital dermatitis lesion in at least one digit. The pilot results demonstrate an increase in expression of TRPV1 receptors in group M4 in comparison with the other groups. Bovine digital dermatitis may cause an increase in expression of TRPV1 receptors in the chronic stages of the disease, possibly contributing to the hyperalgesia described in affected animals; nevertheless, further research is needed to define this relation.

Detection of Treponema spp. in bovine digital dermatitis in the Amazon biome, Brazil / Detecção de Treponema spp. em dermatite digital bovina no bioma amazônico, Brasil

Bovine digital dermatitis (BDD) is a polybacterial claw disease that is endemic to dairy cattle kept in loose house systems, and treponemas are the main bacteria implicated in this disease. The objective of this study was to report the occurrence of Treponema spp. in BDD from crossbred dairy cattle (Holstein x Zebu) kept in a pasture in the Brazilian Amazon biome. The diagnostic of BDD was performed by inspecting the distal extremities of cattle during milking in one or more visits comprising 15 farms. In total, it could be inspected 1,847 cows from August 2016 to July 2017, and 25 lesions of BDD were diagnosed. The feet were scored (System M: M0 = no lesion, M1 = ulcer stage <2cm, M2 = ulcer stage >2cm, M3 = healing stage, M4 = chronic stage, M4.1 = chronic stage with ulcer area). Twenty four biopsy samples were taken from feet with BDD and five biopsy samples from feet with no lesions. The histopathology of stained tissues was performed by hematoxylin and eosin and Warthin-Starry method. The samples were also tested by nested PCR for the three previously isolated BDD Treponema phylogroups (T. medium/T. vincentii-like, T. phagedenis-like and T. putidum/T. denticola-like). Spirochetes were observed in 54.2% (13/24) of the lesions, and in 91.7% (22/24) of the samples were detected the DNA of this spirochete belonging to the treponema phylogroups implicated in BDD. In 25% (6/24) of the lesions were detected all the phylogroups. Forty percent (40%, 2/5) of the M0 samples were also positive for the nested Polymerase Chain Reaction (nested-PCR), as 8.3% (2/24) of the lesions were negative in both techniques employed. Treponema putidum/T. denticola-like was the most detected bacterial in all the stages, and active lesions (M2 and M4.1) presented a greater proportion of T. medium/T. vincentii-like and T. phagedenis-like, but no statistical differences were observed (p>0.05). It could be concluded that BDD lesions in crossbred dairy cattle kept to pasture in the Amazon biome were classified as "polytreponemal" infections and the phylogroup T. putidum/T. denticola-like was the most frequent in the lesions.(AU)

Dermatite digital bovina (DDB) é uma enfermidade polibacteriana dos dígitos endêmica em vacas leiteiras criadas em estábulos e as treponemas são as principais bactérias envolvidas. Este estudo teve como objetivo relatar a ocorrência de Treponema spp. em DDB em bovinos leiteiros mestiços (Holandês x Zebu) criados a pasto no bioma amazônico brasileiro. O diagnóstico da DDB foi realizado pela inspeção, em uma ou mais visitas, das extremidades distais das vacas durante a ordenha em 15 propriedades. No total, foram inspecionadas 1.847 vacas de agosto de 2016 a julho de 2017 e diagnosticou-se 25 lesões de DDB. As extremidades distais inspecionadas foram classificadas em escores (M0 = sem lesão, M1 = estágio ulcerado <2cm, M2 = estágio ulcerado >2cm, M3 = estágio em cicatrização, M4 = estágio crônico, M4.1 = estágio crônico com área ulcerada) e realizada 24 biópsias de dígitos com DDB e cinco biópsias de dígitos em estágio M0. Foram realizadas a histopatologia pelas colorações de hematoxilina e eosina e pelo método de Warthin-Starry, e a nested de reação em cadeia de polimerase (nested-PCR) para os três filogrupos de treponemas previamente isolados de DDB (Treponema medium/T. vincentii-like, T. phagedenis-like e T. putidum/T. denticola-like). Espiroquetas foram observadas em 54,2% (13/24) das lesões e em 91,7% (22/24) detectou-se o DNA de, pelo menos, um dos filogrupos de treponemas pesquisados. Em 25% (6/24) das lesões foram detectados o DNA dos três filogrupos. Em 40% (2/5) das amostras em estágio M0 também foram positivas na nested-PCR, assim como 8,3% (2/24) das lesões foram negativas em ambas as técnicas empregadas. T. putidum/T. denticola-like foi o filogrupo mais detectado em todos os estágios e lesões ativas (M2 e M4.1) apresentaram uma maior proporção para Treponema medium/T. vincentii-like e T. phagedenis-like, mas não se obteve diferença estatística na ocorrência dos filogrupos entre os estágios das lesões (P>0,05). Conclui-se que lesões de DDB em rebanhos leiteiros mestiços criados a pasto no bioma amazônico brasileiro são "politreponemais" e o filogrupo T. putidum/T. denticola-like é o mais frequente nas lesões.(AU)

Bovine digital dermatitis in Victoria, Australia.

AIMS: The objectives of this study were to estimate the prevalence of digital dermatitis (DD) in Victoria, Australia, and to investigate which organisms are consistent with typical DD lesions. The prevalence and causative pathogens of DD are not clear yet in Australia and this paper is one of the first to explore these questions in this country. METHODS: Examination and sampling of limbs was undertaken at three knackeries in Victoria, Australia. Limbs were classified as normal (N), active DD-lesion (A), dried or chronic DD-lesion (D) or suspected case of DD (S). A total of 823 cows were examined. Six skin biopsies were taken at each knackery, from which DNA was extracted for diversity profiling. Histochemical staining of samples was performed on eight of the skin biopsies. RESULTS: DD was detected in 29.8% of all cows. The prevalence of DD was significantly higher in dairy cows (32.2%) than in beef cows (10.8%). The differential abundance of Treponema-species was significantly increased in dried lesions, compared with the normal skin biopsies. Actinobacteria, Proteobacteria, Firmicutes and Tenericutes were found to be significantly different in abundance in the DD lesions compared with normal skin biopsies. Silver staining of samples showed only mild inflammation and in two samples organisms with morphology consistent with Spirochaetes were detected. CONCLUSIONS: The calculated prevalence indicates that DD is present in Victoria, Australia. The results of diversity profiling showed that the presence of Treponema-species was significantly different between the samples of DD lesions and normal skin.

Interobserver agreement of digital dermatitis M-scores for photographs of the hind feet of standing dairy cattle.

Digital dermatitis (DD) is the leading infectious cause of lameness in dairy cattle, and it affects their welfare and productivity worldwide. At the herd level, DD is often assessed while cows are standing in a milking parlor, and lesions are most commonly evaluated using the M-score. The objective of this study was to examine the interobserver agreement for M-scores of the feet of standing cattle, based on digital color photographs of dairy cattle hind feet. A total of 88 photographs and written descriptors of the M-score were sent to 11 scorers working at 10 different institutions in 5 countries. The scorers received no formal training immediately before scoring the photographs; however, all regularly used the M-score to score DD. The answers for 36 photographs were excluded from the analysis because the photograph either had more than 1 M-stage as mode or not all scorers assigned an M-score to it. The M-scores of the 11 scorers from 52 photographs were available for analysis. Interobserver agreement was tested using Gwet's agreement coefficient (AC1) and the mode was assumed correct. Overall, moderate agreement emerged for the M-score (AC1 = 0.48). For the individual M-stages, almost perfect agreement existed for M0 (AC1 = 0.99), M1 (AC1 = 0.92), and M3 (AC1 = 0.82), and substantial agreement for M2 (AC1 = 0.61), M4 (AC1 = 0.65), and M4.1 (AC1 = 0.71). This outcome indicates the degree of individual variation in M-scoring in this context by unstandardized, experienced European observers, particularly for the M2, M4, and M4.1 stages. Standardized training is likely to improve the consistency of M-scoring and thus the generalizability of future DD research results on this important endemic disease.

Inter-observer agreement between two observers for bovine digital dermatitis identification in New Zealand using digital photographs.

AIMS: To assess the inter-observer agreement for detecting bovine digital dermatitis (BDD) lesions in digital colour photographs of the hind feet of cows, which had been taken while the animals were standing to be milked, between two trained observers. METHODS: Thirty-six photographs were selected from a total of 184 photographs held by the first author (R1), who had classified them as negative (n=11) or positive (n=25) for BDD. They were delivered to a technician (R2) who had previously visually inspected cattle for BDD lesions, and who then recorded the photographs as being either BDD-positive or BDD-negative. The percentage agreement between R1 and R2, and two other inter-observer agreement statistics, Cohen's κ and Gwet's first-order chance correction agreement coefficient (AC1), were calculated. The cumulative membership probabilities of Cohen's κ and Gwet's AC1 were then calculated for different benchmark ranges of κ. RESULTS: The percentage agreement between R1 and R2 was 33/36 (92%), Cohen's κ was 0.80 (95% CI=0.57-1.0) and Gwet's AC1 was 0.86 (95% CI=0.69-1.0). Based on the cumulative membership probabilities for Gwet's AC1, there was 75% probability that the two observers had almost perfect agreement (κ≥0.81). For both Cohen's κ and Gwet's AC1, there was >95% probability that the two observers had at least substantial agreement (κ≥0.61). CONCLUSIONS: The two trained observers had at least substantial agreement in identifying from a digital photograph as to whether BDD lesions were present or absent. Therefore results from the two could be used interchangeably. CLINICAL RELEVANCE: Visual assessment for BDD lesions in the milking parlour can be subjective. However a high agreement between these two trained BDD inspectors means BDD prevalence reported from different regions in New Zealand by these two can be directly compared.

The use of infrared thermography for detecting digital dermatitis in dairy cattle: What is the best measure of temperature and foot location to use?

Lameness in dairy cattle is a persistent problem, indicating pain caused by underlying disease states and is associated with reduced milk yields. Digital dermatitis is a common cause of lameness. Thermal imaging is a technique that may facilitate early detection of this disease and has the potential for use in automated detection systems. Previous studies with thermal imaging have imaged either the heels or the coronary band of the foot and typically only used the maximum temperature (Max) value as the outcome measure. This study investigated the utility of other statistical descriptors: 90th percentile (90PCT), 95th percentile (95PCT), standard deviation (SD) and coefficient of variation (CoV) and compared the utility of imaging the heel or coronary band. Images were collected from lame and healthy cows using a high-resolution thermal camera. Analyses were done at the cow and foot level. There were significant differences between lame and healthy feet detectable at the heels (95th percentile: P<0.05; SD: P<0.05) and coronary band (SD: P<0.05). Within lame cows, 95PCT values were higher at the heel (P<0.05) and Max values were higher at the coronary band (P<0.05) in the lame foot compared to the healthy foot. ROC analysis showed an AUC value of 0.72 for Max temperature and 0.68 for 95PCT at the heels. It was concluded that maximum temperature is the most accurate measure, but other statistical descriptors of temperature can be used to detect lameness. These may be useful in certain contexts, such as where there is contamination. Differentiation of lame from healthy feet was most apparent when imaging the heels.

Pathology and bacteria related to digital dermatitis in dairy cattle in all year round grazing system in Brazil.

Digital dermatitis (DD) is one of the main causes of lameness in dairy cattle worldwide, and it is frequently reported in high-yielding, free stall dairy herds from regions with a temperate climate. However, DD is also observed with high prevalence in grazing cattle with a low milk yield in tropical regions. To clarify whether these differences have an impact on the etiology of the disease, we studied DD lesions from all year round grazing cattle of mixed breed in Brazil using high-throughput 16S rRNA gene sequencing and fluorescent in situ hybridization. The study included samples from 66 skin lesions and 5 healthy skins collected from five farms. Both techniques showed Treponema spp. to be the most abundant bacteria, present in all but one of the samples with minimal epidermal alterations. We identified eleven different Treponema strains belonging to the six major phylotypes of Treponema which have all previously been identified in DD lesions. Furthermore, we identify Dichelobacter nodosus in DD lesions by gene sequencing and also by fluorescent in situ hybridization in almost half of biopsy specimens in areas with mild epithelial damage and together with Treponema. The present data support the hypothesis that Treponema constitutes the main pathogen responsible for DD, independent of the environment and region where cows are kept, and it further suggests D. nodosus as another potentially important pathogen.

Evaluation of agreement among digital dermatitis scoring methods in the milking parlor, pen, and hoof trimming chute.

Digital dermatitis (DD) is the most common infectious foot lesion affecting welfare and productivity of dairy cattle. The key to DD control programs is routine and frequent identification of DD lesions. The objective was to evaluate accuracy of detecting and scoring DD lesions in 3 milking parlor designs and in 3 alternative settings compared with scoring in the hoof trimming chute as reference. A total of 552 cows and 1,104 hind feet from 17 freestall farms were scored by 1 observer in the milking parlor and in 1 other setting: pen, headlocks, or management rail. After being scored in the milking parlor and at least 1 other setting, cows were examined in the hoof trimming chute, considered the gold standard. In every setting, all hind feet were inspected visually using a flashlight and without prior washing of feet. Agreement of the scoring settings was assessed using the 5 M-stage scoring system and a dichotomous absence (M0 or M0/M1) or presence (M1 to M4.1 or M2 to M4.1) system. At trimming chute inspection, 44% of feet had a DD lesion, with estimates of 11, 5, 2, 10, and 16% for M1, M2, M3, M4, and M4.1 lesions, respectively. Apparent DD foot-level prevalence at the milking parlor, pen, management rail, and headlocks was 28, 22, 16, and 22%, respectively. M-stages were less discernible in the pen, management rail, and headlocks (apparent prevalence of M1, M2, M3, and M4.1 was ≤1%) compared with the trimming chute and milking parlor. Agreement beyond chance between any scoring setting and trimming chute scoring ranged from 0.48 to 0.70 for the dichotomous scoring system (absence = M0/M1 vs. presence = M2 to M4.1). Diagnostic test performance varied greatly among DD scoring settings but, in general, it had low sensitivity (<70%) and high specificity (>93%) for detecting any DD lesion. Agreement and test characteristics were not affected by the type of milking parlor. Although the milking parlor and headlocks were the most reliable settings in which to detect DD, none of the settings were reliable enough to replace inspection of feet in the trimming chute. However, scoring the presence or absence of DD in the milking parlor, pen, management rail, and headlocks could be used to estimate within-herd DD prevalence, to improve DD surveillance through routine monitoring, and to evaluate effects of interventions at the farm level.

Efficacy of a commercial glutaraldehyde footbath product for the control of bovine digital dermatitis.

OBJECTIVE: To evaluate the relative efficacy of a novel, commercially available disinfecting agent containing ≥ 15 to < 20 % glutaraldehyde in a hydro-colloid matrix ("EIMÜ Klauen Sprint®"; Eimermacher, Nordwalde, Germany, Biozid-Desinfektionsmittel, BAuA Nr. N-57213) as a walkthrough footbath in order to treat digital dermatitis of dairy cattle. MATERIAL AND METHODS: The study was conducted in a dairy farm located in northern Germany from June 1st, 2015 to September 5th, 2015. Eighty-eight cows were enrolled in this study. In both trials of the study, a walkthrough footbath was applied on five consecutive milking times per week. Trial 1 was a blinded, randomised and controlled intervention study. The therapy group (n = 40) received the novel glutaraldehyde product in a 2 % solution while the control group (n = 40) was led through a footbath containing pure water. Trial 2 was a longitudinal study to investigate the development of claw health over a longer time period regarding the use of the new footbath solution. In trial 2 the disinfecting footbath was applied to all trial cows (therapy and control group) of trial 1. The development of claw health and lameness was systematically monitored in the two groups on days 0, 9, 17, 29, 35 (trial 1) and days 42, 68, 94 (trial 2), respectively. The adjusted odds of digital dermatitis lesions (DDL) throughout the study were analysed by mixed logistic regression models. RESULTS: The odds of DDL were 0.15 for the glutaraldehyde foot-bath group compared to the control group. Hence, the disinfecting footbath significantly reduced the DDL prevalence and increased the cure rate of DDL. However, no preventive effect on new digital dermatitis (DD) cases could be identified. CONCLUSION: The efficacy of the novel glutaraldehyde footbath was evaluated and it was concluded that "EIMÜ Klauen Sprint®" was well tolerated over a longer period of time and might therefore be an effective tool to control DD. CLINICAL RELEVANCE: In future, "EIMÜ Klauen Sprint®" could be used within a Mortellaro sanitation concept and thereby might help to solve a major problem of dairy cattle farms.

A clinical and histopathological comparison of the effectiveness of salicylic acid to a compound of inorganic acids for the treatment of digital dermatitis in cattle.

Bovine digital dermatitis (DD) is a painful infectious disease, causing lameness, reduced animal welfare, and production losses in dairy herds. The main factors contributing to DD are an infection with Treponema spp. and poor hygiene. Topical treatment has primarily consisted of antibiotics; however, the demand for effective nonantibiotic alternatives is increasing. The objective was to evaluate the performance of 3 nonantibiotic topical treatments (salicylic acid and a compound of inorganic acids in a 20% solution and in a dry form) on DD in a commercial dairy herd. Within the 30-d test period, 42 DD lesions on 33 Holstein cows were assigned to receive 1 of the 3 treatments. Lesions were biopsied before and after treatment and were clinically evaluated 5 times. Improved lesions were clinically defined as either healed (regeneration of the skin) or healing (dry lesions covered by a scab). Unhealed lesions were defined as either active [with a raw, moist, strawberry-like (granulating) surface] or mature (with a raised papillomatous appearance). The effectiveness of treatment was evaluated histopathologically using the following scores: 0 (no spirochetes present), 1 (small number of spirochetes present in the epidermis), 2 (moderate number of spirochetes present and reaching an intermediary level in the epidermis), and 3 (large number of spirochetes present and reaching the deepest part of the epidermis or the superficial dermis). The improvement rate was 10/14 (71%) for salicylic acid, 11/15 (73%) for the inorganic acid solution, and 8/13 (62%) for the inorganic acid powder. The analysis showed no difference among treatments. The association between clinical score and histopathological score was determined by an odds ratio. The odds ratio of a healed lesion having spirochetes in the epidermis was 0.58 and that of an active DD lesion having spirochetes in the epidermis was 26.5.

Experimental Transmission of Bovine Digital Dermatitis to Sheep: Development of an Infection Model.

Digital dermatitis is an infectious cause of lameness primarily affecting cattle but also described in sheep, goats, and wild elk. Digital dermatitis is a polymicrobial infection, involving several Treponema species and other anaerobic bacteria. Although the exact etiology has not been demonstrated, a number of bacterial, host, and environmental factors are thought to contribute to disease development. To study host-bacterial interactions, a reproducible laboratory model of infection is required. The objective of this study was to demonstrate key aspects of bovine digital dermatitis lesions in an easy-to-handle sheep model. Crossbred sheep were obtained from a flock free of hoof disease. Skin between the heel bulb and dewclaw was abraded before wrapping to emulate a moist, anaerobic environment. After 3 days, abraded areas were inoculated with macerated lesion material from active bovine digital dermatitis and remained wrapped. By 2 weeks postinoculation, experimentally inoculated feet developed erosive, erythematous lesions. At 4 weeks postinoculation, microscopic changes in the dermis and epidermis were consistent with those described for bovine digital dermatitis, including erosion, ulceration, hyperkeratosis, ballooning degeneration of keratinocytes, and the presence of neutrophilic infiltrates. Silver staining of lesion biopsy sections confirmed that spirochetes had penetrated the host epidermis. The model was then perpetuated by passaging lesion material from experimentally infected sheep into naïve sheep. This model of bovine digital dermatitis will allow for future novel insights into pathogenic mechanisms of infection, as well as the development of improved diagnostic methods and therapeutics for all affected ruminants.

Prevalence of digital dermatitis in young stock in Alberta, Canada, using pen walks.

Digital dermatitis (DD), an infectious bacterial foot lesion prevalent in dairy cattle worldwide, reduces both animal welfare and production. This disease was recently identified in replacement dairy heifers, with implications including increased risk of DD and decreased milk production in first lactation, poor reproductive performance, and altered hoof conformation. Therefore, a simple and effective method is needed to identify DD in young stock and to determine risk factors for DD in this group so that effective control strategies can be implemented. The objectives of this study were to (1) determine prevalence of DD in young stock (based on pen walks); and (2) identify potential risk factors for DD in young stock. A cross-sectional study was conducted on 28 dairy farms in Alberta, Canada; pen walks were used to identify DD (present/absent) on the hind feet of group-housed, young dairy stock. A subset of 583 young stock on 5 farms were selected for chute inspection of feet to determine the accuracy of pen walks for DD detection. Pen walks as a means of identifying DD lesions on the hind feet in young stock had sensitivity and specificity at the animal level of 65 and 98%, with positive and negative predictive values of 94 and 83%, respectively, at a prevalence of 37%. At the foot level, pen walks had sensitivity and specificity of 62 and 98%, respectively, with positive and negative predictive values of 92 and 88%, respectively, at a prevalence of 26%. Pen walks identified DD in 79 [2.9%; 95% confidence interval (95% CI): 2.3-3.6%] of 2,815 young stock on 11 (39%; 95% CI: 22-59%) of 28 farms, with all 79 DD-positive young stock ≥309 d of age. Apparent within-herd prevalence estimates ranged from 0 to 9.3%, with a mean of 1.4%. True within-herd prevalence of DD in young stock, calculated using the sensitivity and specificity of the pen walks, ranged from 0 to 12.6%, with a mean of 1.4%. On the 11 DD-positive farms, the proportion of young stock >12 mo of age with DD lesions was 9.9% (95% CI: 7.8-12.0%). Multilevel logistic regression was used to assess associations with potential risk factors for DD lesions, including age, leg cleanliness, and lactating herd DD prevalence. Presence of DD in young stock increased as their age increased and was associated with increased prevalence of DD in the lactating herd. Pen walks can be used to identify specific young stock with DD or groups where management practices can be implemented to prevent disease proliferation and transmission.

Validation of the M-stage scoring system for digital dermatitis on dairy cows in the milking parlor.

A high prevalence of digital dermatitis (DD) and the benefits of early topical treatment highlight the need for simple tools for routine DD detection. The objective of this study was to determine the accuracy of scoring DD lesions using the 5 M-stage scoring system in the milking parlor compared with the trimming chute as the gold standard. Three observers inspected 3,585 cows and 6,991 hind feet from 9 farms in the milking parlor using a mirror (glued to a plastic kitchen spatula) and a headlamp, followed by inspection in a trimming chute within 5 d. Interobserver agreement for scoring DD in various settings was ≥82% (kappa >0.74; weighted kappa >0.76). At trimming chute inspections, 68% of cows had at least 1 DD lesion, 19% had 1 hind leg affected, and 49% had both hind legs affected. Within-herd DD prevalence ranged from 16 to 81% of cows affected. True within-herd prevalence was 2, 6, 0, 36, and 14% for M1, M2, M3, M4, and M4.1 lesions, respectively. At the foot level, DD prevalence was the same (58%) in the milking parlor and trimming chute inspection, but distribution of M-stages differed. Milking parlor inspection as a means of identifying the presence of DD lesions had a sensitivity of 92% and specificity of 88%, with positive and negative predictive values of 91 and 89%, respectively. Agreement between milking parlor and trimming chute inspections was 73% (kappa = 0.59, weighted kappa = 0.65) for the 5 M-stage scoring system and 90% (kappa = 0.80) if only the presence of a lesion was noted. Test characteristics varied greatly among M-stages, with the highest sensitivity for detecting M4 (82%) and M2 (62%) lesions, and the lowest for detecting M4.1 (20%), M1 (7%), and M3 (0%) lesions. In the milking parlor, 20% of M2 lesions were misclassified as M4.1, 8% of M4 lesions were misclassified as M0, and 68% of M4.1 lesions were misclassified as M4. The majority (87%) of DD lesions were located between the heel bulbs; 10 and 2% of DD lesions affected the interdigital space and the front of the foot, respectively. The sensitivity to detect the presence of a lesion when it occurred between the heel bulbs was 93%, but <67% if it occurred elsewhere on the foot. We concluded that inspection of the rear feet in the milking parlor was an inexpensive and simple method of detecting and scoring DD lesions. If the objective is to determine herd-level DD prevalence and routine monitoring, this method was adequately reliable. However, if the objective is to follow up DD in cows with history of interdigital hyperplasia or to detect M1 or M4.1 lesions, this method was not sufficiently reliable. Although DD scoring in the milking parlor as a routine practice should facilitate early detection, prompt treatment interventions, and herd monitoring, it was not sufficiently reliable to replace definitive identification of M-stages in the trimming chute.

A novel approach to probe host-pathogen interactions of bovine digital dermatitis, a model of a complex polymicrobial infection.

BACKGROUND: Polymicrobial infections represent a great challenge for the clarification of disease etiology and the development of comprehensive diagnostic or therapeutic tools, particularly for fastidious and difficult-to-cultivate bacteria. Using bovine digital dermatitis (DD) as a disease model, we introduce a novel strategy to study the pathogenesis of complex infections. RESULTS: The strategy combines meta-transcriptomics with high-density peptide-microarray technology to screen for in vivo-expressed microbial genes and the host antibody response at the site of infection. Bacterial expression patterns supported the assumption that treponemes were the major DD pathogens but also indicated the active involvement of other phyla (primarily Bacteroidetes). Bacterial genes involved in chemotaxis, flagellar synthesis and protection against oxidative and acidic stress were among the major factors defining the disease. CONCLUSIONS: The extraordinary diversity observed in bacterial expression, antigens and host antibody responses between individual cows pointed toward microbial variability as a hallmark of DD. Persistence of infection and DD reinfection in the same individual is common; thus, high microbial diversity may undermine the host's capacity to mount an efficient immune response and maintain immunological memory towards DD. The common antigenic markers identified here using a high-density peptide microarray address this issue and may be useful for future preventive measures against DD.