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Bovine ischaemic teat necrosis (ITN) is a disease affecting the skin of the teats of dairy cows with an unknown aetiopathogenesis. Digital dermatitis (DD)-associated treponemes have previously been suggested as a potential aetiological agent in ITN, although the sample size was small. The current study, using established PCR techniques, aimed to examine the association with the presence of DD-associated treponemes in a large number of ITN samples from a wider geographical area, and surveyed the potential of milk as an infection reservoir. From 95 ITN lesions, 35.8% ( n = 34) were positive for at least one DD-associated treponeme compared with only 5.6% ( n = 1) of 18 non-lesioned teats from cows with ITN lesions on a different teat using a nested PCR approach. All 10 age- and production-matched control cows were negative for DD-associated treponemes via PCR. No DD-associated treponemes could be detected from foremilk of cows with ( n = 19) and without ( n = 31) a DD lesion on the hind feet. DD-associated treponemes could be detected via PCR after incubation in milk for up to 2 h. Therefore, milk does not appear to be a competent reservoir for transmission of DD-associated treponemes. Moreover, in the current study DD-associated treponemes were only detected in a subset of ITN samples, so it is unlikely these opportunistic skin-associated pathogens are the major or sole agent of ITN.

Objectives: Bovine digital dermatitis (BDD), a painful infectious foot disease in dairy cattle, endemic in many countries worldwide, causes substantial economic and welfare impacts. Treponema spp. are considered key to BDD pathogenesis. To aid infection reservoir identification and control measure development, survival of BDD treponemes was investigated in different temperatures (4, 12, 20, 37, 45 and 60 °C), pH values (5-9.0), dairy cattle faeces and bedding types: straw shavings, sand, sand containing 5% lime (w/w) and recycled manure solids (RMS)., Methods: A turbidity microplate methodology was adapted to measure pH impact on growth. Survival of BDD treponemes for the different conditions were assessed by sub-cultures of microcosms over different time points., Results: BDD treponemes remained viable between 4 and 37 °C and pH 5.5 and 9.0 under anaerobic conditions. In sterile faecal microcosms, incubated aerobically at 12 °C, BDD treponemes remained viable for a median of 1 day (15 min - 6 day range). Variation in duration of survival and ability to grow was observed between phylogroups and strains. In aerobic microcosms, T. phagedenis T320A remained viable for the full 7 days in sand, 6 days in sawdust, 5 days in RMS, but was not viable after 15 min in straw or sand containing 5% (w/w) lime., Conclusions: Treponeme survival conditions identified here should enhance future BDD infection reservoir surveys and enable control measures. Of note, straw or sand containing 5% (w/w) lime should be assessed in BDD field trials. Finally, these data indicate BDD treponemes exhibit characteristics of facultative anaerobes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Bovine ischaemic teat necrosis (ITN) is an emerging disease of unknown aetiology that affects the teats of dairy cattle. It causes economic and animal welfare issues with many animals being culled. No effective treatments or epidemiological data to inform control strategies are currently available. The aim of this observational study was to investigate farmer-reported experiences and identify potential farm-level risk factors. In January 2018, a questionnaire was sent to a random sample of 1,855 Great Britain (GB) dairy farmers. A usable response rate of 12.3% was obtained. Fifty-one per cent [95% confidence interval (CI): 44.4-57.8%] of farmers reported having experienced ITN on their farm between 1985 and 2018. Rising numbers of farms indicated that ITN is an emerging disease with 46.3% of farmers reporting the first case in the 3 years up to 2018. At the animal level, 47.3% (95% CI: 38.7-55.9%) of the cases occurred during the first lactation and 78.9% (95% CI: 75.2-82.6%) within the first 90 days in milk. Only 20.8% (95% CI: 15.9-26.4%) of the cases were reported to recover, whereas 22.8% (95% CI: 17.8-28.5%) of the cases required culling. The remaining cases experienced complications such as loss of a teat and/or mastitis. From these data, the cost of ITN, through production losses and expenditure, was estimated to be £1,121 per farm per year. The costs were estimated at £720, £860 and £2,133 for recovered, complicated and culled cases, respectively. Univariable and multivariable logistic regression models were used to explore the associations between the presence of ITN on farm and various risk factors. The presence of udder cleft dermatitis (UCD) (odds ratio 2.80; 95% CI: 1.54-5.07; p < 0.01) and chapped teats (odds ratio 6.07; 95% CI: 1.96-18.76; p < 0.01) in the milking herd was associated with the presence of ITN at the farm level. This is the first national questionnaire of ITN within GB and highlights the association of UCD and chapped teats with ITN at the farm level. While there are many limitations and potential bias around farmer questionnaires, these findings highlight several key areas for further disease investigation and possible intervention., Competing Interests: AM was employed by Quality Milk Management Services Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Crosby-Durrani, Blowey, Manning, Afonso, Carter, Evans and Angell.)

This research communication reports the results from questionnaires used to identify the impact of recent research into the disinfection of cattle foot-trimming equipment to prevent bovine digital dermatitis (BDD) transmission on (a) biosecurity knowledge and (b) hygiene practice of foot health professionals. An initial questionnaire found that more than half of participating farmers, veterinary surgeons and commercial foot-trimmers were not considering hand or hoof-knife hygiene in their working practices. The following year, after the release of a foot-trimming hygiene protocol and a comprehensive knowledge exchange programme by the University of Liverpool, a second survey showed 35/80 (43.8%) farmers, veterinary surgeons and commercial foot-trimmers sampled considered they were now more aware of the risk of spreading BDD during foot- trimming. Furthermore, 36/80 (45.0%) had enhanced their hygiene practice in the last year, impacting an estimated 1383 farms and 5130 cows trimmed each week. Participants who reported having seen both the foot-trimming hygiene protocol we developed with AHDB Dairy and other articles about foot-trimming hygiene in the farming and veterinary press, were significantly more likely to have changed their working practices. Difficulties accessing water and cleaning facilities on farms were identified as the greatest barrier to improving biosecurity practices. Participants' preferred priority for future research was continued collection of evidence for the importance and efficacy of good foot-trimming hygiene practices.

Background: Bovine digital dermatitis (BDD) is an infectious foot disease found commonly in dairy herds. Foot-trimming is an important husbandry procedure for reducing the ensuing lameness; however, epidemiological, and microbiological studies have identified this as a risk activity for transmitting BDD. Three disinfectants have previously been identified in laboratory work as effective for removing viable BDD-associated Treponema spp., from hoof knife blades. The present study enrolled 133 dairy cattle with BDD lesions, and swabbed hoof knife blades before and after foot-trimming, and after knife disinfection with one of three disinfectants (1:100 FAM30®, 2% Virkon® and 2% sodium hypochlorite) to assess their efficacy under field conditions., Results: Detection of BDD treponeme phylogroup DNA was undertaken by direct PCR of swabs, and viable treponemes were detected by PCR of swab cultures after 6 weeks' incubation. Where hoof knives did not contact the lesion, BDD-associated treponemes were detected after foot-trimming in 12/22 (54.5%) cases by direct PCR and 1/22 (4.5%) cases by PCR of cultured organisms. Where contact was made with the lesion, 111/111 (100%) samples taken after trimming were positive by direct PCR and 47/118 (39.8%) were positive by culture PCR. Viable organisms were identified in cultures from lesion stages M2, M3, M4 and M4.1. No viable organisms were detected after disinfection of hoof knives., Conclusions: Hoof knives post-trimming were frequently contaminated with BDD-associated treponeme DNA. Viable organisms were identified in cultures whether contact had been made between hoof knife and lesion or not, although contact clearly increased the frequency of detection of viable organisms. The three disinfectants tested were effective for removing viable organisms. The disinfection protocol used in this study should therefore be considered reliable for adoption as standard industry practice.

Background: Non-healing bovine foot lesions, including non-healing white line disease, non-healing sole ulcer and toe necrosis, are an increasingly important cause of chronic lameness that are poorly responsive to treatment. Recent studies have demonstrated a high-level association between these non-healing lesions and the Treponema phylogroups implicated in bovine digital dermatitis (BDD). However, a polymicrobial aetiology involving other gram-stain-negative anaerobes is suspected., Methods: A PCR-based bacteriological survey of uncomplicated BDD lesions (n=10) and non-healing bovine foot lesions (n=10) targeting Fusobacterium necrophorum , Porphyromonas endodontalis, Dichelobacter nodosus and Treponema pallidum/T. paraluiscuniculi was performed., Results: P. endodontalis DNA was detected in 80.0% of the non-healing lesion biopsies (p=<0.001) but was entirely absent from uncomplicated BDD lesion biopsies. When compared to the BDD lesions, F. necrophorum was detected at a higher frequency in the non-healing lesions (33.3% vs 70.0%, respectively), whereas D. nodosus was detected at a lower frequency (55.5% vs 20.0%, respectively). Conversely, T. pallidum/T. paraluiscuniculi DNA was not detected in either lesion type., Conclusion: The data from this pilot study suggest that P. endodontalis and F. necrophorum should be further investigated as potential aetiological agents of non-healing bovine foot lesions. A failure to detect syphilis treponemes in either lesion type is reassuring given the potential public health implications such an infection would present., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2020. Re-use permitted under CC BY. Published by BMJ.)

Background: Bovine digital dermatitis (BDD) is a painful infectious foot disease of cattle, and much evidence implicates a pathogenic role for treponemes. This study measured the survival of BDD treponemes on hoof knife blades and tested the efficacy of relevant disinfectants under laboratory conditions., Methods: Two strains of BDD treponemes were applied to hoof knife blades under aerobic conditions. Swabs were taken at different time points (10 minutes, one hour, two hours, four hours and 18 hours) and again after 20-second disinfection time with one of five disinfectants. Swabs were used directly for nested PCR to detect treponemes or inoculated for anaerobic growth, and subsequently examined using phase contrast microscopy and PCR., Results: BDD treponeme DNA was detectable by nested PCR at all survival time points, and these organisms were culturable from hoof knives for two hours after exposure under aerobic conditions in the laboratory. Three of the five disinfectants-1 per cent volume per volume (v/v) FAM30 ® , 2 per cent weight per volume (w/v) Virkon ® or 2 per cent (v/v) sodium hypochlorite-were effective at preventing visible growth of treponemes following 20-seconds contact, and 1 per cent (v/v) FAM30 ® also prevented detection of treponemes by PCR., Conclusion: Treponeme viability of two hours under aerobic conditions suggests BDD treponemes could be transmitted between cows on hoof knives. It is therefore important to apply a disinfection protocol during foot-trimming; the authors have identified three common disinfectants that may be suitable., Competing Interests: Competing interests: None declared., (© British Veterinary Association 2019. Re-use permitted under CC BY. Published by BMJ.)

Pressure sores cause severe pain and discomfort in hospitalized people and in farmed cattle and are often infected with unknown bacteria. Pressure sores occur on the upper legs of 6-10% of recumbent cattle and are generally considered to be caused by constant pressure, commonly on bony areas of the limbs. This study analyzed pressure sores taken from the upper limbs of 14 cattle using isolation in culture and nested polymerase chain reaction (PCR) to detect treponemes associated with digital dermatitis (DD). A 100% association of DD treponemes with the pressure sores was demonstrated, but treponemes were shown not to be part of the normal skin microbiota. Immunohistochemistry showed an association of DD treponemes with lesions and particularly with the hair follicles in lesions, identifying the bacteria deep within wounds, thereby suggesting that they could contribute to lesion pathogenesis. The bacteria isolated from the pressure sore lesions were similar or identical on analysis of the 16S rRNA gene to those found in DD foot lesions in cattle, suggesting the same bacteria can infect multiple lesions. Indeed, the results of this study suggest that these spirochaetal bacteria may be expanding in host range and in their ability to colonize different tissues and contribute to a range of disease manifestations in farm animals., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Background: Bovine hock lesions present a serious welfare and production issue on dairy farms worldwide. Current theories suggest that trauma is an important factor in the formation of hock lesions, although infection may also play a role in increasing their severity and duration., Hypothesis: Digital dermatitis (DD) lesions in dairy cows are strongly associated with specific treponeme bacteria which are opportunistic invaders of other skin regions. Hock lesions were tested to ascertain if they too contained treponemes., Animals: Swab and tissue samples were taken from hock lesions from two farms in South West England., Methods: Hock lesions were classified into two categories: open lesions, which were often bleeding and ulcerated, or were encrusted; and closed lesions, which were classified as hair loss with no skin breakage. PCR assays and bacterial isolation were used to detect treponemes in hock lesions., Results: All three phylogroups of digital dermatitis treponemes were detectable and isolated from open hock lesions only, with closed lesions showing no evidence of treponeme infection, either by PCR or bacterial culture. When analysed by 16S rRNA gene sequencing, the cultured treponeme DNA showed complete homology or was very similar to that found in foot lesions. Additionally, skin swabs from near the open hock wounds were also positive by PCR assay and isolation for the DD treponemes., Conclusions and Clinical Importance: Identification of the contribution of these infectious agents will allow for more optimal treatments to be developed that reduce the prevalence and healing times of both hock and DD lesions., (© 2016 ESVD and ACVD.)

Pig skin lesions are common significant welfare issues, and can cause large economic losses, due to culling of severely affected animals or carcass condemnation at slaughter. It was considered that the treponemal bacteria associated with digital dermatitis (DD) lesions in cattle, sheep and goats may have a role in these pig lesions. Specific diagnostic PCR assays for three cultivable DD Treponema phylogroups were used to survey relevant porcine lesion samples. Using these assays, DD treponemes were detected in 88% (22/25), 72% (8/11) and 82% (14/17) of tail, ear and flank lesions, respectively. Mouth swabs from animals kept in enclosures with high prevalence of skin lesions were positive for the DD treponemes, but not in enclosures with low lesion prevalence. Culture of treponemes from skin lesions resulted in pure isolates of all three DD-associated phylogroups. This study shows a strong association of DD treponemes with a range of pig skin lesions., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

A recent outbreak of ischaemic teat necrosis (ITN) on mainland UK has resulted in large economic losses for dairy farmers. Typical cases start as an area of dry, thickened and encrusted skin on the medial aspect of the base of the teat, where the teat joins the udder, often with a fetid odour. The erosion spreads down the teat, often causing intense irritation, which in turn leads to more severely affected animals removing the entire teat. Due to the severity of ITN and the substantial economic costs to the industry, analyses were undertaken to ascertain if an infectious agent might be involved in the pathology. The study has considered a role for digital dermatitis (DD) treponemes in the aetiopathogenesis of ITN because, as well as being the prime bacteria associated with infectious lameness, they have been associated with a number of emerging skin diseases of cattle, including udder lesions. A high association between presence of DD-associated treponemes and incidence of ITN (19/22), compared with absence in the control population is reported. Furthermore, sequencing of the 16S rRNA gene of treponeme isolates supports the hypothesis that the identified treponemes are similar or identical to those isolated from classical foot DD lesions in cattle (and sheep). Further studies are required to allow effective targeted prevention measures and/or treatments to be developed., (British Veterinary Association.)

Bovine digital dermatitis (BDD) is an infective foot disease commonly reported in dairy cattle where Treponema are considered as the primary causative infectious agents. There still remains little definitive information on the etiology of BDD in beef cattle suggesting further investigations are warranted. Beef BDD lesions (n=34) and healthy beef foot tissues (n=38) were analysed by PCR for three BDD-associated Treponema phylogroups and also for Dichelobacter nodosus and Fusobacterium necrophorum. Spirochete culture was attempted on all BDD lesion samples. One or more BDD-associated Treponema phylogroups were detected in 100% of beef BDD lesions. "Treponema medium/Treponema vincentii-like", "Treponema phagedenis-like" and Treponema pedis spirochetes were identified in 27/34 (79%), 31/34 (91%) and 24/34 (71%) of BDD lesions, respectively. No BDD-associated treponeme DNA was amplified from beef healthy foot tissues. D. nodosus and F. necrophorum were present in 24/34 (71%) and 15/34 (44%) of lesions and 10/38 (26%) and 12/38 (32%) of healthy foot tissues, respectively. Twenty spirochetes were isolated from beef BDD lesions; 19 were representatives of the three BDD-associated Treponema phylogroups. One spirochete isolate shared less than 97% 16S rRNA gene similarity to the three cultivable BDD-associated Treponema phylogroups and therefore may represent a novel taxa of Treponema. Upon comparison, sheep contagious ovine digital dermatitis (CODD), dairy cattle and beef cattle BDD lesions appear to have extremely similar bacteriological data and therefore provides evidence of a shared etiopathogenesis posing concerns for cross-species transmission., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Contagious ovine digital dermatitis (CODD) is an important foot disease in sheep, with significant animal welfare and economic implications. It is thought that CODD emerged from bovine digital dermatitis (BDD) via treponemal bacteria. With wildlife species such as elk now suffering a CODD-like disease, it is imperative to clarify these disease etiologies. A large investigation into treponemal association with CODD is warranted. CODD lesions (n = 58) and healthy sheep foot tissues (n = 56) were analyzed by PCR for the three BDD-associated Treponema phylogroups and two other lameness-associated bacteria, Dichelobacter nodosus and Fusobacterium necrophorum. Spirochete culture was also attempted on CODD lesions. "Treponema medium/Treponema vincentii-like," "Treponema phagedenis-like," and Treponema pedis spirochetes were identified in 39/58 (67%), 49/58 (85%), and 41/58 (71%) of CODD lesions, respectively. One or more BDD-associated Treponema phylogroups were detected in 100% of CODD lesions. Healthy foot tissues did not amplify BDD-associated Treponema phylogroup DNA. D. nodosus and F. necrophorum were present in 34/58 (59%) and 41/58 (71%) of CODD lesions and 22/56 (39%) and 5/56 (9%) of healthy foot tissues, respectively. Thirty-two spirochetes were isolated from CODD lesions, with representatives clustering with, and indistinguishable from, each of the three BDD-associated Treponema phylogroups based on 16S rRNA gene comparisons. This study for the first time demonstrates a high-level association for BDD treponeme phylogroups in CODD and their absence from healthy tissues, supporting the hypothesis that BDD treponemes play a primary causative role in CODD and confirming that the specific PCR assays are an effective differential diagnostic tool for CODD., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Since 2008, a large increase in the numbers of cases of lameness have been seen in wild North American elk (Cervus elaphus) from Washington State, USA. The most recent cases manifested as foot lesions similar both clinically and pathologically to those seen in digital dermatitis (DD) in cattle and sheep, a disease with a bacterial etiopathogenesis. To determine whether the same bacteria considered responsible for DD are associated with elk lameness, lesion samples were subjected to bacterial isolation studies and PCR assays for three phylogroups of relevant DD treponemes. The DD treponemes were isolated from lesional tissues but not from control feet or other areas of the diseased foot (including the coronary band or interdigital space), suggesting that the bacteria are strongly associated with DD lesions and may therefore be causal. In addition, PCR analysis revealed that all three unique DD treponeme phylotypes were found in elk hoof disease, and in 23% of samples, all 3 DD-associated treponemes were present in lesions. Sequence analysis of the 16S rRNA gene showed that the elk lesion treponemes were phylogenetically almost identical to those isolated from cattle and sheep DD lesions. The isolates were particularly similar to two of the three culturable DD treponeme phylotypes: specifically, the Treponema medium/Treponema vincentii-like and Treponema phagedenis-like DD spirochetes. The third treponeme culturable phylogroup (Treponema pedis), although detected by PCR, was not isolated. This is the first report describing isolation of DD treponemes from a wildlife host, suggesting that the disease may be evolving to include a wider spectrum of cloven-hoofed animals., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)

Digital dermatitis (DD) is an infectious foot disease causing severe lameness in dairy cattle (worldwide) and sheep (UK). This study investigated whether DD Treponema phylogroups can be present on equipment used to trim ruminant hooves and, therefore, consider this trimming equipment as a possible vector for the transmission of DD. Equipment was tested after being used to trim DD symptomatic and asymptomatic cattle and sheep hooves, and subsequently after disinfection of equipment. After trimming, 'Treponema medium/Treponema vincentii-like', 'Treponema phagedenis-like' and 'Treponema denticola/T putidum-like' DD spirochaetes, were shown to be present on 23/37 (62%), 21/37 (57%) and 20/37 (54%) of knives, respectively. After disinfection, detection rates for the DD treponemes were 9/37 (24%), 6/37 (16%) and 3/37 (8%), respectively. Following culture of a swab, an isolate belonging to the T phagedenis-like spirochaetes was identified from a knife sample after trimming a DD positive cow. No isolates were obtained from knife samples after disinfection. This new data has, for the first time, identified treponemes in the farm environment, and highlighted disinfection of hoof trimming equipment between animals and between farms, as a logical precaution to limit the spread of DD., (British Veterinary Association.)

Digital dermatitis is a painful foot disease compromising welfare in dairy cattle. The disease has a complex multibacterial aetiology, but little is known about its pathogenesis. In this study, gene expression in skin biopsies from five bovine digital dermatitis lesions and five healthy bovine feet was compared using RNA-Seq technology. Differential gene expression was determined after mapping transcripts to the Btau 4.0 genome. Pathway analysis identified gene networks involving differentially expressed transcripts. Bovine digital dermatitis lesions had increased expression of mRNA for α2-macroglobulin-like 1, a protein potentially involved in bacterial immune evasion and bacterial survival. There was increased expression of keratin 6A and interleukin 1β mRNA in bovine digital dermatitis lesions, but reduced expression of most other keratin and keratin-associated genes. There was little evidence of local immune reactions to the bacterial infection present in lesions., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

This study describes the association between bovine digital dermatitis (BDD) treponemes and three 'non-healing' bovine hoof horn lesions, namely, 'toe necrosis' (TN), 'non-healing white line disease' (nhWLD) and 'non-healing sole ulcer' (nhSU), which are disorders that involve penetration through the horn capsule to involve the corium. In this study, these non-healing disorders (n=44) were identified as foot lesions that exhibited a topical granular appearance, exuded a typical pungent smell, were severely painful to the animal involved, and typically originated from farms where BDD is endemic. Given the similarities between these 'non-healing' lesions and BDD, the authors subjected samples of diseased tissue to PCR assays to detect the presence of DNA of BDD treponemes. All the three characterised BDD treponeme groups were identified as present together in 84.2, 81.3 and 55.6 per cent of samples of TN (n=19), nhWLD (n=16) and nhSU (n=9), respectively. In contrast, healthy control horn samples from similar sites (n=16) were PCR-negative for the BDD treponemes. Hence, these non-healing hoof lesions were strongly associated with BDD treponemes. Samples from typical heel horn erosions (n=9) were also subjected to BDD treponeme PCR assays and no association could be identified between the BDD treponemes and this horn manifestation.

Claw lesion treatment records were recorded by farmers on 27 dairy farms (3,074 cows, 36,432 records) in England and Wales between February 2003 and February 2004. These were combined with farm environment and management data collected using a combination of direct observations, interviews with farmers, and milk recording data. Multilevel models were constructed for the 3 most frequently reported lesions related to lameness, namely, sole ulcers, white line disease, and digital dermatitis. Risks associated with an increased incidence of sole ulcers were parity 4 or greater, the use of roads or concrete cow tracks between the parlor and grazing, the use of lime on free stalls, and housing in free stalls with sparse bedding for 4 mo or more. The risks for white line disease were increasing parity and increasing herd size, cows at pasture by day and housed at night, and solid grooved concrete floors in yards or alleys. Solid grooved flooring was also associated with an increased risk of digital dermatitis, and cows 6 or more months after calving had a decreased risk of a first case of digital dermatitis. These results improve our understanding of the specific risks for 3 important lesions associated with bovine lameness and could be used as interventions in future clinical studies targeted at the reduction of specific lesions.

This study used a PCR-based approach targeting 16S rRNA gene fragments to determine the occurrence and association of the three bovine digital dermatitis (BDD) treponeme phylogroups within lesions found in cattle from the United Kingdom. Examination of 51 BDD lesions collected from infected cattle across the United Kingdom revealed that BDD treponeme group 1 (Treponema medium/Treponema vincentii-like), group 2 (Treponema phagedenis-like), and group 3 (Treponema putidum/Treponema denticola-like) were present in 96.1%, 98%, and 76.5% of BDD lesions, respectively. The three phylogroups were present together in 74.5% of lesions. The PCR assays enabled the isolation of further treponeme strains from previously mixed primary BDD lesion cultures. Here a representative from each of the three distinct treponeme phylogroups was isolated from a single BDD lesion for the first time. These data highlight the extent to which this disease is polytreponemal. Immunohistochemistry and electron microscopy were used to investigate lesional hoof tissues, resulting in treponemes being identified copiously in hair follicles and sebaceous glands, suggesting a potential route of exit and/or entry for these pathogens. This study gives further evidence for the importance of the three treponeme groups in BDD pathogenesis and reiterates the value of molecular genetic approaches for isolating and identifying fastidious anaerobes.

Bovine digital dermatitis (BDD) is a severe infectious cause of lameness which has spread through dairy cattle populations worldwide, causing serious welfare and agricultural problems. Spirochetes are the main organisms implicated and have previously proven difficult to isolate. This study aimed to isolate and characterise the range of spirochetes associated with BDD in the UK. Twenty-three spirochete isolates were obtained from 30 BDD lesions, which by 16S rRNA gene and flaB2 gene analysis clustered within the genus Treponema as three phylogroups; groups 1 (Treponema medium/Treponema vincentii-like), 2 (Treponema phagedenis-like) and 3 (Treponema denticola/Treponema putidum-like). The treponemes displayed large genotypic and phenotypic diversity between phylogroups and differed from named treponeme species. A previously isolated contagious ovine digital dermatitis spirochete was located within one of the three phylogroups, group 3, and could also be identified within this group on the basis of phenotype testing, suggesting BDD and contagious ovine digital dermatitis may share the same aetiological agent. A strain isolated from a bovine interdigital dermatitis lesion, could be identified as part of BDD isolate group 2, suggesting bovine interdigital dermatitis and BDD may have the same causative agent. Two common enzyme activities, C4 esterase and C8 esterase lipase, were identified in all BDD associated treponemes suggesting common metabolic pathways for sharing this novel niche or even common virulence traits. Further studies are required to determine whether the three groups of novel treponemes are representative of new treponeme taxa and to delineate how they interact with bovine tissues to cause disease.

The milk yields of 1824 cows were used to investigate the effect of lesion-specific causes of lameness, based on farmer treatment and diagnosis of lame cows, on milk yield. A three-level hierarchical model of repeated test day yields within cows within herds was used to investigate the impact of lesion-specific causes of lameness (sole ulcer, white line disease, digital dermatitis and other causes) on milk yield before and after treatment compared with unaffected cows. Cattle which developed sole ulcer (SU) and white line disease (WLD) were higher yielding cattle before they were diagnosed. Their milk production fell to below that of the mean of unaffected cows before diagnosis and remained low after diagnosis. In cattle which developed digital dermatitis (DD) there was no significant difference in milk yield before treatment and a slightly raised milk yield immediately after treatment. The estimated milk loss attributable to SU and WLD was approximately 570 and 370 kg, respectively. These results highlight that specific types of lameness vary by herds and within herds they are associated with higher yielding cattle. Consequently lesion-specific lameness reduction programmes targeting the cow and farm specific causes of lameness might be more effective than generic recommendations. They also highlight the importance of milk loss when estimating the economic impact of SU and WLD on the farms profitability.

Forty-nine farms in England and Wales were visited on 4 occasions between February 2003 and March 2004. A total of 21,693 scores of locomotion were assigned to 7,722 cattle. Locomotion was assessed on a 3-point scale by observing the posture of a cow's back while standing and walking (1 = sound, 2 = not sound, 3 = lame). Data on measurable factors potentially associated with locomotion were collected from all farms using direct observations of the farm environment and a comprehensive farmer interview. The mean herd locomotion score was 1.77 +/- 0.02. There was no significant difference in mean herd locomotion scores between 5 herds housed in straw yards (1.72 +/- 0.02) and 44 herds housed in free stalls (1.78 +/- 0.02), possibly because of lack of power. A GLM was produced using data from the 44 herds housed in free stalls, with the mean farm locomotion score of all cows examined on all 4 visits as the outcome variable. Factors associated with an elevated locomotion score were dry cows kept in straw yards compared with free stalls (increase in locomotion score = 0.06 +/- 0.03), pregnant heifers kept with milking cows in winter compared with being kept with dry cows (increase in locomotion score = 0.09 +/- 0.03), aisle widths of < 3 m compared with widths of > or = 3 m (increase in locomotion score = 0.06 +/- 0.02), a curb height of < or = 15 cm compared with a height of > 15 cm (increase in locomotion score = 0.07 +/- 0.03), routine trimming of hooves of all cows by a hoof trimmer or by the farmer compared with no routine hoof trimming (increase in locomotion score = 0.18 +/- 0.04 and 0.13 +/- 0.03 respectively), feeding corn silage to milking cows compared with feeding other forage types (increase in locomotion score = 0.10 +/- 0.03), and the use of automatic scrapers in the free-stall barn compared with tractor scrapers (increase in locomotion score = 0.10 +/- 0.03). These variables were correlated with many other management variables. The use of automatic scrapers was correlated with the use of sawdust on rubber mats in free stalls. Curb height was negatively correlated with increasing distance of the neck rail from the front (head end) of the free stall. These putative risk factors support the hypothesis that locomotion score is linked to management factors; in particular, the combination of sawdust on rubber mats with automatic scrapers was associated with elevated locomotion scores.

A 3-point locomotion scoring system was used that incorporated the position of the back of cows while standing and when walking to investigate risk factors for elevated locomotion scores of 1,450 dairy cows on 19 farms in The Netherlands. Each of the farms was visited twice in an 18-mo period from February 2003 to July 2004. At each visit, all milking and dry cows were scored for locomotion by a single observer. Two multivariable regression models were constructed to identify factors associated with elevated mean locomotion score (increased abnormality) and the percentage of cows with the highest score (score 3). Risk factors for increased locomotion score were having a hoof-trimming stall with foot-lifting apparatus compared with not having such apparatus (increase in locomotion score = 0.15), presence of a footbath at the parlor exit or other site compared with not having a footbath on the farm (increase in locomotion score = 0.17 and 0.19, respectively), not providing supplemental vitamins and minerals to lactating cows compared with supplementing animals (increase in locomotion score = 0.17) and feeding corn silage to heifers compared with not doing so (increase in locomotion score = 0.10). The results provide a framework for hypotheses for future investigations of risk factors for high locomotion scores.

The tensile strength of 576 pieces of white line horn collected over 6 mo from 14 dairy cows restricted to parity 1 or 2 was tested. None of the cows had ever been lame. Seven cows were randomly assigned to receive 20 mg/d biotin supplementation, and 7 were not supplemented. Hoof horn samples were taken from zones 2 and 3 (the more proximal and distal sites of the abaxial white line) of the medial and lateral claws of both hind feet on d 1 and on 5 further occasions over 6 mo. The samples were analyzed at 100% water saturation. Hoof slivers were notched to ensure that tensile strength was measured specifically across the white line region. The tensile stress at failure was measured in MPa and was adjusted for the cross-sectional area of the notch site. Data were analyzed in a multilevel model, which accounted for the repeated measures within cows. All other variables were entered as fixed effects. In the final model, there was considerable variation in strength over time. Tensile strength was significantly higher in medial compared with lateral claws, and zone 2 was significantly stronger than zone 3. Where the white line was visibly damaged the tensile strength was low. Biotin supplementation did not affect the tensile strength of the white line. Results of this study indicate that damage to the white line impairs its tensile strength and that in horn with no visible abnormality the white line is weaker in the lateral hind claw than the medial and in zone 3 compared with zone 2. The biomechanical strength was lowest at zone 3 of the lateral hind claw, which is the most common site of white line disease lameness in cattle.

A field study was conducted to examine effects of oral biotin supplementation for up to 18 mo on risks of lameness in dairy cows. The study included a total of 900 cattle from five dairy farms in Gloucestershire, southwest U.K., in a within-herd randomized control trial. The data from this trial were used in this paper to investigate the impact of parity and duration of supplementation with oral biotin at 20 mg/d on white line disease (WLD) lameness. Analysis of the data indicated that WLD increased with increasing parity independent of biotin supplementation from approximately two cases per 100 cow years in primiparous cows to 15.5 cases per 100 cow years in all multiparous cows, but up to 47.7 cases per 100 cow years for cows in parities > or = 5. Supplementation with biotin reduced WLD lameness by 45% in multiparous cows down to 8.5 cases per 100 cow years, whereas the effect of biotin supplementation in primiparous cows was not significant. Although numerical reductions in WLD lameness were observed for shorter periods of supplementation, a supplementation length of at least 6 mo was required to significantly reduce the risk of WLD lameness in multiparous cows. The effect of biotin supplementation in reducing lameness has potential impact for both animal welfare and farm economics.

This paper investigates the impact of lameness on milk yield. The dataset includes approximately 8000 test-day milk yields from 900 cows on five farms in Gloucester, UK, collected over 18 mo from 1997 to 1999. The data were structured to account for repeated measures of test-day yield (1 to 10 per cow) and analyzed to account for this autocorrelation. Factors affecting milk yield included: farm of origin, stage of lactation, parity, and whether a cow ever became lame. In clinically lame cows, milk yield was reduced from up to 4 mo before a case of lameness was diagnosed and treated and for the 5 mo after treatment. The total mean estimated reduction in milk yield per 305-d lactation was approximately 360 kg. We conclude that clinical lameness has a significant impact on milk production. This is important information for assessing the economic impact of clinical lameness and its impact on cow health. It adds weight to the importance of early identification of clinical lameness and the urgency of techniques to improve the definition of this highly subjective diagnosis.

A longitudinal prospective intervention study investigated the effect of biotin supplementation on the incidence (new cases per day) of visible lameness in milking cows and heifers on five commercial farms in Gloucestershire, United Kingdom. The trial lasted from June 1997 to April 1999. Each farm participated in the trial for 18 mo. Within each herd the cows were randomly allocated to either receive a supplement of 20 mg of biotin per day or not. All cows were run as one herd on each farm. When a lame cow was identified, the farmer called one of six veterinarians to examine and treat the affected animal; findings were recorded on a standard form. A veterinarian also carried out a bimonthly locomotion assessment to ensure that all lame cows were diagnosed. There were a total of 900 cows, 1120 cow years, in the trial. The overall incidence rate of lameness (per 100 cows per year) was 68.9, with a range of 31.6 to 111.5 per farm. The incidence rates of the four most frequently reported causes of lameness were sole ulcer, 13.8; white line separation, 12.7; digital dermatitis, 12.0; and interdigital necrobacillosis, 7.1 per 100 cows per year. There was a significant difference in the incidence rate of these four lesions between supplemented and unsupplemented cows on two of the five farms, with a significant decrease in lameness in the cows supplemented with biotin. When all the farms were pooled, the risk of lameness caused by white line separation in cattle supplemented with biotin was approximately halved (Cox proportional hazard survival analysis hazard ratio = 0.57).

A potentially infectious aetiology for digital dermatitis in dairy cattle was investigated and centred on the possible involvement of spirochaetes. An enzyme-linked immunosorbent assay (ELISA) was developed to detect bovine anti-Borrelia burgdorferi (B31) and anti-Treponeme (USA bovine isolates) antibodies in the sera of cows; sera were further tested for antigen specificity by Western blotting. Compared to normal cows, those with digital dermatitis had a much higher seropositivity rate to B. burgdorferi and the treponemes. Significant correlations were shown between antibodies to B. burgdorferi and to Treponemes (P < 0.001), suggesting strong cross-reacting epitopes shared by these spirochaetes. In Western blotting of B. burgdorferi antigens, the main band detected by ELISA positive sera was the 41 kDa flagellar protein; lesser frequency of staining was seen with 34 (OspB), 39 and 55 kDa bands. For the USA treponeme antigens, ELISA positive sera gave reactions to the 34-kDa band and also bands at 41 and 55 kDa. Polyclonal antibodies to Treponema denticola and T. vincentii showed reactions with the bovine treponemes which were predominantly to the 34-kDa antigen. Monoclonal antibodies to B. burgdorferi flagella (41 kDa) antigen and OspA (31 kDa) did not detect any treponeme bands in Western blotting. The study has provided serological evidence that spirochaetes (which are related to human treponemes) may be involved in the pathogenesis of digital dermatitis.