Multilocus sequence typing of pathogenic treponemes isolated from contagious ovine digital dermatitis stage five lesions: Implications for disease transmission dynamics.

Contagious ovine digital dermatitis (CODD) causes a severe, infectious foot disease and lameness of sheep, is common within the UK and is now also emerging in other countries. As well as causing severe animal welfare issues, huge economic losses emerge from the disease due to weight loss/lack of weight gain, and veterinary treatments. CODD lesion progress is measured, with a scoring system from 1 (early lesions) to 5 (healed). Here, using samples from an experimental flock infected by natural means, samples were taken from CODD stage 5 lesions, post treatment, and subjected to bacterial isolation and MLST using previously published methods. Sequences were compared to others from the same flock, and those from previous studies. All CODD 5 lesions produced viable Treponema spp. bacteria. High levels of variation of bacteria were seen, with 12 sequence types (STs) for T. medium phylogroup (11 new), 15 STs for T. phagedenis phylogroup (9 new) and six T. pedis STs, of which two were new. This study shows that CODD stage 5 lesions still contain viable bacteria, representing all three known pathogenic Treponema spp. phylogroups, and these may thus play a role in disease transmission and epidemiology despite appearing healed after treatment. The high level CODD treponeme variability within an infected flock where sheep were bought from different sources, as might occur in common agricultural practice, may suggest reasons as to why the bacterial disease is difficult to treat, control and eradicate, and adds further complexity to the polybacterial pathogenesis of these lesions.

Short communication: Screening stable flies and house flies as potential vectors of digital dermatitis in dairy cattle.

Digital dermatitis is a bacterial infection that causes lesions above the heel bulbs on cattle hooves, and several bacterial species from the genus Treponema are suspected to be causative agents of this polymicrobial condition. Transmission of the bacteria to healthy cows is understudied, particularly with regard to potential insect vectors. Therefore, the objective of this research was to determine if flies captured from a dairy farm known to have digital dermatitis are contaminated with Treponema bacteria. The DNA-based assays did not detect any Treponema phagedenis from stable flies and house flies collected at a dairy experiencing an outbreak of digital dermatitis. Other potential means of bacterial transmission are discussed.

Removal of bovine digital dermatitis-associated treponemes from hoof knives after foot-trimming: a disinfection field study.

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.

Modelling the transmission dynamics of bovine digital dermatitis in New Zealand pastoral dairy production systems.

Bovine digital dermatitis (DD) is an important infectious cause of cattle lameness worldwide that has become increasingly prevalent in New Zealand pastoral dairy herds. In this study, a simplified DD scoring system after considering both M and Iowa DD scoring systems was applied to explore the transmission dynamics of DD in a typical spring-calving pastoral New Zealand dairy herd. The modified model only included three compartments: normal skin, early stage lesions and advanced lesions. Lesions regressing after treatment were excluded as DD lesions are rarely treated in New Zealand. Furthermore, sub-classes within each lesion class were not defined due to the lack of variability in DD lesion presentations within New Zealand. The model was validated based on longitudinal field data from three dairy herds in the Waikato region during one lactation season (2017-18). The model suggested that in infected dairy herds, although DD prevalence will tend to increase year-on-year it is likely to remain relatively low (< 18%) even after 10 years of within-herd transmission. It is likely that the low transmission rate during the late lactation (model assumption) results in more cases resolving than developing during this period and therefore results in the low prevalence of infectious cattle at the start of each subsequent lactation. Cattle with advanced lesions had a stronger influence on the establishment and maintenance of DD than cattle with early stage lesions highlighting the importance of targeting these animals for intervention. On-going monitoring of DD is highly recommended to assess the long-term progression of the disease in affected dairy herds.

A genome-wide association study for susceptibility and infectivity of Holstein Friesian dairy cattle to digital dermatitis.

Selection and breeding can be used to fight transmission of infectious diseases in livestock. The prevalence in a population depends on the susceptibility and infectivity of the animals. Knowledge on the genetic background of those traits would facilitate efficient selection for lower disease prevalence. We investigated the genetic background of host susceptibility and infectivity for digital dermatitis (DD), an endemic infectious claw disease in dairy cattle, with a genome-wide association study (GWAS), using either a simple linear mixed model or a generalized linear mixed model based on epidemiological theory. In total, 1,513 Holstein-Friesian cows of 12 Dutch dairy farms were scored for DD infection status and class (M0 to M4.1) every 2 wk for 11 times; 1,401 of these cows were genotyped with a 75k SNP chip. We performed a GWAS with a linear mixed model on 10 host disease status traits, and with a generalized linear mixed model with a complementary log-log link function (GLMM) on the probability that a cow would get infected between 2 scorings. With the GLMM, we fitted SNP effects for host susceptibility and host infectivity, while taking the variation in exposure of the susceptible cow to infectious herd mates into account. With the linear model we detected 4 suggestive SNP (false discovery rate < 0.20), 2 for the fraction of observations a cow had an active lesion on chromosomes 1 and 14, one for the fraction of observations a cow had an M2 lesion on at least one claw on chromosome 1 (the same SNP as for the fraction of observations with an active lesion), and one for the fraction of observations a cow had an M4.1 lesion on at least one claw on chromosome 10. Heritability estimates ranged from 0.09 to 0.37. With the GLMM we did not detect significant nor suggestive SNP. The SNP effects on disease status analyzed with the linear model had a correlation coefficient of only 0.70 with SNP effects on susceptibility of the GLMM, indicating that both models capture partly different effects. Because the GLMM better accounts for the epidemiological mechanisms determining individual disease status and for the distribution of the y-variable, results of the GLMM may be more reliable, despite the absence of suggestive associations. We expect that with an extended GLMM that better accounts for the full genetic variation in infectivity via the environment, the accuracy of SNP effects may increase.

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.

Digital Dermatitis in dairy cattle: The contribution of different disease classes to transmission.

Digital Dermatitis (DD) is a claw disease mainly affecting the hind feet of dairy cattle. Digital Dermatitis is an infectious disease, transmitted via the environment, where the infectious "agent" is a combination of bacteria. The standardized classification for DD lesions developed by Döpfer et al. (1997) and extended by Berry et al. (2012) has six distinct classes: healthy (M0), an active granulomatous area of 0-2 cm (M1), an ulcerative lesion of >2 cm (M2), an ulcerative lesion covered by a scab (M3), alteration of the skin (M4), and a combination of M4 and M1 (M4.1). We hypothesize that classes M1, M2, M3, M4, and M4.1 are the potentially infectious classes that can contribute to the basic reproduction ratio (R0), the average number of new infections caused by a typical infected individual. Here, we determine differences in infectivity between the classes, the sojourn time in each of the classes, and the contribution of each class to R0. The analysis is based on data from twelve farms in the Netherlands that were visited every two weeks, eleven times. We found that 93.89% of the transitions from M0 was observed as a transition to class M4, and feet with another class-at-infection rapidly transitioned to class M4. As a consequence, about 70% of the infectious time was spent in class M4. Transmission rate parameters of class-at-infection M1, M2, M3, and M4 were not significantly different from each other, but differed from class-at-infection M4.1. However, due to the relative large amount of time spend in class M4, regardless of the class-at-infection, R0 was almost completely determined by this class. The R0 was 2.36, to which class-at-infection M4 alone contributed 88.5%. Thus, M4 lesions should be prevented to lower R0 to a value below one, while painful M2 lesions should be prevented for animal welfare reasons.

Associations between biosecurity practices and bovine digital dermatitis in Danish dairy herds.

The relationship between biosecurity and digital dermatitis (DD) was evaluated in 8,269 cows from a convenience sample of 39 freestall dairy herds. The hypothesis was that poor implementation of biosecurity was associated with higher within-herd prevalence of DD. All lactating cows were scored as negative or positive for DD at the hind legs during milking in the milking parlor. Information about biosecurity was obtained through questionnaires addressed to farmers, on-farm observations, and information from the Danish Cattle Database (www.seges.dk). These assessment tools covered potential infection sources of DD pathogens to susceptible cows (e.g., via animals, humans, manure, vehicles, equipment, and facilities). External and internal biosecurity measures were explanatory variables in 2 separate logistic regression models, whereas within-herd DD prevalence was the outcome. Overall DD prevalence among cows and herds were 24 and 97%, respectively; the within-herd DD prevalence ranged from 0 to 56%. Poor external biosecurity measures associated with higher prevalence of DD were recent animal purchase, access to pasture, lack of boots available for visitors, farm staff working at other dairy farms as well, hoof trimming without a professional attending, and animal transporters having access to cattle area. For internal biosecurity, higher DD prevalence were associated with infrequent hoof bathing, manure scraping less than 8 times a day, manure removal direction from cows to heifers, animal pens' exit without water hoses, manure-handling vehicle used in other activities, and water troughs contaminated with manure. These findings showed that improvements on biosecurity may be beneficial for controlling DD in dairy herds. The study is relevant for farmers facing problems with DD, as well as hoof trimmers, advisors, and veterinarians, who can use the results for optimized recommendations regarding biosecurity in relation to DD. Furthermore, our results might be considered by future studies investigating DD pathogen reservoirs and transmission routes.

Experimental infection of cattle with ovine Dichelobacter nodosus isolates.

BACKGROUND: Dichelobacter nodosus is the main causative agent of ovine footrot, and there are strong indications that the bacterium can be transferred to cattle grazing on the same pasture as sheep. The aim of this study was to investigate if benign and virulent D. nodosus strains isolated from sheep can be transferred to the interdigital skin of cattle under experimental conditions. Further, we wanted to observe the impact of such infection on bovine foot health, and test the effect of topical chlortetracycline (Cyclo spray(®): Eurovet) on the infection. FINDINGS: Six heifers were included in the study. After an initial 18-day maceration period, three heifers were inoculated on one single foot with a benign strain and three with a virulent strain by adding bacterial suspension in a bandage. The bandages were left on for 17 days, and when removed, D. nodosus was isolated from all six heifers. All six heifers developed interdigital dermatitis. In five of the heifers D. nodosus organisms were demonstrated within the epidermis. Twenty-four days after treatment with chlortetracycline all heifers were negative by cultivation, but tested positive for D. nodosus by polymerase chain reaction (PCR). Two of the six heifers still tested positive for D. nodosus by PCR 49 days after treatment. After 70 days, all heifers tested negative for D. nodosus. CONCLUSIONS: This study shows that both virulent and benign D. nodosus strains originating from sheep can be transferred to naïve heifers under experimental conditions. Further, the study supports the hypothesis that infections with virulent D. nodosus in cattle are associated with interdigital dermatitis. No conclusion regarding the treatment of D. nodosus infection with chlortetracycline was possible.

Treponemes detected in digital dermatitis lesions in Brazilian dairy cattle and possible host reservoirs of infection.

The main pathogenic treponemes causing bovine digital dermatitis were identified from 17 infected herds in southern Brazil for the first time in this study using PCR. We did not find a relationship between treponeme phylogroup composition and clinical classification. Treponema phagedenis was present in all lesions. Rumen fluid was implicated as a reservoir location for these pathogens.

Presence of digital dermatitis treponemes on cattle and sheep hoof trimming equipment.

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.

Infection dynamics of digital dermatitis in first-lactation Holstein cows in an infected herd.

Digital dermatitis (DD) refers to painful lesions primarily affecting the skin in the interdigital region of dairy cattle. The purpose of this study was to evaluate the dynamics of DD in 39 cows, observed at approximately 3-d intervals, for the first 6 mo of lactation. Specifically, the study aimed at evaluating different levels of DD susceptibility in cows, identifying the bacterial colonization of the interdigital skin, and exploring the relationship between clinical DD diagnosis and laboratory findings. Three different susceptibility categories were identified for DD: 1=consistently healthy cow; 2=intermittently infected cow; and 3=consistently infected cow. Susceptibility categories were associated with age at calving. The average age at calving was 775 d (SD ±43.4), with the youngest heifer calving at age 669 d and the oldest heifer at 858 d. Advancing age at calving was associated with greater odds of being intermittently or consistently infected. This corresponded with an odds ratio of 2.02 over a period of 30 d. During the study period, 161 DD lesions were identified in 28 of the 39 cows (72%). Of those 28 cows, 13 cows were consistently infected. The remaining 11 of the 39 cows (28%) showed slight thickening of the skin with no pain (5 cows) and no signs of skin changes (6 cows). Histopathology and fluorescence in situ hybridization were possible to perform on 132 biopsy samples. A clinical diagnosis of DD was confirmed in 70% of the lesions by histopathology, and colonization of Treponema spp. Dichelobacter nodosus was found in 35 samples (29%).

The dynamics of digital dermatitis in populations of dairy cattle: model-based estimates of transition rates and implications for control.

Five groups of dairy cows affected by digital dermatitis were subjected to five different footbath strategies and evaluated at regular 3-weekly intervals. A standard protocol was used to record five different stages of disease from early (M1), acute ulcerative (M2), healing (M3) and chronic lesions (M4) in addition to the negative stage of disease (M0). The effect of the footbathing was evaluated using mathematical modelling for the transmission dynamics of infections and summarized using the reproduction ratio R(0). Sensitivity analysis for a range of parameters in the mathematical model showed that the speed of detecting acute lesions and the efficiency with which those lesions were treated were the key parameters which determined whether lesions became more severe or whether they healed.

Host and environmental reservoirs of infection for bovine digital dermatitis treponemes.

Bovine digital dermatitis (BDD) is a global infectious disease causing lameness of cattle and is responsible for substantial animal welfare issues and economic losses. The causative agents are considered to be spirochetal bacteria belonging to the genus Treponema, which have consistently been identified in BDD lesions worldwide. One potential means of controlling infection is the disruption of transmission; however, the infection reservoirs and transmission routes of BDD treponemes have yet to be elucidated. To address these issues, we surveyed for evidence of BDD treponeme presence in the dairy farm environment, in bovine tissues and in bovine gastrointestinal (GI) tract contents. A total of 368 samples were tested using PCR assays specific for each of three currently recognised, isolated phylotypes of BDD treponemes. All environmental samples, together with insects and GI tract content samples were negative for BDD treponeme DNA from the three phylotypes. However, we identified BDD treponemes in two non-pedal bovine regions: the oral cavity (14.3% of cattle tested) and the rectum (14.8% of cattle tested). Whilst only single phylotypes were detected in the oral cavity, two of the rectal tissues yielded DNA from more than one phylotype, with one sample yielding all three BDD treponeme phylotypes. Whilst it might be considered that direct skin to skin contact may be a major transmission route of BDD treponemes, further studies are required to characterise and determine the potential contribution of oral and rectal carriage to BDD transmission.