Papillomatous Digital Dermatitis Research Articles

Transcellular penetration of Treponema phagedenis isolated from papillomatous digital dermatitis in polarized normal human epidermal keratinocytes in vitro.

Papillomatous digital dermatitis (PDD) is a polymicrobial infection causing lameness in dairy cattle. Culture-independent analysis has shown that Treponema phagedenis is present consistently and predominantly in the lesions. However, the pathogenesis of PDD, especially the tissue penetration pathway, has not been examined. In the present study, we investigated whether T. phagedenis strains isolated from PDD produce proteolytic enzyme (s) for disruption of the epithelial cell barrier and have the ability to translocate in polarized normal human epidermal keratinocytes (NHEK) in vitro. Ten strains of T. phagedenis isolated from lesions did not show proteolytic activity on modified skim milk agar, although a human strain of T. denticola used as a control showed such activity. The integrity of tight junctions was monitored by measurement of transepithelial electrical resistance (TER). The TER values after inoculation of the T. phagedenis strains examined did not change during the experimental period; however, apical to basolateral translocation of T. phagedenis was confirmed after 24 hr by microscopy and Treponema-specific PCR. We further confirmed that translocation of T. phagedenis was accelerated by co-inoculation with live T. denticola, but not with heat-killed organisms. Furthermore, tight junction ZO-1 protein was not lost intensity after inoculation with T. phagedenis and the organism was observed in NHEK cells using a florescence microscope. These results suggest that T. phagedenis strains may translocate via a transcellular route in vitro and that the invasion is accelerated by other bacteria, such as T. denticola, producing proteolytic activity.

65 Health, management, and future directions for indoor cattle feeding facilities

Abstract The use of soil-surfaced pens with limited over-head or enclosed structures have declined in popularity in the Midwest over the last few decades. Various facility types that utilize complete or partial concrete flooring and side walls have increased in popularity. These changes are driven by 1) readily observable impacts that precipitation and pen conditions have on cattle performance and cost of gain and 2) continual pressure to be compliant with environmental regulations. Cost of indoor cattle feeding facilities can easily approach 2-4x the cost of soil-surfaced pens but for many, such pens are not an option. Management of production and animal health varies within these facilities and likely interacts with cattle types and their associated mortality risk. Direct animal health and performance comparisons among facility types are incredibly difficult to perform due to the multiple confounders (diet, risk, cattle type) that usually exist. Morbidity that frequently occurs in enclosed systems typically revolves around management of papillomatous digital dermatitis (PDD), foot and leg issues, and stiffness in cattle. Rolled compacted concrete (RCC) has been a new development that may provide a cost-effective option to curb mud in traditionally soil-surfaced facilities. RCC may also substantially reduce the cost of enclosed facilities that rely on traditional concrete.

16S rRNA-based amplicon analysis of changes in the bacterial population in the lesions of papillomatous digital dermatitis in dairy cattle after topical treatment with allyl isothiocyanate.

Papillomatous digital dermatitis (PDD) is a foot disease causing lameness in dairy cattle. It is regarded as a polymicrobial infection, although its etiology is not fully understood. PDD is treated by the topical or systemic administration of antibiotics such as lincomycin (LCM); however, the milk of the cows cannot be marketed during the treatment and withdrawal period due to the residual antibiotics in milk. Allyl isothiocyanate (AITC), an extract of Wasabia japonica (known as wasabi or Japanese horseradish) widely employed as a food additive, can be used as an alternative antimicrobial agent that overcomes this problem. We previously showed that AITC is as effective as LCM in PDD treatment. Here, using the samples obtained in the previous clinical study, we analyzed changes in the bacterial population in the PDD-associated microbiota after AITC treatment and compared those with that following LCM treatment by 16S ribosomal RNA (rRNA)-based amplicon analysis. Both treatments induced major changes in the bacterial population, and Treponema species, which have been regarded as the major causative agents of PDD, were efficiently eliminated by both agents. However, the AITC-treated samples exhibited higher diversity compared with pretreatment samples, but this trend was not observed for LCM treatment, probably reflecting different antibacterial activities of the two agents. Importantly, this analysis detected population changes before morphological changes in PDD lesions (clinical signs of healing) became evident, indicating that 16S rRNA-based amplicon analysis represents an efficient strategy for analyzing and monitoring the treatment efficiency of PDD as well as other polymicrobial diseases.

Prospective Randomized Field Study to Determine the Efficacy of a Serpens spp Vaccine Combined with Topical Treatment with Lincomycin HCl for Treatment of Papillomatous Digital Dermatitis (Footwarts) on a California Dairy

Fifty-nine cows with active, painful Papillomatous Digital Dermitis (PDD) were randomly assigned to control (n=29) or vaccinate (n=30) groups on day 1. Cows were enrolled in the study based on the presence of a visible lesions on one or both rear heels and a pain response.

Diagnosis and therapy of feedlot lameness

While not at an incidence level comparative to bovine respiratory disease, lameness in feedlots still presents a significant challenge and results in significant economic loss. These proceedings address the basics of diagnosis and therapy of infectious pododermatitis (foot rot), septic arthritis, toe abscesses (toe tip necrosis syndrome), and papillomatous digital dermatitis (hairy heel wart). The benefits of surgical intervention in selected lameness cases are recognized but are beyond the scope of this presentation. Understanding of the causal factors for each disease is pivotal in minimizing additional cases.

Immunohistochemical and Zymographic Analyses of Matrix metalloproteinase-2 (MMP-2) and Matrix Metalloproteinase-9 (MMP-9) in Bovine Papillomatous Digital Dermatitis

Bovine digital dermatitis (BDD) is an important health issue of dairy cattle. Painful papillomatous tissue proliferations are observed in hind feet of affected animals. The pathogenesis of this papillomatous lesion in BDD is still poorly understood. Matrix metalloproteinases are a group of enzymes for tissue remodeling in both health and disease. In the submitted study, immune expressions of MMP-2 and MMP-9 enzymes and their zymographic activities were evaluated. Higher immune staining of MMP-2 and MMP-9 were found in BDD samples when compared with healthy skin samples. The differences observed in MMP-2 and MMP-9 staining intensity and distributions were statistically significant (p< 0.05, p<0.05 respectively). Zymographic analyses revealed skin samples from both healthy and affected animals had pro-active and active forms of MMP-2 and MMP-9 enzymes. Immunohistochemistry results indicate that significant increase in MMP-2 and MMP-9 enzymes in BDD samples. These results suggest MMP-2 and MMP-9 might have roles in the pathogenesis of BDD.

Biochemical and molecular characterization of Treponema phagedenis-like spirochetes isolated from a bovine digital dermatitis lesion

BackgroundBovine papillomatous digital dermatitis (DD) is the leading cause of lameness in dairy cattle and represents a serious welfare and economic burden. Found primarily in high production dairy cattle worldwide, DD is characterized by the development of an often painful red, raw ulcerative or papillomatous lesion frequently located near the interdigital cleft and above the bulbs of the heel. While the exact etiology is unknown, several spirochete species have been isolated from lesion material. Four isolates of Treponema phagedenis-like spirochetes were isolated from dairy cows in Iowa. Given the distinct differences in host, environmental niche, and disease association, a closer analysis of phenotypic characteristics, growth characteristics, and genomic sequences of T. phagedenis, a human genitalia commensal, and the Iowa DD isolates was undertaken.ResultsPhenotypically, these isolates range from 8.0 to 9.7 μm in length with 6–8 flagella on each end. These isolates, like T. phagedenis, are strictly anaerobic, require serum and volatile fatty acids for growth, and are capable of fermenting fructose, mannitol, pectin, mannose, ribose, maltose, and glucose. Major glucose fermentation products produced are formate, acetate, and butyrate. Further study was conducted with a single isolate, 4A, showing an optimal growth pH of 7.0 (range of 6–8.5) and an optimal growth temperature of 40°C (range of 29°C-43°C). Comparison of partial genomic contigs of isolate 4A and contigs of T. phagedenis F0421 revealed > 95% amino acid sequence identity with amino acid sequence of 4A. In silico DNA-DNA whole genome hybridization and BLAT analysis indicated a DDH estimate of >80% between isolate 4A and T. phagedenis F0421, and estimates of 52.5% or less when compared to the fully sequenced genomes of other treponeme species.ConclusionUsing both physiological, biochemical and genomic analysis, there is a lack of evidence for difference between T. phagedenis and isolate 4A. The description of Treponema phagedenis should be expanded from human genital skin commensal to include being an inhabitant within DD lesions in cattle.

An experimental infection model to induce digital dermatitis infection in cattle

Bovine digital dermatitis (DD), also known as papillomatous digital dermatitis (foot warts), has been recognized as a major cause of lameness in cattle, with important economic and welfare consequences. The evaluation of therapeutic and preventive interventions aiming to control DD infections in dairy cattle is often challenged by the complex multifactorial etiology of the disease. An experimental infection model to induce acute DD lesions in a controlled environment is proposed. The goal was to provide a standard way of reproducing DD infections independent of external factors that could confound the natural course of the disease, such as management practices or infection pressure, resulting in transmission of DD between animals. A group of 4 yearling Holstein heifers free of any clinical evidence of hoof disease was recruited from a commercial dairy farm and housed in an experimental facility in 1 pen with slatted flooring. The hind feet were wrapped to mimic conditions of prolonged moisture (maceration) and reduced access to air (closure) and inoculated at the heel and dewclaw areas with a homogenate of a naturally occurring DD lesion skin biopsy or a culture broth of Treponema spp. After a period of 12 to 25 d, 4 of 6 and 1 of 4 dewclaw areas inoculated with biopsied DD lesion or a Treponema spp. culture, respectively, had gross lesions compatible with DD. Histopathology confirmed the gross diagnosis in the sites inoculated with tissue homogenate. In the site inoculated with Treponema spp. culture broth, histopathology revealed an incipient DD lesion. Treponema spp. were detected by PCR in both naturally occurring DD homogenate and Treponema spp. culture broth inoculation sites. An experimental infection model to induce acute DD in cattle was developed, which may be used to evaluate interventions to control DD and study the pathogenesis of this infectious hoof disease in a controlled manner.

Microbial diversity in bovine papillomatous digital dermatitis in Holstein dairy cows from upstate New York

Papillomatous digital dermatitis (PDD) is one of the most prevalent diseases of cattle, adversely affecting the dairy industry by its negative effect on milk production and reproductive performance. Our objective was to use culture-independent methods to determine the microbial diversity in different strata of PDD lesions of three Holstein dairy cows, analyzing whether major differences exist compared to foot skin of three non-infected cows. Both group-specific 16S rRNA gene PCR-denaturing gradient gel electrophoresis and clone library sequencing of broad-range 16S rRNA gene showed differences between the microbial composition of healthy dairy cows and the different strata of the lesion. The predominant bacterial community in the lesion, regardless of the stratum, consisted of 166 specific phylotypes belonging to seven bacterial phyla. Spirochetes (particularly, treponemes) was the most prominent group detected in PDD deep biopsies and was only found in samples from the lesion. Additionally, one phylotype phylogenetically affiliated with uncultured Euryarchaeota was detected in two strata of the lesion. Sequences from healthy foot skin samples revealed 86 specific phylotypes that were affiliated with Firmicutes and Proteobacteria. Our study corroborates the theory that treponemes are involved in PDD disease etiology and suggests, for the first time, the presence of archaeal members in this particular bovine infection.

Prevalencia y factores de riesgo para dermatitis papilomatosa digital en ganado lechero en la X Region, Chile.

In order to determine herd-level risk factors for papillomatous digital dermatitis (PDD) among dairy cows in Region X, Chile, 3884 lactating cows on 23 dairy farms were screened for PDD lesions. Herd prevalence was determined for each participating farm by examining the feet of all lactating cows during milking. Overail prevalence of PDD for all cows examined was 8.5 % (SE:1.1). Introduction of replacement heifers from another premises was a statistically significant (p

Effect of Paste or Wrap Oxytetracycline Treatment on Papillomatous Digital Dermatitis

Papillomatous digital dermatitis (PDD) is a contagious and painful foot lesion causing lameness in cattle. One of the common treatments for PDD is oxytetracycline powder held onto the hoof with a bandage wrap. The objective of this study was to determine if application of oxytetracycline in a topical paste without bandaging would be as effective as the wrap for the treatment of PDD.

Pharmacokinetics of tetracycline in plasma, synovial fluid and milk using single intravenous and single intravenous regional doses in dairy cattle with papillomatous digital dermatitis

The purpose of this study was to compare the pharmacokinetics of tetracycline in plasma, synovial fluid, and milk following either a single systemic intravenous (i.v.) injection or a single i.v. regional antibiosis (IVRA) administration of tetracycline hydrochloride to dairy cattle with papillomatous digital dermatitis (PDD). To this end, plasma and synovial fluid tetracycline concentrations were compared with the minimal inhibitory concentration (MIC) values of the major bacteria, which are known to cause digital diseases and thus assess its efficacy in PDD. Residual tetracycline concentrations in milk from cows treated by both methods were also determined. Twelve Holstein cows with various stages of PDD were randomly assigned to two groups of six animals. Group 1 received a single systemic i.v. injection of 10 mg/kg of tetracycline hydrochloride. Group 2 received 1000 mg of tetracycline hydrochloride by IVRA of the affected limb. Blood, synovial fluid and milk samples were taken prior to tetracycline administration (time 0 control), and then at 22, 45 and 82 min, and 2, 3, 4, 6, 8, 12, 24, 48, 72, 96, and 120 h following drug administration. Tetracycline concentrations were determined by high-performance liquid chromatography. Mean tetracycline plasma and milk concentrations in Group 1 were higher than Group 2. The opposite was observed for synovial fluid concentrations. Group 2 synovial fluid concentrations were higher than the MIC value over 24 h for the bacteria most frequently responsible for claw disease. Compared with i.v. administration, IVRA administration of tetracycline produced very high synovial fluid and low plasma and milk concentrations.

Detection of antibodies against Fusobacterium necrophorum and Porphyromonas levii-like species in dairy cattle with papillomatous digital dermatitis

Bovine digital epidermitis involves different pathologies, including PDD, interdigital dermatitis, and foot rot. Bacteriological and molecular biological studies suggest that these are multimicrobial infections. During our study on the isolation of treponemes from biopsies of PDD, colonies producing black pigment were isolated frequently from the primary cultures, suggesting that Porphyromonas species were present. Moreover, 16S rRNA genes of Fusobacterium necrophorum and Porphyromonas levii-like species were detected in the lesions. We therefore determined whether an immunological response could be elicited by a P. levii-like organism isolated from a PDD lesion, as well as two subspecies of F. necrophorum in the sera from cows with and without PDD. A total of 151 serum samples were collected from 85 cows with PDD lesions and 33 cows without lesions on 12 PDD-positive farms and from 33 cows on two PDD-free farms. ELISA data showed that IgG antibody levels against antigens of P. levii-like species and F. necrophorum subsp. necrophorum were significantly higher in cows on PDD-positive farms than in cows on PDD-free farms, regardless of the presence of PDD lesions in the cows on the PDD-positive farms. However, F. necrophorum subsp. funduliforme was present at low levels in both groups. The ELISA results were confirmed by western blot analysis. Furthermore, antigens of these bacteria were detected in PDD-biopsy sections examined by immunohistochemical staining. F. necrophorum subsp. necrophorum and P. levii-like species may be involved in the pathogenesis of PDD.

Corn Silage Growth and Yield after Application of Dairy Manure with High Copper Concentrations

Dairy producers use footbaths containing CuSO4 for the prevention of papillomatous digital dermatitis. After cows (Bos taurus) have passed through the footbath, waste is usually disposed of into manure storages and applied to fields. The objectives of this research were to evaluate the effects of the application of high‐Cu dairy manure on growth and yield of corn (Zea mays L.) for silage and to evaluate the accumulation of Cu in the soil. Manure was collected, CuSO4 was added, and all plots received approximately 105.9 L of manure (53,572 L ha−1). A control (0.38 kg Cu ha−1), medium Cu (9.5 kg Cu ha−1), and high Cu (18.6 kg Cu ha−1) treatments were applied to the same plots in 2006 and 2007. An early maturity and late maturity corn hybrid was used each year. Soil samples from 0 to 15 cm were analyzed for solution, extractable, and total Cu. Soil samples from 15 to 46 cm were analyzed for solution and total Cu. The growth, yield, forage quality, and Cu concentration of corn were not affected by treatments. Soil extractable and total soil Cu increased linearly as Cu application increased (P < 0.001). Soil solution Cu increased as Cu treatment rate increased, but was not linear (P = 0.04). Solution and total soil Cu were not different as sampling depth increased. Two annual applications of high‐Cu dairy manure at rates relative to agronomic practices did not affect corn grown for silage and resulted in no vertical movement of Cu in the soil.

Comparação entre as administrações tópica e sistêmica de oxitetraciclina no tratamento de vacas com dermatite digital papilomatosa

Comparou-se a eficiência dos tratamentos tópico e sistêmico com oxitetraciclina em vacas com dermatite digital papilomatosa (DDP) e determinaram-se a presença de resíduos desse antimicrobiano no leite e sua concentração no líquido sinovial e no plasma. Utilizaram-se o tratamento tópico com oxitetraciclina em pó (grupo 1) e o sistêmico de longa ação (grupo 2) em 16 vacas holandesas em lactação, acometidas por DDP. Obtiveram-se amostras de plasma, líquido sinovial e leite nos momentos: M0, antes dos tratamentos; M1, seis horas após o tratamento e em intervalos de 12 horas até M23 (264 horas pós-tratamentos). Avaliaram-se o grau de claudicação, a extensão da lesão e a concentração da oxitetraciclina pela cromatografia líquida de alta eficiência. Nas vacas do grupo 1, ocorreu redução das lesões e da claudicação, quando comparadas com as do grupo 2. Nenhuma das amostras de leite, de líquido sinovial e de plasma nos animais do grupo 1 foi positiva para oxitetraciclina. As amostras de leite dos animais do grupo 2, entre M1 e M23, apresentaram valores acima do limite máximo residual permitido para esse antimicrobiano. O tratamento tópico foi eficiente no tratamento de DDP, sem produzir resíduos no leite ou concentrações no plasma e no líquido sinovial. O tratamento sistêmico foi ineficiente para DDP, resultando em resíduos no leite, durante a avaliação.

Analysis of the IgG Immune Response toTreponema phagedenis-Like Spirochetes in Individual Dairy Cattle with Papillomatous Digital Dermatitis

Papillomatous digital dermatitis (PDD) is a major infectious disease of the foot skin in dairy cattle. Treponema phagedenis-like spirochetes have been consistently detected in PDD lesions, and antibodies against these organisms have been demonstrated in affected cattle. However, little is known about the dominant antigens recognized by the immune system of affected cattle. Here, we investigated the IgG immune response to T. phagedenis-like isolates by Western blotting with different sera using whole-cell lysates and extracted glycolipid from 18 and 8 isolates, respectively, including those from different cattle on the same or different farms, isolates from different lesions affecting a single cow, and different isolates from the same lesion affecting a single cow. The reactivity of sera in Western blot assays revealed different banding patterns or showed no bands, suggesting that considerable antigenic variations, including glycolipid, may exist among the isolates, even in those from single individuals. With use of a total of 151 serum samples collected from three groups of cattle, i.e., PDD-positive cows on PDD-positive farms (group A), PDD-negative cows on PDD-positive farms (group B), and cows on PDD-free farms (group C), the levels of IgG antibodies against four T. phagedenis-like isolates were measured by enzyme-linked immunosorbent assay (ELISA). The optical density in groups A and B was significantly higher than that in group C, even though the value varied among the antigens used. Therefore, combinations of multiple Treponema species should be used for serological analysis and the development of a suitable vaccine because of antigenic variations.

Antimicrobial Susceptibility of Treponema phagedenis-Like Spirochetes Isolated from Dairy Cattle with Papillomatous Digital Dermatitis Lesions in Japan

The minimal inhibitory concentrations (MICs) of 23 Treponema phagedenis-like spirochetes isolated from dairy cattle with papillomatous digital dermatitis (PDD) lesions in Japan were investigated by a broth microdilution method using 15 antimicrobial agents. Although all MIC values showed a monomodal distribution, the MICs of the antimicrobial agents for 90% (MIC(90)) of the isolates tested varied among the agents examined. The MIC(90) values for penicillin G, ampicillin, and erythromycin were <0.06 microg/ml. In contrast, the MIC(90) values for kanamycin, streptomycin, rifampicin, sulfamethoxazole, trimethoprim, and colistin were >128 microg/ml. Oxytetracycline, lincomycin, enrofloxacin, chloramphenicol, ceftiofur, and gentamicin showed intermediate values, i.e., 0.5~32 microg/ml. The present study suggested that no isolate had acquired resistance to the antimicrobial agents examined, although they may have natural resistance to some agents. Furthermore, the in vitro antimicrobial susceptibility data would provide helpful information for PDD treatment and the development of a selective medium for isolating the organism effectively.

Detection of Treponemes in Canker Lesions of Horses by 16S rRNA Clonal Sequencing Analysis

Equine canker is a chronic pododermatitis of the hoof in horses. Although spirochetes are detectable histopathologically in the lesions, the precise etiology remains unclear. This study reports the 16S rRNA gene sequencing of randomly selected clones based on PCR with Treponema-specific primers, using the canker lesions from two horses and healthy frog and sole from a horse. A total of 114 clones were obtained from the lesions, but no clones were detected in the healthy hoof tissues. The clones from the canker lesions examined were grouped into 19 operational taxonomic units, such as treponemal phylotypes originating from papillomatous digital dermatitis lesions of dairy cattle and as-yet uncultured human oral treponemes, indicating the presence of multiple treponemes in the lesions.

Identification of candidate pathogens of papillomatous digital dermatitis in dairy cattle from quantitative 16S rRNA clonal analysis

Although it is suspected that papillomatous digital dermatitis (PDD), an infectious foot disease of cattle, is caused by multiple bacteria, it remains unclear precisely which ones are involved in the etiology. To study the bacterial community, we used 16S rRNA gene sequencing of randomly selected clones based on PCR with minimum amplification cycles to search for organisms present in PDD lesions but not in healthy foot skin. The nucleotide sequences of 1525 clones from 5 PDD lesions (836 clones) and 4 samples of healthy foot skin (689 clones) were determined and grouped into 316 operational taxonomic units (OTUs) with a cut-off value of >99% sequence identity. Two OTUs, P-01 (143 clones; 100% nucleotide sequence identity with Treponema phagedenis) and P-02 (112 clones; 86% identity with Bacteroidetes), were detected most frequently in all PDD samples examined. In contrast, OTU N-01 (87 clones), showing 99% nucleotide sequence identity with Moraxella phenylpyruvica, was the most prevalent in the normal samples examined. Spirochaetes were detected in only 1 sample. Phylogenetic analysis showed that T. denticola-like and T. phagedenis-like spirochetes were the predominant groups in the PDD lesions. Detection of multiple treponemes and an unknown bacterium close to Bacteroides sp. at high rates by a culture-independent approach could be evidence of the association of these organisms with PDD.

Serum haptoglobin concentrations in dairy cattle with lameness due to claw disorders

In cattle, elevated blood serum concentrations of haptoglobin, an acute phase protein, have been demonstrated in association with several diseases, but not with lameness. Serum haptoglobin was measured in 60 Holstein dairy cattle diagnosed with lameness due to four claw disorders, pododermatitis septica (PS; n=41), pododermatitis circumscripta (PC; n=8), interdigital necrobacillosis (IN; n=7), papillomatous digital dermatitis (PDD; n=4). Haptoglobin was measured on day 1 (0-3 days after lameness was observed but before treatment) and on days 3 and 5. A total of 10 healthy cows served as controls (haptoglobin values <1.0 mg/dL). Each of the claw disorders was associated with elevated haptoglobin on day 1 (PS, PC, IN and PDD: 65.9%, 37.5%, 71.4% and 25.0%, respectively). Trimming and antibiotic treatment led to a reduction in the number of PS and IN cows with increased haptoglobin concentrations, respectively (P<0.05), but trimming did not lead to any reduction in cows with PC. The study showed that lameness due to claw disorders can be associated with a systemic acute phase response and elevated serum haptoglobin in dairy cattle. Based on the course of haptoglobin, treatments seemed effective for all claw disorders except for PC.