Bovine digital dermatitis: natural lesion development and experimental induction

Bovine Digital Dermatitis (DD) is a leading cause of lameness in dairy cattle in the US with more than 70% of herds affected. Despite 40 years of research, the definitive etiologic agent(s) associated with the disease process is unknown. While clinical lesions have been well described, little is known about the macroscopic, microscopic, and bacterial changes associated with the early stages of lesion development from normal skin to clinical lesions. The goal of this dissertation was to describe the temporal changes associated with lesion development in Holstein dairy cattle, particularly early stage lesions, and develop a model for lesion induction. By following a cohort of Holstein dairy cows for a three year period, several important epidemiologic findings were recognized. In the absence of control measures, DD lesions developed at a rate of 4 lesions per 100 cow feet-months, with the average time for a lesion to develop being 133 days. From the recognition of the macroscopic changes that preceded clinical DD lesions, a novel scoring system was developed. While 20% of the feet observations had clinical DD lesions, an additional 55% of observations had lesions that were indicative of early DD lesion development. Biopsies from these different stages of lesion development were submitted for metagenomic analysis using next generation sequencing. The bacterial microbiota of these biopsies was found to progress through a systematic series of changes that correlate with the macroscopic lesion scoring system with the microbiota of each stage being statistically different from other stages. As one of the major goals for these studies was to gain a better understanding of the etiology of disease, an experimental model was needed to test candidate pathogens. Four preliminary studies were conducted to optimize conditions and methodologies for induction of DD lesions that led to a final consensus protocol that was able to induce DD lesions in 95% of Holstein calves within 28 days. The results of these studies support the hypothesis that DD is a polybacterial disease process that undergoes systematic macroscopic, microscopic, and bacterial changes as lesions develop

Evaluating corellations in Salmonella serotypes in swine in four longitudinal dataset

S. enterica serovars surveillance program have been established for many years in the United State of America (USA). Data from long running surveillance programs provides the opportunity to compare prevalence of serotypes within and across surveillance programs, observed patterns and generate hypothesis. To this end, the aim of this project was to estimate the correlation between changes in the yearly changes in serotypes proportions in concurrent years and lagged years from swine, beef and avian longitudinal datasets: (The Iowa State University Veterinary Diagnostic Laboratory (VDL), The National Antimicrobial Resistance Monitoring System ( NARMS animal-based isolates only), compared to data from The Centers for Disease Control (CDC) Laboratory-based Enteric Disease Surveillance (LEDS) Program. The lagged correlations were as follows: a) 1-year lag with the animal data preceding the human data and b) The correlation across a 2-year lag with the human data preceding the animal data

Deep Sequencing Analysis Reveals the Temporal Microbiota Changes Associated with the Development of Bovine Digital Dermatitis

Bovine digital dermatitis (DD) is a leading cause of lameness in dairy cattle throughout the world. Despite 35 years of research, the definitive etiologic agent associated with the disease process is still unknown. Previous studies have demonstrated that multiple bacterial species are associated with lesions, with spirochetes being the most reliably identified organism. This study details the deep sequencing-based metagenomic evaluation of 48 staged DD biopsy specimens collected during a 3-year longitudinal study of disease progression. Over 175 million sequences were evaluated by utilizing both shotgun and 16S metagenomic techniques. Based on the shotgun sequencing results, there was no evidence of a fungal or DNA viral etiology. The bacterial microbiota of biopsy specimens progresses through a systematic series of changes that correlate with the novel morphological lesion scoring system developed as part of this project. This scoring system was validated, as the microbiota of each stage was statistically significantly different from those of other stages (P\u3c 0.001). The microbiota of control biopsy specimens were the most diverse and became less diverse as lesions developed. Although Treponema spp. predominated in the advanced lesions, they were in relatively low abundance in the newly described early lesions that are associated with the initiation of the disease process. The consortium of Treponema spp. identified at the onset of disease changes considerably as the lesions progress through the morphological stages identified. The results of this study support the hypothesis that DD is a polybacterial disease process and provide unique insights into the temporal changes in bacterial populations throughout lesion development

Bacterial Causes of Digital Dermatitis (DD) in Dairy Cattle

Bovine digital dermatitis (DD) is a leading cause of lameness in dairy cattle throughout the world. Despite 35 years of research, the definitive cause of the disease process is still unknown. Previous studies have demonstrated that multiple bacterial species are associated with lesions, with spirochetes being the most reliably identified organism. This study utilized total DNA sequencing of 48 staged DD biopsy specimens collected during a 3-year longitudinal study of disease progression in dairy cattle. Over 175 million sequences were obtained and used to identify the bacterial species that were present in the biopsies. There was no evidence of a fungal or DNA viral causes. The bacterial communities present in the biopsy specimens was found to progress through a systematic series of changes that correlate with the novel visual lesion scoring system developed as part of this project. Although Treponema spp. predominated in the advanced lesions, they were in relatively low abundance in the newly described early lesions that are associated with the initiation of the disease process. The results of this study support the hypothesis that DD is a polybacterial disease process and provide unique insights into the temporal changes in bacterial populations throughout lesion development. These insights are expected to be critical in the development of new treatment strategies and the potential development of effective vaccines. In addition, the development and validation of a lesion scoring system will assist with determining the prognosis of a lesion

A Highly Effective Protocol for the Rapid and Consistent Induction of Digital Dermatitis in Holstein Calves

Bovine Digital Dermatitis (DD) is a leading cause of lameness in dairy cattle. DD is reportedly increasing in prevalence in beef cattle feedlots of the US. The exact etiologic agent(s) responsible for the disease have yet to be determined. Multiple studies have demonstrated the presence of a variety of Treponema spp. within lesions. Attempts to reproduce clinically relevant disease using pure cultures of these organisms has failed to result in lesions that mirror the morphology and severity of naturally occurring lesions. This manuscript details the systematic development of an experimental protocol that reliably induces digital dermatitis lesions on a large enough scale to allow experimental evaluation of treatment and prevention measures. In total, 21 protocols from five experiments were evaluated on their effectiveness in inducing DD lesions in 126 Holstein calves (504 feet). The protocols varied in the type and concentration of inoculum, frequency of inoculation, duration the feet were wrapped, and type of experimental controls need to validate a successful induction. Knowledge gained in the first four experiments resulted in a final protocol capable of inducing DD lesions in 42 of 44 (95%) feet over a 28 day period. All induced lesions were macroscopically and microscopically identified as clinical DD lesions by individuals blinded to protocols. Lesions were also located at the site of inoculation in the palmer aspect of the interdigital space, and induced clinically measurable lameness in a significant portion of the calves. Collectively these results validate the model and provide a rapid and reliable means of inducing DD in large groups of calves

Clinical presentations and antimicrobial susceptibilities of Corynebacterium cystitidis associated with renal disease in four beef cattle

Background Renal disease caused by Corynebacterium cystitidis in beef cattle may be misclassified as Corynebacterium renale, and limited information about C. cystitidis infections in beef cattle currently is available. Objective To describe clinical presentation, diagnosis, minimum inhibitory concentrations (MICs), and outcome of renal disease caused by C. cystitidis in beef cattle. Methods Retrospective case series. Animals Four client-owned beef cattle. Results All affected cattle had anorexia as a primary complaint. Of the 3 that had ante-mortem diagnostic tests performed, all had pyelonephritis based on azotemia in combination with urinalysis and ultrasonographic findings. Cultures yielded C. cystitidis which was identified by biochemical testing, 16S RNA sequencing, and mass spectrometry. All affected cattle deteriorated despite aggressive treatment, indicating that C. cystitidis infections in beef cattle may carry a poor prognosis. Bacterial isolates collected from the 4 cattle showed similarities in MICs for ampicillin, florfenicol, gentamicin, neomycin, sulfadimethoxine, trimethoprim sulfonamide, and tylosin. Conclusions and clinical importance Corynebacterium cystitidis should be considered in the differential diagnosis of cattle with renal disease. Definitive diagnosis of C. cystitidis as compared to C. renale may be challenging

Thermal Emission and Tidal Heating of the Heavy and Eccentric Planet XO-3b

We determined the flux ratios of the heavy and eccentric planet XO-3b to its parent star in the four IRAC bands of the Spitzer Space Telescope: 0.101% +- 0.004% at 3.6 micron; 0.143% +- 0.006% at 4.5 micron; 0.134% +- 0.049% at 5.8 micron and 0.150% +- 0.036% at 8.0 micron. The flux ratios are within [-2.2,0.3, -0.8, -1.7]-sigma of the model of XO-3b with a thermally inverted stratosphere in the 3.6 micron, 4.5 micron, 5.8 micron and 8.0 micron channels, respectively. XO-3b has a high illumination from its parent star (Fp ~(1.9 - 4.2) x 10^9 ergs cm^-2 s^-1) and is thus expected to have a thermal inversion, which we indeed observe. When combined with existing data for other planets, the correlation between the presence of an atmospheric temperature inversion and the substellar flux is insufficient to explain why some high insolation planets like TrES-3 do not have stratospheric inversions and some low insolation planets like XO-1b do have inversions. Secondary factors such as sulfur chemistry, atmospheric metallicity, amounts of macroscopic mixing in the stratosphere or even dynamical weather effects likely play a role. Using the secondary eclipse timing centroids we determined the orbital eccentricity of XO-3b as e = 0.277 +- 0.009. The model radius-age trajectories for XO-3b imply that at least some amount of tidal-heating is required to inflate the radius of XO-3b, and the tidal heating parameter of the planet is constrained to Qp < 10^6 .Comment: Accepted for publications in The Astrophysical Journa

Association between antimicrobial drug class for treatment and retreatment of bovine respiratory disease (BRD) and frequency of resistant BRD pathogen isolation from veterinary diagnostic laboratory samples

Although 90% of BRD relapses are reported to receive retreatment with a different class of antimicrobial, studies examining the impact of antimicrobial selection (i.e. bactericidal or bacteriostatic) on retreatment outcomes and the emergence of antimicrobial resistance (AMR) are deficient in the published literature. This survey was conducted to determine the association between antimicrobial class selection for treatment and retreatment of BRD relapses on antimicrobial susceptibility of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni. Pathogens were isolated from samples submitted to the Iowa State University Veterinary Diagnostic Laboratory from January 2013 to December 2015. A total of 781 isolates with corresponding animal case histories, including treatment protocols, were included in the analysis. Original susceptibility testing of these isolates for ceftiofur, danofloxacin, enrofloxacin, florfenicol, oxytetracycline, spectinomycin, tilmicosin, and tulathromycin was performed using Clinical and Laboratory Standards Institute guidelines. Data were analyzed using a Bayesian approach to evaluate whether retreatment with antimicrobials of different mechanistic classes (bactericidal or bacteriostatic) increased the probability of resistant BRD pathogen isolation in calves. The posterior distribution we calculated suggests that an increased number of treatments is associated with a greater probability of isolates resistant to at least one antimicrobial. Furthermore, the frequency of resistant BRD bacterial isolates was greater with retreatment using antimicrobials of different mechanistic classes than retreatment with the same class. Specifically, treatment protocols using a bacteriostatic drug first followed by retreatment with a bactericidal drug were associated with a higher frequency of resistant BRD pathogen isolation. In particular, first treatment with tulathromycin (bacteriostatic) followed by ceftiofur (bactericidal) was associated with the highest probability of resistant M. haemolytica among all antimicrobial combinations. These observations suggest that consideration should be given to antimicrobial pharmacodynamics when selecting drugs for retreatment of BRD. However, prospective studies are needed to determine the clinical relevance to antimicrobial stewardship programs in livestock production systems

Determination of reference intervals for fluid analysis and cytologic evaluation variables in synovial fluid samples obtained from carpal and tarsal joints in commercial nonlame growing swine

OBJECTIVE To determine reference intervals for total nucleated cell count, total protein concentration, pH, RBC count, and percentages of neutrophils, lymphocytes, and large mononuclear cells in synovial fluid samples (SFSs) obtained from the carpal and tarsal joints of healthy swine. ANIMALS 54 healthy commercial finisher pigs that had no evidence of lameness or gross joint swelling. PROCEDURES Each pig was anesthetized, and SFSs were collected from 1 carpal and 1 tarsal joint for fluid analysis, cytologic evaluation, bacterial culture, and PCR analyses for common swine joint pathogens. Each pig was euthanized after SFS collection, and synovial tissue samples were collected for histologic assessment. If necessary, postmortem SFSs were collected. RESULTS Overall, 37 of 50 tarsal and 46 of 53 carpal SFSs met inclusion criteria of sufficient volume, no gross blood contamination, and negative results of bacterial culture and PCR analyses, and were from joints with histologically normal synovial tissues. For the carpal and tarsal joints, upper reference limits were as follows: total nucleated cell count, 3,281 cells/μL and 2,368 cells/μL, respectively; total protein concentration, 3.6 g/dL and 3.6 g/dL, respectively; pH, 7.2 and 7.0, respectively; RBC count, 0.8 X 106 cells/μL and 0.1 X 106 cells/μL, respectively; and percentage of neutrophils, 46.5% and 33.7%, respectively; percentage of lymphocytes, 40.6% and 56.3%, respectively; and percentage of large mononuclear cells, 92.0% and 95.3%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE Results have provided reference intervals for selected variables in SFSs obtained from the carpal and the tarsal joints of healthy swine, which should be useful in diagnostic investigations of swine lameness and arthritis