This study quantified the effect of peripartum reproductive disorders and parity on repeat breeder status and involuntary culling of dairy cows. Reproductive data of 418 383 lactations were taken from a computerized databank of health records for dairy cows. A logistic regression model was used with dystocia, retained placenta (RP), metritis complex, and parity as fixed effect risk factors and herd entered as the random effect. Of the peripartum problems studied, dystocia had the greatest effect on future fertility. Dystocia increased the odds of a cow being a repeat breeder by 44% [odds ratio (OR): 1.44; confidence interval (CI): 1.37 to 1.51]. Compared to first-parity cows, cows in second, third, and fourth parities had significantly higher odds of being a repeat breeder: 18% (OR: 1.18; CI: 1.16 to 1.20); 24% (OR: 1.24; CI: 1.21 to 1.26); and 42% (OR: 1.42; CI: 1.39 to 1.45), respectively. The odds for second-, third-, or fourth-parity repeat breeders being culled were 24% (OR: 1.24; CI: 1.20 to 1.28); 39% (OR: 1.39; CI: 1.35 to 1.43); and 67% (OR: 1.67; CI: 1.62 to 1.71) respectively, while peripartum reproductive problems had less of an effect.

Escherichia coli O157:H7 remains a threat to humans via cattle-derived fecal contamination of food and water. Preharvest intervention strategies represent a means of reducing the pathogen burden before harvest. In this study, the efficacy of a commercially produced type III secreted protein (TTSP) vaccine was evaluated with the use of a commingled experimental calf infection model (30 placebo-treated animals and 30 vaccinates). The calves were vaccinated on days 0, 21, and 42 and infected with 109 colony-forming units (CFU) of E. coli O157 by oral–gastric intubation on day 56. Fecal shedding was monitored daily for 14 d. Serologic assessment revealed a robust immune response to vaccination; the serum titers of antibodies against EspA, Tir, and total TTSPs were significantly higher in the vaccinates than in the placebo-treated animals on days 21, 42, 56, and 70. Significantly less (P = 0.011) of the challenge organism was shed by the vaccinates than by the placebo-treated animals on days 3 to 10. Peak shedding occurred in both groups on days 3 to 6; during this period the vaccinates showed a mean log reduction of 1.4 (P = 0.002) and a mitigated fraction of 51%. The number of animals shedding was significantly lower among the vaccinates compared with the placebo group on days 3 to 6 (P ≤ 0.05), with a mean prevented fraction of 21%. No differences in the duration of shedding were observed. Owing to the low challenge shedding in both groups on days 11 to 14 (mean CFU/g < 10; median = 0), no significant differences were observed. These data indicate that TTSP vaccination had protective effects through significant reductions in the number of animals shedding and the number of challenge organisms shed per animal and provides evidence that TTSP vaccination is an effective preharvest intervention strategy against E. coli O157.

Objective-To evaluate effects of extracorporeal shock wave therapy (ESWT) and polysulfated glycosaminoglycan treatment (PSGAGT) on subchondral bone (SCB), serum biomarkers, and synovial fluid biomarkers in horses with induced osteoarthritis. Animals-24 healthy 2- to 3-year-old horses. Procedures-An osteochondral fragment was created on the distal aspect of the radial carpal bone in 1 middle carpal joint of each horse. Horses were randomly allocated to receive local application of ESWT (days 14 and 28; n = 8), PSGAGT (IM, q 4 d for 28 days; 8), or a sham ESWT probe (placebo; days 14 and 28; 8). Serum biomarkers were measured every 7 days, and synovial fluid biomarkers were measured every 14 days. Bone density was measured by use of computed tomography on days 0 and 70, and microdamage and bone formation variables were compared among groups at the end of the study (day 70). Results-There was no significant effect of ESWT or PSGAGT on any bone variable. Serum osteocalcin concentration was significantly greater in horses that received ESWT, compared with placebo-treated horses, and serum concentration of the C-terminal telopeptide of type I collagen was significantly higher in horses that received ESWT, compared with placebo- and PSGAG-treated horses. Concentrations of the synovial fluid epitope CS846 were significantly higher in joints with osteoarthritis treated with ESWT Conclusions and Clinical Relevance-Treatment of osteoarthritis with ESWT had no effect on SCB but did induce increases in serum biomarkers indicative of bone remodeling. Treatment of osteoarthritis with PSGAG had no effect on SCB or biomarkers.

Objective-To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses. Sample-Cartilage explants from 5 one-month-old and 3 adult guinea pigs and 5 adult clinically normal horses. Procedures-Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1micromolar [1×]). Additionally, monolayer chondrocytes were exposed to 2× the suggested amount of a proprietary siRNA molecule. Full-thickness cartilage explants were treated with 1× (1micromolar), 2× (2micromolar), and 4× (4micromolar) or 1× (0.13micromolar), 4× (0.52micromolar), and 8× (1.04micromolar) the recommended concentrations of the proprietary siRNA and the cationic liposome siRNA, respectively, in equivalent media volumes. Use of fluorescent siRNA duplexes allowed quantification of transfected cells via flow cytometry and direct visualization of the depth and distribution of in situ transfection via fluorescent microscopy. Results-With both transfection reagents, > 90% of monolayer chondrocytes were transfected. In explants, only use of the proprietary molecule achieved > 50% transfection efficiency, whereas use of the cationic liposome achieved < 20%. Only the proprietary molecule-treated cartilage consistently contained fluorescent cells throughout all zones; the cationic liposome-transfected chondrocytes were restricted to explant surfaces. Conclusions and Clinical RelevanceRobust transfection of chondrocytes in monolayer was achieved with both reagents, but only use of the proprietary molecule attained effective full-thickness transfection of explants that may allow relevant transcript reduction via RNAi.

Objective-To determine the accuracy of pressure plate kinetic asymmetry indices (ASIs) for diagnosis of unilateral hind limb lameness in dogs and their correlation with visual gait assessment (VGA) scores. Animals-9 healthy dogs and 16 dogs with previously diagnosed unilateral rupture of the cranial cruciate ligament and concurrent unilateral hind limb lameness. Procedures-Dogs were walked over a pressure plate to determine paw contact area (PCA), peak vertical pressure (PVP), peak vertical force (PVF), and vertical impulse (VI) of both hind limbs. An ASI was calculated for each gait variable. Simultaneously, gait was assessed visually and scored by use of a numeric rating scale (0 to 10). The ASI of each variable was tested for its usefulness in discrimination between lame and nonlame dogs and for correlation with VGA scores. Results-Sensitivity and specificity of ASIs to discriminate between lame and nonlame dogs were excellent for PVF, VI, and PCA; these values were substantially lower for ASI of PVP. Cutoff values to discriminate between lame and nonlame dogs were determined by use of ASIs for PVF, VI, and PCA; however, this could not be done for ASI of PVP. Correlations between ASIs of PVF, VI, and PCA and VGA scores were higher than correlation between the ASIs of PVP and VGA scores. Conclusions and Clinical Relevance-Results indicated that ASIs of PVF and VI determined via analysis of pressure plate measurements were reliable indicators of clinical lameness in dogs, but the ASI of PVP was not. The ASI of PCA is an interesting new variable for assessment of limb loading symmetry.

Objective—To determine the Helicobacter spp present in the oral cavity of dogs and the relationship of those organisms with gastric Helicobacter spp to better define the potential for dog-human and dog-dog transmission. Sample—Saliva and dental plaque from 28 dogs and gastric biopsy specimens from a subset of 8 dogs. Procedures—PCR-based screening for Helicobacter spp was conducted on samples obtained from the oral cavity of 28 dogs. Comparative analysis was conducted on Helicobacteraceae 16S rDNA clone libraries from the oral cavity and stomach of a subset of 8 dogs (5 vomiting and 3 healthy) that had positive PCR results for Helicobacter spp. Results—Helicobacteraceae DNA was identified in the oral cavity of 24 of 28 dogs. Analysis of cloned 16S rDNA amplicons from 8 dogs revealed that Wolinella spp was the most common (8/8 dogs) and abundant (52/57 [91%] clones) member of the Helicobacteraceae family in the oral cavity. Only 2 of 8 dogs harbored Helicobacter spp in the oral cavity, and 1 of those was coinfected with Helicobacter heilmannii and Helicobacter felis in samples obtained from the stomach and saliva. Evaluation of oral cavity DNA with Wolinella-specific PCR primers yielded positive results for 16 of 20 other dogs (24/28 samples were positive for Wolinella spp). Conclusions and Clinical Relevance—Wolinella spp rather than Helicobacter spp were the predominant Helicobacteraceae in the oral cavity of dogs. The oral cavity of dogs was apparently not a zoonotically important reservoir of Helicobacter spp that were non–Helicobacter pylori organisms.

Objective—To identify a causative mutation for dilated cardiomyopathy (DCM) in Doberman Pinschers by sequencing the coding regions of 10 cardiac genes known to be associated with familial DCM in humans. Animals—5 Doberman Pinschers with DCM and congestive heart failure and 5 control mixed-breed dogs that were euthanized or died. Procedures—RNA was extracted from frozen ventricular myocardial samples from each dog, and first-strand cDNA was synthesized via reverse transcription, followed by PCR amplification with gene-specific primers. Ten cardiac genes were analyzed: cardiac actin, α-actinin, α-tropomyosin, β-myosin heavy chain, metavinculin, muscle LIM protein, myosinbinding protein C, tafazzin, titin-cap (telethonin), and troponin T. Sequences for DCM-affected and control dogs and the published canine genome were compared. Results—None of the coding sequences yielded a common causative mutation among all Doberman Pinscher samples. However, 3 variants were identified in the α-actinin gene in the DCM-affected Doberman Pinschers. One of these variants, identified in 2 of the 5 Doberman Pinschers, resulted in an amino acid change in the rod-forming triple coiled-coil domain. Conclusions and Clinical Relevance—Mutations in the coding regions of several genes associated with DCM in humans did not appear to consistently account for DCM in Doberman Pinschers. However, an α-actinin variant was detected in some Doberman Pinschers that may contribute to the development of DCM given its potential effect on the structure of this protein. Investigation of additional candidate gene coding and noncoding regions and further evaluation of the role of α-actinin in development of DCM in Doberman Pinschers are warranted.

Objective—To investigate the density of the primary epidermal lamellae (PEL) around the solar circumference of the forefeet of near-term fetal feral and nonferal (ie, domesticated) horses. Sample—Left forefeet from near-term Australian feral (n = 14) and domesticated (4) horse fetuses. Procedures—Near-term feral horse fetuses were obtained from culled mares within 10 minutes of death; fetuses that had died in utero 2 weeks prior to anticipated birth date and were delivered from live Thoroughbred mares were also obtained. Following disarticulation at the carpus, the left forefoot of each fetus was frozen during dissection and data collection. In a standard section of each hoof, the stratum internum PEL density was calculated at the midline center (12 o'clock) and the medial and lateral break-over points (11 and 1 o'clock), toe quarters (10 and 2 o'clock), and quarters (4 and 6 o'clock). Values for matching lateral and medial zones were averaged and expressed as 1 density. Density differences at the 4 locations between the feral and domesticated horse feet were assessed by use of imaging software analysis. Results—In fetal domesticated horse feet, PEL density did not differ among the 4 locations. In fetal feral horse feet, PEL density differed significantly among locations, with a pattern of gradual reduction from the dorsal to the palmar aspect of the foot. The PEL density distribution differed significantly between fetal domesticated and feral horse feet. Conclusions and Clinical Relevance—Results indicated that PEL density distribution differs between fetal feral and domesticated horse feet, suggestive of an adaptation of feral horses to environment challenges.

Objective—To evaluate the impact of oxytetracycline exposure on horizontal transfer of an antimicrobial resistance plasmid. Sample—Populations of Salmonella enterica subsp enterica serovar Typhimurium and Escherichia coli. Procedures—Mixed populations of plasmid donor (Salmonella Typhimurium) and recipient (E coli) bacteria were assigned to 1 of 2 simulated oxytetracycline dosing regimens (high peak concentration-short elimination half-life [HC-SHL] or low peak concentration—long elimination half-life [LC-LHL]) or served as untreated control replicates. Donor, recipient, and transconjugant (E coli that acquired the plasmid) bacteria populations were quantified at 12, 24, and 36 hours after oxytetracycline administration by use of culture on selective bacterial growth media. Results—The ratio of transconjugant to donor bacteria was significantly reduced in the oxytetracycline-exposed replicates, compared with the ratio for the control replicates, at 12 hours. At 24 and 36 hours, results for the HC-SHL regimens were not significantly different from results for the respective control replicates, and results for the LC-LHL regimens also were not significantly different from results for the respective control replicates. The oxytetracycline concentration at these time points (12 hours in the HC-SHL regimen and all 3 time points in the LC-LHL regimen) were in excess of the minimum inhibitory concentration of the recipient bacteria. Conclusions and Clinical Relevance—Transfer of antimicrobial resistance plasmids may be suppressed in vitro by oxytetracycline exposure at concentrations greater than the minimum inhibitory concentration of the recipient bacteria.

Objective—To evaluate changes in the cortical bone of the proximal phalanx of the fore-limbs of Thoroughbreds in response to training. Animals—Twenty-seven 2-year-old Thoroughbreds (20 females, 2 males, and 5 geldings). Procedures—Horses were principally in training for races in a straight line and in a clockwise direction. Lateromedial and dorsopalmar radiographic views of each metacarpophalangeal joint were obtained before the horses started training and 1 year after starting exercise and racing. Width of the dorsal, palmar, lateral, and medial cortex and the width and thickness of the medulla were measured. Ratios (rather than absolute values) were used to remove the effect of differences in bone size among horses. Results—10 horses were lost from the study. Radiographs were obtained for 17 horses 1 year after starting training (9 horses raced in a clockwise direction, and 8 raced in clockwise and counterclockwise directions). There was no difference between the cortical bone in the right and left forelimbs at the start of the study. After training for 1 year, the palmar cortex in the right forelimb was significantly thicker than that in the left forelimb. Conclusions and Clinical Relevance—The strain patterns, biomechanics of rapid exercise, and type of training most probably determined differences in the adaptive responses of the proximal phalanx. The data reported here can be used in the evaluation of weight-bearing distribution along the proximal phalanx and evaluation of the relationship between exercise and bone remodelling of the proximal phalanx.