OBJECTIVE To determine optimal sample preparation conditions with potassium triiodide (I2KI) and optimal imaging settings for microfocus CT (micro-CT) of excised cat hearts. SAMPLE 7 excised hearts (weight range, 10 to 17.6 g) obtained from healthy adult cats after euthanasia by IV injection of pentobarbital sodium. PROCEDURES Following excision, the hearts were preserved in 10% formaldehyde solution. Six hearts were immersed in 1.25% I2KI solution (n = 3) or 2.5% I2KI solution (3) for a 12-day period. Micro-CT images were acquired at time 0 (prior to iodination) then approximately every 24 and 48 hours thereafter to determine optimal sample preparation conditions (ie, immersion time and concentration of I2KI solution). Identified optimal conditions were then used to prepare the seventh heart for imaging; changes in voltage, current, exposure time, and gain on image quality were evaluated to determine optimal settings (ie, maximal signal-to-noise and contrast-to-noise ratios). Images were obtained at a voxel resolution of 30 μm. A detailed morphological assessment of the main cardiac structures of the seventh heart was then performed. RESULTS Immersion in 2.5% I2KI solution for 48 hours was optimal for sample preparation. The optimal imaging conditions included a tube voltage of 100 kV, current of 150 μA, and exposure time of 354 milliseconds; scan duration was 12 minutes. CONCLUSIONS AND CLINICAL RELEVANCE Results provided an optimal micro-CT imaging protocol for excised cat hearts prepared with I2KI solution that could serve as a basis for future studies of micro-CT for high resolution 3-D imaging of cat hearts.

OBJECTIVE To assess the effect of transrectal palpation (TRP) performed with the fetal membrane slip (FMS) technique for early pregnancy diagnosis on the proportion and type of associated pregnancy losses (PLs) in dairy cattle. ANIMALS 580 healthy pregnant cattle. PROCEDURES Data for artificially inseminated females with 1 or 2 viable embryos detected by transrectal ultrasonography (TRUS) at approximately 30 days of gestation were retrospectively assessed. Cattle were assigned to 1 of 2 groups on the basis of whether they did or did not undergo TRP once between 34 and 41 days of gestation (palpation and control group, respectively). At approximately 45 and 60 days of gestation, all cattle were reevaluated by TRUS; PL was categorized as type I (FMS detectable by TRP and TRUS-confirmed evidence of embryo or fetus degeneration and a functional corpus luteum) or type II (FMS undetectable by TRP and no TRUS-confirmed evidence of an embryo or fetus or of a functional corpus luteum). RESULTS Of the 580 healthy pregnant cattle, 271 underwent TRP and 309 did not. In the palpation and control groups, PL occurred in 40 (14.8%) and 47 (15.2%) cattle, respectively. Among the palpation group's PLs, 17 (43%) were type I and 23 (58%) were type II. Among the control group's PLs, 27 (57%) were type I and 20 (43%) were type II. The prevalance and type of PL did not differ between groups. CONCLUSIONS AND CLINICAL RELEVANCE TRP with the FMS technique for early pregnancy diagnosis did not increase the prevalence of PL in dairy cattle or alert the proportion of type I versus type II PL.

OBJECTIVE To evaluate histologic changes and gene expression patterns in body and limb wounds in horses in response to bacterial inoculation. SAMPLE Wound biopsy specimens from 6 horses collected on days 7, 14, 21, and 27 after excisional wounds (20 wounds/horse) were created over the metacarpal and metatarsal region and lateral thoracic region (body) and then inoculated or not inoculated on day 4 with Staphylococcus aureus and Pseudomonas aeruginosa. PROCEDURES Specimens were histologically scored for the amount of inflammation, edema, angiogenesis, fibrosis organization, and epithelialization. Quantitative PCR assays were performed to quantify gene expression of 10 inflammatory, proteolytic, fibrotic, and hypoxia-related markers involved in wound healing. RESULTS Except for gene expression of interleukin-6 on day 27 and tumor necrosis factor-α on day 14, bacterial inoculation had no significant effect on histologic scores and gene expression. Gene expression of interleukin-1β and -6, serum amyloid A, and matrix metalloproteinase-9 was higher in limb wounds versus body wounds by day 27. Gene expression of cellular communication network factor 1 was higher in limb wounds versus body wounds throughout the observation period. CONCLUSIONS AND CLINICAL RELEVANCE The lack of clear markers of wound infection in this study reflected well-known difficulties in detecting wound infections in horses. Changes consistent with protracted inflammation were evident in limb wounds, and gene expression patterns of limb wounds shared similarities with those of chronic wounds in humans. Cellular communication network factor warrants further investigation and may be useful in elucidating the mechanisms underlying poor limb wound healing in horses.

Salmonella enterica serovar Typhimurium (S. Typhimurium) is one of the most significant zoonotic pathogens that poses a threat to humans. Previous studies have identified that Salmonella-secreted effector K3 (SseK3) is a novel translated and secreted protein of S. Typhimurium. The objective of this study was to determine whether deletion of the sseK3 gene can attenuate the virulence of S. Typhimurium. To do this, we constructed an sseK3 deletion mutant using the double-exchange allele of the suicide plasmid pRE112ΔsseK3 and assessed the virulence and intracellular proliferation of the mutant. The sseK3 deletion mutant exhibited adhesion and invasion properties similar to those of wild-type (WT) S. Typhimurium, although the virulence and intracellular proliferation of the mutant were significantly reduced compared to that of the WT strain. Furthermore, the observed increase in the median lethal dose (LD(50)) reflects a decrease in the pathogenicity of the sseK3 deletion mutant in a murine model. In summary, we concluded that disruption of sseK3 can attenuate the intracellular proliferation and reduce the virulence of S. Typhimurium.

OBJECTIVE To compare analgesic efficacy and fetal effects between transdermal administration of fentanyl and IM administration of buprenorphine in pregnant sheep. ANIMALS 12 healthy pregnant ewes. PROCEDURES Before study initiation, each ewe was confirmed pregnant with a single fetus between 113 and 117 days of gestation. Ewes were randomly assigned to receive buprenorphine (0.01 mg/kg, IM, q 8 h for 48 hours beginning 1 hour before anesthesia induction; n = 6) or fentanyl (a combination of transdermal fentanyl patches sufficient to deliver a dose of 2 μg of fentanyl/kg/h applied between the dorsal borders of the scapulae 24 hours before anesthesia induction; 6). Ewes were anesthetized and underwent a surgical procedure to instrument the fetus with an arterial catheter and place a catheter in utero for collection of amniotic fluid samples. Physiologic variables and behavioral changes indicative of pain were assessed, and amniotic fluid and blood samples from ewes and fetuses were collected for determination of drug concentrations at predetermined times. RESULTS Both protocols provided acceptable postoperative analgesia with no adverse effects observed in the ewes or fetuses. Compared with the buprenorphine protocol, the fentanyl protocol induced more profound analgesia, decreased the requirement for isoflurane during surgery, and was associated with a shorter anesthesia recovery time. Fetal indices did not differ significantly between the 2 analgesic protocols. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that both protocols provided acceptable analgesia. However, the fentanyl protocol was superior in regard to the extent of analgesia induced, inhalant-sparing effects, and anesthesia recovery time.

OBJECTIVE To evaluate the analgesic and tissue effects of liposomal bupivacaine administered SC as an abaxial sesamoid nerve block in horses with experimentally induced lameness. ANIMALS 6 healthy mature light-breed horses. PROCEDURES In a randomized crossover study, a circumferential hoof clamp was applied to a forelimb to induce reversible lameness. An abaxial sesamoid nerve block of the lame forelimb was performed by SC perineural injection of 10 mg of liposomal bupivacaine or bupivacaine HCl/site. Quantitative gait data were objectively obtained with a body-mounted inertial sensor system before (baseline) and at 30-minute intervals after treatment. Time to return to 85% of baseline lameness was determined. After a minimum 4-day washout period, procedures were repeated with the alternate limb and treatment. Lastly, the palmar digital nerves and perineural tissues were collected and examined histologically. RESULTS SC perineural injection of liposomal bupivacaine ameliorated forelimb lameness in 5 of 6 horses. The median duration of analgesia was not significantly different between liposomal bupivacaine (4.5 hours) and bupivacaine HCl (3.0 hours). Histologically, mild inflammation was noted in 3 of 10 sites injected with liposomal bupivacaine and in none of the sites injected with bupivacaine HCl. CONCLUSIONS AND CLINICAL RELEVANCE SC perineural injection of 10 mg of liposomal bupivacaine/site ameliorated experimentally induced forelimb lameness in some horses. At milligram-equivalent doses, liposomal bupivacaine had a similar duration of analgesia to that of bupivacaine HCl. Further investigation is required before recommending clinical use of liposomal bupivacaine for nerve blocks in horses.

OBJECTIVE To evaluate the effect of suture caliber on the tensile strength of tenorrhaphies performed with a locking-loop technique in cadaveric canine tendons SAMPLE 60 superficial digital flexor tendons (SDFTs) from 30 cadaveric adult dogs. PROCEDURES Transverse tenotomy was performed, and SDFTs were repaired with a locking-loop technique and polypropylene suture of 5 randomly assigned calibers: size-0, 2-0, 3-0, 4-0, or 5-0 (n = 12 SDFTs/suture caliber). Tendon constructs were tested to failure. Yield, peak, and failure forces and causes of failure were compared between groups. RESULTS Mean ± SD failure force for the constructs was significantly greater with large-caliber suture (size-0: 73.5 ± 3.1 N; size 2-0: 54.4 ± 7.1 N; size 3-0: 28.7 ± 4.9 N; size 4-0: 18.7 ± 3.4 N; and size 5-0: 8.8 ± 2.8 N). The likelihood of construct failure by suture pullout through the tendon substance increased with large-caliber suture (size-0: 12/12), whereas the likelihood of construct failure by suture breakage increased with small-caliber suture (2-0: 10/12; 3-0, 4-0, and 5-0: 12/12 each). CONCLUSIONS AND CLINICAL RELEVANCE Large-caliber suture had greater tensile strength for tenorrhaphies performed with a locking-loop technique in cadaveric canine tendons. Prior to the use of large-caliber suture in patients requiring tenorrhaphy, however, in vivo studies are required to confirm the results obtained here.

OBJECTIVE To compare load-to-failure results for laparoscopic absorbable fixation straps (AFSs) deployed at various angles and for AFSs versus absorbable knotless (barbed) suture when used in simulated total laparoscopic gastropexy (TLG) in specimens from cadaveric dogs. SAMPLE 30 stomach and abdominal body wall specimens. PROCEDURES Specimens were assigned to 1 of 3 groups for use in simulated TLG constructs for comparisons of load-to-failure results for single AFSs deployed at 30°, 60°, or 90° (AFS-angle group; n = 10) or for a gastropexy span of 4 to 5 cm achieved with 3-0 absorbable knotless (barbed) monofilament suture applied in a simple continuous pattern (TLG-1; 10) versus 8 AFSs applied with a deployment angle > 30° (TLG-2; 10). A 1-way ANOVA was used to compare results among AFS deployment angles (30°, 60°, or 90°) and between TLG-1 and TLG-2. RESULTS Mean ± SD load to failure for the AFS-angle group was significantly higher for the AFS deployment angles of 60° (8.00 ± 3.90 N) and 90° (12.71 ± 8.00 N), compared with 30° (5.17 ± 1.90 N). However, no substantial difference was detected in the mean ± SD load to failure for TLG-1 (39.18 ± 7.1 N) versus TLG-2 (31.43 ± 10.86 N). CONCLUSIONS AND CLINICAL RELEVANCE Results of the present study supported the potential use of AFSs in gastropexy in dogs; however, prospective clinical research with adequate long-term follow-up is warranted before recommendations can be made.

Achyranthes japonica Nakai (A. japonica) is a medicinal herb found widely distributed throughout Korea. The biological activities of A. japonica are well-documented and include anti-fungal, anti-inflammatory, and immunity enhancement. The objective of the present study was to investigate the immune-related activities of A. japonica extract in dogs. The extract was acquired by ethanol extraction and purified by filtration. To examine the effect of A. japonica extract on immune cell viability, human lymphocytes, such as Jurkat T-cells and Ramos B-cells, were exposed to the extract. After treatment with the extract, the number of Ramos B-cells was increased, whereas Jurkat T-cells remained unaffected. Griess assay revealed decreased nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated mouse macrophage Raw 264.7 cells after exposure to A. japonica extract. To evaluate the in-vivo effect in dogs, feed containing A. japonica extract was provided to 8 dogs for 2 months. Blood samples were collected before, during, and after consumption of the feed. Peripheral blood mononuclear cells (PBMCs) were isolated from the blood samples and the number of T-cells and B-cells were assessed using flow cytometry with anti-dog fluorescein isothiocyanate (FITC)-conjugated CD3 and anti-dog phycoerythrin (PE)-conjugated CD21 antibodies, respectively. We observed a significant increase in the average number of B-cells in the PBMCs during ingestion of the feed containing A. japonica. In addition, enzyme-linked immunosorbent assay (ELISA) revealed a decrease in the levels of tumor necrosis factor-alpha (TNF-α), a pro-inflammatory cytokine, in 3 out of 8 dogs and increased levels of interleukin-10 (IL-10), an anti-inflammatory cytokine, in 4 out of 8 dogs. Taken together, we believe that these changes indicate that A. japonica extract is beneficial in improving the immunity of dogs by stimulating B-cells and inducing production of anti-inflammatory responses.

OBJECTIVE To characterize antimicrobial prescribing patterns of clinicians and clinical services at a large animal veterinary teaching hospital and identify factors associated with antimicrobial prescribing. ANIMALS All large animals (ie, equids, bovids, sheep, goats, camelids, swine, and cervids) evaluated at the New Bolton Center hospital at the University of Pennsylvania from 2013 through 2018. PROCEDURES In a cross-sectional study design, data on antimicrobial use by clinicians and clinical services were collected from administrative and billing records. Multivariable regression modeling was performed to identify factors associated with antimicrobial prescribing patterns. RESULTS Antimicrobials and critically important antimicrobials of the highest priority were dispensed in 42.1% (9,853/23,428) and 24.0% (2,360/9,853) of visits, respectively, and these proportions differed significantly among clinicians. Per visit, the median (interquartile [25th to 75th percentile] range) number of animal-defined daily doses dispensed was 3.6 (0.8 to 11.1) and the mean (SD) number of antimicrobial classes dispensed was 2.0 (1.3). Patient species, age, affected body system, and duration of hospitalization as well as submission of specimens for bacterial culture were significantly associated with prescribing patterns. CONCLUSIONS AND CLINICAL RELEVANCE The frequency and quantity of antimicrobials prescribed differed significantly among clinicians within and across services, even for animals with clinical signs affecting the same body system. Patient- and visit-level factors explained some but not all of the heterogeneity in prescribing patterns, suggesting that other clinician-specific factors drove such practices. More research is needed to better understand antimicrobial prescribing patterns of clinicians, particularly in situations for which no antimicrobial use guidelines have been established.