OBJECTIVE To investigate the effect of lipopolysaccharide (LPS) on type VII collagen– cleaving matrix metalloproteinases (MMPs) in the lamellar tissue of extracorporeally perfused equine limbs. SAMPLE 10 right forelimbs and 3 left forelimbs collected from 10 adult horses after slaughter at a licensed abattoir. PROCEDURES Extracorporeal perfusion of the isolated equine limbs was performed for 10 hours under physiologic conditions (control-perfused limbs; n = 5) and with the addition of 80 ng of LPS/L of perfusate (LPS-perfused limbs; 5). Lamellar tissue specimens were then collected from the dorsal aspect of the hooves. Additionally, corresponding control specimens were collected from the 3 nonperfused left forelimbs. Immunohistochemical analysis was performed on paraffin-embedded tissue blocks with antibodies against total (latent and active) MMP-1, MMP-2, MMP-8, and MMP-9 as well as antibody against active MMP-9. Intensity of immunohistochemical staining was scored, and stain distribution in the lamellar tissue was noted. RESULTS Staining intensity of total and active MMP-9 was significantly increased in LPS-perfused versus control-perfused limbs. No such difference was identified for MMP-1, MMP-2, and MMP-8. CONCLUSIONS AND CLINICAL RELEVANCE Of the 4 MMPs that are capable of degrading type VII collagen, MMP-9 was the only one for which production increased in the lamellar tissue of isolated equine limbs perfused with versus without a clinically relevant concentration of LPS. These results suggested that MMP-9 may be involved in initiation of pathological changes in lamellar tissue in endotoxin-induced laminitis, whereas MMP-1, MMP-2, and MMP-8 may be less relevant.

Although mesenchymal stem cells (MSCs) are now regarded as a promising cell resource for tissue repair and regeneration, the optimal source of MSCs has not yet been determined. The objective of this study was to provide a theoretical basis for the clinical application of umbilical cord mesenchymal stem cells (UCMSCs) in the future. Umbilical cord is an easily obtainable tissue resource, which is one reason that it has become a candidate resource for mesenchymal stem cells. In this study, we analyzed the biological characteristics of UCMSCs, such as their multiple differentiation and clone-forming ability, through morphological observation, reverse transcription polymerase chain reaction (RT-PCR), growth curve, positive rate test, and immunophenotype. Umbilical cord MSCs were successfully isolated and passaged to 29 generations. The results from RT-PCR showed that UCMSCs were positive for CD29, CD44, CD73, but negative for CD34. The expression of the stem cell marker nucleostemin and tenocyte-related markers showed similar positive results with CD44, CD73, and CD90. In addition, UCMSCs can be induced to differentiate into osteoblasts, adipocytes, or chondrocytes. Our study showed that UCMSCs not only have the ability to self-renew, but also have the potential to differentiate into multiple lineages. In general, we concluded that UCMSCs are a reliable source for use in cell therapy.

OBJECTIVE To investigate the effects of respiratory phase, body position, beam center location, and gastric distention on radiographic assessment of liver size in dogs. ANIMALS 12 Beagles. PROCEDURES Liver length and the ratio of liver length to T11 length were determined on lateral radiographic views obtained with various techniques. Images were acquired at maximal expiration or maximal inspiration, with dogs in right or left recumbency, with the beam centered on the caudal border of the scapula or the 13th rib, and after food was withheld or with gastric distention. Effects on organs adjacent to the liver were assessed with CT. Changes of the thoracic cavity during the respiratory cycle were investigated with fluoroscopy. RESULTS Liver length was significantly greater on radiographs obtained at maximal expiration than at maximal inspiration, but there was no increase in the ratio of liver length to T11 length. Body position, beam center location, and gastric distention did not significantly affect liver size. For CT, location of the spleen and stomach and location or size of the liver did not change markedly between right and left recumbency. Fluoroscopy revealed that thoracic width was less at maximal expiration than maximal inspiration. CONCLUSIONS AND CLINICAL RELEVANCE Liver length was greater at maximal expiration than at maximal inspiration because of a smaller thoracic width. Body position, beam center location, and gastric distention did not affect liver length. The ratio of liver length to T11 length was not significantly affected by any of the factors investigated.

OBJECTIVE To evaluate the effects of drinking nutrient-enriched water (NW) on water intake and indices of hydration in healthy domestic cats fed a dry kibble diet ad libitum. ANIMALS 18 domestic shorthair cats. PROCEDURES Group-housed cats were assigned to tap water (TW; n = 9) or NW (9) groups. All cats received TW at baseline (days −7 to −1). No changes were made to the food-water regimen for the TW group. The NW group received NW instead of TW from days 0 through 10, then received TW and NW in separate bowls (days 11 through 56). Food intake was measured through day 10; liquid consumed by drinking was measured throughout the study. Blood and urine samples were collected at predetermined times for analyses; 48-hour urine collection (days 28 through 30 or 31 through 33) was performed to assess output volume and aid endogenous creatinine-based glomerular filtration rate (GFR) determination. Data were analyzed with linear mixed-effects models. RESULTS Baseline TW and calorie intake were similar between groups. The NW treatment was significantly associated with increased liquid consumption during the treatment phase. Mean urine output was significantly higher in the NW group (15.2 mL/kg/d) than in the TW group (10.3 mL/kg/d). Mean GFR (1.75 vs 1.87 mL/min/kg, respectively) did not differ between groups. Effects of treatment and time were each significant for urine specific gravity and osmolality and urine creatinine, phosphate, and urea nitrogen concentrations, with lower values for the NW group. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that consumption of the NW can increase liquid intake and improve measures of hydration in healthy cats. These effects may offer health benefits to some cats in need of greater water consumption.

The objective of this study was to evaluate the impact of a trap-neuter-return (TNR) event on the size of free-roaming rural cat colonies in Quebec. This prospective randomized, controlled study included 18 cat colonies around barns and stables that were randomly assigned to either a TNR group (10 colonies of 7 to 27 cats; 14.3 cats on average) or a control group (8 colonies of 7 to 26 cats; 14.5 cats on average). The number of cats in each colony was calculated from the images obtained by camera-trapping at: baseline (T0), 7.5 mo (T7), and 12 mo (T12). At baseline, the TNR group was subjected to a TNR event. When taking into account adults only, a significant growth difference was observed in the number of cats between the TNR group and the control group at T7 (P = 0.03). When including kittens as well as adults, a trend towards a lower growth of the TNR group compared to the control group was noted at T7 (P = 0.06). There was no difference in the number of kittens between the 2 groups at T7 (P = 0.49) or at T12 (P = 0.36). There was a trend towards more emigration in the control group at T12 (P = 0.095). Isolated TNR events have a low and temporary impact on colony size in Quebec's rural cat colonies.

OBJECTIVE To determine plasma concentrations of lidocaine after laryngeal administration or laryngeal and intratesticular administration in cats. ANIMALS 14 healthy adult sexually intact male cats (7 cats/treatment). PROCEDURES Cats were randomly allocated to receive 0.1 mL of 2% or 10% lidocaine hydrochloride solution (treatments L2 and L10, respectively) sprayed on the larynx for laryngeal desensitization, followed by endotracheal intubation and isoflurane anesthesia. After a 7-day washout period, cats were again randomly allocated to receive treatment L2 or L10, and castration was performed under isoflurane anesthesia following intratesticular administration of 2% lidocaine solution (0.1 mL/kg). In both experiments, a blood sample for measurement of plasma lidocaine concentration was obtained before (0 minutes) and 3, 5, 10, 15, 20, 30, 45, 60, and 75 minutes after laryngeal administration of lidocaine solution. Anesthesia was discontinued at 60 minutes. Plasma lidocaine concentrations were measured with high-performance liquid chromatography. RESULTS After treatments L2 and L10, median maximum plasma lidocaine concentrations were 34.1 ng/mL (range, 0 to 279.4 ng/mL) and 93.6 ng/mL (range, 79.3 to 182.2 ng/mL), respectively. Time to maximum plasma concentration was 10 minutes (range, 0 to 20 minutes) for each treatment. When cats received intratesticular lidocaine administration following L2 or L10 treatment, median maximum plasma concentration was 181.0 ng/mL (range, 103.7 to 600.2 ng/mL) and 301.2 ng/mL (range, 265.8 to 1,770.0 ng/mL), respectively. CONCLUSIONS AND CLINICAL RELEVANCE On the basis of these data, combined laryngeal and intratesticular administration of lidocaine solution at a total dose of approximately 5 mg/kg appears to be safe for use in healthy adult cats.

Salmonella is an important cause of foodborne illnesses in humans. Food-producing animals, including swine, are a major source of Salmonella in food products. This study investigated on farm Salmonella fecal shedding in pigs at different production stages - from weaning to marketing - and its association with the presence of Salmonella in tissues at slaughter. Fourteen groups from 8 commercial farrowing sources (N = 809 pigs) were monitored 5 times from birth to slaughter. Fecal and tissue samples were collected from pigs and cultured for Salmonella. A survey was conducted to collect farm management information. A multi-level mixed-effects logistic regression modelling method was used to analyze Salmonella shedding over time and the association between Salmonella shedding and the presence of Salmonella in tissue samples. Salmonella was recovered from 13% (421/3339) of fecal samples collected from 809 pigs over the course of the study. Overall, 35% (284) of pigs shed Salmonella at least once, while 12% (99) shed more than once. Salmonella shedding increased as pigs aged (P = 0.01) and increased in the summer months (P < 0.01). Salmonella was isolated from tissue samples collected from 23% (134/580) of pigs; however, the presence of Salmonella at slaughter was not associated with on farm shedding. The seasonal trend in Salmonella shedding and its association with age may be used to identify high-risk groups and implement more effective control measures accordingly. The identification of repeat shedders warrants interventions that target this source of infection on swine farms.

OBJECTIVE To assess effects of photobiomodulation, silver sulfadiazine, and a topical antimicrobial product for the treatment of experimentally induced full-thickness skin wounds in green iguanas (Iguana iguana). ANIMALS 16 healthy subadult green iguanas. PROCEDURES Iguanas were anesthetized, and three 5-mm cutaneous biopsy specimens were obtained from each iguana (day 0). Iguanas were randomly assigned to 2 treatment groups, each of which had a control treatment. Wounds in the topical treatment group received silver sulfadiazine, a topical antimicrobial product, or no treatment. Wounds in the laser treatment group received treatment with a class 4 laser at 5 or 10 J/cm2 or no treatment. Wound measurements were obtained daily for 14 days. Iguanas were euthanized, and treatment sites were evaluated microscopically to detect ulceration, bacterial contamination, reepithelialization, necrosis, inflammation, fibrosis, and collagen maturity. RESULTS On day 14, wounds treated with a laser at 10 J/cm2 were significantly smaller than those treated with silver sulfadiazine, but there were no other significant differences among treatments. Histologically, there were no significant differences in ulceration, bacterial infection, reepithelialization, necrosis, inflammation, fibrosis, and collagen maturity among treatments. CONCLUSIONS AND CLINICAL RELEVANCE Photobiomodulation at 10 J/cm2 appeared to be a safe treatment that was tolerated well by green iguanas, but it did not result in substantial improvement in histologic evidence of wound healing, compared with results for other treatments or no treatment.

OBJECTIVE To evaluate in vitro phagocytosis and bactericidal activity of circulating blood neutrophils in horses with severe equine asthma and control horses and to determine whether circulating blood neutrophils in horses with severe equine asthma have an increase in expression of the proinflammatory cytokine tumor necrosis factor (TNF)-α and the chemokine interleukin (IL)-8 and a decrease in expression of the anti-inflammatory cytokine IL-10 in response to bacteria. ANIMALS 6 horses with severe equine asthma and 6 control horses. PROCEDURES Circulating blood neutrophils were isolated from horses with severe equine asthma and control horses. Phagocytosis was evaluated by use of flow cytometry. Bactericidal activity of circulating blood neutrophils was assessed by use of Streptococcus equi and Streptococcus zooepidemicus as targets, whereas the cytokine mRNA response was assessed by use of a quantitative PCR assay. RESULTS Circulating blood neutrophils from horses with severe equine asthma had significantly lower bactericidal activity toward S zooepidemicus but not toward S equi, compared with results for control horses. Phagocytosis and mRNA expression of TNF-α, IL-8, and IL-10 were not different between groups. CONCLUSIONS AND CLINICAL RELEVANCE Impairment of bactericidal activity of circulating blood neutrophils in horses with severe equine asthma could contribute to an increased susceptibility to infections.

OBJECTIVE To quantitatively analyze brain perfusion parameters in dogs with idiopathic epilepsy (IE) by use of MRI and to compare those findings with brain perfusion parameters for healthy dogs. ANIMALS 12 client-owned dogs with IE. PROCEDURES For each dog, standard MRI and perfusion-weighted imaging (before and after injection of gadoteric acid contrast medium) sequences of the brain were obtained during the interictal period by means of the same protocol used in a comparable study of healthy dogs. Time of contrast medium arrival, time to peak contrast enhancement, mean contrast transit time, and cerebral blood flow were calculated for the caudate nucleus, thalamus, piriform lobe, hippocampus, semioval center, and temporal cerebral cortex. Parameters for each structure were compared between dogs with IE and healthy dogs. RESULTS Dogs with IE had a significantly greater mean time of contrast arrival and lower mean cerebral blood flow than healthy dogs. Differences in cerebral blood flow between dogs with IE and healthy dogs were most pronounced in the piriform lobe, thalamus, and temporal cerebral cortex. The mean contrast transit time did not differ between dogs with IE and healthy dogs. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that, compared with healthy dogs, dogs with IE have decreased blood perfusion of the brain. Findings of this study can be used as a basis for further research into functional changes within the brains of epileptic dogs during the interictal phase.