Humoral immune responses and protective efficacy by various doses of Salmonella ghost cells carrying enterotoxigenic Escherichia coli (ETEC) fimbrial antigens for protection against piglet colibacillosis were studied. All groups were orally primed and boosted at 11 and 14 wk of pregnancy, respectively. Group A sows were inoculated with phosphate-buffered saline (PBS), and groups B, C, and D sows were immunized with 2 × 10(9) 2 × 10(10), and 2 × 10(11) ghost cells, respectively. Serum immunoglobulin (Ig)G, and colostrum (Ig)G and (Ig)A levels of groups C and D sows were significantly higher than those of group A sows. In addition, serum (Ig)G and (Ig)A levels in group C and D piglets were significantly increased compared to those of group A piglets. After challenge with wild-type ETEC, diarrhea and mortality were not observed in group C and D piglets, while diarrhea was observed in 88.9% and 58.8% of groups A and B piglets, respectively, and 16.7% mortality was observed in group A piglets. These findings indicate that oral immunization of sows with 2 × 1010 or 1011 ghost cells can effectively protect their offspring from colibacillosis.

OBJECTIVE To compare antibacterial effects among 3 types of foam used with negative-pressure wound therapy (NPWT) in an ex vivo equine perfused wound model. SAMPLES Abdominal musculocutaneous flaps from 6 equine cadavers. PROCEDURES Each musculocutaneous flap was continuously perfused with saline (0.9% NaCl) solution. Four 5-cm circular wounds were created in each flap and contaminated with 106 CFUs of both Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus (MRSA). After a 1-hour incubation period, 1 of 4 treatments (NPWT with silver-impregnated polyurethane foam [NPWT-AgPU], polyurethane foam [NPWT-PU], or polyvinyl alcohol foam [NPWT-PVA] or a nonadherent dressing containing polyhexamethylene biguanide without NPWT [control]) was randomly applied to each wound. An 8-mm punch biopsy specimen was obtained from each wound immediately before and at 6, 12, 18, and 24 hours after treatment application to determine the bacterial load for both P aeruginosa and MRSA. RESULTS The bacterial load of P aeruginosa for the NPWT-PVA treatment was significantly lower than that for the other 3 treatments at each sampling time after application, whereas the bacterial load for the NPWT-AgPU treatment was significantly lower than that for the NPWT-PU and control treatments at 12 hours after application. The bacterial load of MRSA for the NPWT-PVA treatment was significantly lower than that for the other 3 treatments at each sampling time after application. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that wounds treated with NPWT-PVA had the greatest decrease in bacterial load; however, the effect of that treatment on wound healing needs to be assessed in vivo.

OBJECTIVE To assess the ability to regenerate an equine meniscus by use of a collagen repair patch (scaffold) seeded with mesenchymal stem cells (MSCs) derived from bone marrow (BM) or adipose tissue (AT). SAMPLE 6 female Hispano-Breton horses between 4 and 7 years of age; MSCs from BM and AT were obtained for the in vitro experiment, and the horses were subsequently used for the in vivo experiment. PROCEDURES Similarities and differences between MSCs derived from BM or AT were investigated in vitro by use of cell culture. In vivo assessment involved use of a meniscus defect and implantation on a scaffold. Horses were allocated into 2 groups. In one group, defects in the medial meniscus were treated with MSCs derived from BM, whereas in the other group, defects were treated with MSCs derived from AT. Defects were created in the contralateral stifle joint but were not treated (control samples). RESULTS Both types of MSCs had universal stem cell characteristics. For in vivo testing, at 12 months after treatment, treated defects were regenerated with fibrocartilaginous tissue, whereas untreated defects were partially repaired or not repaired. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that MSCs derived from AT could be a good alternative to MSCs derived from BM for use in regenerative treatments. Results also were promising for a stem cell-based implant for use in regeneration in meniscal lesions. IMPACT FOR HUMAN MEDICINE Because of similarities in joint disease between horses and humans, these results could have applications in humans.

OBJECTIVE To evaluate optimal isolation of endothelial colony-forming cells (ECFCs) from peripheral blood of horses. SAMPLE Jugular and cephalic venous blood samples from 17 adult horses. PROCEDURES Each blood sample was divided; isolation was performed with whole blood adherence (WBA) and density gradient centrifugation (DGC). Isolated cells were characterized by uptake of 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine perchlorate–labeled acetylated low-density lipoprotein (DiI-Ac-LDL), vascular tubule formation, and expression of endothelial (CD34, CD105, vascular endothelial growth factor receptor-2, and von Willebrand factor) and hematopoietic (CD14) cell markers by use of indirect immunofluorescence assay (IFA) and flow cytometry. RESULTS Colonies with cobblestone morphology were isolated from 15 of 17 horses. Blood collected from the cephalic vein yielded colonies significantly more often (14/17 horses) than did blood collected from the jugular vein (8/17 horses). Of 14 cephalic blood samples with colonies, 13 were obtained with DGC and 8 with WBA. Of 8 jugular blood samples with colonies, 8 were obtained with DGC and 4 with WBA. Colony frequency (colonies per milliliter of blood) was significantly higher for cephalic blood samples and samples isolated with DGC. Cells formed vascular tubules, had uptake of DiI-Ac-LDL, and expressed endothelial markers by use of IFA and flow cytometry, which confirmed their identity as ECFCs. CONCLUSIONS AND CLINICAL RELEVANCE Maximum yield of ECFCs was obtained for blood samples collected from both the jugular and cephalic veins and use of DGC to isolate cells. Consistent yield of ECFCs from peripheral blood of horses will enable studies to evaluate diagnostic and therapeutic uses.

OBJECTIVE To evaluate the effect of IM administration of a tiletamine hydrochloride–zolazepam hydrochloride (TZ) combination with either dexmedetomidine hydrochloride or saline (0.9% NaCl) solution (SS) on the motor response to claw clamping, selected cardiorespiratory variables, and quality of recovery from anesthesia in alpacas. ANIMALS 5 adult sexually intact male alpacas. PROCEDURES Each alpaca was given the TZ combination (2 mg/kg) with dexmedetomidine (5 [D5], 10 [D10], 15 [D15], or 20 [D20] µg/kg) or SS IM at 1-week intervals (5 experiments); motor response to claw clamping was assessed, and characteristics of anesthesia, recovery from anesthesia, and selected cardiorespiratory variables were recorded. RESULTS Mean ± SEM duration of lack of motor response to claw clamping was longest when alpacas received treatments D15 (30.9 ± 5.9 minutes) and D20 (40.8 ± 5.9 minutes). Duration of lateral recumbency was significantly longer with dexmedetomidine administration. The longest time (81.3 ± 10.4 minutes) to standing was observed when alpacas received treatment D20. Following treatment SS, 4 alpacas moved in response to claw clamping at the 5-minute time point. Heart rate decreased from pretreatment values in all alpacas when dexmedetomidine was administered. Treatments D10, D15, and D20 decreased Pao2, compared with treatment SS, during the first 15 minutes. During recovery, muscle stiffness and multiple efforts to regain a sternal position were observed in 3 SS-treated and 1 D5-treated alpacas; all other recoveries were graded as excellent. CONCLUSIONS AND CLINICAL RELEVANCE In TZ-anesthetized alpacas, dexmedetomidine (10, 15, and 20 µg/kg) administered IM increased the duration of lack of motor response to claw clamping, compared with the effect of SS.

The only way to diagnose hepatic fibrosis in dogs is by histological assessment of a liver biopsy specimen. As this technique is invasive and susceptible to sampling variation, serum biomarkers are used to detect hepatic fibrosis in humans. The objective of this study was to assess the utility of hyaluronic acid (HA), procollagen type III N-terminal peptide (PIIINP), and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) as serum markers of canine hepatic fibrosis. Serum samples were collected from 47 dogs with histologically confirmed hepatobiliary disease and 24 healthy dogs in order to measure concentrations of HA, PIIINP, and TIMP-1. Hepatic fibrosis was staged using a 5-point scoring scheme. There was no correlation between serum concentrations of HA or PIIINP and the severity of hepatic fibrosis. There was a negative correlation between serum concentration of TIMP-1 and the severity of hepatic fibrosis (r s = −0.33; P = 0.036). It was not possible to use serum concentrations of HA, PIIINP, or TIMP-1 to discriminate between dogs with absent-to-moderate hepatic fibrosis and those with marked-to-very-marked fibrosis. The results of this study do not support the utility of measuring serum concentrations of HA, PIIINP, or TIMP-1 for diagnosing canine hepatic fibrosis. Further studies are needed to support this finding.

OBJECTIVE To develop and assess the feasibility, repeatability, and safety of an ultrasound-guided technique to stimulate the first cervical nerve (FCN) at the level of the alar foramen of the atlas of horses. ANIMALS 4 equine cadavers and 6 clinically normal Standardbreds. PROCEDURES In each cadaver, the FCN pathway was determined by dissection, and any anastomosis between the first and second cervical nerves was identified. Subsequently, each of 6 live horses underwent a bilateral ultrasound-guided stimulation of the FCN at the alar foramen 3 times at 3-week intervals. After each procedure, horses were examined daily for 5 days. RESULTS In each cadaver, the FCN passed through the alar foramen; a communicating branch between the FCN and the accessory nerve and anastomoses between the ventral branches of the FCN and second cervical nerve were identified. The anastomoses were located in the upper third of the FCN pathway between the wing of the atlas and the nerve's entry in the omohyoideus muscle. Successful ultrasound-guided electrical stimulation was confirmed by twitching of the ipsilateral omohyoideus muscle in all 6 live horses; this finding was observed bilaterally during each of the 3 experimental sessions. No complications developed at the site of stimulation. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that ultrasound-guided stimulation of the FCN at the alar foramen appears to be a safe and straightforward procedure in horses. The procedure may have potential for use in horses with naturally occurring recurrent laryngeal neuropathy to assess reinnervation after FCN transplantation or nerve-muscle pedicle implantation in the cricoarytenoideus dorsalis muscle.

The sedative effect of acepromazine combined with 2 doses of tramadol [3 and 5 mg/kg body weight (BW)] was compared with the sedative effect of acepromazine alone in dogs and the effects of each sedative protocol on cardiorespiratory variables were examined. This was a prospective, randomized, blinded, crossover study. Each of 6 dogs received 3 treatments at 1-week intervals. During all anesthetic episodes, dogs received 0.05 mg/kg BW acepromazine. Approximately 25 min later, dogs were given physiological saline (control) or tramadol [3 mg/kg BW (TR3) or 5 mg/kg BW (TR5)]. All drugs were administered intravenously. Variables evaluated included heart rate (HR), respiratory rate (RR), systolic, mean, and diastolic blood pressures (SAP, MAP, and DAP), and sedation [by use of a simple descriptive scale (SDS, range: 0 to 3) and a numeric rating scale (NRS, range: 0 to 10)]. Variables were recorded 25 min after acepromazine and for 80 min after saline or tramadol. Acepromazine administration resulted in mild sedation in most dogs and decreased RR, SAP, MAP, and DAP in all treatments. Tramadol administration did not significantly increase SDS or NRS scores compared to acepromazine alone. The only exception to this rule was observed at 20 min after TR3, when NRS was higher in this group than in the control treatment. Administration of tramadol (TR3 and TR5) decreased HR. Under the conditions of this study, sedation induced by acepromazine with tramadol was similar to that of acepromazine alone. The main adverse effects of the combination were a decrease in blood pressure and HR, without clinical significance.

OBJECTIVE To compare tear cortisol concentrations between horses and ponies with pituitary pars intermedia dysfunction (PPID) and healthy nonaged (≤ 15 years old) and aged (≥ 20 years old) horses and to determine whether serum and tear cortisol concentrations were correlated. ANIMALS 11 horses and ponies with PPID and 20 healthy control horses and ponies (11 nonaged and 9 aged). PROCEDURES Paired tear and serum samples were obtained from PPID and control animals. All animals were free of active ocular disease. Tear and serum cortisol concentrations were measured with an ELISA and chemiluminescent assay, respectively. Groups were compared with Kruskal-Wallis and Mann-Whitney U tests, and Spearman correlation analysis was used to examine relationships between tear and serum cortisol concentrations within groups. RESULTS Median tear cortisol concentration was significantly higher in PPID animals than in aged control animals, despite comparable serum cortisol concentrations in PPID and aged control animals. Median tear-to-serum cortisol concentration ratios were also significantly higher in PPID animals than in aged control animals. Serum and tear cortisol concentrations were not significantly correlated in PPID or control animals. CONCLUSIONS AND CLINICAL RELEVANCE Some horses and ponies with PPID had increased tear cortisol concentrations, compared with concentrations in healthy aged animals. Localized cortisol production in the tear film or altered cortisol binding dynamics could have contributed to this increase. Further studies are warranted to evaluate these mechanisms and to determine whether increased tear cortisol concentrations are associated with delays in corneal wound healing in horses and ponies with and without PPID.

The probability of detecting influenza A virus (IAV) in oral fluid (OF) specimens was calculated for each of 13 assays based on real-time reverse-transcription polymerase chain reaction (rRT-PCR) and 7 assays based on virus isolation (VI). The OF specimens were inoculated with H1N1 or H3N2 IAV and serially diluted 10-fold (10(−1) to 10(−8)). Eight participating laboratories received 180 randomized OF samples (10 replicates × 8 dilutions × 2 IAV subtypes plus 20 IAV-negative samples) and performed the rRT-PCR and VI procedure(s) of their choice. Analysis of the results with a mixed-effect logistic-regression model identified dilution and assay as variables significant (P < 0.0001) for IAV detection in OF by rRT-PCR or VI. Virus subtype was not significant for IAV detection by either rRT-PCR (P = 0.457) or VI (P = 0.101). For rRT-PCR the cycle threshold (Ct) values increased consistently with dilution but varied widely. Therefore, it was not possible to predict VI success on the basis of Ct values. The success of VI was inversely related to the dilution of the sample; the assay was generally unsuccessful at lower virus concentrations. Successful swine health monitoring and disease surveillance require assays with consistent performance, but significant differences in reproducibility were observed among the assays evaluated.