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Development and assessment of a novel ex vivo corneal culture technique involving an agarose-based dome scaffold for use as a model of in vivo corneal wound healing in dogs and rabbits
2020
Berkowski, William M. Jr | Gibson, Daniel J. | Craft, Serena L. | Whitley, Robert D. | Schultz, Gregory S. | Plummer, Caryn E.
OBJECTIVE To develop and assess a novel ex vivo corneal culture technique involving an agarose-based dome scaffold (ABDS) for use as a model of in vivo corneal wound healing in dogs and rabbits. SAMPLE Corneas from clinically normal dogs (paired corneas from 8 dogs and 8 single corneas) and rabbits (21 single corneas). PROCEDURES 8 single dog corneas (DCs), 1 DC from each pair, and 10 rabbit corneas (RCs) were wounded with an excimer laser; 1 DC from each pair and 11 RCs remained unwounded. Corneas were cultured for 21 days on ABDSs (8 pairs of DCs and all RCs) or on flat-topped scaffolds (8 single DCs). The surface area of corneal fluorescein retention was measured every 6 (DCs) or 12 (RCs) hours until full corneal epithelialization was detected. Changes in corneal clarity were evaluated at 0, 7, 14, and 21 days. RESULTS Median time to full epithelialization for wounded dog and rabbit corneas was 48 and 60 hours, respectively; among wounded DCs, time to full epithelization did not differ by scaffold type. After 21 days of culture on ABDSs, all DCs and RCs that epithelialized developed a circular, diffuse, cloud-like pattern of optical haze, whereas DCs cultured on flat-topped scaffolds developed a focal, crater-like region of optical haze. All corneas on the ABDSs maintained convex curvature throughout the study. CONCLUSIONS AND CLINICAL RELEVANCE Wounded ex vivo DCs and RCs cultured on ABDSs reliably epithelialized, formed optical haze (consistent with in vivo wound healing), and maintained convex curvature. This culture technique may be adaptable to other species.
Show more [+] Less [-]Comparison of the use of a vessel-sealing device versus ligatures for occlusion of uterine tissues during ovariohysterectomy or ovariectomy in rabbits (Oryctolagus cuniculus)
2020
McLean, Euan J. | Woodward, Andrew P. | Ryan, Stewart D.
OBJECTIVE To compare the bursting strength of the uterine horns (UHs) and cervical-vestibule junction (CVJs) of rabbits following sealing with a vessel-sealing device (VSD) or encircling ligatures. SAMPLE UHs and CVJs collected from 30 rabbit (Oryctolagus cuniculus) cadavers. PROCEDURES UHs and CVJs were randomly assigned to sealing with encircling Miller knot ligatures (LIG; n = 10 CVJs and 20 UHs) or a VSD (12 CVJs and 24 UHs). Lumens were infused with saline (0.9% NaCl) solution under pressure until seals burst or to a maximum pressure of 300 mm Hg. RESULTS For CVJs, median (range) bursting pressure of the LIG and VSD groups was > 300 mm Hg (224 to > 300 mm Hg) and 35 mm Hg (0 to 60 mm Hg), respectively. Five of 12 CVJs in the VSD group failed at pressures < 33 mm Hg. For UHs, median (range) bursting pressure of the LIG and VSD groups was 255 mm Hg (120 to > 300 mm Hg) and 154 mm Hg (range, 44 to 202 mm Hg), respectively. CONCLUSION AND CLINICAL RELEVANCE The evaluated VSD was effective in sealing UHs at bursting pressures well in excess of expected physiologic pressures, indicating that the VSD may be useful for ovariectomy procedures in rabbits. However, CVJ seals created with the VSD were ineffective and could potentially burst at low pressures, which could predispose to urine entering the abdomen. Given these results, we do not recommend sealing of the CVJ with a VSD for ovariohysterectomy in rabbits.
Show more [+] Less [-]Establishment of a method for evaluation of the efficacy of a classical swine fever virus subunit vaccine in rabbits
2020
Cao, Zhi | Zhang, Heng | Yang, Qian | Zhang, Hui | Fan, Gencheng
OBJECTIVE To establish a method for evaluation of the efficacy of a classical swine fever virus (CSFV) subunit vaccine in rabbits as determined via humoral immune responses to the virus. ANIMALS 40 specific pathogen–free rabbits. PROCEDURES Rabbits were randomly assigned to 4 groups (10 rabbits/group) for SC injection of 0.05, 0.1, and 0.2 mL of a CSFV subunit E2 vaccine (representing 1.15, 2.3, or 4.6 μg of E2 protein/dose, respectively) or saline (0.9% NaCl) solution. Blood samples were collected 21 days after vaccination for measurement of the antibody response against CSFV via ELISA and virus neutralization methods. On the same day, the CSFV Chinese (C) strain was injected into an ear vein. Vaccine efficacy was determined by monitoring of rabbits for pyrexia for 4 days and measurement of viral copies in spleen lysates at the end of the study. Reproducibility of the antibody response was tested with 2 other batches of the vaccine at the minimum immunization dose identified for the initially tested batch. RESULTS The E2 protein dose of the initially tested vaccine was positively correlated with the antibody response and protection rate in rabbits. The identified minimum immunization dose per rabbit was 0.1 mL, representing an E2 protein content of approximately 2.3 μg, and reproducibility of the antibody response to vaccination with the 2 other batches at this dose was good. CONCLUSIONS AND CLINICAL RELEVANCE A method was established in rabbits for evaluation of the efficacy of a CSFV subunit vaccine that could help in the optimization of later large-scale vaccine production and quality control processes as well as in the clinical application of the vaccine.
Show more [+] Less [-]Assessment of luteinizing hormone receptor expression in structural support tissues of canine hip and femorotibial joints
2020
Kiefel, Claire A. | Kutzler, Michelle A.
OBJECTIVE To determine whether luteinizing hormone receptors (LHRs) are expressed in canine femoral head subchondral bone (FHSB), hip joint round ligament (RL), cranial cruciate ligament (CCL), and femorotibial joint synovium (FJS) specimens. SAMPLE 1 specimen each of the FHSB, RL, CCL, and FJS obtained from the left hind limbs of 19 fresh canine cadavers. PROCEDURES 1 section of each FHSB, RL, CCL, and FJS specimen was processed with rabbit polyclonal IgG anti-human LHR antibody, and 1 section was treated with negative control reagents. Percentage immunoexpression of LHRs in FHSB and FJS sections was analyzed by assessment of 100 bone marrow cells or synoviocytes in 3 adjacent hpf (400×). In each RL and CCL section, immunoexpression of LHRs in fibrocytes was semiquantitatively analyzed on the basis of the mean of the product of percentage staining score (from 0 [no staining] to 3 [> 50% of cells stained]) and staining intensity score (from 0 [no staining] to 2 [moderate to strong staining]) for 3 adjacent hpf. RESULTS All tissues examined had variable LHR expression. Expression of LHRs in FHSB, CCL, or FJS specimens did not differ between sexes or between sexually intact and gonadectomized dogs. However, RL specimens from female dogs had significantly greater LHR expression scores, compared with findings for male dogs. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that LHRs are expressed in structural support tissues of canine hip and femorotibial joints. Further research is required to determine the LHRs' function, mechanism of action, and potential contribution to the pathogenesis of hip dysplasia or CCL rupture in dogs.
Show more [+] Less [-]Investigation for correction formulas on the basis of packed cell volume for blood glucose concentration measurements obtained with portable glucometers when used in rabbits
2020
Cutler, Daniel C. | Koenig, Amie | Di Girolamo, Nicola | Mayer, Jörg
OBJECTIVE To determine effects of PCV on blood glucose (BG) concentration measurements obtained with a human portable blood glucometer (HPBG) and a veterinary portable blood glucometer (VPBG) on canine (cVPBG) and feline (fVPBG) settings (test methods) when used in rabbits and to develop correction formulas to mitigate effects of PCV on such measurements. SAMPLE 48 resuspended blood samples with known PVCs (range, 0% [plasma] to 92% [plasma and packed RBCs]) from 6 healthy research rabbits (experimental sample set) and 252 historic measurements of BG concentration and PCV in 84 client-owned rabbits evaluated at a veterinary hospital (validation data set). PROCEDURES Duplicate measurements of BG concentration with each test method and of PCV were obtained for each sample in the experimental sample set, and the mean results for each variable for each test method and sample were compared with results from a clinical laboratory analyzer (reference method) used to determine the true BG concentration for each sample. Mean ± SD differences in measurements between the reference and test methods were calculated. Linear regression and modified Clarke error grid analysis were used to develop correction formulas for the test methods given known PCVs, and these formulas were evaluated on the validation data set with linear regression and a modified Clarke error grid. RESULTS Blood glucose concentrations were falsely low for cVPBG and fVPBG used on samples with PCV < 31% and were falsely high for all test methods used on samples with PCV > 43%. Compared with original measurements, formula-corrected measurements overall had better agreement with reference method measurements for the experimental sample set; however, only the formula-corrected HPBG measurements had improved agreement for the validation data set. CONCLUSIONS AND CLINICAL RELEVANCE Findings indicated that, in rabbits, HPBG measurements had improved accuracy with the use of the correction formula HPBG measurement of BG concentration + ([0.75 × PCV] − 15); however, the correction formulas did not improve the accuracy of VPBG measurements, and we believe that neither the cVPBG nor fVPBG should be used in rabbits.
Show more [+] Less [-]Cardiovascular and respiratory effects of carbon dioxide pneumoperitoneum in the domestic rabbit (Oryctolagus cuniculus)
2020
Kabakchiev, Claudia | Valverde, Alex | Singh, Ameet | Beaufrere, Hugues
The objective of this study was to evaluate the effect of intra-abdominal pressure (IAP) on cardiorespiratory parameters during pneumoperitoneum with carbon dioxide in domestic rabbits. Six juvenile female New Zealand white rabbits were assigned to randomized sequences of IAP (0, 4, 8 mmHg) in a crossover study. The following parameters were measured at each IAP: direct arterial blood pressure (ABP); cardiac output, (CO), cardiac index, and stroke volume index (CI, SVI); heart rate; end-tidal carbon dioxide (ETCO(2)); arterial blood gases (PaCO(2), PaO(2)); peak inspiratory pressure (PIP); and peripheral oxygen saturation (SpO(2)). Between IAPs, the abdomen was desufflated for a 5-minute washout period. Mixed linear regression models were used for statistical analysis. Heart rate, SpO(2), and ABP were not significantly affected by IAP, although there was a positive increase in ABP with IAP. Partial pressure of carbon dioxide (PaCO(2)) was increased at an IAP of 8 mmHg and ETCO(2) and PIP were greater with each IAP applied. Cardiac output and CI were significantly decreased with IAP and, although SVI showed the same trend, it was not statistically significant. In conclusion, pneumoperitoneum with carbon dioxide causes an increase in ETCO(2), PaCO(2), and PIP, whereas cardiac output and CI decrease. These cardiorespiratory changes should be considered when determining the optimal IAP for laparoscopic procedures in rabbits.
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