In vitro comparison of equine cancellous bone graft donor sites and tibial periosteum as sources of viable osteoprogenitors

OBJECTIVE: To compare the osteogenic potential of cancellous bone of conventional graft sites with that of one nonconventional site (fourth coccygeal vertebra) and to investigate the tibial periosteum as a donor site with respect to osteogenic potential. STUDY DESIGN: In vitro osteogenic cell culture system. SAMPLE POPULATION: Eight adult horses. METHODS: Cancellous bone or tibial periosteum was aseptically collected and cut into bone chips or periosteal strips of 1 to 2 mm(3) for primary explant cultures. After 2 weeks, primary tissue cultures that yielded a population of osteogenic cells were counted and subcultured at 1 x 10(5) cells/35-mm dish in osteogenic media. After 7 to 10 days, subcultures were stained with Von Kossa (VK) to assess mineralized bone nodule formation. VK-positive bone nodules were counted as osteoprogenitors and compared among 3 donor sites, which provided consistent primary osteogenic cells (tuber coxae, fourth coccygeal vertebra, periosteum) using ANOVA (P <.05). RESULTS: Sternal and tibial bone yielded viable osteogenic cells from 25% and 50% of horses, respectively, whereas yields from tuber coxae, coccygeal vertebra, and periosteum were 75%, 100%, and 100%, respectively. Tuber coxae and periosteum had significantly greater numbers of osteoprogenitors compared with fourth coccygeal vertebra. CONCLUSIONS: Among the conventional donor sites, tuber coxae most consistently yielded viable osteogenic cells with an acceptable percentage of osteoprogenitors. Sternal and tibial sites were unreliable in providing osteogenic cells. Two new donor sites, the fourth coccygeal vertebra and tibial periosteum, were tissues with good osteogenic potential. CLINICAL RELEVANCE: When a source of transplantable viable osteoprogenitor cells is desired, use of the tuber coxae as a conventional donor site is warranted. Use of tibial periosteum or fourth coccygeal vertebra as reliable sources of transplantable osteoprogenitors should be considered.     

Association between initial systolic blood pressure and risk of developing a uremic crisis or of dying in dogs with chronic renal failure

OBJECTIVE: To determine whether high systolic blood pressure (SBP) at the time of initial diagnosis of chronic renal failure in dogs was associated with increased risk of uremic crisis, risk of dying, or rate of decline in renal function. DESIGN: Prospective cohort study. ANIMALS: 45 dogs with spontaneous chronic renal failure. PROCEDURE: Dogs were assigned to 1 of 3 groups on the basis of initial SBP (high, intermediate, low); Kaplan-Meier and Cox proportional hazards methods were used to estimate the association between SBP and development of a uremic crisis and death. The reciprocal of serum creatinine concentration was used as an estimate of renal function. RESULTS: Dogs in the high SBP group were more likely to develop a uremic crisis and to die than were dogs in the other groups, and the risks of developing a uremic crisis and of dying increased significantly as SBP increased. A greater decrease in renal function was observed in dogs in the high SBP group. Retinopathy and hypertensive encephalopathy were detected in 3 of 14 dogs with SBP > or = 180 mm Hg. Systolic blood pressure remained high in 10 of 11 dogs treated with antihypertensive drugs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that initial high SBP in dogs with chronic renal failure was associated with increased risk of developing a uremic crisis and of dying. Further studies are required to determine whether there is a cause-and-effect relationship between high SBP and progressive renal injury and to identify the risks and benefits of antihypertensive drug treatment.     

Antebrachial fractures in four captive polar bears (Ursus maritimus)

OBJECTIVE: To identify common risk factors for antebrachial fractures of captive polar bears and to evaluate outcome after fracture repair. STUDY DESIGN: Retrospective study. ANIMALS: Four captive polar bears. METHODS: United States zoological collections were surveyed to determine the prevalence of fractures in captive polar bears. Medical records of captive polar bears that had antebrachial fractures were reviewed for signalment, history, physical and radiographic findings, fracture management, postoperative care, and outcome. Serum samples from healthy bears and bears with antebrachial fractures were assayed for 25-hydroxyvitamin D (25-OHD) concentrations. RESULTS: Nineteen fractures (12 polar bears) occurred from 1974 to 2002; 12 fractures involved the antebrachium. Management of 4 antebrachial fractures was reviewed; 3 were repaired by internal fixation and 1 by external coaptation. Fractures healed and bears were returned to exhibit on average 3 months postfracture. Of 11 serum samples assayed for 25-OHD concentrations, 6 were below normal, 1 was low normal and 4 were within normal reference intervals. The 7 bears with subnormal or low normal values were housed in 2 zoos. Subnormal vitamin D concentrations were identified in 2 of 3 bears with fractures. CONCLUSIONS: Fracture disease is not uncommon in captive polar bears. Additional research is necessary to explore the role of nutrition in polar bear fracture disease. CLINICAL RELEVANCE: Internal fixation of antebrachial fractures is feasible and reasonably well tolerated in captive polar bears.     

Evaluation of serologic methods for the detection of antibodies to encephalomyocarditis virus in swine fetal thoracic fluids.

Hemagglutination inhibition (HI) and agar gel immunodiffusion (AGID) tests were compared to the serum neutralization (SN) test to evaluate their ability to detect antibodies to encephalomyocarditis virus (EMCV). Swine fetal thoracic fluids of known EMCV SN antibody titers (200 samples greater than or equal to 1:2, 100 samples less than 1:2) were selected from a collection of field cases. The thoracic fluids were tested for EMCV antibodies by HI and AGID, and the results were compared to those of the SN test. Of 200 SN antibody-positive samples, 183 (91.5%) and 173 (86.5%) were positive in HI and AGID tests, respectively. Of 100 SN-negative samples, 81 (81%) and 94 (94%) were negative in HI and AGID tests, respectively. Agreement between the tests was analyzed by calculating Kappa values. The values were 0.73 between SN and HI tests and 0.77 between SN and AGID tests, indicating very good to excellent agreement for HI and AGID tests with the SN test. Of 200 SN-positive samples, 19 samples with low SN titers (1:2-1:16) were further tested by Western immunoblotting, and all were confirmed as positive. Interpretation of the present results suggests that both HI and AGID tests can be used as alternatives to the SN test.     

Experimental reproduction of swine infertility and respiratory syndrome in pregnant sows.

The purpose of this study was to experimentally reproduce swine infertility and respiratory syndrome (SIRS). Six multiparous sows were intranasally inoculated at 93 days of gestation with lung homogenates from clinically affected pigs, and 3 additional sows were similarly inoculated with a virus isolated in cell culture from the lung homogenate (SIRS virus, isolate ATCC VR-2332). Inoculated sows developed transient anorexia, farrowed up to 7 days prematurely, and delivered a mean of 5.8 live pigs and 6.0 dead fetuses/litter. Clinical signs of disease were not observed in 3 sham-inoculated control sows that delivered a mean of 12.7 live pigs and 0.3 stillborn fetuses/litter. The SIRS virus was isolated from 50 of 76 live-born and stillborn fetuses from the 9 infected litters. Virus was not isolated from 26 autolyzed fetuses or 15 control pigs. Six of 9 inoculated sows developed neutralizing antibodies to SIRS virus. The reproductive effects found in these experiments were identical to those found in field cases. On the basis of our findings, virus isolate ATCC VR-2332 causes the reproductive failure associated with SIRS.     

Seroprevalence of Mycoplasma alligatoris among free-ranging alligators (Alligator mississippiensis) in Florida--2003.

Mycoplasma alligatoris causes acute lethal infection of alligators (Alligator mississippiensis). The objective of this study was to assess the current seroprevalence of M. alligatoris among free-ranging, juvenile and subadult alligators in Florida. Thirty-two of 592 (5.4%) plasma samples from alligators at 12 of 20 sites (60%) in April and October 2003 were tested seropositive (titer 1: > or = 32) by enzyme-linked immunosorbent assay for anti-M. alligatoris antibodies. These results show that alligators throughout Florida have a recent history of exposure to M. alligatoris and suggest that contact with free-ranging alligators may constitute a risk of lethal infection of susceptible crocodilians.