Objective-To compare aesthesiometer-determined corneal sensitivity between diabetic and nondiabetic dogs and to investigate the correlation between corneal sensitivity and duration of diabetes or status of glycemic control, as estimated by use of glycated blood protein concentrations. Animals-23 diabetic and 29 nondiabetic normoglycemic dogs. Procedure-A Cochet-Bonnet aesthesiometer was used to measure corneal touch threshold (CTT) in 5 corneal regions of each dog. At the time of ocular examination, duration of diabetes mellitus was estimated from the history, and blood was drawn for assessment of blood glycosylated hemoglobin and serum fructosamine concentrations. Results-Median CTT for central, nasal, dorsal, temporal, and ventral corneal regions in nondiabetic dogs (1.6, 2.3, 2.8, 2.8, and 5.1 g/mm2, respectively) was significantly lower than in diabetic dogs (2.8, 4.0, 5.1, 5.1, and 6.6 g/mm2, respectively). Median regional CTT in diabetic dogs was not significantly correlated with estimated duration of diabetes mellitus or blood glycated protein concentrations. No significant difference was found in regional CTT between eyes of normoglycemic dogs with unilateral cataracts. Conclusion and Clinical Relevance-Diabetic dogs have significantly reduced corneal sensitivity in all regions, compared with nondiabetic normoglycemic dogs. Regional variation in corneal sensitivity is similar in diabetic and normoglycemic dogs. Neither glycemic control nor duration of diabetes, as estimated, is significantly correlated with corneal hyposensitivity. Corneal nerve dysfunction may be associated with recurrent or nonhealing ulcers in diabetic dogs for which no other underlying cause can be found.

Objective-To determine the prevalence of biofilm formation under long-term cell culture conditions in serum samples of dairy cattle, goats, cats, and dogs, and to determine whether there is an association between nanobacteria and biofilm formation. Sample Population-Serum samples of clinically normal animals (313 dairy cattle, 48 goats, 140 dogs, and 44 cats) and animals with various medical conditions (60 dogs and 116 cats). Procedure-Serum was incubated under cell culture conditions and observed for biofilm formation by use of light microscopy, electron microscopy, and spectroscopy. A polymerase chain reaction assay was developed to identify 16S rRNA gene sequences of nanobacteria. Results-Biofilm formation developed in serum samples of 304 of 313 (97%) cattle, 44 of 48 (92%) goats, 44 of 44 (100%) cats, and 126 of 140 (90%) dogs. Prevalence of serum samples with positive results for biofilm formation was not significantly different between cats or dogs with and without medical conditions associated with pathologic extraskeletal calcification processes. Scanning electron microscopy and spectroscopy of biofilm samples revealed small coccoid particles consisting mainly of calcium and phosphate. Polymerase chain reaction assay failed to amplify sequences of nanobacteria. Conclusions and Clinical Relevance-Under longterm cell culture conditions, biofilm made up of aggregates of calcium and phosphate crystals does form in serum samples of clinically normal dairy cattle, goats, cats, and dogs. Disease, however, does not predispose to biofilm formation in serum samples of dogs and cats. Our findings did not support the existence of nanobacteria in serum samples of cattle, goats, cats, and dogs.

Nursing sickness, the largest single cause of mortality in adult female mink (Mustela vison), is an example of a metabolic disorder, which develops when the demands for lactation require extensive mobilization of body energy reserves. The condition is characterized by progressive weight loss, emaciation, and dehydration with high concentrations of glucose and insulin in the blood. Morbidity due to nursing sickness can be as high as 15% with mortality around 8%, but the incidence is known to vary from year to year. Stress has been shown to trigger the onset of the disease and old females and females with large litters are most often affected. Increasing demand for gluconeogenesis from amino acids due to heavy milk production may be a predisposing factor. Glucose metabolism is inextricably linked to that of protein and fats. In obesity (or lipodystrophy), the ability of adipose tissue to buffer the daily influx of nutrients is overwhelmed (or absent), interfering with insulin-mediated glucose disposal and leading to insulin resistance. Polyunsaturated fatty acids of the n-3 family play an important role in modulating insulin signalling and glucose uptake by peripheral tissue. The increasing demand on these fatty acids for milk fat synthesis towards late lactation may result in deficiency in the lactating female, thus impairing glucose disposal. It is suggested that the underlying cause of mink nursing sickness is the development of acquired insulin resistance with 3 contributing key elements: obesity (or lipodystrophy), n-3 fatty acid deficiency, and high protein oxidation rate. It is recommended that mink breeder females be kept in moderate body condition during fall and winter to avoid fattening or emaciation. A dietary n-3 fatty acid supplement during the lactation period may be beneficial for improved glycemic control. Lowering of dietary protein reduces (oxidative) stress and improves water balance in the nursing females and may, therefore, prevent the development and help in the management of nursing sickness. It is also surmised that other, thus far unexplained, metabolic disorders seen in male and female mink may be related to acquired insulin resistance.

The use of naturally-farrowed, artificially-reared piglets as an alternative model to study Haemophilus parasuis infections was evaluated. Two trials were performed in order to evaluate the proposed model. In trial 1, animals were vaccinated and challenged with H. parasuis. Results showed that the proposed model was effectively used to evaluate protective immunity against this organism. In trial 2, animals were challenged with different doses of H. parasuis. Results showed that the severity of clinical signs and lesions tended to increase with higher doses. The reproduction of clinical signs and lesions characteristic of H. parasuis systemic infection was successful in both trials, proving that this model is a viable alternative to specific-pathogen free and cesarean-derived, colostrum-deprived pigs.

Multiplex nested polymerase chain reactions (PCRs) were developed for the simultaneous detection and differentiation of genomic material of porcine circovirus 1 (PCV1), porcine circovirus 2 (PCV2), and porcine parvovirus (PPV) in formalin-fixed, paraffin-embedded tissues. Multiplex conventional and nested PCR and in situ hybridization were compared for their ability to detect the 3 viruses in such tissues. Xylene deparaffinization followed by proteinase K digestion yielded DNA of sufficient quality for reliable and consistent PCR analyses. The DNA from PCV1, PCV2, and PPV was detected by both multiplex nested PCR and in situ hybridization in lymph-node tissue from 12 pigs experimentally co-infected with the 3 viruses, as well as in formalin-fixed, paraffin-embedded lymph-node tissue from 30 pigs with naturally occurring postweaning multisystemic wasting syndrome; the agreement rates for the 2 methods were 100% in both groups of pigs. Thus, multiplex nested PCR could be applied successfully to formalin-fixed, paraffin-embedded tissues for simultaneous detection of these 3 porcine viruses.

Objective-To compare the minimum alveolar concentration (MAC) of isoflurane required to prevent corticocerebral activation, autonomic responses, and purposeful movements after somatic or visceral stimulation in cats anesthetized with isoflurane. Animals-17 healthy spayed female cats. Procedure-Bispectral index (BIS), autonomic parameters, and purposeful movements were monitored before and after somatic or visceral stimuli in cats anesthetized with isoflurane. End-tidal (ET) isoflurane concentration was varied to determine MAC values for cortical arousal (MACBIS), autonomic responsiveness (MACBAR), and purposeful movement (MAC). Bispectral index values greater than 60 were considered to represent corticocerebral activation. Results-Minimum alveolar concentration for purposeful movement was significantly less than MACBIS and MACBAR for both somatic and visceral stimulation. Individual MAC values for somatic stimulation were not significantly different from respective MAC values for visceral stimulation. The percentage of cats that had a BIS response greater than 60 was inversely related to the end-tidal isoflurane concentration. Conclusions and Clinical Relevance-Corticocerebral arousal and subcortical autonomic reflexes occured at isoflurane anesthetic concentrations at which reflexive or purposeful movements were absent. These results suggested that isoflurane had a preferential effect on voluntary motor output at low end-tidal isoflurane concentrations, and that sensory pathways, subcortical sympathetic output, and cortical responsiveness are less susceptible to the anesthetic effects of isoflurane. Bispectral index values obtained after somatic or visceral stimulation were sensitive for the detection of early changes in cortical excitability.

Objective-To compare measurements of blood flow in the common femoral artery obtained by duplex Doppler ultrasonography (DDU) and a reference ultrasonic transit-time flow (TTF) method and to examine the impact of Doppler spectral waveform measurement techniques on volumetric estimates. Animals-5 healthy female pigs. Procedure-Femoral arterial blood flow was measured simultaneously in anesthetized pigs by use of a TTF probe (left femoral artery) and transcutaneous DDU (right femoral artery). A range of flow states was induced pharmacologically by using xylazine, bradykinin, dobutamine, and isoflurane. Volumetric blood flow was calculated from DDU waveforms, using the product of the flow velocity integral (FVI), the cross-sectional vessel area, and heart rate. Three calculations of FVI were obtained by manually tracing the Doppler spectral envelopes at the outer envelope, the modal, and the inner envelope of the spectral dispersion pattern. Data analysis included calculation of Pearson correlation coefficients and Bland-Altman limits of agreement. Results-Blood flow measured by DDU was more closely correlated with TTF measurements when the modal or inner envelope tracing method was used ( r, 0.76 and 0.78; limits of agreement, -100 to 54.2 and -48.5 to 77.0 mL/min, respectively). Limits of agreement for the outer envelope tracing method were -238.5 to 64 mL/min. Conclusion and Clinical Relevance-Transcutaneous DDU is a reliable noninvasive technique for measuring blood flow in the femoral artery of pigs over a range of flow states. Tracing the inner envelope of the Doppler spectral dispersion pattern provided the best estimate of blood flow in this study.

Objective-To evaluate a modified digital imaging technique for quantitative assessment of the grade of osteoarthritis across the proximal articular surface of the first phalanx in horses. Sample Population-6 metacarpophalangeal (fetlock) joint specimens from 6 horses with various stages of osteoarthritis. Procedure-First phalanx specimens, together with 4 gray scale reference calibration targets, were positioned in a bath with the proximal articular cartilage surface submerged in saline (0.9% NaCl) solution. Digital images were obtained from the articular surface before and after staining with Indian ink. Computer-controlled gray level analysis of the nonstained and Indian ink-stained cartilage surfaces and gray scale reference calibration targets was performed by use of the mean pixel value (based on 255-gray scale). An increase in the mean pixel value after staining was used to calculate the cartilage degeneration index (CDI). Results-The CDI of the proximal articular cartilage surface of the first phalanx specimens ranged from 9.2 +/- 5.7 (early stage osteoarthritis) to 41.5 +/- 3.6% (late stage osteoarthritis). The effect of repeating the measurement 6 times in nonstained (including repositioning) and stained specimens (including repositioning and restaining) was not significant. Up to 10 measurements of nonstained specimens could be made without refreshing the bath solution. In stained specimens, mean gray level increased significantly after the sixth measurement. Conclusion and Clinical Relevance-The modified digital imaging technique allowed quantitative assessment of cartilage degeneration across the articular cartilage surface. The CDI is the first quantitative measure for osteoarthritis-induced cartilage degeneration over an entire joint surface in horses.

The objectives of the study were to determine the duration of porcine reproductive and respiratory syndrome virus (PRRSV) survival in houseflies (Musca domestica Linnaeus) following feeding on an infected pig, and to determine whether the virus was present on the exterior surface or within the internal viscera of the fly. A total of 210 laboratory-colonized houseflies were allowed to feed to repletion on a pig, experimentally infected with PRRSV on day 7 postinoculation, and then maintained alive under laboratory conditions (27°C). Two subsets (A and B) of 30 flies were collected at each of the following sampling points; 0, 6, and 12 hours post feeding (pf). Subset A contained an extra group of 30 flies collected at 24 hours pf due to the availability of extra flies. Flies in subset A were processed as whole fly homogenates, while the exterior surface washes and digestive organs were collected from flies in subset B. Whole fly homogenates, collected at 0, 6, and 12 hours pf, were positive by both polymerase chain reaction (PCR) and swine bioassay. Digestive organs, collected at 0 and 12 hours pf, were positive by PCR and swine bioassay. The PRRSV RNA was detected by PCR from the exterior surface wash of subset B flies collected at 0, 6, and 12 hours pf; however, only the subset collected at 0 hour pf was swine bioassay-positive. This study indicates that infectious PRRSV can survive within the intestinal tract of houseflies for up to 12 hours following feeding on an infected pig, but only for a short period on the exterior surface of the flies.

Objective-To compare isolates of Rhodococcus equi on the basis of geographic source and virulence status by use of pulsed-field gel electrophoresis (PFGE). Sample Population-290 isolates of R equi(218 virulent isolates from foals and 72 avirulent isolates from feces, soil, and respiratory tract samples) obtained between 1985 and 2000 from horses and horse farms from 4 countries. Procedure-DNA from isolates was digested with the restriction enzyme AseI and tested by use of PFGE. Products were analyzed for similarities in banding patterns by use of dendrograms. A similarity matrix was constructed for isolates, and the matrix was tested for nonrandom distributions of similarity values with respect to groupings of interest. Results-There was little grouping of isolates on the basis of country, virulence status, or region within Texas. Isolates of R equi were generally < 80% similar, as determined by use of PFGE. Isolates from the same farm generally were rarely of the same strain. Conclusions and Clinical Relevance-Considerable chromosomal variability exists among isolates of R equi obtained from the same farm, sites within Texas, or among countries from various continents. Only rarely will it be possible to link infections to a given site or region on the basis of analysis of isolates by use of PFGE of chromosomal DNA.