Objective-To determine the effects of ketamine hydrochloride, xylazine hydrochloride, and lidocaine hydrochloride after subarachnoid administration in goats. Animals-6 healthy goats. Procedure-In each goat, ketamine (3 mg/kg), xylazine (0.1 mg/kg), lidocaine (2.5 mg/kg), and saline (0.9% NaCl) solution were injected into the subarachnoid space between the last lumbar vertebra and first sacral vertebra (time 0). Analgesic, ataxic, sedative, cardiovascular, and respiratory effects and rectal temperature were evaluated before (baseline) and 2, 5, 10, 15, and 30 minutes after administration and at 30-minute intervals thereafter as needed. Results-Administration of anesthetics induced varying degrees of analgesia. Onset of the analgesic effect was more delayed for xylazine (mean +/- SD, 9.5 +/- 2.6 minutes) than for ketamine (6.7 +/- 2.6 minutes) or lidocaine (3.5 +/- 1.2 minutes). Duration of analgesia induced by xylazine (88.3 ± 15 minutes) was twice as long as the duration of analgesia induced by ketamine (48.8 ± 13.5 minutes) but similar to that induced by lidocaine (66.5 ± 31 minutes). Xylazine induced bradycardia, whereas ketamine caused a nonsignificant increase in heart rate. Xylazine induced a reduction in arterial pressure, whereas ketamine or lidocaine did not affect arterial pressure. Conclusion and Clinical Relevance-Subarachnoid administration of xylazine in goats resulted in longer duration of analgesia of the tail, perineum, hind limbs, flanks, and caudodorsal rib areas than administration of ketamine or lidocaine. However, xylazine caused bradycardia and respiratory depression. Additional studies are needed to determine whether the analgesia would be sufficient to allow clinicians to perform surgical procedures.

Objective-To evaluate biocompatibility and effects of implantation of 3-dimensional chondrocyte-agarose autografts in tibial defects in rabbits and to compare in vitro and in vivo chondrocyte-agarose constructs with respect to cell viability, differentiation, and matrix production. Animals-24 adult New Zealand White rabbits. Procedure-Three-dimensional constructs with (grafted group) or without (control group) autogenous chondrocytes were implanted into tibial defects of rabbits and cultured in vitro. During an 8-week period, defects were evaluated radiographically, grossly, histologically, biochemically, and immunohistochemically. In vitro constructs were evaluated histologically, biochemically, and immunohistochemically. Results-Tibial defects had significantly higher radiographic densitometry values at 4 and 6 weeks after implantation in grafted group rabbits, compared with control group rabbits. Number of observed centers of endochondral ossification was significantly greater in defects of grafted group rabbits, compared with control group rabbits. On day 14, glycosaminoglycan concentration was significantly higher in tibial defects of grafted group rabbits, compared to defects of control group rabbits or in vitro constructs. At weeks 2, 4, and 8, glycosaminoglycan concentrations were significantly lower in the in vitro control constructs, compared with other groups. Collagen type I was present in bone and bony callous in defects of grafted and control group rabbits. Collagen type II was identified in cartilaginous tissues of grafted and control group rabbits. Collagen type X was associated with hypertrophic chondrocytes. Only type II collagen was found in the in vitro chondrocyte constructs. Conclusion and Clinical Relevance-Chondrocyte-agarose grafts are biocompatible in large tibial defects and appear to provide a cell source for augmenting endochondral ossification.

The purpose of this study was to determine the relationship between the serum level of C-reactive protein (CRP) and lactation and health status. Blood samples were collected every 2 wk for 12 mo from 29 randomly selected dairy cattle on 3 farms. At the time the blood samples were collected, the stage of pregnancy, lactation status, breeding records, general health condition, reproductive status, and body condition score were recorded for each cow. Serum CRP was detected with sodium dodecyl sulfate polyacrylamide gel electrophoresis and western immunoblotting. C-reactive protein levels were measured with a densitometer and expressed as an optimal dose value. C-reactive protein levels were correlated with the body condition score, lactation status, and animal health (P < 0.05), but not with ambient temperature, animal age, or parity. C-reactive protein levels increased with milk production, peaking during high lactation (2 to 4 mo of pregnancy), and decreased when lactation ceased. In addition, the CRP level was highest during naturally occurring infections, such as mastitis and other tissue inflammation. Thus, the CRP level can confirm the presence of inflammation. The stress effect of taking blood samples as measured by the CRP level, was also examined. The CRP level became rapidly elevated 12 h after the blood samples were taken but returned to normal 36 h later. In conclusion, the stresses resulting from overall poor health, heavy lactation, and blood sampling caused the elevation of serum CRP. C-reactive protein is a marker or tool for evaluating the health status of a herd. C-reactive protein should also be considered as a useful criteria to assess the stress levels and may be useful in early surveillance of disease conditions in a dairy herd.

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.

The purpose of this study was to collect initial data to determine the potential clinical usefulness of a 13C-aminopyrine demethylation blood test, and whether additional clinical investigation is warranted. Six dogs, initially suspected of having hepatic disease based on their history, physical examination, imaging studies, general laboratory parameters, or any combination of the above, were enrolled in the study. A baseline blood sample was collected, 2 mg/kg 13C-aminopyrine was administered intravenously, and another blood sample was collected 45 min afterwards. Carbon dioxide was extracted from the blood samples and analyzed using fractional mass spectrometry. Results from the 13C-aminopyrine demethylation blood test were compared to clinical data and histologic findings. Intravenous administration of 13C-aminopyrine leads to a decrease in the percent dose of 13C recovered from dogs with histologically confirmed liver disease. Based on our results, a full-scale investigation of the potential clinical usefulness of a 13C-aminopyrine demethylation blood test for assessment of hepatic function in dogs is warranted.

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.

The objective of this study was to determine whether mosquitoes, Aedes vexans (Meigen), could serve as biological vectors of porcine reproductive and respiratory syndrome virus (PRRSV). Specifically, the study assessed the duration of viability and the site of PRRSV within mosquitoes, and evaluated whether PRRSV could be transmitted to a susceptible pig by mosquitoes following a 7- to 14-day incubation period after feeding on an infected pig. For the first experiment, a total of 100 mosquitoes were allowed to feed on a pig, experimentally infected with PRRSV (day 7 post-inoculation) and were then maintained alive under laboratory conditions. A set of 10 mosquitoes were collected at 0 hour (h), 6 h, 12 h, 24 h, 48 h, 72 h, 5 days (d), 7 d, 10 d, and 14 d post-feeding (pf). Samples of exterior surface washes, salivary glands, thorax carcasses, and gut homogenates were collected from each set of mosquitoes, and tested for PRRSV. Infectious PRRSV was detected by polymerase chain reaction and swine bioassay only from the gut homogenates of mosquitoes collected at 0 h and 6 h pf. For the second experiment, a total of 30 mosquitoes were allowed to feed on a pig, experimentally infected with PRRSV and the mosquitoes were then maintained under laboratory conditions. On each of day 7, 10, and 14 pf, a set of 10 mosquitoes were allowed to feed on a susceptible pig. Transmission of PRRSV to susceptible pigs did not occur, and PRRSV was not detected from the mosquitoes. These findings indicate that mosquitoes are not likely to serve as biological vectors of PRRSV.

The time available to implement successful control measures against epidemics was estimated. Critical response time (CRT), defined as the time interval within which the number of epidemic cases remains stationary (so that interventions implemented within CRT may be the most effective or least costly), was assessed during the early epidemic phase, when the number of cases grows linearly over time. The CRT was calculated from data of the 2001 foot-and-mouth disease (FMD) epidemic that occurred in Uruguay. Significant regional CRT differences (ranging from 1.4 to 2.7 days) were observed. The CRT may facilitate selection of control measures. For instance, a CRT equal to 3 days would support the selection of measures, such as stamping-out, implementable within 3 days, but rule out measures, such as post-outbreak vaccination, because intervention and immunity building require more than 3 days. Its use in rapidly disseminating diseases, such as FMD, may result in regionalized decision-making.

Objective-To determine pharmacokinetics and tissue concentrations of azithromycin in ball pythons ( Python regius ) after IV or oral administration of a single dose. Animals-2 male and 5 female ball pythons. Procedures-Using a crossover design, each snake was given a single dose of azithromycin (10 mg/kg) IV. After a 4-week washout period, each snake was given a single dose of azithromycin (10 mg/kg) orally. Blood samples were collected prior to dose administration and 1, 3, 6, 12, 24, 48, 72, and 96 hours after azithromycin administration. Azithromycin was quantitated by use of liquid chromatography-mass spectrometry. Results-After IV administration, azithromycin had an apparent volume of distribution of 5.69 L/kg and a plasma clearance of 0.19 L/h/kg. Harmonic means for the terminal half-life were 17 hours following IV administration and 51 hours following oral administration. Mean residence times were 37 and 94 hours following IV and oral administration, respectively. Following oral administration, azithromycin had a peak plasma concentration (Cmax) of 1.04 micrograms/mL, a time to Cmax of 8.4 hours, and a prolonged mean absorption time of 57 hours. Mean oral bioavailability was 77%. Tissue concentrations ranged from 4 to 140 times the corresponding plasma concentration at 24 and 72 hours after azithromycin administration. Conclusions and Clinical Relevance-Azithromycin is well absorbed and tolerated by ball pythons. On the basis of plasma pharmacokinetics and tissue concentration data, we suggest an azithromycin dosage in ball pythons of 10 mg/kg, orally, every 2 to 7 days, depending upon the site of infection and susceptibil ity of the infective organism.

Objective-To evaluate response of euthyroid cats to administration of recombinant human thyroid-stimulating hormone (rhTSH). Animals-7 healthy cats. Procedure-Each cat received each of 5 doses of rhTSH (0, 0.025, 0.050, 0.100, and 0.200 mg), IV, at 1-week intervals. Serum concentration of total thyroxine (TT4) and free thyroxine (fT4) was measured immediately before each injection (time 0) and 2, 4, 6, and 8 hours after administration of each dose. Results-Overall TT4 response did not differ significantly among cats when administered doses were ≥ 0.025 mg. Serum TT4 concentrations peaked 6 to 8 hours after administration for all doses greater than 0.025 mg. For all doses greater than 0.025 mg, mean +/- SEM TT4 concentration at 0, 6, and 8 hours was 33.9 +/- 1.7, 101.8 +/- 5.9, and 101.5 +/- 5.7 nmol/L, respectively. For all doses greater than 0.025 mg, mean fT4 concentration at 0, 6, and 8 hours was 38.7 +/- 2.9, 104.5 +/- 7.6, and 100.4 +/- 8.0 pmol/L, respectively. At 8 hours, the fT4 response to 0.025 and 0.050 mg was less than the response to 0.100 and 0.200 mg. Adverse reactions after rhTSH administration were not detected. Conclusions and Clinical Relevance-The TSH stimulation test can be performed in cats by IV administration of 0.025 to 0.200 mg of rhTSH and measurement of serum TT4 concentrations at time of injection and 6 or 8 hours later. Clinical validation of the TSH stimulation test would facilitate development of additional tests of thyroid gland function, such as a TSH assay.