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.

Objective-To evaluate effects of sedation on stability of resistance of the respiratory system (RRS) and measures of resting energy expenditure (REE) by use of open-flow indirect calorimetry (IC) and treatment with aerosolized albuterol on REE in horses with recurrent airway obstruction (RAO). Animals-9 clinically normal horses and 8 horses with RAO. Procedure-In phase 1, RRS was measured by using forced oscillometry (FOT) in 5 clinically normal horses before and after sedation with xylazine. In phase 2, REE was measured in 4 clinically normal horses between 20 and 25 minutes and again 35 to 40 minutes after sedation with xylazine. In phase 3, IC was performed between 20 and 25 minutes and FOT was performed between 30 and 35 minutes after xylazine administration in 8 horses with RAO; after administration of 450 µg of albuterol, IC and FOT were repeated. Results-In phase 1, RRS values were significantly lower 5 and 10 minutes after sedation. In phase 2, diminishing sedation did not significantly affect REE. In phase 3, there was a significant decrease in mean RRS (1.15 +/- 0.25 vs 0.84 +/- 0.14 cm H20/L/s) and REE (30.68 +/- 17.89 vs 27.46 +/- 16.54 kcal/kg/d) after albuterol administration. Conclusions and Clinical Relevance-FOT and IC are useful in obtaining repeatable measurements of RRS and REE, respectively, in sedated horses. Concurrent bronchodilation and decreased REE after albuterol administration suggest that increased work of breathing as a result of airway obstruction may contribute to increased energy demands in horses with RAO.

Six tests for the detection of West Nile virus (WNV) antibodies in the serum of experimentally infected chickens were compared. The tests included the hemagglutination-inhibition test (HIT), immunoglobulin M (IgM)-capture enzyme-linked immunosorbent assay (ELISA) with WNV-infected mouse brain antigen, immunoglobulin G (IgG) indirect ELISA with tickborne encephalitis viral antigen, the microtitre virus neutralization test, the standard plaque reduction neutralization test (PRNT), and the microtitre PRNT (micro-PRNT). Thirty adult chickens, intravenously and intramuscularly inoculated with 107 plaque-forming units (PFU) of WNV strain Egypt 101, were bled and given a booster of 107 PFU at 7, 15, and 21 d postinoculation; the final blood collection was on day 28. Although the micro-PRNT is capable of detecting the highest antibody titres during both early and late infection, because of the technical complexity and time requirements of this test a combination of IgM and IgG ELISAs is recommended for serologic screening. Serum samples that give positive results in the ELISAs can then be tested by the micro-PRNT to determine the specificity of antibodies to WNV.

Cross-reactivity indices (CRIs) of 28 isolates of Moraxella bovis recovered from outbreaks of infectious bovine keratoconjunctivitis in Argentina (A, 11 isolates), Brazil (B, 7), and Uruguay (U, 10) between 1983 and 2000 were estimated. Hyperimmune sera were produced in rabbits and antibody titres determined with each isolate. Isolates showing CRIs3 70 were placed in the same group. Group I had 13 isolates (A, 1; B, 6; U, 6); group II had 6 isolates (A, 4; U, 2); groups III, IV, and V had 2 isolates each, recovered in Argentina; group VI had 2 isolates, from Uruguay; and group VII had 1 isolate, from Brazil. The CRIs3 70 between vaccine strains and isolates recovered before and after 1990 were 58% and 42%, 50% and 50%, and 33% and 67% with vaccine strains 2419, 2358, and 2439, respectively. Isolate 273, from Uruguay, showed CRIs > 70 with 78% of the isolates and is recommended as the vaccine strain.

Objective-To investigate telomere lengths in tissues of domestic shorthair (DSH) cats of various ages, evaluate the relationship between telomere length and age of cats, and investigate telomerase activity in the somatic tissues of cats. Sample Population-Tissues obtained from 2 DSH cats and blood samples obtained from 30 DSH cats. Procedure-DNA isolated from blood cells and somatic tissue samples was subjected to terminal restriction fragment (TRF) analysis to determine mean telomere repeat lengths. Protein samples were subjected to analysis by use of a telomeric repeat-amplification protocol to assess telomerase activity. Results-Mean TRF values of cats ranged from 4.7 to 26.3 kilobase pairs, and there was significant telomeric attrition with increasing age of cat. Telomerase activity was not found in a wide range of normal tissues obtained from 2 cats. Conclusions and Clinical Relevance-Analysis of these results clearly indicates that telomeres are shorter in older cats, compared with young cats; therefore, telomeres are implicated in the aging process. The analysis of telomerase activity in normal somatic tissues of cats reveals a pattern of expression similar to that found in human tissues. Impact for Human Medicine-Fundamental differences in the biological characteristics of telomeres and telomerase exist between humans and the other most widely studied species (ie, mice). The results reported here reveal similarities in telomere and telomerase biologic characteristics between DSH cats and humans. Hence, as well as developing our understanding of aging in cats, these data may be usefully extrapolated to aging in humans.

Objective-To evaluate lactoferrin and lysozyme content in various ocular glands of bison and cattle and in tears of bison. Sample Population-Tissues of ocular glands obtained from 15 bison and 15 cattle and tears collected from 38 bison. Procedure-Immunohistochemical analysis was used to detect lysozyme and lactoferrin in formalin-fixed, paraffin-embedded sections of the ocular glands. Protein gel electrophoresis was used to analyze ocular glands and pooled bison tears by use of a tris-glycine gel and SDS-PAGE. Western blotting was used to detect lactoferrin and lysozyme. Results-Immunohistochemical staining for lactoferrin was evident in the lacrimal gland and gland of the third eyelid in cattle and bison and the deep gland of the third eyelid (Harder's gland) in cattle. Equivocal staining for lactoferrin was seen for the Harder's gland in bison. An 80-kd band (lactoferrin) was detected via electrophoresis and western blots in the lacrimal gland and gland of the third eyelid in cattle and bison, Harder's glands of cattle, and bison tears. An inconsistent band was seen in Harder's glands of bison. Lysozyme was not detected in the lacrimal gland of cattle or bison with the use of immunohistochemical analysis or western blots. Western blots of bison tears did not reveal lysozyme. Conclusion and Clinical Relevance-Distribution of lactoferrin and a lack of lysozyme are similar in the lacrimal gland of cattle and bison. Differences in other tear components may be responsible for variability in the susceptibility to infectious corneal diseases that exists between bison and cattle.

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.

Objective-To explore whether early analysis of spatial data may result in identification of variables associated with epidemic spread of foot and mouth disease. Sample Population-37 farms with infected cattle (ie, case farms) reported within the first 6 days of the 2001 Uruguayan foot-and-mouth disease epidemic. Procedure-A georeferenced database was created and retrospective analysis was performed on case farm location in relation to farm density, cattle density, farm type (ie, beef vs dairy cattle production), road density, case farm distance to the nearest road, farm size, farm ownership, and day of infection. Mean or median results of 1 to 3 day versus 4 to 6 day spatial data were compared. Spatial-temporal associations were investigated by correlation analysis. Results-Comparison of mean or median values between the first 3 days and days 4 to 6 of the epidemic and results of correlation analysis indicated a significant increase in road density, cattle density, and dairy cattle production and a significant decrease in farm size and case farm distance to the nearest road that developed over time. A route that linked most case farms by the shortest possible distance and also considered significantly associated variables was created. It included 86.1% of all case farms reported by 60 days into the epidemic. Conclusions and Clinical Relevance-Epidemic direction can be assessed on the basis of road density and other spatial variables as early as 6 days into an epidemic. Epidemic control areas may be more effectively identified if local and regional georeferenced data are considered.

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.

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.