Cytokine expression by neutrophils of adult horses stimulated with virulent and avirulent Rhodococcus equi in vitro

Rhodococcus equi is an intracellular pathogen of macrophages that causes rhodococcal pneumonia in foals and immunocompromised people. Evidence exists that neutrophils play a vital role in resistance to infection with R. equi; however, the means by which neutrophils exert their effects have not been clearly defined. In addition to directly killing bacteria, neutrophils also may exert a protective effect by linking innate and adaptive immune responses. In the present study we evaluated the cytokine expression profiles of adult equine neutrophils in response to stimulation with isogenic strains of virulent and avirulent R. equi in vitro. After 2 and 4 h incubation with virulent or avirulent R. equi, adult equine neutrophils expressed significantly (P < 0.05) greater tumor necrosis factor alpha (TNFα), interleukin (IL)-12p40, IL-6, IL-8 and IL-23p19 mRNA, but not interferon gamma (IFNγ) or IL-12p35 mRNA than unstimulated neutrophils. Furthermore, virulent R. equi induced significantly greater IL-23p19 mRNA than avirulent R. equi. These results demonstrate that R. equi-stimulated neutrophils are a source of many proinflammatory cytokines. Furthermore, these results suggest that IL-23 may be preferentially expressed over IL-12 in response to exposure with R. equi, and that this response may be more strongly induced by virulent R. equi than avirulent R. equi. Collectively, the data presented herein suggest a non-phagocytic role for neutrophils that may influence the type of adaptive immune response to R. equi.     

Assessment of Best-Management Practice Effects on Rangeland Stream Water Quality Using Multivariate Statistical Techniques

Quantifying the effects of watershed improvement efforts is critical to agencies responsible for protecting water resources of the semiarid western United States. A complex water quality data set collected from 1994 to 2004 of upper Muddy Creek Basin was subjected to cluster analysis, discriminant analysis, and canonical correlation analysis to improve understanding of basin fluvial processes and to investigate whether livestock grazing best-management practices (BMPs) improved the water quality of the watershed. Hierarchical agglomerative cluster analysis grouped nine sampling sites into two clusters based on similarity of biological indices, separating the clusters into aquatic communities more and less tolerant of degraded stream conditions. Discriminant analysis yielded strong spatial and temporal distinctions, providing important data reduction by rendering seven key parameters (total dissolved solids [TDS], temperature, elevation, slope, 10-dominant taxa, percent collector-gatherers, and percent Plecoptera) for the spatial variation and four parameters (TDS, dissolved oxygen, total taxa, and community tolerance quotient) for the temporal variation. Canonical correlation analysis identified strong negative relationships among Plecoptera taxa and total taxa with TDS and turbidity in addition to strong positive associations with elevation, slope, and channel substrate weighted embeddedness value. Despite the onset of severe drought midway through the study period, overall reductions of 13% for TDS and a 30% increase in macroinvertebrate total taxa occurred across years, strongly suggesting that improvements in water quality were correlated to BMPs that stabilized stream channels and improved the condition of riparian areas.     

Comparison of the Antibody Response in Adult Cattle Against Different Epitopes of Brucella abortus Lipopolysaccharide

The comparison of serological responses in a sample of adult, vaccinated and field‐infected bovines with Brucella abortus is reported. Indirect enzyme immunoassay (EIA) titration curves and Western blotting tests for smooth‐type lipopolysaccharide (S‐LPS), rough‐type LPS (R‐LPS) and lipid A were performed. In the initial screening of sera, an overall prevalence of 20.5 % was found, which corresponds to a country with a high incidence of brucellosis. End‐point EIA titres against LPS antigens from vaccinated and field‐infected cows were not significantly different. However, the absorbance values in the titration curves were significantly higher for S‐LPS as compared with the other antigens. A high correlation coefficient (r=0.933) was obtained when the titres to R‐LPS versus lipid A were compared. Western blotting reactions of vaccinated and field‐infected animals were indistinguishable. S‐LPS, R‐LPS and lipid A epitopes were recognized in a heterogeneous manner. In general, the number of bovines that reacted against LPS was higher in the field‐infected group, with a stronger binding to S‐LPS. Based on our observations, the vaccinated and field‐infected bovines are capable of producing similar antibody responses to the Brucella main outer surface antigen, LPS. It should be emphasized that the humoral response of cattle to Brucella LPS contains significant amounts of antibodies to other antigenic moieties of this important surface molecule, which may contribute to the immunity to brucellosis.     

Site and extent of nutrient digestion by sheep fed alkaline hydrogen peroxide-treated wheat straw-alfalfa hay combinations at restricted intakes.

Five crossbred wethers (58 kg) fitted with cannulas in the rumen, duodenum, and ileum were used in a 5 x 5 Latin square design to study effects of feeding combinations of alkaline hydrogen peroxide-treated wheat straw (AHP-WS) and alfalfa hay at restricted intakes on site and extent of nutrient digestion. Additionally, flows and disappearance of N and amino acids (AA) in the small intestine were regressed on alfalfa nitrogen intake (ANI) to estimate alfalfa's contribution to postruminal N and AA supplies. Diets consisted of 80:20 forage:concentrate mixtures; diet designations were 80:0, 80% AHP-WS and no alfalfa; 60:20, 60% AHP-WS and 20% alfalfa; 40:40, 40% AHP-WS and 40% alfalfa; 20:60, 20% AHP-WS and 60% alfalfa; and 0:80, no AHP-WS and 80% alfalfa. A modest positive quadratic (P less than .05) response was noted for total tract digestibility (TTD) of OM; values were 76.5% for diets 80:0 and 0:80 vs 78% for diet 40:40. Ruminal digestibility (percentage of intake) of NDF and ADF increased in a quadratic manner from 43 and 30%, respectively, for diet 0:80 to 71 and 70%, respectively, for diet 80:0. Ruminal digestibility of fiber may have been enhanced due to linear (P less than .05) decreases in liquid and particulate dilution rates, resulting in increased ruminal residence time of fiber as alfalfa hay replaced AHP-WS. Liquid and particulate dilution rates decreased linearly from 6.4 and 5.2%/h, respectively, for diet 80:0 to 5.4 and 3.4%/h, respectively, for diet 0:80. Regression analysis of N data indicated that alfalfa N had a ruminal escape value of 26%.     

Escape protein for beef cows: III. Performance of lactating beef cows grazing smooth brome or big bluestem.

Grazing trials were conducted utilizing individually supplemented lactating beef cows in a 2-yr study to determine the effect of supplemental escape protein (EP) on cow/calf performance during the active growth periods of smooth brome and big bluestem. Graded levels (0, .11, .23, and .34 kg of EP/animal from an equal-protein-basis mixture of blood meal and corn gluten meal were offered daily, replacing corn starch and molasses, which was used as the energy control. All cows received 582 g of supplemental DM/d. Analyses for the smooth brome study indicated a cubic response (P less than .05) to milk production and calf daily gain. No response to EP supplementation (P greater than .10) was observed for the big bluestem study. Analysis of esophageal extrusa samples collected throughout the grazing seasons for both years demonstrated that quality of forage selected by the animal decreased as the trial progressed. Using a 16-h modified dacron bag technique that did not adjust for microbial attachment, protein escaping degradation for smooth brome and big bluestem was estimated to range from 7.0 to 14.5% and 22.2 to 30.3%, respectively. There was limited response to a moderate level of EP supplementation of lactating cows grazing smooth brome, but calf gains were depressed by both the low and the high levels of EP. Lactating cows grazing big bluestem did not respond to EP supplementation, but added energy supplement depressed calf gains.     

A histomorphologic and immunohistochemical study of chordoma in twenty ferrets (Mustela putorius furo).

The histomorphologic and immunohistochemical features of chordoma in 20 ferrets were evaluated. The mean age was 3.4 years, and, in the cases for which sex was known, females (n = 10) outnumbered males (n = 5) two to one. All 20 tumors occurred on the tip of the tail. Nineteen of 20 tumors (95%) were composed of three tissue components, often arranged concentrically with lobules of physaliferous cells at the periphery, trabecular bone in the center, and cartilage in between. The bone often contained marrow and hematopoietic cells. One tumor lacked chondromatous or osseous tissue. Immunohistochemical results were consistent with previous studies of chordoma. All 20 tumors (100%) were positive for keratin and vimentin intermediate filaments; 15 (75%) were positive for S-100 protein; and 17 (85%) were positive for neuron specific enolase. This neoplasm shares morphologic and immunohistochemical features with "classic," as well as chondroid chordoma, of human beings, making it a potential animal model.     

Clinical and pathologic features of prostatic adenocarcinoma in sexually intact and castrated dogs: 31 cases (1970-1987).

The medical records of 31 dogs diagnosed with prostatic carcinoma at the teaching hospital between January 1970 and October 1987 were reviewed to determine whether gender status had an effect on the clinical features or biologic behavior of the disease. The only significant difference between sexually intact and castrated dogs was increased prevalence of pulmonary metastasis in castrated dogs.     

Use of a flexed dorsoplantar radiographic view of the talocrural joint to evaluate lameness in two dogs.

A flexed dorsoplantar radiographic view of the talocrural joint was a useful additional view to diagnose abnormalities of the lateral trochlear ridge of the talus of 2 dogs. This view outlined the subchondral bone of both trochlear ridges of the talus and the apposing cochlea tibiae of the distal portion of the tibia. The tarsus was flexed at the level of the talocrural joint, and an x-ray beam was centered on the joint. With this additional view, fractures of the lateral trochlear ridge were readily diagnosed. This view would help to demonstrate osteochondral lesions of the lateral trochlear ridge.     

Evaluation of a guarded bronchoscopic method for microbial sampling of the lower airways in foals.

A novel method to reduce contamination of the bronchoscope during microbial sampling of the lower airways of foals was evaluated. Methylene blue (MB) was used as a nasopharyngeal dye marker to assess the relative contamination from the upper airways of bronchoalveolar lavage (BAL) specimens obtained by standard bronchoscopy (SB) and a "guarded" bronchoscopic method (GB). For GB, a clear sterile cellulose sheath was fitted over the bronchoscope in an effort to protect the endoscope tip and channel from contamination. Methylene blue was detected visually in seven of eight BAL samples from foals following SB, but in none of the samples recovered by GB (p less than 0.001). Significantly less MB was detected in BAL by spectrophotometry in the GB group as well (p less than 0.02). The GB was next employed to study the microbial flora in the lower airways of healthy weaned foals (n = 30). Bacteria were isolated from 29 of 30 (97%) BAL samples, and in moderate or large numbers from 26 of 30 (87%) of the foals. Potential pathogens, including Bordetella bronchiseptica, Streptococcus zooepidemicus, Staphylococcus aureus, Mycoplasma felis and Streptococcus pneumoniae, were cultured from the lower airways of foals. In conclusion, the bronchoscope and bronchoalveolar lavage specimens were readily contaminated by a dye marker placed in the nasopharynx of foals, and the degree of contamination was significantly reduced by sheathing the endoscope. This contamination during bronchoscopy may obscure the interpretation of isolates from BAL specimens from foals, which may possess a bacterial flora in the lower airways without cytological evidence of inflammation.