Evaluation of lipopolysaccharide-induced activation of equine neutrophils

OBJECTIVE: To evaluate lipopolysaccharide (LPS)-induced activation of equine neutrophils in blood. SAMPLE POPULATION: Blood samples from 5 healthy adult Thoroughbreds. PROCEDURES: Neutrophil integrin (CD11/CD18) expression, size variation, degranulation, and deformability were measured with and without incubation with LPS. Time and concentration studies were done. The mechanism of endotoxin-induced neutrophil activation was investigated by inactivating complement or preincubating neutrophils with inhibitors of tumor necrosis factor-alpha (TNF-alpha) synthesis, prostaglandin-leukotriene synthesis, or platelet-activating factor. RESULTS: Incubation of equine neutrophils with LPS increased cell surface expression of CD11/CD18, decreased neutrophil deformability, increased and decreased neutrophil size, and induced neutrophil degranulation. The LPS-induced neutrophil activation was attenuated by addition of inhibitors of TNF-alpha and prostaglandin-leukotriene synthesis. CONCLUSIONS AND CLINICAL RELEVANCE: Equine neutrophils are readily activated in vitro by LPS, resulting in increased expression of integrin adhesion molecules, decreased deformability, variation in neutrophil size, and degranulation. The tests used to detect activated neutrophils in this study may be useful in detecting in vivo neutrophil activation in horses with sepsis and endotoxemia.     

Multiple-center study of reduced-concentration triamcinolone topical solution for the treatment of dogs with known or suspected allergic pruritus

OBJECTIVE: To determine the efficacy of triamcinolone acetonide topical solution (TTS) in dogs for use in reduction of clinical signs of pruritic inflammatory skin diseases of a known or suspected allergic basis and to evaluate adverse effects associated with TTS administration. ANIMALS: 103 pruritic adult dogs with known or suspected allergic skin disease. PROCEDURE: Dogs were treated for 4 weeks with TTS or with vehicle solution (control dogs) in a multiple-center study. Clinical signs were scored by owners and by examining veterinarians before and after treatment. Blood samples obtained before and after treatment were subjected to routine hematologic and serum biochemical analyses. RESULTS: Treatment success, as defined by an improvement of at least 2 of 6 grades in overall clinical score, was evident in 35 of 52 (67%) TTS-treated dogs (mean improvement, 1.98) and 12 of 51 (24%) control dogs (mean improvement, 0.29). For several criteria, TTS was significantly more effective than vehicle in reducing clinical signs. Minor alterations in hematologic determinations in TTS-treated dogs were limited to slightly lower total leukocyte, lymphocyte, and eosinophil counts after treatment. Minor adverse effects were reported by owners in 6 of 52 (12%) TTS-treated and 9 of 51 (18%) control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Triamcinolone used as a spray solution at a concentration approximately one-sixth the concentration of triamcinolone topical preparations currently available for veterinary use is effective for short-term alleviation of allergic pruritus in dogs. Adverse effects are few and mild and, thus, do not preclude prolonged treatment with the solution.     

Effect of vaccination on serum concentrations of total and antigen-specific immunoglobulin E in dogs

OBJECTIVE: To determine the effect of vaccination on serum concentrations of total and antigen-specific IgE in dogs. ANIMALS: 20 female Beagles. PROCEDURE: Groups of 5 dogs each were vaccinated repeatedly between 8 weeks and 4 years of age with a multivalent and rabies vaccine, a multivalent vaccine only, or a rabies vaccine only. A fourth group of 5 dogs served as unvaccinated controls. Serum concentrations of total immunoglobulins and antigen-specific IgE were determined following vaccination. RESULTS:-The multivalent vaccine had little effect on serum total IgE concentrations. The concentration of IgE increased slightly following vaccination for rabies at 16 weeks and 1 year of age and increased greatly after vaccination at 2 and 3 years of age in most dogs, with a distinct variation between individual dogs. Vaccination had no effect on serum concentrations of IgA, IgG, and IgM as measured at 2 and 3 years of age. The rabies vaccine contained aluminum adjuvant in contrast to the multivalent vaccine. An increase of IgE that was reactive with vaccine antigens, including bovine serum albumin and bovine fibronectin, was detected in some of the dogs vaccinated for rabies. There was no significant correlation between serum concentrations of total IgE and antigen-specific IgE following vaccination. Serum total IgE concentration rapidly returned to preimmunization concentrations in most dogs, but high concentrations of antigen-specific IgE persisted. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination of dogs for rabies increases serum concentrations of total IgE and induces IgE specific for vaccine antigens, including tissue culture residues. Vaccination history should be considered in the interpretation of serum total IgE concentrations.     

Evaluation of a commercially available immunoassay for assessing adequacy of passive transfer in calves

OBJECTIVE: To evaluate diagnostic utility of a commercially available immunoassay for assessing adequacy of passive transfer of immunity in neonatal calves. DESIGN: Prospective study. ANIMALS: 123 calves. PROCEDURE: Blood and serum samples were obtained from the calves prior to 2 weeks of age. The immunoassay was performed, along with refractometry and an 18% sodium sulfite turbidity test. Serum IgG concentration was determined with a radial immunodiffusion assay. Sensitivity and specificity of the immunoassay, refractometry, and the sodium sulfite test were calculated by comparing results with results of the radial immunodiffusion assay. RESULTS: Sensitivity and specificity of the blood IgG immunoassay were 0.93 and 0.88, respectively, compared with 1.00 and 0.53 for the sodium sulfite test. For refractometry, sensitivity and specificity were 0.71 and 0.83, respectively, when a serum total solids concentration of 5.2 g/dl was used as the cutoff between positive and negative test results. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the immunoassay performs well in detecting calves with inadequate passive transfer of immunity.     

Effect of background serum lithium concentrations on the accuracy of lithium dilution cardiac output determination in dogs

OBJECTIVES: To assess the effect of increasing serum lithium concentrations on lithium dilution cardiac output (LiDCO) determination and to determine the ability to predict the serum lithium concentration from the cumulative lithium chloride dosage. ANIMALS: 10 dogs (7 males, 3 females). PROCEDURE: Cardiac output (CO) was determined in anesthetized dogs by measuring LiDCO and thermodilution cardiac output (TDCO). The effect of the serum lithium concentration on LiDCO was assessed by observing the agreement between TDCO and LiDCO at various serum lithium concentrations. Also, cumulative lithium chloride dosage was compared with the corresponding serum lithium concentrations. RESULTS: 44 paired observations were used. The linear regression analysis for the effect of the serum lithium concentration on the agreement between TDCO and LiDCO revealed a slope of -1.530 (95% confidence interval [CI], -2.388 to -0.671) and a y-intercept of 0.011 (r2 = 0.235). The linear regression analysis for the effect of the cumulative lithium chloride dosage on the serum lithium concentration revealed a slope of 2.291 (95% CI, 2.153 to 2.429) and a y-intercept of 0.008 (r2 = 0.969). CONCLUSIONS AND CLINICAL RELEVANCE: The LiDCO measurement increased slightly as the serum lithium concentration increased. This error was not clinically relevant and was minimal at a serum lithium concentration of 0.1 mmol/L and modest at a concentration of 0.4 mmol/L. The serum lithium concentration can be reliably predicted from the cumulative lithium dosage if lithium chloride is administered often within a short period.     

Safety and immunologic effects after inoculation of inactivated and combined live-inactivated dermatophytosis vaccines in cats

OBJECTIVE: To determine antidermatophyte immunologic effects of an experimental combined live-inactivated dermatophytosis vaccine (CLIDV) and a commercial inactivated dermatophytosis vaccine (IDV) in cats and to evaluate adverse effects associated with administration of these vaccines. ANIMALS: 20 healthy juvenile domestic shorthair cats. PROCEDURE: Cats were injected with 2 doses of CLIDV at the standard dosage or 1 dose of CLIDV at 10 times the standard dosage; IDV was administered at the manufacturer-recommended dosage. Cats were observed for illness and reactions at inoculation sites. Periodically, samples were obtained for fungal culture, lymphocyte blastogenesis test (LBT) as an indicator of cell-mediated immunity against dermatophyte antigens, and antidermatophyte IgG titers. Following vaccination, cats were challenge-exposed by topical application of Microsporum canis macroconidia and examined weekly for clinical signs of dermatophytosis. RESULTS: of 10 cats given CLIDV developed focal crusts at the injection site that resolved without treatment; these were areas of dermatophyte infection with the vaccine strain. Antidermatophyte IgG titers increased significantly with all vaccination protocols. Cellular immunity against M canis increased slightly and variably during the vaccination period and did not differ significantly between vaccinated and control cats. All cats developed dermatophyte infection after challenge exposure. Vaccination with CLIDV or IDV was associated with slightly reduced severity of initial infection. CONCLUSIONS AND CLINICAL RELEVANCE: Noculation with IDV or CLIDV did not provide prophylactic immunity against topical challenge exposure with M canis. Inoculation with either vaccine did not provide a more rapid cure of an established infection.     

An exercise in leadership training for veterinary students aiming for careers in biomedical research

A group discussion on the theme of "leadership" has been a central event in the annual Cornell Leadership Program for Veterinary Students since 1990. However, these discussions were often unfocused and did not readily demonstrate the leadership skills of distinguished guests who were invited to participate. Since 1998, a new format for this session has been developed in which students and guests are assigned individual roles in a scenario that is unfolded by a moderator over two to three hours. This role-playing exercise ensures that every student is obliged to participate and has an opportunity to practice such leadership skills as critical thinking, verbal communication, and decision making under pressure and with inadequate information. The distinguished guests, in their assigned roles, are able to interact freely with the student fellows and thus demonstrate their expertise as experienced leaders. This challenging experience has become an enjoyable part of the 10-week Leadership Program and one that shows the importance of leadership skills for those who aspire to careers in the biomedical sciences.     

Flow cytometric evaluation of canine bone marrow differential cell counts

Abstract: Three flow cytometric techniques were evaluated for determination of differential cell counts on canine clinical bone marrow specimens. Techniques included staining bone marrow specimens with 2'7'-dichlo-rofluorescein (DCF) or 3,3'-dihexyloxacarbocyanine iodide (DiOC6) and evaluation of forward-angle light scatter vs. side-angle light scatter plots. Flow cytometric evaluation of bone marrow cells stained with DCF failed to separate bone marrow cells into distinct cell populations. Staining with DiOC6 resulted in separation of bone marrow cells into populations of mature and immature erythroid cells, mature and immature myeloid cells, and lymphocytes. The scatter plot method resulted in identification of mature and immature erythroid cells, immature myeloid cells, metamyelocytes, and bands and segmenters. Lymphocytes could not be differentiated from mature erythroid cells by the scatter plot method. When the results of the DiOC6 method and the scatter plot method were compared with manual bone marrow differential cell counts, the scatter plot method had more similar mean values and higher correlation coefficients. The scatter plot method has the potential of providing rapid semiquantitative assessment of bone marrow differential cell counts in dogs for specimens that contain low numbers of lymphocytes.     

Genomic typing of Streptococcus uberis isolates from cases of mastitis, in New Zealand dairy cows, using pulsed-field gel electrophoresis

Three hundred and forty-two Streptococcus uberis isolates were cultured from milk samples from subclinical and clinical cases of dairy cattle mastitis. The samples were collected from 15 different New Zealand farming regions, including eight specific farms, during field research trials and veterinary diagnostic investigations. Pulsed-field gel electrophoresis was used to determine and compare the degree of genetic dissimilarity between the restriction endonuclease fragment pattern of the 342 New Zealand and a single United States S. uberis isolate. The 343 isolates exhibited 330 different restriction endonuclease fragment patterns. The United States isolate had a pattern unlike any of the New Zealand isolates. Most of the isolates were genetically different strains (pattern deferred by at least 33%), but identical patterns were noted within the same or different quarters of an individual cow, different cows within the same farm, and from different cows from the same or different districts, farming regions or islands. Seven of the eight selected farms had at most only one pair of isolates with banding patterns, which differed by less than 33%. A high degree of dissimilarity was noted in individual herds in which all the samples were collected on the same day or over a 2-year period. The high degree of dissimilar isolates is an indication that S. uberis infections in New Zealand dairy cattle are largely due to the opportunistic nature of the organism in the cows' environment. Prevention and treatment of S. uberis mastitis will therefore need to be directed at a multitude of different strains present throughout the country as well as in individual herds.