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Summary

A murine IgM monoclonal antibody causing bacterial agglutination was used in an immunoaffinity procedure to purify a serotype 1-specific polysaccharide epitope from Pasteurella haemolytica. The P haemolytica serotype 1-specific antibody was precipitated from peritoneal ascitic fluid, dialyzed, and covalently attached to cyanogen bromide-activated Sepharose 4B beads. Retention of purified antibody activity and coupling efficiency were > 99% when evaluated by elisa, agglutination testing, and protein determination. Potassium thiocyanate was selected as an eluant on the basis of reversible dissociation of bacterial agglutination and was titrated for the lowest effective concentration. Immunobead activity was observed microscopically by immobilization of encapsulated P haemolytica serotype 1 and its reversible dissociation after elution with 0.4M potassium thiocyanate. Specificity of immobilization was visualized, using P haemolytica serotypes 2 and 5, which were not bound, and by blocking serotype-1 binding with homologous capsular material. Saline-extractable capsular material from P haemolytica serotype 1 was used as an antigen source. After elution of the serotype 1-specific polysaccharide epitope, the product was dialyzed and analyzed, using chemical and immunologic methods. The immunoaffinity product contained no detectable protein and greater than half the original hexosamine content. Using defined monoclonal antibodies in elisa, titration of the original capsular material and the immunoaffinity product revealed specific retention of lipopolysaccharide, a 10- to 30-kd polysaccharide antigen common to all P haemolytica and P multocida serotypes, and serotype 1-specific capsular polysaccharide, indicating possible epitope sharing among polysaccharide antigens of P haemolytica serotype 1.

Free access
in American Journal of Veterinary Research

Summary

Virologic and pathologic investigations were done on prednisolone-treated bitches with a history of canine herpesvirus (chv) infection. Reactivation of chv was demonstrated in 5 Beagle bitches after daily administration of 600 mg of prednisolone for 5 days. The reactivation was confirmed in 4 of 5 bitches. Canine herpesvirus was recovered from nasal, oral, vaginal, and ocular secretions on the 5th to 21st days after initiation of treatment with prednisolone, and also from nasal mucosa and tonsil tissues. Results indicated that latent chv infections develop and that the virus may be reactivated, without clinical signs, in dogs with a history of chv infection.

Free access
in American Journal of Veterinary Research

Summary

Some interrelations between physical stress and immune competence were studied in pigs. One group of pigs underwent 2 intense short-term treadmill exercise tests, separated by an interval of 1 week, and another group served as controls. In vitro production of interferon α by blood mononuclear cells and the ability of lymphocytes to proliferate and produce interleukin 2 were chosen as markers of immune competence, plasma concentrations of cortisol, lactate, and purines were used as markers of physical stress. Blood samples were drawn from a catheter in situ 60 minutes before, immediately after, and at 10, 30, and 60 minutes, and 7, 24, and 72 hours after exercise. Physical stress resulted in immediate increase in the plasma concentrations of cortisol, lactate, and hypoxanthine, but had no effect on the blastogenic capability of lymphocytes or on their interleukin-2 production on either of the test occasions. Ability of blood mononuclear cells to produce interferon α in vitro was not affected by exercise stress.

Free access
in American Journal of Veterinary Research

Summary

Neutrophil migration through bovine teat tissues into the teat cistern, after endotoxin infusion into the teat cistern, was determined in vivo by 2 experimental procedures, indium-111 labeling of blood neutrophils, and obtaining multiple biopsy specimens from the teat cistern tissues. In both experiments, the number of leukocytes in the teat cistern flushing samples was continuously measured.

A lag phase of approximately 1 hour was required between endotoxin infusion into the teat cistern and the first observed neutrophil accumulation in the teat tissues. The rate of neutrophil accumulation in the teat tissues was highest between postinfusion (pi) hours 1 and 2, and the accumulation process ceased after pi hour 3. Neutrophils migrated toward the epithelium, and intraepithelial neutrophils were observed beginning approximately 2 hours after infusion, which coincided with the first influx of cells into the teat cistern. The cell influx into the teat cistern increased continuously up to pi hour 3, peaked between pi hours 3 and 5, and was close to preinfusion value at pi hour 22.

Use of indium-111-labeled neutrophils in the study of the inflammatory process provides a reliable noninvasive method to quantify cell migration in vivo. Use of biopsies allows quantification of the number of cells in different tissue areas, but has the disadvantage of being invasive. These 2 procedures complement each other, and could be of use in future studies of the local inflammatory process.

Free access
in American Journal of Veterinary Research

SUMMARY

The purpose of this study was to evaluate the effectiveness of an aromatic-dependent mutant of Salmonella typhimurium as a parenteral vaccine for prevention of fecal shedding of Salmonella spp. Pigs and chickens were vaccinated im, with 1 × 109 and 1 × 108 organisms, respectively, followed by a second identical vaccination 2 weeks later. Salmonella organisms were not detected by analysis of fecal or cloacal swab specimens from any animal after vaccination. Deleterious side effects were not noticed after vaccination. Pigs were challenge-inoculated po with 1 × 1012 virulent S typhimurium 1 week after the second vaccination. Chickens were challenge-inoculated po with 3 × 108 organisms of either S enteritidis or the virulent parent strain of S typhimurium 3 weeks after the second vaccination. Vaccinated pigs shed Salmonella spp significantly less frequently than did nonvaccinated pigs. Vaccinated chickens challenge-inoculated with either S enteritidis or S typhimurium also shed Salmonella less frequently than the corresponding nonvaccinated control birds; however, the difference was not significant.

Free access
in American Journal of Veterinary Research
in Journal of the American Veterinary Medical Association

Abstract

Objective

To test the immunocompetence of isogenic families of rainbow trout by measuring their ability to accept or reject skin grafts.

Animals

3 families of isogenic rainbow trout (Oncorhynchus mykiss), produced by mating homozygous females and homozygous males, plus 4 Chinook salmon (O tshawytscha) were used in these experiments.

Procedure

Grafts (allografts, members of the same family; autografts, donor and recipient were the same fish; and xenografts, O tshawytscha as donor) were exchanged. Grafts were applied on day 0 and removed on day 21, placed in neutral-buffered formalin, and embedded in paraffin. Lymphocytes and nuclei were counted in representative stained sections in the epidermis, dermis, and hypodermis. Results were analyzed by univariate analysis, using the Shapiro-Wilk statistic.

Results

Autografts were retained and minimal histologic changes were apparent. Allografts were histologically similar to autografts. Xenografts were rejected.

Conclusions

Results indicate that the immune system of isogenic rainbow trout is unable to distinguish between family members within isogenic families, but that a vigorous response is mounted against Chinook salmon xenografts. The isogenic rainbow trout are immunocompetent with respect to the phenomenon of graft rejection. (Am J Vet Res 1996;57:1576–1579)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To investigate ovarian responses and kinetics of gonadotropin-binding immunoglobulin production in domestic cats repeatedly treated with equine chorionic gonadotropin (eCG) and human chorionic gonadotropin (hCG) at short or long treatment intervals.

Design

Queens were treated 3 or 4 times with a standard eCG/hCG regimen at short (49 to 57 days) or long (130 to 135 days) intervals and subjected to laparoscopy after each treatment to evaluate ovarian follicular development. Serial serum samples were assessed by ELISA for the presence of eCG-binding immunoglobulins.

Animals

11 clinically normal sexually mature female cats.

Results

Queens repeatedly stimulated with eCG/hCG at long intervals typically had no decrease (P > 0.05) in ovarian follicle production or in maturity of recovered oocytes, whereas queens treated at short intervals had reduced (P < 0.05) follicular development and compromised oocyte maturity by the third stimulation. For both interval groups, ELISA data indicated individual variability in seroconversion after eCG/hCG challenge exposure. In general, queens treated at short intervals had higher peak anti-eCG immunoglobulin titer than did queens treated at long intervals; high titer at the time of eCG/hCG injection, or rapid increases in titer immediately after injection were predictive (P < 0.05) of poor ovarian responses.

Conclusions

Results suggest that individual variability in immune responses and intervals between repeated gonadotropin treatments determine whether queens develop immunologically mediated ovarian refractoriness to exogenous gonadotropins. Intervals of at least 4 months between successive eCG/hCG treatments are recommended for assisted reproductive procedures in domestic and nondomestic cats.(Am J Vet Res 1996;57:302-307)

Free access
in American Journal of Veterinary Research

Objective

To monitor the prevailing viral respiratory tract infections in cattle after transportation to feedlots.

Animals

100 cattle with signs of respiratory tract disease on arrival at 2 feedlots.

Procedures

Nasal swab samples were obtained from each animal and were used for inoculation of defined cell culture systems that detected bovine viruses known to cause respiratory tract infections, as well as viruses previously not recognized as respiratory pathogens for cattle.

Results

Bovine respiratory coronaviruses were isolated from 38 of the 100 cattle, including 6 of 50 cattle from California, 22 of 31 cattle from Oklahoma, 6 of 11 cattle from Texas, and 4 of 8 cattle of unknown origin. Parainfluenza 3 viruses also were isolated from 4 California cattle, but other bovine viruses were not detected.

Clinical Implications

The high rate of coronavirus isolations from feedlot cattle with signs of respiratory tract disease implied wide distribution and high susceptibility among cattle to this infection, which had not been detected by use of viral isolation systems in previous etiologic evaluations of feedlot cattle affected with bovine respiratory disease complex. (J Am Vet Med Assoc 1996; 208:1452-1455)

Free access
in Journal of the American Veterinary Medical Association