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Summary

Supernatant preparations containing antigens produced by Pasteurella haemolytica serotype 1 grown in nonsupplemented RPMI 1640 medium and 3 grown in supplemented RPMI 1640 media were evaluated. Antigens were detected by immunoblotting each supernatant preparation with sera from 20 cattle with various degrees of resistance to experimentally induced pneumonic pasteurellosis. Antigen antibody bands at 49 and 30 kd were correlated with resistance in all 4 media. A 66-kd antigen-antibody band was correlated with resistance in 2 media, and antigen-antibody bands at 100 and 16 kd were correlated with resistance in 1 medium each. These results indicated that the number and relative amount of resistance-associated antigens in P haemolytica supernatants can be optimized on the basis of type of growth medium used.

Free access
in American Journal of Veterinary Research

Summary

Biological and biochemical characteristics of the leukotoxin of Fusobacterium necrophorum were determined. Culture supernatant of F necrophorum was toxic to polymorphonuclear neutrophilic, leukocytes from cattle and sheep, but not to those from pigs and rabbits. Culture supernatant and sonicated bacterial cell fractions had low hemolytic activity and did not cause dermonecrosis in a guinea pig. Supernatant derived leukotoxin was inactivated at 56 C for 5 minutes and became unstable at pH > 7.8 or < 6.6. Chemical treatment with 0.1% sodium dodecyl sulfate, 0.25% sodium deoxycholate, 5.2% sodium sulfide, or 0.25 mM titanium (III) citrate markedly decreased leukotoxicity. Enzymatic treatment with protease, trypsin, and chymotrypsin inactivated the toxin completely, whereas amylase had no effect. Use of protease inhibitors failed to prevent loss of leukotoxin activity. Using membrane partition chromatography and gel filtration, the estimated molecular weight of the toxin was > 300,000. On reduction and denaturation, the toxin dissociated into several components by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Free access
in American Journal of Veterinary Research

SUMMARY

Objective

To determine the resistance and susceptibility to antimicrobial compounds of Fusobacterium necrophorum isolates from bovine hepatic abscesses.

Procedure

37 isolates of F necrophorum (21 subsp necrophorum and 16 subsp funduliforme) isolated from bovine hepatic abscesses were obtained from cultures grown and maintained in anaerobic brain heart infusion broth. A broth dilution method was used as an initial screening to determine general susceptibility to 31 antimicrobial compounds. The minimal inhibitory concentrations (MIC) of 19 of the antimicrobial compounds that inhibited growth in the initial test were determined by use of the broth microdilution method.

Results

Fusobacterium necrophorum isolates were generally susceptible to penicillins, tetracyclines (chlortetracycline and oxytetracycline), lincosamides (clindamycin and lincomycin), and macrolides (tylosin and erythromycin), and were resistant to aminoglycosides (kanamycin, neomycin, gentamicin, and streptomycin), ionophores (except narasin), and peptides (avoparcin, polymyxin, and thiopeptin). The 5 antimicrobials (bacitracin, chlortetracycline, oxytetracycline, tylosin, and virginiamycin) that have FDA approval for prevention of liver abscesses in feedlot cattle were inhibitory to F necrophorum. Differences in antimicrobial susceptibility patterns were observed between the 2 subspecies only for clindamycin and lincomycin. The MIC of F necrophorum isolates from antibiotic-fed cattle were similar to those for isolates from nonantibiotic-fed cattle.

Conclusions

The MIC of FDA-approved antibiotics were not reflective of the efficacy of antibiotics in preventing liver abscesses in feedlot cattle. Also, continuous feeding of tylosin did not appear to select resistant F necrophorum. (Am J Vet Res 1998;59:44–47)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To compare results of polymerase chain reaction (PCR) testing of urine samples, serologic testing, and bacteriologic culture of urine to determine prevalence of urinary shedding of leptospires in dogs.

Design—Serial case study.

Animals—500 dogs evaluated serially without regard to health status.

Procedure—Urine samples were examined via PCR assay and bacteriologic culture for leptospires. Blood samples were analyzed for antibodies against serovars canicola, bratislava, pomona, icterohemorrhagiae, grippotyphosa, and hardjo.

Results—Titers ≥ 1:100 against at least 1 serovar were detected in 104 (20.8%) dogs, and titers ≥ 1:400 were detected in 41 (8.2%) dogs. High titers were detected most commonly to serovar grippotyphosa, followed by icterohemorrhagiae, canicola, pomona, bratislava, and hardjo. High titers to > 1 serovar were detected in 14 dogs. A positive PCR assay result was obtained in 41 (8.2%) dogs, only 9 of which had a titer ≥ 1:100. Leptospires were not cultured from the urine of any dog. Only 4 dogs had clinical leptospirosis. Overall disease prevalence was 0.8% for the 6-month evaluation period. Compared with PCR assay, serologic testing for predicting shedding had a sensitivity of 22%, specificity of 79%, positive predictive value of 9%, and negative predictive value of 92%.

Conclusions and Clinical Relevance—Irrespective of health status, 8.2% of dogs were shedding pathogenic leptospires. Serologic testing was a poor predictor of urinary shedding. Clinically normal dogs that shed leptospires may pose a zoonotic risk to their owners. (J Am Vet Med Assoc 2003;222:1230–1233)

Full access
in Journal of the American Veterinary Medical Association

Abstract

Objective

To determine the efficacy of leukotoxin-based Fusobacterium necrophorum vaccines and dietary tylosin in providing protection against experimentally induced hepatic abscesses in steers.

Design

30 steers assigned randomly to 6 treatment groups of 5 steers each: 1, phosphate-buffered saline solution (PBSS; control); 2, PBSS control, fed tylosin (100 mg/steer) daily; 3, inactivated whole-cell culture with oil emulsion adjuvant; 4, culture supernatant (crude toxoid) with oil emulsion adjuvant; 5, semipurified leukotoxoid with oil emulsion adjuvant; and 6, semipurified leukotoxoid with saponin adjuvant.

Procedure

Steers were inoculated SC with emulsified antigen or PBSS on days 0 and 21. Blood samples were collected at weekly intervals to monitor serum antileukotoxin antibody titer. On day 42, all steers were challenge exposed intraportally with F necrophorum culture. Three weeks later (day 63), steers were euthanatized and necropsied to examine liver and assess protection.

Results

Antileukotoxin antibody titers of all vaccinated groups markedly increased from baseline values, and mean titers of vaccinated groups were higher than those of the control and tylosin-treated groups. Steers vaccinated with culture supernatant with oil emulsion adjuvant or semipurified leukotoxoid with saponin adjuvant had the highest mean antibody titers. All 5 steers in the control group developed liver abscesses. Tylosin feeding did not protect steers challenge exposed with F necrophorum intraportally.

Conclusions

Culture supernatant was more protective than whole-cell culture or semipurified leukotoxin against experimentally induced hepatic abscesses. Partial purification of leukotoxin appeared to reduce its protective immunity. (Am J Vet Res 1996;57:483–488)

Free access
in American Journal of Veterinary Research

Objective

To establish ocular characteristics, determine nature and prevalence of ocular lesions, and identify representative bacterial flora from the conjunctiva of North American bison (Bison bison).

Design

Prospective study.

Animals

63 bison; 45 males and 18 females.

Procedure

Ophthalmic examinations were performed on 1 group of 38 bison in December 1997 and on a second group of 25 in March 1998. Eyes were examined with a penlight, magnification loop, and indirect ophthalmoscope. Two culture swabs were used to obtain samples from the inferior conjunctival sac. One swab was submitted for isolation of bacteria and the second was submitted for isolation of Mycoplasma organisms.

Results

15 ocular abnormalities were observed in 13 of the 63 bison. These included minor ocular discharge in 5 animals, 1 eyelid laceration, 1 periocular Demodex spp infection, 6 corneal abnormalities, 1 anterior synechia, and 1 cataract. Seventeen species of bacteria were isolated from the 63 swabs submitted for culture. The most prevalent bacteria were of the genus Bacillus (74.6%). Mycoplasma organisms were not observed.

Conclusions and Clinical Relevance

Corneal abnormalities were the most frequently identified ocular lesions in bison. Bacterial flora of the conjunctiva and ocular characteristics were similar to those reported for cattle. (J Am Vet Med Assoc 1999;215:1142–1144)

Free access
in Journal of the American Veterinary Medical Association

Abstract

Objective

To isolate Actinomyces pyogenes and A pyogenes-like (APL) organisms from the ruminal wall and ruminal contents of cattle and compare them with isolates from liver abscesses from the same animals, using ribosomal DNA restriction fragment length polymorphism analysis or ribotyping.

Procedure

Specimens of liver abscesses, ruminal walls, and ruminal contents were collected from 59 cattle at slaughter. All β-hemolytic, pinpoint colonies that were gram positive, pleomorphic rod-shaped, and catalase negative, and that hydrolyzed casein and gelatin were presumptively identified as A pyogenes and were characterized biochemically, using an identification kit. The isolates that resembled A pyogenes but fermented mannitol or raffinose, or both, were called APL organisms. Isolates from the ruminal wall and ruminal contents were compared with liver abscess isolates from the same animal by use of ribotyping.

Results

Actinomyces pyogenes and APL organisms were isolated more frequently from the ruminal wall than from ruminal contents. Ruminal isolates of A pyogenes and APL had biochemical characteristics similar to those of the isolates from liver abscesses. Among 6 sets of isolates (4 A pyogenes and 2 APL), 2 isolates from liver abscesses had ribopatterns identical to the corresponding ruminal wall isolates. Also, the APL organisms isolated from the ruminal content matched with the corresponding liver abscess isolates for both sets of specimens tested.

Conclusions

The ruminal wall may be the niche for A pyogenes and APL organisms in the rumen. The genetic similarity, on the basis of ribotyping among isolates from liver abscesses, the ruminal wall, and ruminal contents of the same animal suggests that A pyogenes and APL organisms that cause liver abscesses originate from the rumen. (Am J Vet Res 1998;59:271–276)

Free access
in American Journal of Veterinary Research

Summary

Conjunctival swab specimens from healthy pigs were cultured to determine normal microbial population. Four commercial swine operations were selected for study. Pigs of 4 age groups were tested: nursing pigs, nursery pigs, feeder pigs, and sows. Swab specimens were taken from the conjunctival sac of each pig. Bacterial, fungal, and mycoplasmal growth was determined separately. Chlamydia sp was detected by use of an elisa. Bacteria were recovered from 98% of specimens evaluated. α-Streptococcus sp (89%) was the most commonly recovered organism, followed by Staphylococcus epidermidis (39%) and Staphylococcus sp (39%). Mycoplasma sp was not detected in any of the specimens. Chlamydia sp was identified in 28% of all specimens evaluated. These results are similar to reports of normal conjunctival flora in other domestic animals.

Free access
in American Journal of Veterinary Research

Summary

Eighteen 4-week-old pigs were used in a study to evaluate tiamulin in drinking water for control of experimentally induced Streptococcus suis type-2 infection. Pigs in groups A and B (n = 6 pigs/group) were aerosolized with a logarithmic-growth phase culture of S suis type 2, whereas pigs in group C (n = 6 pigs) served as noninfected and nonmedicated controls. After exposure to S suis, pigs in group B were given 180 mg of tiamulin/L of drinking water for 5 days. Pigs in group B consumed more feed (P = 0.009) and gained body weight faster (P = 0.02) than did pigs in group A. Pigs in group A had higher rectal temperature (P = 0.05) for up to 7 days after S suis exposure, higher clinical sign scores (P = 0.008), higher serum cortisol concentration on days 7 and 14, higher gross lesion scores (P = 0.03), and higher microscopic lesion scores (P = 0.01) than did pigs in groups B and C. Gross and microscopic lesions in pigs of groups A and B included meningitis, pneumonia, pleuritis, pericarditis, peritonitis, and synovitis of variable severity. Streptococcus suis type 2 was recovered from tissue specimens of 2 group-A pigs and 1 group-B pig. Data indicated that tiamulin administered via drinking water significantly reduced the effects of S suis type-2 infection.

Free access
in Journal of the American Veterinary Medical Association

SUMMARY

Neutrophils were purified from blood of dexamethasone-treated (0.04 mg/kg of body weight) and untreated calves. Cells were untreated (controls) or cultured in media containing 5 or 10 ng of bovine recombinant granulocyte-macrophage colony-stimulating factor (rbgm-csf)/ml for 10 to 12 hours before being tested for various functions. Dexamethasone treatment of calves decreased luminol-dependent chemiluminescence, decreased phagocytosis of Pasteurella multocida and several Staphylococcus spp by various degrees, and decreased antibody-dependent cell-mediated cytotoxocity against bovine herpesvirus-infected cells by 26 to 32%. The percentage phagocytosis of coagulase-positive S aureus and S intermedius was higher than that of coagulase-negative S epidermidis for neutrophils from all calves. Culture of neutrophils with rbgm-csf significantly increased (P < 0.05) all of the aforementioned functions, compared with control neutrophils; however, rbgm-csf-induced increases in function tended to be higher in neutrophils from dexamethasone-treated calves than in neutrophils from untreated calves.

Free access
in American Journal of Veterinary Research