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Abstract

Objectives

To determine whether the vaccine Brucella abortus strain RB51 (SRB51) would infect dogs, be shed in urine or feces, or cause placentitis and abortion.

Animals

18 Beagles.

Procedure

Males (n = 3), nonpregnant females (n = 3), and pregnant females (n = 4) were inoculated orally with SRB51; control dogs (n = 2) were fed sterile saline solution. A separate group of pregnant females (n = 5) received SRB51 IV, and their controls (n = 1) received sterile saline solution IV. Dogs were observed twice daily for evidence of abortion. Urine and feces were collected periodically for bacteriologic culture, and blood was collected for bacteriologic culture and serologic analysis. At full gestation (oral and IV inoculated pregnant females) or on postinoculation day 49 (nonpregnant females and males), dogs were euthanatized and samples were collected for bacteriologic culture and microscopic examination.

Results

Abortion was not apparent during the study, and SRB51 was not found in samples of urine or feces from any dog. Strain RB51 was isolated from retropharyngeal lymph nodes from all orally inoculated dogs (9/9). One orally inoculated and 1 IV inoculated pregnant dog had SRB51 in placental tissues. Strain RB51 was also isolated from 1 fetus from the orally inoculated female dog with placentitis, but lesions were not detected in the fetus.

Conclusions and Clinical Relevance

Oral inoculation of nonpregnant female or male dogs with SRB51 did not result in shedding in urine or feces, although oropharyngeal lymph nodes became infected; in pregnant females, it caused infection of the placenta, with resulting placentitis and fetal infection, but abortion was not apparent. Intravenous inoculation resulted in infection of maternal spleen, liver, and placenta; however, fetal infection and abortion were not observed. Infected canine placental membranes or fluids may be a source of infection for other animals and human beings. (Am J Vet Res 1997;58:851–856)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine effects of intranasal inoculation with porcine reproductive and respiratory syndrome virus (PRRSV) or Bordetella bronchiseptica on challenge with nontoxigenic Pasteurella multocida in pigs.

Animals—Seventy 3-week-old pigs.

Procedure—In experiment 1, pigs were not inoculated (n= 10) or were inoculated with PRRSV (10), P multocida (10), or PRRSV followed by challenge with P multocida (10). In experiment 2, pigs were not inoculated (n = 10) or were inoculated with B bronchiseptica (10) or PRRSV and B bronchiseptica (10); all pigs were challenged with P multocida. Five pigs from each group were necropsied 14 and 21 days after initial inoculations.

ResultsPasteurella multocida was not isolated from tissue specimens of pigs challenged with P multocida alone or after inoculation with PRRSV. However, in pigs challenged after inoculation with B bronchiseptica, P multocida was isolated from specimens of the nasal cavity and tonsil of the soft palate. Number of bacteria isolated increased in pigs challenged after coinoculation with PRRSV and B bronchiseptica, and all 3 agents were isolated from pneumonic lesions in these pigs.

Conclusion and Clinical Relevance—Infection of pigs with B bronchiseptica but not PRRSV prior to challenge with P multocida resulted in colonization of the upper respiratory tract and tonsil of the soft palate with P multocida. Coinfection with PRRSV and B bronchiseptica predisposed pigs to infection of the upper respiratory tract and lung with P multocida. Porcine reproductive and respiratory syndrome virus and B bronchiseptica may interact to adversely affect respiratory tract defense mechanisms, leaving pigs especially vulnerable to infection with secondary agents such as P multocida. (Am J Vet Res 2001; 62:521–525)

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in American Journal of Veterinary Research

Abstract

Objective—To determine whether Mycobacterium bovis can be transmitted from experimentally infected deer to uninfected in-contact deer.

Animals—Twenty-three 6-month-old white-tailed deer.

Procedure—On day 0, M bovis (2 × 108 colony-forming units) was administered by intratonsillar instillation to 8 deer; 3 control deer received saline (0.9% NaCl) solution. Eight in-contact deer were comingled with inoculated deer from day 21. On day 120, inoculated deer were euthanatized and necropsied. On day 180, 4 in-contact deer were euthanatized, and 4 new incontact deer were introduced. On day 360, all in-contact deer were euthanatized. Rectal, oral, and nasal swab specimens and samples of hay, pelleted feed, water, and feces were collected for bacteriologic culture. Tissue specimens were also collected at necropsy for bacteriologic culture and histologic analysis.

Results—On day 90, inoculated and in-contact deer developed delayed-type hypersensitivity (DTH) reactions to purified protein derivative of M bovis. Similarly, new in-contact deer developed DTH reactions by 100 days of contact with original in-contact deer. Tuberculous lesions in in-contact deer were most commonly detected in lungs and tracheobronchial and medial retropharyngeal lymph nodes. Mycobacterium bovis was isolated from nasal secretions and saliva from inoculated and in-contact deer, urine and feces from in-contact deer, and hay and pelleted feed.

Conclusions and Clinical RelevanceMycobacterium bovis is efficiently transmitted from experimentally infected deer to uninfected in-contact deer through nasal secretions, saliva, or contaminated feed. Wildlife management practices that result in unnatural gatherings of deer may enhance both direct and indirect transmission of M bovis. (Am J Vet Res 2001;62:692–696)

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in American Journal of Veterinary Research

Abstract

Objective—To examine effects of co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and Bordetella bronchiseptica in pigs.

Animals—Forty 3-week-old pigs.

Procedure—30 pigs (10 pigs/group) were inoculated with PRRSV, B bronchiseptica, or both. Ten noninoculated pigs were control animals.

Results—Clinical signs, febrile response, and decreased weight gain were most severe in the group inoculated with both organisms. The PRRSV was isolated from all pigs in both groups inoculated with virus. All pigs in both groups that received PRRSV had gross and microscopic lesions consistent with interstitial pneumonia. Bordetella bronchiseptica was cultured from all pigs in both groups inoculated with that bacterium. Colonization of anatomic sites by B bronchiseptica was comparable between both groups. Pigs in the group that received only B bronchiseptica lacked gross or microscopic lung lesions, and B bronchiseptica was not isolated from lung tissue. In the group inoculated with B bronchiseptica and PRRSV, 3 of 5 pigs 10 days after inoculation and 5 of 5 pigs 21 days after inoculation had gross and microscopic lesions consistent with bacterial bronchopneumonia, and B bronchiseptica was isolated from the lungs of 7 of those 10 pigs.

Conclusions and Clinical Relevance—Clinical disease was exacerbated in co-infected pigs, including an increased febrile response, decreased weight gain, and B bronchiseptica-induced pneumonia. Bordetella bronchiseptica and PRRSV may circulate in a herd and cause subclinical infections. Therefore, co-infection with these organisms may cause clinical respiratory tract disease and leave pigs more susceptible to subsequent infection with opportunistic bacteria. (Am J Vet Res 2000;61:892–899)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To investigate the infection of calves with Mycobacterium bovis through oral exposure and transmission of M bovis from experimentally infected white-tailed deer to uninfected cattle through indirect contact.

Animals—24 11-month-old, white-tailed deer and 28 6-month-old, crossbred calves.

Procedure—In the oral exposure experiment, doses of 4.3 × 106 CFUs (high dose) or 5 × 103 CFUs (low dose) of M bovis were each administered orally to 4 calves; as positive controls, 2 calves received M bovis (1.7 × 105 CFUs) via tonsillar instillation. Calves were euthanatized and examined 133 days after exposure. Deer-to-cattle transmission was assessed in 2 phases (involving 9 uninfected calves and 12 deer each); deer were inoculated with 4 × 105 CFUs (phase I) or 7 × 105 CFUs (phase II) of M Bovis. Calves and deer exchanged pens (phase I; 90 days' duration) or calves received uneaten feed from deer pens (phase II; 140 days' duration) daily. At completion, animals were euthanatized and tissues were collected for bacteriologic culture and histologic examination.

Results—In the low- and high-dose groups, 3 of 4 calves and 1 of 4 calves developed tuberculosis, respectively. In phases I and II, 9 of 9 calves and 4 of 9 calves developed tuberculosis, respectively.

Conclusions and Clinical Relevance—Results indicated that experimentally infected deer can transmit M bovis to cattle through sharing of feed. In areas where tuberculosis is endemic in free-ranging white-tailed deer, management practices to prevent access of wildlife to feed intended for livestock should be implemented. (Am J Vet Res 2004;65:1483–1489)

Full access
in American Journal of Veterinary Research

SUMMARY

Objective

To determine the influence of brucellosis vaccination on tumor necrosis factor-α (TNF-α) con-centrations in pregnant cattle and the possible role of the bovine placenta in TNF-α production.

Animals

Polled Hereford heifers obtained from a nonvaccinated, brucellosis-free herd and bred at 16 to 27 months at age. All cattle were seronegative for Brucella abortus by results of the standard tube agglutination test.

Procedure

At 6 months' gestation, cattle were vaccinated IV with B abortus strain RB51 (n = 10), SC with B abortus strain RB51 (n = 5), or SC with B abortus strain 19 (n = 5); controls received pyrogen-free saline solution SC (n = 2). Blood samples were collected periodically for TNF-α assays. At necropsy, 8 to 12 weeks after vaccination, placental fluids and fetal blood were collected for TNF-α analysis and placental tissues were collected for immunohistochemical detection of TNF-α.

Results

Radioimmunoassays indicated no increase in TNF-α concentration in blood from IV or SC vaccinated cattle, compared with controls. Similarly, TNF-α concentrations in amniotic and allantoic fluids from SC vaccinated cattle were not different from values for controls. Although only IV vaccinated cattle developed placentitis, immunohistochemical analysis for TNF-α revealed increased immunoreactivity within placental trophoblastic epithelial cells of SC and IV vaccinated cattle.

Conclusions

SC vaccination for prevention of brucellosis, using recommended adult dosages, does not result in increase of TNF-α concentration in plasma, serum, or placental fluids; however, vaccination of pregnant cattle stimulates trophoblastic epithelial cells to express TNF-α, although the physiologic and quantitative importance of this expression remains unknown. (Am J Vet Res 1998;59:153–156)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To evaluate clearance of the vaccine strain, immunologic responses, and potential shedding of Brucella abortus strain RB51 organisms after vaccination of bison calves.

Animals

Fourteen 7-month-old female bison calves.

Procedure

10 bison calves were vaccinated SC with 1.22 × 1010 colony-forming units of B abortus strain RB51. Four bison calves were vaccinated SC with 0.15M NaCl solution. Rectal, vaginal, nasal, and ocular swab specimens were obtained to evaluate potential shedding by vaccinated bison. The superficial cervical lymph node was biopsied to evaluate clearance of the vaccine strain. Lymphocyte proliferative responses to strain RB51 bacteria were evaluated in lymph node cells obtained from biopsy specimens and also in peripheral blood mononuclear cells.

Results

Strain RB51 was recovered from superficial cervical lymph nodes of vaccinates examined 6, 12, and 18 weeks after vaccination (4/4, 3/4, and 1/4, respectively) but not in vaccinates examined at 24 weeks (0/3) after vaccination or nonvaccinates examined at all sample collection times (n = 1 bison/sample period). Serologic, immunologic, and bacterial culture techniques failed to reveal shedding of strain RB51 by vaccinates or infection of nonvaccinated bison. Lymphocyte proliferative responses were evident in lymph node cells and blood mononuclear cells from strain RB51-vaccinated bison beginning 12 weeks after vaccination.

Conclusion

Strain RB51 was cleared from bison by 18 to 24 weeks after vaccination. Bison vaccinated with strain RB51 did not shed the vaccine strain to nonvaccinated bison housed in close proximity. Strain RB51 did not induce antibody responses in bison that would interfere with brucellosis surveillance tests, but did stimulate cell-mediated immunity. (Am J Vet Res 1998;59:410–415)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To determine the safety and immunogenicity of Brucella abortus strain RB51 as a vaccine in pregnant cattle.

Animals

12 Polled Hereford heifers obtained from a brucellosis-free herd and bred on site at 16 months of age to a brucellosis-free bull.

Procedure

Pregnant heifers were vaccinated at 6 months’ gestation with 109 colony-forming units of B abortus strain RB51 (n = 5), 3 × 108 colony-forming units of B abortus strain 19 (n = 5), or sterile pyrogen-free saline solution (n = 2). Samples were periodically collected for serologic testing and lymphocyte blastogenesis assays. At full gestation, heifers were euthanatized and specimens were collected for bacteriologic culture, histologic analysis, and lymphocyte blastogenesis assay, using various antigenic stimuli.

Results

None of the strain RB51- or strain 19-vaccinates aborted or had gross or microscopic lesions at necropsy that were consistent with brucellosis. Maternal blood mononuclear cells from strain RB51- and strain 19-vaccinates had proliferative responses to γ-irradiated strain RB51 and strain 19 that were greater than responses by cells from nonvaccinated controls. In contrast, maternal superficial cervical lymph node cells from strain 19-vaccinates had proliferative responses to γ-irradiated strain RB51 or strain 19 bacteria greater than those of cells from RB51-vaccinates and nonvaccinated controls. None of the heifers vaccinated with strain RB51 developed antibodies detected by use of the standard tube agglutination test, but all developed antibodies to strain RB51 that reacted in a dot ELISA, using irradiated strain RB51 as antigen.

Conclusions

Pregnant cattle can be safely vaccinated with strain RB51 without subsequent abortion or placentitis. Furthermore, strain RB51 is immunogenic in pregnant cattle, resulting in humoral and cell-mediated immune responses, but does not interfere with serologic diagnosis of field infections. (Am J Vet Res 1997;58:472–477)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To inoculate white-tailed deer (Odocoileus virginianus) during the sixth or seventh week of gestation with bovine viral diarrhea virus (BVDV) and observe for signs of reproductive tract disease during a 182-day period.

Animals—10 pregnant white-tailed deer (8 seronegative and 2 seropositive [control deer] for BVDV).

Procedures—Deer were inoculated with 1 of 2 deer-derived BVDV strains (RO3-20663 or RO3-24272). Serum anti-BVDV antibody titers were determined prior to and 21 or 35 days after inoculation. Virus isolation (VI) procedures were performed on tissues from fetuses and does that died and on blood samples collected from live fawns. Ear notch specimens obtained from live fawns were assessed by use of BVDV antigen-capture ELISA (ACE).

Results—Both RO3-20663–inoculated seropositive deer gave birth to apparently normal fawns. Among the RO3-24272–inoculated seronegative deer, 1 died, and 1 aborted and 1 resorbed their fetuses; among the RO3-20663–inoculated seronegative deer, 3 died, 1 aborted its fetus, and 1 gave birth to 2 fawns that were likely persistently infected. On the basis of VI and ACE results, those 2 fawns were positive for BVDV; both had no detectable neutralizing anti-BVDV antibodies in serum.

Conclusions and Clinical Relevance—Reproductive tract disease that developed in pregnant white-tailed deer following BVDV inoculation was similar to that which develops in BVDV-exposed cattle. Methods developed for BVDV detection in cattle (VI, immunohistochemical evaluations, and ACE) can be applied in assessments of white-tailed deer. Fawns from does that had serum anti-BVDV antibodies prior to inoculation were protected against BVDV infection in utero.

Full access
in American Journal of Veterinary Research

Abstract

Objective

To examine the temporal development of tuberculous lesions in cattle inoculated with Mycobacterium bovis.

Animals

15 mature crossbred cows obtained from a herd with no history of M bovis infection.

Procedure

Inoculation of cattle was done by intratonsilar instillation of 1.48 × 105 to 5.4 × 107 colony-forming units of M bovis strain 2045T. At 3 to 4 hours, 4 weeks, 6 weeks, and 8 weeks after inoculation, tissues were examined for gross and microscopic lesions and processed for isolation of M bovis.

Results

Retropharyngeal lymph nodes from cattle examined 4 weeks after inoculation contained microgranulomas consisting of aggregates of macrophages with few neutrophils. Retropharyngeal lymph nodes from all cattle examined 6 and 8 weeks after inoculation contained multiple, large, coalescing granulomas consisting of central areas of necrosis with mild fibrosis, numerous macrophages, lymphocytes, plasma cells, multinucleated giant cells, and neutrophils. Three of 8 cattle examined 6 or 8 weeks after inoculation had lesions in nonretropharyngeal sites with morphologic characteristics similar to that seen in retropharyngeal lymph node granulomas from cattle examined 4 weeks after inoculation.

Conclusion

Granulomas can develop in draining lymph nodes of cattle in as little as 4 weeks after inoculation via intratonsilar instillation of M bovis. Intralesional morphologic changes between 4 and 6 weeks after inoculation indicate an increase in cellular chemotaxis and differentiation. Dissemination of bacteria to distant sites most likely was by lymphatic and hematogenous routes after establishment of the primary infection in retropharyngeal lymph nodes. (Am J Vet Res 1999;60:310–315)

Free access
in American Journal of Veterinary Research