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Sample records for pasteurella multocida bordetella

  1. Serum haptoglobin concentration in growing swine after intranasal challenge with Bordetella bronchiseptica and toxigenic Pasteurella multocida type D.

    PubMed Central

    Francisco, C J; Shryock, T R; Bane, D P; Unverzagt, L

    1996-01-01

    The acute phase reaction, in association with progressive atrophic rhinitis (AR), was monitored for 3 wk using serum haptoglobin (HPT) quantification in thirty-six, 15 kg swine after intranasal challenge with varying doses of Pasteurella multocida type D (toxigenic strain) and Bordetella bronchiseptica. The challenge doses were administered alone or in combination with pigs divided into 9 isolated treatment groups. Increasing doses of B. bronchiseptica were associated with lower serum HPT (P < 0.05), whereas increasing doses of P. multocida tended to increase serum HPT (0.05 < P < 0.10). Significant and positive correlation of mean HPT and AR score was found in these pigs; increased AR scores were associated with elevated mean HPT concentration (r = 0.41, P < 0.01). A significant interaction between P. multocida and B. bronchiseptica dose indicated that increasing the dose of B. bronchiseptica, for a fixed P. multocida dose, was associated with less AR (P < 0.05). The AR scores were greater in pigs given P. multocida, than B. bronchiseptica alone. These results indicate that a complex interaction between Pasteurella multocida and Bordetella bronchiseptica causes progressive atrophic rhinitis and alters serum HPT concentration in swine. Images Figure I. Figure II. Figure III. Figure IV. PMID:8809387

  2. Association between Pneumocystis spp. and co-infections with Bordetella bronchiseptica, Mycoplasma hyopneumoniae and Pasteurella multocida in Austrian pigs with pneumonia.

    PubMed

    Kureljušić, B; Weissenbacher-Lang, C; Nedorost, N; Stixenberger, D; Weissenböck, H

    2016-01-01

    In this retrospective study, 218 pig lung tissue samples were analyzed to examine a possible association between Pneumocystis spp. using in situ hybridization, Bordetella bronchiseptica (B.b.) using immunohistochemistry (IHC), Mycoplasma hyopneumoniae (M.h.) by quantitative PCR, and Pasteurella multocida (P.m.; IHC). Compared to the bacterial agents (B.b., 5%; M.h., 30%; P.m., 23%), Pneumocystis occurred with a higher prevalence (51%). Co-infections with two or three pathogens were present in 28% of the examined cases. Those of Pneumocystis and M.h. were most commonly seen, followed by Pneumocystis and P.m. and M.h. and P.m. Histologically, interstitial pneumonia was found in both the Pneumocystis positive lungs and lungs with a mild M.h. infection. The B.b. and P.m. positive lungs were mainly associated with suppurative bronchopneumonia and severe M.h. cases with fibrinous or fibrino-haemorrhagic pneumonia. In suckling piglets, the number of samples positive for Pneumocystis predominated, whereas samples from fattening pigs were mainly positive for bacteria or Pneumocystis and bacteria.

  3. 9 CFR 113.69 - Pasteurella Multocida Vaccine, Bovine.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Pasteurella Multocida Vaccine, Bovine... REQUIREMENTS Live Bacterial Vaccines § 113.69 Pasteurella Multocida Vaccine, Bovine. Pasteurella Multocida Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  4. 9 CFR 113.69 - Pasteurella Multocida Vaccine, Bovine.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Multocida Vaccine, Bovine... REQUIREMENTS Live Bacterial Vaccines § 113.69 Pasteurella Multocida Vaccine, Bovine. Pasteurella Multocida Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  5. 9 CFR 113.69 - Pasteurella Multocida Vaccine, Bovine.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Pasteurella Multocida Vaccine, Bovine... REQUIREMENTS Live Bacterial Vaccines § 113.69 Pasteurella Multocida Vaccine, Bovine. Pasteurella Multocida Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  6. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or...

  7. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or...

  8. 9 CFR 113.69 - Pasteurella Multocida Vaccine, Bovine.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Pasteurella Multocida Vaccine, Bovine... REQUIREMENTS Live Bacterial Vaccines § 113.69 Pasteurella Multocida Vaccine, Bovine. Pasteurella Multocida Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  9. 9 CFR 113.69 - Pasteurella Multocida Vaccine, Bovine.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Pasteurella Multocida Vaccine, Bovine... REQUIREMENTS Live Bacterial Vaccines § 113.69 Pasteurella Multocida Vaccine, Bovine. Pasteurella Multocida Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  10. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or...

  11. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or...

  12. 9 CFR 113.70 - Pasteurella Multocida Vaccine, Avian Isolate.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Pasteurella Multocida Vaccine, Avian... REQUIREMENTS Live Bacterial Vaccines § 113.70 Pasteurella Multocida Vaccine, Avian Isolate. Pasteurella Multocida Vaccine, Avian Isolate, shall be prepared as a desiccated live culture of an avirulent or...

  13. Intranasal immunization of mice against Pasteurella multocida.

    PubMed Central

    Smith, R H; Babiuk, L A; Stockdale, P H

    1981-01-01

    A potassium thiocyanate (KSCN) extract of Pasteurella multocida serotype III:A was shown to protect mice from an intranasal challenge with up to 300 50% lethal doses of P. multocida. In addition to preventing death, bacteria were rapidly cleared from the lungs of immunized mice so that by 72 to 96 h postchallenge no bacteria were present in the lungs of immunized mice, whereas up to 10(9) bacteria were present in lungs of nonimmunized mice. Immunization by the intranasal route was slightly better than that by the intramuscular route. Protection was considered specific, since immunization with P. multocida protected only against P. multocida and not against Salmonella agona. Furthermore, a similar KSCN extract from P. haemolytica did not protect against P. multocida challenge. A comparison of the KSCN extract with a Formalin-killed bacterin suggested that the KSCN extract may be superior to the bacterin. PMID:7216441

  14. Pasteurella multocida meningitis in an adult: case report

    PubMed Central

    Armstrong, G; Sen, R; Wilkinson, J

    2000-01-01

    Pasteurella multocida is known to form part of the normal flora in the nasopharynx or gastrointestinal tract in many domestic and wild animals. Most human P multocida infections are soft tissue infections caused by dog or cat bites. Less commonly this bacterium is associated with infections affecting other organ systems of man. A case of fatal P multocida meningitis discovered at the necropsy of a 52 year old man is described. P multocida is an unusual causative agent of meningitis which tends to affect those at the extremes of age. Key Words: Pasteurella multocida • meningitis PMID:10823146

  15. Pasteurella multocida Bacteremia in an Immunocompromised Patient

    PubMed Central

    Parekh, Jai; Townley, Theresa

    2016-01-01

    We present the case of a 61-year-old Caucasian gentleman who presented with a one-day history of fever, chills, and altered mental status. His symptoms were initially thought to be secondary to cellulitis. Blood cultures grew Pasteurella multocida, a rare pathogen to cause bacteremia. Our patient was treated with ciprofloxacin for two weeks and made a complete and uneventful recovery. Our patient's uncontrolled diabetes mellitus and chronic kidney disease put him at a higher risk for developing serious P. multocida infection. The patient's dog licking the wounds on his legs was considered as the possible source of infection. As P. multicoda bacteremia is rare, but severe with a high mortality rate, it is imperative to have a high index of suspicion for this infection especially in the vulnerable immunocompromised population. PMID:27847521

  16. [Biochemical tests for identifying Pasteurella multocida].

    PubMed

    Karaivanov, L

    1984-01-01

    Studied was the biochemical activity of a total of 168 strains of Pasteurella--73 isolated from birds (48 from cases of acute fowl cholera, and 25--of chronic cholera), and 95 isolated from mammals (3 from lambs, 24 from pigs, 36 from cattle, and 32 from rabbits) with regard to the tests determining the hemolytic activity, production of indol, reduction of nitrates, breakdown of urea, beta galactosidase activity, production of hydrogen sulfide, ornitin-, arginine-, lysine-decarboxylase-, and phosphatase activity, and the fermentation of substrates such as manite, glucose, galactose, saccharose, manose, levulose, dulcite, lactose, maltose, rafinose, trechalose, salicin, melobiose, icelobiose, arabinose, xylose, and sorbite. To differentiate Pasteurella multocida strains isolated from mamals from those isolated from birds the phosphatase activity test on solid media with sodium phenolphtalein diphosphate had to be employed Pasteurella organisms isolated from mammals showed positive phosphatase activity, while those isolated from birds exhibited a negative one. Arabinose and xylose fermentation tests could simultaneously be used. Pasteurellae isolated in cases of acute fowl cholera showed positive reaction for arabinose and a negative one for xylose, while the strains isolated from mammals showed the reverse activity. The strains isolated in cases of chronic fowl cholera were shown to belong to this group.

  17. Pasteurella multocida: from Zoonosis to Cellular Microbiology

    PubMed Central

    Ho, Mengfei

    2013-01-01

    SUMMARY In a world where most emerging and reemerging infectious diseases are zoonotic in nature and our contacts with both domestic and wild animals abound, there is growing awareness of the potential for human acquisition of animal diseases. Like other Pasteurellaceae, Pasteurella species are highly prevalent among animal populations, where they are often found as part of the normal microbiota of the oral, nasopharyngeal, and upper respiratory tracts. Many Pasteurella species are opportunistic pathogens that can cause endemic disease and are associated increasingly with epizootic outbreaks. Zoonotic transmission to humans usually occurs through animal bites or contact with nasal secretions, with P. multocida being the most prevalent isolate observed in human infections. Here we review recent comparative genomics and molecular pathogenesis studies that have advanced our understanding of the multiple virulence mechanisms employed by Pasteurella species to establish acute and chronic infections. We also summarize efforts being explored to enhance our ability to rapidly and accurately identify and distinguish among clinical isolates and to control pasteurellosis by improved development of new vaccines and treatment regimens. PMID:23824375

  18. Hydrogen Peroxide as an Effective Disinfectant for Pasteurella multocida

    PubMed Central

    Jung, In-Soo; Kim, Hyun-Jung; Jung, Won-Yong

    2014-01-01

    Pasteurella multocida (P. multocida) infections vary widely, from local infections resulting from animal bites and scratches to general infections. As of yet, no vaccine against P. multocida has been developed, and the most effective way to prevent pathogenic transmission is to clean the host environment using disinfectants. In this study, we identified which disinfectants most effectively inhibited environmental isolates of P. multocida. Three readily available disinfectants were compared: 3% hydrogen peroxide (HP), 70% isopropyl alcohol, and synthetic phenol. In suspension tests and zone inhibition tests, 3% HP was the most promising disinfectant against P. multocida. PMID:24954350

  19. Community-acquired pneumonia due to Pasteurella multocida.

    PubMed

    Marinella, Mark A

    2004-12-01

    Most cases of community-acquired pneumonia result from infection with predictable common pathogens. However, rare patients develop pneumonia from unusual bacterial species such as Pasteurella multocida, a Gram-negative oral commensal of most dogs and cats. The majority of P. multocida infections involve skin and soft tissue and complicate a bite or scratch. I report the case of an elderly man who owned 16 cats and developed bacteremic pneumonia with P. multocida. .

  20. Pasteurella multocida bacterial meningitis caused by contact with pigs

    PubMed Central

    López, C.; Sanchez-Rubio, P.; Betrán, A.; Terré, R.

    2013-01-01

    Pasteurella multocida belongs to the normal flora of the respiratory and digestive tract of many animals. Animal exposure is a considerable risk factor for Pasteurella infection. P. multocida is the most common cause of local infection after an animal bite but is an unusual cause of meningitis. We present a case of bacterial meningitis by P. multocida in a 37-year-old man who worked in a pig farm and was bitten by a pig. The patient had a defect located in the lamina cribosa and this lesion could be the gateway of the infection, although in this case the infection could also be acquired through the pig bite. The bacteria was identified as P. multocida with the biochemical test API 20E (bioMérieux). In agreement with findings in the literature, the strain was susceptible in vitro to penicillin, ampicillin, cefotaxime, ceftriaxone ciprofloxacin, levofloxacin, imipenem and tetracycline. PMID:24294240

  1. A cryopreservation method for Pasteurella multocida from wetland samples

    USGS Publications Warehouse

    Moore, Melody K.; Shadduck, D.J.; Goldberg, D.R.; Samuel, M.D.

    1998-01-01

    A cryopreservation method and improved isolation techniques for detection of Pasteurella multocida from wetland samples were developed. Wetland water samples were collected in the field, diluted in dimethyl sulfoxide (DMSO, final concentration 10%), and frozen at -180 C in a liquid nitrogen vapor shipper. Frozen samples were transported to the laboratory where they were subsequently thawed and processed in Pasteurella multocida selective broth (PMSB) to isolate P. multocida. This method allowed for consistent isolation of 2 to 18 organisms/ml from water seeded with known concentrations of P. multocida. The method compared favorably with the standard mouse inoculation method and allowed for preservation of the samples until they could be processed in the laboratory.

  2. A Case of Polyarticular Pasteurella multocida Septic Arthritis

    PubMed Central

    Nitoslawski, Sarah; McConnell, Todd M.; Semret, Makeda; Stein, Michael A.

    2016-01-01

    A 76-year-old man with a history of osteoarthritis presents with right leg erythema and inability to weight-bear and pain in his right shoulder. Synovial fluid cell count of the knee and shoulder showed abundant neutrophils, and cultures of the knee showed growth of Pasteurella multocida. The patient owned four cats with which he had frequent contact, but history and physical examination elicited no evidence of scratches or bites. This case highlights the invasive potential of Pasteurella multocida in an immunocompetent individual and its capacity to cause septic arthritis in the setting of frequent animal contact. PMID:27366169

  3. Pasteurella multocida Toxin Triggers RANKL-Independent Osteoclastogenesis.

    PubMed

    Chakraborty, Sushmita; Kloos, Bianca; Harre, Ulrike; Schett, Georg; Kubatzky, Katharina F

    2017-01-01

    Bone remodeling is a continuous process to retain the structural integrity and function of the skeleton. A tight coupling is maintained between osteoclast-mediated resorption of old or damaged bones and osteoblast-mediated formation of new bones for bone homeostasis. While osteoblasts differentiate from mesenchymal stem cells, osteoclasts are hematopoietic in origin and derived from myeloid precursor cells. Osteoclast differentiation is driven by two cytokines, cytokine receptor activator of NF-κB ligand (RANKL), and macrophage colony-stimulating factor. Imbalances in the activity of osteoblasts and osteoclasts result in the development of bone disorders. Bacterially caused porcine atrophic rhinitis is characterized by a loss of nasal ventral conche bones and a distortion of the snout. While Bordetella bronchiseptica strains cause mild and reversible symptoms, infection of pigs with toxigenic Pasteurella multocida strains causes a severe and irreversible decay. The responsible virulence factor Pasteurella multocida toxin (PMT) contains a deamidase activity in its catalytical domain that constitutively activates specific heterotrimeric G proteins to induce downstream signaling cascades. While osteoblasts are inhibited by the toxin, osteoclasts are activated, thus skewing bone remodeling toward excessive bone degradation. Still, the mechanism by which PMT interferes with bone homeostasis, and the reason for this unusual target tissue is not yet well understood. Here, we show that PMT has the potential to differentiate bone marrow-derived macrophages into functional osteoclasts. This toxin-mediated differentiation process is independent of RANKL, a cytokine believed to be indispensable for triggering osteoclastogenesis, as addition of osteoprotegerin to PMT-treated macrophages does not show any effect on PMT-induced osteoclast formation. Although RANKL is not a prerequisite, toxin-primed macrophages show enhanced responsiveness to low concentrations of RANKL

  4. Pasteurella multocida Toxin Triggers RANKL-Independent Osteoclastogenesis

    PubMed Central

    Chakraborty, Sushmita; Kloos, Bianca; Harre, Ulrike; Schett, Georg; Kubatzky, Katharina F.

    2017-01-01

    Bone remodeling is a continuous process to retain the structural integrity and function of the skeleton. A tight coupling is maintained between osteoclast-mediated resorption of old or damaged bones and osteoblast-mediated formation of new bones for bone homeostasis. While osteoblasts differentiate from mesenchymal stem cells, osteoclasts are hematopoietic in origin and derived from myeloid precursor cells. Osteoclast differentiation is driven by two cytokines, cytokine receptor activator of NF-κB ligand (RANKL), and macrophage colony-stimulating factor. Imbalances in the activity of osteoblasts and osteoclasts result in the development of bone disorders. Bacterially caused porcine atrophic rhinitis is characterized by a loss of nasal ventral conche bones and a distortion of the snout. While Bordetella bronchiseptica strains cause mild and reversible symptoms, infection of pigs with toxigenic Pasteurella multocida strains causes a severe and irreversible decay. The responsible virulence factor Pasteurella multocida toxin (PMT) contains a deamidase activity in its catalytical domain that constitutively activates specific heterotrimeric G proteins to induce downstream signaling cascades. While osteoblasts are inhibited by the toxin, osteoclasts are activated, thus skewing bone remodeling toward excessive bone degradation. Still, the mechanism by which PMT interferes with bone homeostasis, and the reason for this unusual target tissue is not yet well understood. Here, we show that PMT has the potential to differentiate bone marrow-derived macrophages into functional osteoclasts. This toxin-mediated differentiation process is independent of RANKL, a cytokine believed to be indispensable for triggering osteoclastogenesis, as addition of osteoprotegerin to PMT-treated macrophages does not show any effect on PMT-induced osteoclast formation. Although RANKL is not a prerequisite, toxin-primed macrophages show enhanced responsiveness to low concentrations of RANKL

  5. Pasteurella multocida serotype 1 isolated from a lesser snow goose

    USGS Publications Warehouse

    Samuel, M.D.; Goldberg, D.R.; Shadduck, D.J.; Price, J.I.; Cooch, E.G.

    1997-01-01

    Pharyngeal swabs were collected from 298 lesser snow geese (Chen caerulescens caerulescens) at Banks Island (Northwest Territories. Canada) in the summer of 1994. Pasteurella multocida serotype 1 was isolated from an adult male bird and P. multocida serotype 3 was isolated from an adult female goose. Pathogenicity of the serotype 1 isolate was confirmed by inoculation in Pekin ducks (Anas platyrhynchos). The serotype 3 isolate was non-pathogenic in Pekin ducks. This is the first documented isolation of pathogenic P. multocida serotype 1 from apparently healthy wild snow geese.

  6. Pasteurella multocida subsp. multocida and P. multocida subsp. septica Differentiation by PCR Fingerprinting and α-Glucosidase Activity

    PubMed Central

    Hunt Gerardo, Sharon; Citron, Diane M.; Claros, Marina C.; Fernandez, Helen T.; Goldstein, Ellie J. C.

    2001-01-01

    Pasteurella multocida is composed of three subspecies that are often differentiated by fermentation of sorbitol and dulcitol. We studied 35 dulcitol-negative P. multocida isolates from infected dog and cat bite wounds, 16 of which yielded weak and/or conflicting fermentation reactions in Andrades sorbitol, thus making it difficult to distinguish between the two dulcitol-negative subspecies of P. multocida, i.e., P. multocida subsp. multocida and P. multocida subsp. septica. All isolates and two control strains were further analyzed using a PCR fingerprinting technique with a single primer (M13 core) and assessed for α-glucosidase (α-Glu) activity. Although the PCR fingerprint patterns and α-Glu activity did not correlate well with the sorbitol fermentation reactions, they did correlate well with each other. All strains identified as P. multocida subsp. septica were positive for α-Glu activity and exhibited the group I PCR fingerprint profile. All strains categorized as P. multocida subsp. multocida displayed either the group II or group III PCR fingerprint profile; 9 of 11 of these isolates were α-Glu negative. These data suggest that both PCR fingerprinting and α-Glu activity provide reliable means for differentiating P. multocida subsp. multocida from P. multocida subsp. septica, particularly in strains that produce weak and/or discrepant sorbitol fermentation reactions. PMID:11427568

  7. Genome Sequence of Pasteurella multocida Strain Razi_Pm0001

    PubMed Central

    Tadayon, Keyvan

    2017-01-01

    ABSTRACT We report here the genome sequence of Pasteurella multocida Razi_Pm0001 from bovine origin, isolated in Iran in 1936. The genome has a size of 2,360,663 bp, a G+C content of 40.4%, and is predicted to contain 2,052 coding sequences. PMID:28153892

  8. Persistence of Pasteurella multocida in wetlands following avian cholera outbreaks

    USGS Publications Warehouse

    Blanchong, Julie A.; Samuel, M.D.; Goldberg, D.R.; Shadduck, D.J.; Lehr, M.A.

    2006-01-01

    Avian cholera, caused by Pasteurella multocida, affects waterbirds across North America and occurs worldwide among various avian species. Once an epizootic begins, contamination of the wetland environment likely facilitates the transmission of P. multocida to susceptible birds. To evaluate the ability of P. multocida serotype-1, the most common serotype associated with avian cholera in waterfowl in western and central North America, to persist in wetlands and to identify environmental factors associated with its persistence, we collected water and sediment samples from 23 wetlands during winters and springs of 1996a??99. These samples were collected during avian cholera outbreaks and for up to 13 wk following initial sampling. We recovered P. multocida from six wetlands that were sampled following the initial outbreaks, but no P. multocida was isolated later than 7 wk after the initial outbreak sampling. We found no significant relationship between the probability of recovery of P. multocida during resampling and the abundance of the bacterium recovered during initial sampling, the substrate from which isolates were collected, isolate virulence, or water quality conditions previously suggested to be related to the abundance or survival of P. multocida. Our results indicate that wetlands are unlikely to serve as a long-term reservoir for P. multocida because the bacterium does not persist in wetlands for long time periods following avian cholera outbreaks.

  9. Pasteurella Multocida Peritonitis After Cat Scratch in a Patient with Cirrhotic Ascites

    PubMed Central

    Gunathilake, Roshan; Verma, Ajay; Caffery, Michael; Sowden, Sowden

    2015-01-01

    Pasteurella multocida, a zoonotic agent transmitted by canines and felines, has been very rarely reported to cause bacterial peritonitis in humans. Pasteurella multocida peritonitis is associated with high mortality even with appropriate treatment, therefore its early recognition is essential. We report a case of Pasteurella multocida peritonitis following cat scratch in a patient with Child Pugh Class C alcoholic cirrhosis, culminating in multiple organ failure and death PMID:26294953

  10. Sepsis-induced purpura fulminans caused by Pasteurella multocida

    PubMed Central

    Borges, Lisa; Oliveira, Nelson; Cássio, Isabel; Costa, Humberto

    2014-01-01

    A 52-year-old man was admitted with a cutaneous rash associated with septic shock and multiorganic failure, 6 days after a dog bite. He was started on empiric antibiotherapy and supportive measures. The patient's condition aggravated, with need for invasive mechanical ventilation and intermittent haemodialysis, and evolution from a petechiae-like rash to purpura and gangrene, culminating in bilateral lower limb amputation. The blood cultures revealed only Pasteurella multocida, after 10 days of incubation. P multocida infection is a rare cause of soft tissue infection that subsides with oral antibiotherapy. Infections causing sepsis are rare and appear in immunocompromised patients. Purpura fulminans induced by sepsis is a rare, life-threatening disorder. This syndrome should be recognised promptly, so early treatment is instituted. We found no case reports of purpura fulminans caused by Pasteurella infections in our literature review. PMID:24554680

  11. Atrophic rhinitis caused by Pasteurella multocida type D: morphometric analysis.

    PubMed Central

    Martineau-Doizé, B; Dumas, G; Larochelle, R; Frantz, J C; Martineau, G P

    1991-01-01

    In order to study the distribution and the extent of atrophy caused by Pasteurella multocida in the nasal conchae, experimental piglets were injected intramuscularly at seven days of age with either two or four 50% mouse lethal doses per kg body weight of P. multocida type D dermonecrotoxin. Experimental and control piglets were killed four, six and ten days postinjection. Serial transverse paraffin embedded sections of the noses were cut throughout the entire length of the nasal conchae. The area of the nasal ventral conchae was measured and the morphometric index of the nasal cavity was calculated. It was observed that P. multocida type D dermonecrotoxin induced severe atrophy of the nasal ventral conchae. This atrophy was present along the entire conchae. However, it was most severe at the level of the first and second premolar teeth. Images Fig. 1. Fig. 4. PMID:1889032

  12. Antibiotic sensitivity patterns among Indian strains of avian Pasteurella multocida.

    PubMed

    Shivachandra, S B; Kumar, A A; Biswas, A; Ramakrishnan, M A; Singh, Vijendra P; Srivastava, S K

    2004-11-01

    An investigation was carried out to study the antibiotic sensitivity of avian strains of Pasteurella multocida and to select an effective antimicrobial agent for control of avian pasteurellosis in India. A total of 123 strains of P. multocida recently isolated from different avian species (chicken, duck, turkey, quail, and goose), from different regions of India were subjected to antibiotic sensitivity tests using 20 different antibiotics. Absolute resistance was observed against sulfadiazine. The studies indicated that the strains were most sensitive to chloramphenicol (73.98%), followed by enrofloxacin (71.54%), lincomycin (64.23%), norfloxacin (61.79%) and doxycycline-HCl (56.91%). The majority of the strains were found to exhibit intermediate sensitivity. Chloramphenicol was selected and suggested for treatment. Antibiogram studies also revealed the emergence of multidrug-resistant strains of P. multocida among Indian poultry.

  13. Pasteurella multocida infected total knee arthroplasty: a case report and review of the literature

    PubMed Central

    Ferguson, KB; Bharadwaj, R; MacDonald, A; Syme, B

    2014-01-01

    Pasteurella multocida is a rare cause of prosthetic joint infection. This infection generally follows significant animal contact, usually licks and scratches. We report a case of P multocida infection that was treated with linezolid with salvage of the implant. Linezolid is generally active against Gram-positive organisms only with the exception of Pasteurella, which is Gram-negative. We extensively review the previous reported cases of implant infection with P multocida. PMID:24780653

  14. Characteristics and biotypes of Pasteurella multocida isolated from humans.

    PubMed Central

    Oberhofer, T R

    1981-01-01

    Fifty-two isolates of Pasteurella (48 strains of Pasteurella multocida and 4 strains of atypical Pasteurella) were identified by conventional and commercial test systems. All strains fermented glucose, sucrose, and fructose in purple broth base (Difco Laboratories) with bromocresol purple as indicator, although the atypical Pasteurella produced fermentation reactions that were barely perceptible. Eleven different biotypes were identified by fermentation reactions in maltose, mannitol, xylose, sorbitol, and trehalose media. There was a correlation of biotypes to cat bites, with 61% of cat bite isolates falling into biotype A and B. A correlation of biotype and dog bite isolates was not seen. The choice of medium used for fermentation tests was critical as evidenced by the inability of the organisms to grow in a second commercially purchased preparation of purple broth base. The reliability of commercial test systems in identifying Pasteurella was 81% for Oxi/Ferm (Roche Diagnostics, Div. Hoffmann-La Roche, Inc., Nutley, N.J.), 68% for API (Analytab Products, Plainview, N.Y.), and 11% for Minitek (BBL Microbiology Systems, Cockeysville, MD.). PMID:7240390

  15. The Pasteurella multocida toxin is encoded within a lysogenic bacteriophage.

    PubMed

    Pullinger, Gillian D; Bevir, Thomas; Lax, Alistair J

    2004-01-01

    Toxigenic strains of Pasteurella multocida produce a 146 kDa toxin (PMT) that acts as a potent mitogen. Sequence analysis of the structural gene for PMT, toxA, previously suggested it was horizontally acquired, because it had a low G + C content relative to the P. multocida genome. To address this, the sequence of DNA flanking toxA was determined. The sequence analysis showed the presence of homologues to bacteriophage tail protein genes and a bacteriophage antirepressor, suggesting that the toxin gene resides within a prophage. In addition to phage genes, the toxA flanking DNA contained a homologue of a restriction/modification system that was shown to be functional. The presence of a bacteriophage was demonstrated in spent medium from toxigenic P. multocida isolates. Its production was increased by mitomycin C addition, a treatment that is known to induce the lytic cycle of many temperate bacteriophages. The genomes of bacteriophages from three different toxigenic P. multocida strains had similar but not identical restriction profiles, and were approximately 45-50 kb in length. The prophages from two of these had integrated at the same site in the chromosome, in a tRNA gene. Southern blot analysis confirmed that these bacteriophages contained the toxA gene.

  16. Clinical Features and Outcomes of Pasteurella multocida Infection

    PubMed Central

    Giordano, Antonio; Dincman, Toros; Clyburn, Benjamin E.; Steed, Lisa L.; Rockey, Don C.

    2015-01-01

    Abstract Pasteurella multocida, a zoonotic infectious organism, has most often been described in patients after an animal bite. Here, we characterize the clinical features and outcomes of P multocida infection in a large cohort of patients according to the presence or absence of an animal bite. We retrospectively searched MUSC's laboratory information system for all patients with positive P multocida cultures from 2000 to 2014. Extensive data were abstracted, including clinical and outcome data. The Charlson comorbidity index (CCI) was used to assess comorbidities among patients. We identified 44 patients with P multocida infections, including 25 with an animal bite. The average age was 64 years and the majority of patients were women (N = 30). There was no difference in age and sex distribution among those with and without a bite (P = 0.38 and 0.75, respectively). A CCI ≥1 was significantly associated with the absence of a bite (P = 0.006). Patients presenting without a bite were more frequently bacteremic (37% vs 4%, respectively, P = 0.001), and were hospitalized more often (84% vs 44%, respectively, P = 0.012). Of the 8 patients who required intensive care unit (ICU)-based care, 7 were non-bite-related. There were 4 deaths, all occurring in patients not bitten. P multocida infections not associated with an animal bite were often associated with bacteremia, severe comorbidity(ies), immune-incompetent states, the need for ICU management, and were associated with substantial mortality. PMID:26356688

  17. Cellular and Molecular Action of the Mitogenic Protein-Deamidating Toxin from Pasteurella multocida

    PubMed Central

    Wilson, Brenda A.; Ho, Mengfei

    2011-01-01

    Summary The mitogenic toxin from Pasteurella multocida (PMT) is a member of the dermonecrotic toxin family, which includes toxins from Bordetella, E. coli and Yersinia. Members of the dermonecrotic toxin family modulate G-protein targets in host cells through selective deamidation and/or transglutamination of a critical active site glutamine residue in the G-protein target, which results in activation of the intrinsic GTPase activity. Structural and biochemical data point to the uniqueness of PMT among these toxins in its structure and action. Whereas the other dermonecrotic toxins act on small Rho GTPases, PMT acts on the α subunits of heterotrimeric Gq, Gi and G12/13 protein families. To date, experimental evidence support a model whereby PMT potently stimulates various mitogenic and survival pathways through activation of Gq and G12/13 signaling, ultimately leading to cellular proliferation, while strongly inhibiting pathways involved in cellular differentiation through activation of Gi signaling. The resulting cellular outcomes account for the global physiological effects observed during infection with toxinogenic P. multocida, as well as hint at potential long-term sequelae that may result from PMT exposure. PMID:21569202

  18. Comparison of methods to detect Pasteurella multocida in carrier waterfowl

    USGS Publications Warehouse

    Samuel, M.D.; Shadduck, D.J.; Goldberg, D.R.; Johnson, W.P.

    2003-01-01

    We conducted laboratory challenge trials using mallard ducks (Anas platyrhynchos) to compare methods for detecting carriers of Pasteurella multocida, the bacterium that causes avian cholera, in wild birds. Birds that survived the initial infection were euthanized at 2-4 wk intervals up to 14 wk post challenge. Isolates of P. multocida were obtained at necropsy from 23% of the birds that survived initial infection. We found that swab samples (oral, cloacal, nasal, eye, and leg joint) were most effective for detecting carrier birds up to 14 wk post infection. No detectable differences in isolation were observed for samples stored in either 10% dimethysulfoxide or brain heart infusion broth. The frequency of detecting carriers in our challenge trials appeared to be related to mortality rates observed during the trial, but was not related to a number of other factors including time after challenge, time delays in collecting tissues postmortem, and route of infection. In our trials, there was little association between antibody levels and carrier status. We concluded that swabs samples collected from recently dead birds, stored in liquid nitrogen, and processed using selective broth provide a feasible field method for detecting P. multocida carriers in wild waterfowl.

  19. Transcriptional Response of Pasteurella multocida to Nutrient Limitation

    PubMed Central

    Paustian, Michael L.; May, Barbara J.; Kapur, Vivek

    2002-01-01

    Bacteria often encounter environments where nutrient availability is limited, and they must adapt accordingly. To identify Pasteurella multocida genes that are differentially expressed during nutrient limitation, we utilized whole-genome microarrays to compare levels of gene expression during growth in rich and minimal media. Our analysis showed that the levels of expression of a total of 669 genes, representing approximately one-third of the genome, were detectably altered over the course of the experiment. A large number (n = 439) of genes, including those involved in energy metabolism, transport, protein synthesis, and binding, were expressed at higher levels in rich medium, suggesting that, upon exposure to a rich environment, P. multocida immediately begins to turn on many energy-intensive biosynthetic pathways or, conversely, turns these genes off when it is exposed to a nutrient-deficient environment. Genes with increased expression in minimal medium (n = 230) included those encoding amino acid biosynthesis and transport systems, outer membrane proteins, and heat shock proteins. Importantly, our analysis also identified a large number (n = 164) of genes with unknown functions whose expression was altered during nutrient limitation. Overall, the results of our study show that a wide repertoire of genes, many of which have yet to be functionally classified, undergo transcriptional regulation in P. multocida in response to growth in minimal medium and provide a strong foundation to investigate the transcriptional response of this multispecies pathogen to growth in a nutrient-limited environment. PMID:12057970

  20. Antigenicity of partial fragments of recombinant Pasteurella multocida toxin.

    PubMed

    Lee, Jeongmin; Woo, Hee-Jong

    2010-12-01

    Pasteurella multocida serogroup D strain, which produces P. multocida toxin (PMT), is a widespread and harmful pathogen of respiratory diseases such as pneumonia and progressive atrophic rhinitis (PAR) in swine. Vaccination has been considered the most desirable and effective approach for controlling the diseases caused by toxigenic P. multocida. To investigate the antigenicity and immunogenicity of partial fragments of recombinant PMT, recombinant proteins of the N-terminal (PMT-A), middle (PMT-B), Cterminal (PMT-C), and middle-C-terminal (PMT2.3) regions of PMT were successfully produced in an Escherichia coli expression system. The molecular masses of PMT-A, PMT-B, PMT-C, and PMT2.3 were ca. 53, 55, 35, and 84 kDa, respectively, purified by nickel-nitrilotriacetic acid (Ni-NTA) affinity column chromatography. All the recombinant proteins except for PMT-A showed immune responses to antisera obtained from a swine showing symptoms of PAR. Moreover, high titers of PMT-specific antibodies were raised from mice immunized with each of the recombinant proteins; however, the immunoreactivities of the antibodies to authentic PMT and heat-inactivated whole bacteria were different, respectively. In the protection study, the highest protection against homologous challenge was shown in the case of PMT2.3; relatively poor protections occurred for the other PMT fragments.

  1. Host Response in Rabbits to Infection with Pasteurella multocida Serogroup F Strains Originating from Fowl Cholera

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The ability of two avian Pasteurella multocida serogroup F strains to induce disease in rabbits was investigated in this study. Two groups of 18 Pasteurella-free rabbits each were intranasally challenged with strains isolated from chicken and turkey, respectively. Half the animals in each challenge ...

  2. Effect of Pasteurella multocida toxin on bone resorption in vitro.

    PubMed Central

    Felix, R; Fleisch, H; Frandsen, P L

    1992-01-01

    Pasteurella multocida toxin (PMT), which is the primary etiologic factor in the pathogenesis of progressive atrophic rhinitis in pigs, was found to stimulate bone resorption in vitro. This stimulation was observed both in cultures of murine calvaria by measuring the release of calcium and of the lysosomal enzyme beta-glucuronidase and in murine long bone cultures by measuring the release of calcium. Both systems showed the same dose response curve, with the maximal effect at a concentration of 5 ng/ml. The effect on calvaria was studied in more detail. PMT increased bone resorption 24 h after its addition and always had to be present to express an effect. Calcitonin was able to inhibit this increase of resorption completely, and inhibitors of prostaglandin synthesis suppressed it partially. Although the data show an effect of PMT on bone tissue, the results do not exclude an action on cells in the nasal cavity, which could indirectly stimulate bone resorption. PMID:1452328

  3. Pasteurella multocida-associated sinusitis in khaki Campbell ducks (Anas platyrhynchos).

    PubMed

    Songserm, Thaweesak; Viriyarampa, A Srisamai; Sae-Heng, Namdee; Chamsingh, Wilairat; Bootdee, Orawan; Pathanasophon, Pornpen

    2003-01-01

    Pasteurella multocida group B, serotype 3, was isolated from sinusitis-affected khaki Campbell ducks. To study the role of P. multocida in sinusitis, commercial khaki Campbell ducks were experimentally infected with P. multocida alone or combined with Escherichia coli. In Expt. 1, experimental ducks were infected with P. multocida intranasally or ocularly. A comparison was done by intranasal inoculation with pooled nasal discharge from the affected ducks or phosphate-buffered saline. The ducks intranasally inoculated with the nasal discharge or P. multocida showed sinusitis. In Expt. 2, E. coli alone or a combination of P. multocida and E. coli was intranasally inoculated into experimental ducks. The ducks intranasally inoculated with the combination of P. multocida and E. coli had sinusitis, the same as found in the field but less severe than that of the field cases. Pasteurella multocida was already present in litter/floor of duck farms. We concluded that P. multocida played a role in induction of sinusitis. However, the sinusitis in ducks may be initiated by poor management, especially in the brooding period of ducks.

  4. Pasteurella multocida infection in a cirrhotic patient: case report, microbiological aspects and a review of literature.

    PubMed

    Migliore, E; Serraino, C; Brignone, C; Ferrigno, D; Cardellicchio, A; Pomero, F; Castagna, E; Osenda, M; Fenoglio, L

    2009-01-01

    Pasteurellosis is a zoonosis often caused by cat or dog bites or scratches, or by direct exposure to their secretions. Pasteurella multocida is the main pathogen involved in infections through domestic animal bites; generally a local infection characterized by its particular virulence with consequent rapid onset. Serious infection has also been reported in persons affected by comorbidity without domestic animal bite injuries. Here we report the case of a woman with lower limb exudating vesicular skin ulcers affected by liver cirrhosis, bilateral knee arthritis, septicemia with positive blood culture and synovial fluid culture for Pasteurella multocida. The etiology of Pasteurella multocida must be borne in mind in cases of sepsis in immunodeficient individuals, such as the cirrhotic patient, as well as exposure to domestic animals.

  5. Outer membrane vesicles of Pasteurella multocida contain virulence factors

    PubMed Central

    Fernández-Rojas, Miguel A; Vaca, Sergio; Reyes-López, Magda; de la Garza, Mireya; Aguilar-Romero, Francisco; Zenteno, Edgar; Soriano-Vargas, Edgardo; Negrete-Abascal, Erasmo

    2014-01-01

    Pasteurella multocida (Pm) is a gram-negative bacterium able to infect different animal species, including human beings. This bacterium causes economic losses to the livestock industry because of its high morbidity and mortality in animals. In this work, we report the characterization of outer membrane vesicles (OMVs) released into the culture medium by different Pm serogroups. Purified OMVs in the range of 50–300 nm were observed by electron microscopy. Serum obtained from chickens infected with Pm recognized several proteins from Pm OMVs. Additionally, rabbit antiserum directed against a secreted protease from Actinobacillus pleuropneumoniae recognized a similar protein in the Pm OVMs, suggesting that OMVs from these bacterial species contain common immunogenic proteins. OmpA, a multifunctional protein, was identified in OMVs from different Pm serogroups, and its concentration was twofold higher in OMVs from Pm serogroups B and D than in OMVs from other serogroups. Three outer membrane proteins were also identified: OmpH, OmpW, and transferrin-binding protein. Three bands of 65, 110, and 250 kDa with proteolytic activity were detected in Pm OMVs of serogroups A and E. Additionally, β-lactamase activity was detected only in OMVs from Pm 12945 Ampr (serogroup A). Pm OMVs may be involved in different aspects of disease pathogenesis. PMID:25065983

  6. Membrane interaction of Pasteurella multocida toxin involves sphingomyelin.

    PubMed

    Brothers, Michael C; Ho, Mengfei; Maharjan, Ram; Clemons, Nathan C; Bannai, Yuka; Waites, Mark A; Faulkner, Melinda J; Kuhlenschmidt, Theresa B; Kuhlenschmidt, Mark S; Blanke, Steven R; Rienstra, Chad M; Wilson, Brenda A

    2011-12-01

    Pasteurella multocida toxin (PMT) is an AB toxin that causes pleiotropic effects in targeted host cells. The N-terminus of PMT (PMT-N) is considered to harbor the membrane receptor binding and translocation domains responsible for mediating cellular entry and delivery of the C-terminal catalytic domain into the host cytosol. Previous studies have implicated gangliosides as the host receptors for PMT binding. To gain further insight into the binding interactions involved in PMT binding to cell membranes, we explored the role of various membrane components in PMT binding, utilizing four different approaches: (a) TLC-overlay binding experiments with (125) I-labeled PMT, PMT-N or the C-terminus of PMT; (b) pull-down experiments using reconstituted membrane liposomes with full-length PMT; (c) surface plasmon resonance analysis of PMT-N binding to reconstituted membrane liposomes; (d) and surface plasmon resonance analysis of PMT-N binding to HEK-293T cell membranes without or with sphingomyelinase, phospholipase D or trypsin treatment. The results obtained revealed that, in our experimental system, full-length PMT and PMT-N did not bind to gangliosides, including monoasialogangliosides GM(1) , GM(2) or GM(3) , but instead bound to membrane phospholipids, primarily the abundant sphingophospholipid sphingomyelin or phosphatidylcholine with other lipid components. Collectively, these studies demonstrate the importance of sphingomyelin for PMT binding to membranes and suggest the involvement of a protein co-receptor.

  7. A new selective enrichment procedure for isolating Pasteurella multocida from avian and environmental samples

    USGS Publications Warehouse

    Moore, M.K.; Cicnjak-Chubbs, L.; Gates, R.J.

    1994-01-01

    A selective enrichment procedure, using two new selective media, was developed to isolate Pasteurella multocida from wild birds and environmental samples. These media were developed by testing 15 selective agents with six isolates of P. multocida from wild avian origin and seven other bacteria representing genera frequently found in environmental and avian samples. The resulting media—Pasteurella multocida selective enrichment broth and Pasteurella multocida selective agar—consisted of a blood agar medium at pH 10 containing gentamicin, potassium tellurite, and amphotericin B. Media were tested to determine: 1) selectivity when attempting isolation from pond water and avian carcasses, 2) sensitivity for detection of low numbers of P. multocida from pure and mixed cultures, 3) host range specificity of the media, and 4) performance compared with standard blood agar. With the new selective enrichment procedure, P. multocida was isolated from inoculated (60 organisms/ml) pond water 84% of the time, whereas when standard blood agar was used, the recovery rate was 0%.

  8. Interaction between Ascaris suum and Pasteurella multocida in the lungs of mice.

    PubMed

    Tjørnehøj, K; Eriksen, L; Aalbaek, B; Nansen, P

    1992-01-01

    In an experiment including 8 groups of 15 mice, the effect of migrating Ascaris suum larvae in the lungs on the establishment and pathogenicity of aerosol exposure to Pasteurella multocida was investigated. Following aerosol exposure to P. multocida, mice with migrating A. suum in their lungs developed more severe pneumonia and septicaemia than did parasite-free mice. The parasite-induced effect on bacterial pathogenicity was more marked for a non-toxin-producing P. multocida as compared with a toxin-producing strain of P. multocida, possibly due to the higher spontaneous pathogenicity of the non-toxigenic strain of P. multocida. The present results should encourage controlled experiments on possible interactions between A. suum and various airborne microbial infections in pigs.

  9. The actions of Pasteurella multocida toxin on neuronal cells.

    PubMed

    Surguy, Susan M; Duricki, Denise A; Reilly, Joanne M; Lax, Alistair J; Robbins, Jon

    2014-02-01

    Pasteurella multocida toxin (PMT) activates the G-proteins Gαi(₁₋₃), Gα(q), Gα₁₁, Gα₁₂ and Gα₁₃ by deamidation of specific glutamine residues. A number of these alpha subunits have signalling roles in neurones. Hence we studied the action of this toxin on rat superior cervical ganglion (SCG) neurones and NG108-15 neuronal cells. Both Gα(q) and Gα₁₁ could be identified in SCGs with immunocytochemistry. PMT had no direct action on Kv7 or Cav2 channels in SCGs. However PMT treatment enhanced muscarinic receptor mediated inhibition of M-current (Kv7.2 + 7. 3) as measured by a 19-fold leftward shift in the oxotremorine-M concentration-inhibition curve. Agonists of other receptors, such as bradykinin or angiotensin, that inhibit M-current did not produce this effect. However the amount of PIP₂ hydrolysis could be enhanced by PMT for all three agonists. In a transduction system in SCGs that is unlikely to be affected by PMT, Go mediated inhibition of calcium current, PMT was ineffective whereas the response was blocked by pertussis toxin as expected. M1 muscarinic receptor evoked calcium mobilisation in transformed NG108-15 cells was enhanced by PMT. The calcium rises evoked by uridine triphosphate acting on endogenous P2Y₂ receptors in NG108-15 cells were enhanced by PMT. The time and concentration dependence of the PMT effect was different for the resting calcium compared to the calcium rise produced by activation of P2Y₂ receptors. PMT's action on these neuronal cells would suggest that if it got into the brain, symptoms of a hyperexcitable nature would be seen, such as seizures.

  10. Sialic Acid Uptake Is Necessary for Virulence of Pasteurella multocida in turkeys

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many pathogenic bacteria employ systems to incorporate sialic acid into their membranes as a means of protection against host defense mechanisms. Pasteurella multocida is an opportunistic pathogen which causes diseases of economic importance in a wide range of animal species and sialic acid uptake p...

  11. Proximity-dependent inhibition of growth of mannheimia haemolytica by pasteurella multocida.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Mannheimia haemolytica, Pasteurella multocida, and Bibersteinia trehalosi have been identified in the lungs of pneumonic bighorn sheep (BHS; Ovis canadensis). Of these pathogens, M. haemolytica has been shown to consistently cause fatal pneumonia in BHS under experimental conditions. However, M. hae...

  12. Molecular Identification and Epidemiological Tracing of Pasteurella multocida Meningitis in a Baby

    PubMed Central

    Boerlin, Patrick; Siegrist, Hans H.; Burnens, André P.; Kuhnert, Peter; Mendez, Purita; Prétat, Gérard; Lienhard, Reto; Nicolet, Jacques

    2000-01-01

    We report a case of Pasteurella multocida meningitis in a 1-month-old baby exposed to close contact with two dogs and a cat but without any known history of injury by these animals. 16S rRNA gene sequencing of the isolate from the baby allowed identification at the subspecies level and pointed to the cat as a possible source of infection. Molecular typing of Pasteurella isolates from the animals, from the baby, and from unrelated animals clearly confirmed that the cat harbored the same P. multocida subsp. septica strain on its tonsils as the one isolated from the cerebrospinal fluid of the baby. This case stresses the necessity of informing susceptible hosts at risk of contracting zoonotic agents about some basic hygiene rules when keeping pets. In addition, this study illustrates the usefulness of molecular methods for identification and epidemiological tracing of Pasteurella isolates. PMID:10699029

  13. An atypical presentation of a Pasteurella multocida infection following a cat bite: a case report.

    PubMed

    Collins, Chris; Flanagan, Brigitte; Henning, J Scott

    2012-06-01

    Pasteurella multocida is a bacterial organism that commonly causes cellulitis after animal bites, especially cat bites. We report an unusual vesiculopustular infection of the hand following a domestic cat bite. Pasteurella multocida and Staphylococcus aureus were cultured from the wound and the patient was treated with amoxicillin-clavulanate potassium. Further history revealed that the patient's cat had nibbled on her hand. Pasteurella usually is resistant to many of the typical empiric antibiotics used to treat skin infections. Amoxicillin-clavulanate potassium (500 mg 3 times daily) is the treatment of choice for patients who have an infected cat or dog bite with no known bacterial cause. A thorough patient history is needed to promptly arrive at a proper diagnosis for an atypical presentation of a common disease.

  14. Using amplified fragment length polymorphism analysis to differentiate isolates of Pasteurella multocida serotype 1

    USGS Publications Warehouse

    Blehert, D.S.; Jefferson, K.L.; Heisey, D.M.; Samuel, M.D.; Berlowski, B.M.; Shadduck, D.J.

    2008-01-01

    Avian cholera, an infectious disease caused by the bacterium Pasteurella multocida, kills thousands of North American wild waterfowl annually. Pasteurella multocida serotype 1 isolates cultured during a laboratory challenge study of Mallards (Anas platyrhynchos) and collected from wild birds and environmental samples during avian cholera outbreaks were characterized using amplified fragment length polymorphism (AFLP) analysis, a whole-genome DNA fingerprinting technique. Comparison of the AFLP profiles of 53 isolates from the laboratory challenge demonstrated that P. multocida underwent genetic changes during a 3-mo period. Analysis of 120 P. multocida serotype 1 isolates collected from wild birds and environmental samples revealed that isolates were distinguishable from one another based on regional and temporal genetic characteristics. Thus, AFLP analysis had the ability to distinguish P. multocida isolates of the same serotype by detecting spatiotemporal genetic changes and provides a tool to advance the study of avian cholera epidemiology. Further application of AFLP technology to the examination of wild bird avian cholera outbreaks may facilitate more effective management of this disease by providing the potential to investigate correlations between virulence and P. multocida genotypes, to identify affiliations between bird species and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission. ?? Wildlife Disease Association 2008.

  15. Associations between water quality, Pasteurella multocida, and avian cholera at Sacramento National Wildlife Refuge

    USGS Publications Warehouse

    Lehr, M.A.; Botzler, R.G.; Samuel, M.D.; Shadduck, D.J.

    2005-01-01

    We studied patterns in avian cholera mortality, the presence of Pasteurella multocida in the water or sediment, and water chemistry characteristics in 10 wetlands at the Sacramento National Wildlife Refuge Complex (California, USA), an area of recurrent avian cholera epizootics, during the winters of 1997 and 1998. Avian cholera outbreaks (a?Y50 dead birds) occurred on two wetlands during the winter of 1997, but no P. multocida were recovered from 390 water and 390 sediment samples from any of the 10 wetlands. No mortality events were observed on study wetlands during the winter of 1998; however, P. multocida was recovered from water and sediment samples in six of the 10 study wetlands. The pH levels were higher for wetlands experiencing outbreaks during the winter of 1997 than for nonoutbreak wetlands, and aluminum concentrations were higher in wetlands from which P. multocida were recovered during the winter of 1998. Water chemistry parameters (calcium, magnesium, sodium, and dissolved protein) previously linked with P. multocida and avian cholera mortality were not associated with the occurrence of avian cholera outbreaks or the presence of P. multocida in our study wetlands. Overall, we found no evidence to support the hypothesis that wetland characteristics facilitate the presence of P. multocida and, thereby, allow some wetlands to serve as long-term sources (reservoirs) for P. multocida.

  16. Neonatal Pasteurella multocida subsp. septica Meningitis Traced to Household Cats: Molecular Linkage Analysis Using Repetitive-Sequence-Based PCR

    PubMed Central

    Freij, Bishara J.; Robinson-Dunn, Barbara; Makin, Jacob; Runge, Jessica K.; Luna, Ruth Ann

    2015-01-01

    Pasteurella multocida is a rare cause of neonatal bacterial meningitis. We describe such a case and verify two household cats as the source of infection using repetitive-element PCR (rep-PCR) molecular fingering. PMID:26491173

  17. Pasteurella multocida urinary tract infection in a patient with cervical cancer.

    PubMed

    Singh, Manmeet B; Harrington, Amanda T

    2017-01-01

    Introduction. Infections caused by Pasteurella species are commonly associated with contact with dogs and cats, typically involving bites and scratches, but casual contact with household pets can also be a risk factor. Urinary tract infection (UTI) caused by Pasteurella species is rare and a significant majority of cases have some known risk factor associated with an underlying chronic illness or structural and/or functional urological abnormality. Case presentation. Here, we present a case of a UTI due to Pasteurella multocida in a patient with squamous cell carcinoma of the cervix who also had a household cat. Conclusion. Providers and laboratorians should be aware of risk factors associated with UTIs caused by Pasteurella species.

  18. Pasteurella multocida urinary tract infection in a patient with cervical cancer

    PubMed Central

    Singh, Manmeet B.

    2017-01-01

    Introduction. Infections caused by Pasteurella species are commonly associated with contact with dogs and cats, typically involving bites and scratches, but casual contact with household pets can also be a risk factor. Urinary tract infection (UTI) caused by Pasteurella species is rare and a significant majority of cases have some known risk factor associated with an underlying chronic illness or structural and/or functional urological abnormality. Case presentation. Here, we present a case of a UTI due to Pasteurella multocida in a patient with squamous cell carcinoma of the cervix who also had a household cat. Conclusion. Providers and laboratorians should be aware of risk factors associated with UTIs caused by Pasteurella species. PMID:28348800

  19. Pasteurella multocida septic shock after a cat scratch in an elderly otherwise healthy woman: a case report.

    PubMed

    Fernández-Valencia, Jenaro A; García, Sebastian; Prat, Salvio

    2008-03-01

    Pasteurella multocida, a gram-negative coccobacillus, is a commensal in the nasopharynx of many animals. P. multocida infections most commonly involve the skin, soft tissues, and respiratory tract, particularly in immunosuppressed patients. The present case illustrates a severe articular infection caused by this bacterium, leading to septic shock, in an elderly, otherwise healthy woman, after a simple scratch of a cat.

  20. Pasteurella multocida infections. Report of 34 cases and review of the literature.

    PubMed

    Weber, D J; Wolfson, J S; Swartz, M N; Hooper, D C

    1984-05-01

    Pasteurella multocida, a small, gram-negative coccobacillus , is part of the normal oral flora of many animals, including the dog and cat. P. multocida is the etiologic agent in a variety of infectious disease syndromes. We have reported 34 cases of infection caused by P. multocida and have reviewed the English literature. P. multocida infections may be divided into three broad groups: 1. Infections resulting from animal bites and scratches : The most common infections caused by P. multocida are local wound infections following animal bites or scratches . Cats are the source of infection in 60 to 80% of cases and dogs in the great majority of the remainder. Local infections are characterized by the rapid appearance of erythema, warmth, tenderness, and frequently purulent drainage. The most common local complications are abscess formation and tenosynovitis. Serious local complications include septic arthritis proximal to bites or scratches , osteomyelitis resulting from direct inoculation or extension of cellulitis, and the combination of septic arthritis and osteomyelitis, most commonly involving a finger or hand after a cat bite. 2. Isolation of P. multocida from the respiratory tract: The isolation of P. multocida from the respiratory tract must be interpreted differently than its isolation from other systemic sites. Most commonly P. multocida found in the respiratory tract is a commensal organism in patients with underlying pulmonary disease, but serious respiratory tract infections including pneumonia, empyema, and lung abscesses may develop. Most patients with respiratory tract colonization or infection have a history of animal exposure. 3. Other systemic infections: P. multocida is recognized as a pathogen in a variety of systemic infections including bacteremia, meningitis, brain abscess, spontaneous bacterial peritonitis, and intra-abdominal abscess. P. multocida often acts as an opportunistic pathogen with a predilection for causing bacteremia in patients

  1. Development of OMP based indirect ELISA to gauge the antibody titers in bovines against Pasteurella multocida

    PubMed Central

    Dogra, V; Verma, S; Singh, G; Wani, A. H; Chahota, R; Dhar, P; Verma, L; Sharma, M

    2015-01-01

    Pasteurella multocida (P. multocida) is an important pathogen of various domestic animals. The outer membrane proteins (OMPs) play a major role in pathogenesis and immunogenicity of P. multocida. The aim of the study was to develop indirect enzyme linked immuno sorbant assay (ELISA) based on OMPs to ascertain the antibody titers in animals post-infection or to gauge the potency of vaccine. The OMPs were extracted and purified from P. multocida P:52 (vaccine strain) and P. multocida B:2 isolated from natural outbreak of Haemorrhagic septicaemia (HS) and analyzed on SDS PAGE and through western blot. The OMPs profile of the vaccine strain and the isolate from the natural outbreak of HS were found to be similar. Optimization of various components viz. coating antigens, anti-species conjugate, etc. were carried out against both anti-P. multocida hyper immune and pre immune serum. Validation of OMP based indirect ELISA assay to measure immune response against P. multocida in bovine revealed 91% diagnostic sensitivity (DSN) and about 100% diagnostic specificity (DSP) at 25% cut off. OMP based indirect ELISA was found to be more specific, but less sensitive as compared to WCL based assay. PMID:27175202

  2. In-silico analysis of Pasteurella multocida to identify common epitopes between fowl, goat and buffalo.

    PubMed

    Ghaffar, Ammarah; Tariq, Aamira

    2016-04-10

    Pasteurella multocida represents a highly diverse group of bacteria infecting various hosts like the fowl, goat and buffalo leading to huge economic loss to the poultry and cattle industry. Previous reports indicated that the outer membrane proteins contribute significantly to the pathogenesis of Pasteurella multocida. The comparative in-silico genome wide analysis of four pathogenic Pasteurella multocida strains (Anand1-poultry, Anand1-goat, PMTB and VTCCBAA264) with their respective hosts was performed. A pipeline was developed to identify the list of non-homologous proteins of Pasteurella multocida strains and their hosts. The list was further analyzed for the identification of the essential outer membrane proteins responsible for the pathogenicity. Outer membrane proteins were further selected from these antigenic proteins on the basis of their pathogenic potential. A common B-cell epitope (TDYRNRDRS, ARRSVTSKEN, and KINDQWRW) determined via sequential and structural approach from the lipopolysaccharide (LPS) assembly outer membrane complex protein was predicted from fowl, goat and buffalo. Furthermore, we identified T-cell epitopes based on the lipopolysaccharide (LPS) assembly outer membrane complex protein via docking studies which were either similar to the B-cell epitopes or were occurring in the same patch except for MHC class II M fowl. We propose that this difference in epitope sequence is due to different interacting MHC class II protein predicted from the fowl. Hence, in the current study we found that a unique epitope based on the common antigenic lipopolysaccharide (LPS) outer membrane complex protein present in fowl, goat and buffalo can be a suitable target for vaccine development against the two economic devastating diseases; fowl cholera (FC) and hemorrhagic septicemia (HS).

  3. Isolation and Sequencing of a Temperate Transducing Phage for Pasteurella multocida

    PubMed Central

    Campoy, Susana; Aranda, Jesús; Àlvarez, Gerard; Barbé, Jordi; Llagostera, Montserrat

    2006-01-01

    A temperate bacteriophage (F108) has been isolated through mitomycin C induction of a Pasteurella multocida serogroup A strain. F108 has a typical morphology of the family Myoviridae, presenting a hexagonal head and a long contractile tail. F108 is able to infect all P. multocida serogroup A strains tested but not those belonging to other serotypes. Bacteriophage F108, the first P. multocida phage sequenced so far, presents a 30,505-bp double-stranded DNA genome with cohesive ends (CTTCCTCCCC cos site). The F108 genome shows the highest homology with those of Haemophilus influenzae HP1 and HP2 phages. Furthermore, an F108 prophage attachment site in the P. multocida chromosome has been established to be inside a gene encoding tRNALeu. By using several chromosomal markers that are spread along the P. multocida chromosome, it has been demonstrated that F108 is able to perform generalized transduction. This fact, together with the absence of pathogenic genes in the F108 genome, makes this bacteriophage a valuable tool for P. multocida genetic manipulation. PMID:16672452

  4. Deadly case of Pasteurella multocida aortitis and mycotic aneurysm following a cat bite

    PubMed Central

    Cho, Dennis Dane; Berliner, Yaniv; Carr, David

    2016-01-01

    Animal bites are frequently encountered in the emergency department (ED). Aortitis leading to mycotic abdominal aortic aneurysm is a rare and potentially deadly complication of Pasteurella multocida (P. multocida) following an animal bite. We present the case of a 68-year-old male who presented to the ED after falling at home. He complained of weakness and abdominal pain. He was in septic shock and was treated empirically with broad-spectrum antibiotics and intravenous fluids. He reported previous antibiotic treatment of a cellulitis secondary to a cat bite injury to his right thumb four weeks prior. Abdominal ultrasound and subsequent computed tomography scan revealed a leaking mycotic abdominal aneurysm that was surgically repaired. Blood cultures and aortic wall tissue cultures grew P. multocida. Given how common animal bite presentations are in the ED, this case highlights the need to consider aortitis and mycotic abdominal aortic aneurysm in an unwell patient with an animal bite. PMID:27326399

  5. Pasteurella multocida septicemia caused by close contact with a domestic cat: case report and literature review.

    PubMed

    Kimura, Ryosuke; Hayashi, Yoshinari; Takeuchi, Toyo; Shimizu, Manabu; Iwata, Masaru; Tanahashi, Junji; Ito, Makoto

    2004-08-01

    We report here a case of Pasteurella multocida infection caused by cat exposure presenting with septic shock, sinusitis, and pneumonia. The patient was a febrile 20-year-old woman who had been experiencing disturbed consciousness progressively. She had close contact with a domestic cat and had received some scratches on both arms. A magnetic resonance imaging (MRI) scan of the head showed a high intensity in the paranasal cavity, and a computed tomographic (CT) scan of the chest showed bilateral lung consolidations. The pathogen was identified as P. multocida by the cultures from blood and nasal discharge. She was given intensive antibiotic therapy with ceftriaxone and piperacillin, continuous hemodiafiltration (CHDF) therapy, and anticoagulation therapy. Owing to these therapeutic regimens, the septic shock was successfully treated without complications. We also review the literature on P. multocida septicemia.

  6. Deadly case of Pasteurella multocida aortitis and mycotic aneurysm following a cat bite.

    PubMed

    Cho, Dennis Dane; Berliner, Yaniv; Carr, David

    2016-06-16

    Animal bites are frequently encountered in the emergency department (ED). Aortitis leading to mycotic abdominal aortic aneurysm is a rare and potentially deadly complication of Pasteurella multocida (P. multocida) following an animal bite. We present the case of a 68-year-old male who presented to the ED after falling at home. He complained of weakness and abdominal pain. He was in septic shock and was treated empirically with broad-spectrum antibiotics and intravenous fluids. He reported previous antibiotic treatment of a cellulitis secondary to a cat bite injury to his right thumb four weeks prior. Abdominal ultrasound and subsequent computed tomography scan revealed a leaking mycotic abdominal aneurysm that was surgically repaired. Blood cultures and aortic wall tissue cultures grew P. multocida. Given how common animal bite presentations are in the ED, this case highlights the need to consider aortitis and mycotic abdominal aortic aneurysm in an unwell patient with an animal bite.

  7. Differentiation of toxigenic from nontoxigenic isolates of Pasteurella multocida by PCR.

    PubMed Central

    Nagai, S; Someno, S; Yagihashi, T

    1994-01-01

    A PCR assay was developed for the differentiation of toxigenic Pasteurella multocida subsp. multocida strains, the major etiologic agent for progressive atrophic rhinitis in pigs, from nontoxigenic strains. The PCR targeted a toxA gene encoding a 143-kDa dermonecrotic toxin that is considered to be the central etiologic factor in progressive atrophic rhinitis. toxA fragments were amplified from toxigenic P. multocida isolates but not from nontoxigenic isolates or other bacteria isolated from pigs. The sensitivity of the reaction was as low as 10 pg of chromosomal DNA from a toxigenic strain. The results obtained by PCR of the DNAs of 187 field isolates of P. multocida were consistent with those obtained by the guinea pig skin test and Western blot (immunoblot) analysis. Restriction fragment analysis of the PCR-amplified fragments from 67 field isolates and comparison of the DNA sequences of fragments from capsular serotype A and D strains suggest that the PCR-amplified region, which is considered to encode the major immunologic determinants of the toxin, would be the same among P. multocida strains. The PCR that we describe should be useful for the diagnosis and the etiologic survey of progressive atrophic rhinitis. Images PMID:8027302

  8. Plasmid and restriction endonuclease patterns in Pasteurella multocida isolated from a swine pyramid.

    PubMed

    Rúbies, Xavier; Casal, Jordi; Pijoan, Carlos

    2002-01-03

    Restriction endonuclease analysis (REA) and plasmid profile were used to study the epidemiology of Pasteurella multocida in a swine pyramid structure. The studied pyramid was comprised of a group of 12 swine farrow-to-finish farms related by unidirectional animal movement. P. multocida isolates were obtained from the lungs of 275 slaughtered pigs. Serotyping was performed by hyaluronidase sensitivity test and toxicity was investigated by the ELISA test. HpaII was used to cleave the P. multocida extracted DNA. REA patterns relationships were studied using the Sokal-Michener coefficients, and the dendrogram was built using the UPGMA system. The 218 P. multocida isolates obtained were distributed in 17 REA patterns. In 9 of the 12 farms studied only 2-3 REA patterns were detected, with one clearly predominant pattern. The 81 strains with plasmids were assigned to six plasmid profiles. REA and plasmid profiles proved to be good epidemiological tools for identifying different strains of P. multocida with the same phenotype.

  9. A 5-year retrospective report of Gallibacterium anatis and Pasteurella multocida isolates from chickens in Mississippi.

    PubMed

    Jones, K H; Thornton, J K; Zhang, Y; Mauel, M J

    2013-12-01

    A 5-yr retrospective study (November 2006-December 2011) was conducted to determine the isolation frequency of Pasteurella multocida and Gallibacterium anatis and their antibiograms from chickens submitted to the Mississippi Poultry Research and Diagnostic Laboratory. The number of isolations of G. anatis increased over the last 5 yr in broiler and broiler breeder type chickens. For P. multocida, the number of isolations increased from 2006 to 2010, but decreased through 2011 with all isolations being from boiler breeder type chickens. Gallibacterium anatis demonstrated almost complete resistance to novobiocin, tylosin, lincosamide, and tetracycline antimicrobials with moderate to high sensitivity to sulfonamides, fluoroquinolones, and florfenicol. There was intermediate sensitivity for spectinomycin and erythromycin and variable resistance to β-lactam and aminoglycoside antimicrobials. In sharp contrast, P. multocida showed moderate to high sensitivity to β-lactam, novobiocin, and tetracycline antimicrobials, but had antibiograms similar to G. anatis for the other antimicrobials. Sensitivities were determined using minimum inhibitory concentration. This study examines the trends over a 5-yr period of the number of isolates of P. multocida and G. anatis and their sensitivities. These 2 pathogens produce very similar clinical signs and lesions (fowl cholera-like) in breeders despite having extremely antagonistic sensitivity patterns. This study shows the necessity for producers to attempt culture and sensitivity in suspect fowl cholera-like flocks before initiating antimicrobial treatment commonly used with P. multocida for fear that the culprit may actually be the more antimicrobial-resistant G. anatis.

  10. Prevalence of Pasteurella multocida and other respiratory pathogens in the nasal tract of Scottish calves.

    PubMed

    Hotchkiss, E J; Dagleish, M P; Willoughby, K; McKendrick, I J; Finlayson, J; Zadoks, R N; Newsome, E; Brulisauer, F; Gunn, G J; Hodgson, J C

    2010-10-09

    The prevalence of Pasteurella multocida, a cause of bovine respiratory disease, was studied in a random sample of beef suckler and dairy farms throughout Scotland, by means of a cross-sectional survey. A total of 637 calves from 68 farms from six geographical regions of Scotland were sampled between February and June 2008. Deep nasal swabs were taken, and samples that were culture-positive for P multocida were confirmed by PCR. Prevalence of P multocida was 17 per cent (105 of 616 calves); 47 per cent of farms had at least one positive animal. A higher prevalence was detected in dairy calves than beef calves (P=0.04). It was found that P multocida was associated with Mycoplasma-like organisms (P=0.06) and bovine parainfluenza type 3 virus (BPI-3) (P=0.04), detected by culture and quantitative PCR of nasal swabs, respectively. Detection of P multocida was not associated with bovine respiratory syncytial virus (BRSV), bovine herpesvirus type 1 (BoHV-1) or bovine viral diarrhoea virus (BVDV). Mycoplasma-like organisms, BPI-3, BRSV, BoHV-1 and BVDV were detected in 58, 17, four, 0 and eight calves, on 25, five, two, 0 and five of the 68 farms, respectively.

  11. Aortic Endograft Infection by Pasteurella multocida: A Rare Case.

    PubMed

    Jayakrishnan, Thejus T; Keyashian, Brian; Amene, Juliet; Malinowski, Michael

    2016-08-01

    Infection of an aortic endograft is a rare complication following endovascular aneurysm repair. These patients have been treated with explantation of the graft to obtain source control followed by an extra-anatomic bypass to restore circulation. The present case study describes an interesting case of Pasteurella infection involving an aortic endograft managed nonoperatively by percutaneous drainage and graft preservation.

  12. Comparative genome analysis of an avirulent and two virulent strains of avian Pasteurella multocida reveals candidate genes involved in fitness and pathogenicity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fowl cholera is a highly contagious systemic disease affecting wild and domestic birds, frequently resulting in high morbidity and mortality. The causative agent is Pasteurella multocida (P. multocida). The completed genome of P. multocida strain Pm70 has been available for over eleven years and has...

  13. A serotype-specific polymerase chain reaction for identification of Pasteurella multocida serotype 1

    USGS Publications Warehouse

    Rocke, Tonie E.; Smith, Susan R.; Miyamoto, Amy; Shadduck, Daniel J.

    2002-01-01

    A serotype-specific polymerase chain reaction (PCR) assay was developed for detection and identification of Pasteurella multocida serotype 1, the causative agent of avian cholera in wild waterfowl. Arbitrarily primed PCR was used to detect DNA fragments that distinguish serotype 1 from the other 15 serotypes of P. multocida (with the exception of serotype 14). Oligonucleotide primers were constructed from these sequences, and a PCR assay was optimized and evaluated. PCR reactions consistently resulted in amplification products with reference strains 1 and 14 and all other serotype 1 strains tested, with cell numbers as low as 2.3 cells/ml. No amplification products were produced with other P. multocida serotypes or any other bacterial species tested. To compare the sensitivity and further test the specificity of this PCR assay with traditional culturing and serotyping techniques, tissue samples from 84 Pekin ducks inoculated with field strains of P. multocida and 54 wild lesser snow geese collected during an avian cholera outbreak were provided by other investigators working on avian cholera. PCR was as sensitive (58/64) as routine isolation (52/64) in detecting and identifying P. multocida serotype 1 from the livers of inoculated Pekins that became sick or died from avian cholera. No product was amplified from tissues of 20 other Pekin ducks that received serotypes other than type 1 (serotype 3, 12 × 3, or 10) or 12 control birds. Of the 54 snow geese necropsied and tested for P. multocida, our PCR detected and identified the bacteria from 44 compared with 45 by direct isolation. The serotype-specific PCR we developed was much faster and less labor intensive than traditional culturing and serotyping procedures and could result in diagnosis of serotype 1 pasteurellosis within 24 hr of specimen submission.

  14. Inhibition of growth and alteration of host cell interactions of Pasteurella multocida with natural byproducts.

    PubMed

    Salaheen, S; Almario, J A; Biswas, D

    2014-06-01

    Pasteurella multocida is a leading cause of fowl cholera in both free-range pasture and conventional/commercially raised poultry. Its infection is a serious threat to poultry health and overall flock viability. Organic poultry is comparatively more vulnerable to this pathogen. It is a significant cause of production loss and price increase of poultry products, specifically organic poultry products. Some plant products are well documented as sources of natural antimicrobials such as polyphenols found in different berry pomaces and citrus oil. Pomace, a byproduct (primarily of seeds and skins) of fruits used for juice and wine production, and citrus oil, the byproduct of citrus juice production, show promising antimicrobial activity against various pathogens. Here, we showed for the first time that blackberry and blueberry pomace extracts and citrus oil inhibited P. multocida growth. Minimum bactericidal concentrations were determined as 0.3 and 0.4 mg/mL gallic acid equivalent for blackberry and blueberry pomace extracts, respectively. Similarly, only 0.05% citrus oil (vol/vol) completely inhibited P. multocida growth. Under shaking conditions, the antimicrobial activity of both pomace extracts and citrus oil was more intensive. Even citrus oil vapor also significantly reduced the growth of P. multocida. In addition, cell surface hydrophobicity of P. multocida was increased by 2- to 3-fold and its adherence to chicken fibroblast (DF1) and bovine mammary gland (MacT) cells was reduced significantly in the presence of pomace extracts only. This study indicates that these natural products might be good alternatives to conventional antimicrobial agents, and hence, may be used as feed or water supplements to control fowl cholera and reduce production loss caused by P. multocida.

  15. Immunological roles of Pasteurella multocida toxin (PMT) using a PMT mutant strain.

    PubMed

    Kim, Tae Jung; Toan, Nguyen Tat; Jang, Eun Jin; Jung, Bock Gie; Lee, Jae Il; Lee, Bong Joo

    2007-08-01

    The immunological role of the Pasteurella multocida toxin (PMT) in mice was examined using a PMT mutant strain. After a nasal inoculation, the mutant strain failed to induce interstitial pneumonia. Moreover, PMT had no significant effect on the populations of CD4+, CD8+, CD3+, and CD19+ immunocytes in blood or on the populations of CD4+ and CD8+ splenocytes (P<0.01). However, there was a significant increase in the total number of cells in the BAL samples obtained from the wild-type P. multocida-inoculated mice. On the other hand, the level of IL-1 expression decreased when the macrophages from the bronchio-alveolar lavage were stimulated with PMT. Overall, PMT appears to play some role (stimulating and/or inhibiting) in the immunological responses but further studies will be required to confirm this.

  16. Persistence of Pasteurella multocida in Nebraska (USA) wetlands under epizootic conditions

    USGS Publications Warehouse

    Price, J.I.; Brand, C.J.

    1984-01-01

    Gleason Basin, a marsh located in the western part of the Rainwater Basin in Nebraska, was selected during the 1980 spring waterfowl migration as a study site to determine the presence and persistence of virulent Pasteurella multocida. Avian cholera mortality in migratory waterfowl using the Basin increased during a 2-wk period of a die-off beginning the first week of March when 2,409 carcasses were collected from the marsh. Study sites within the marsh were established for sampling water associated with and not associated with intact and scavenged carcasses. Isolations of virulent P. multocida were made from five of six study sites associated with either intact or scavenged carcasses for 3 days and from three of five non-carcass-associated study sites for 2 days. Recovery of these bacteria from this environment suggested a possible source of infection for susceptible waterfowl using the contaminated site.

  17. Evaluation of transport media for Pasteurella multocida isolates from rabbit nasal specimens.

    PubMed

    Kawamoto, E; Sawada, T; Maruyama, T

    1997-08-01

    A suitable medium for the transport of Pasteurella multocida in nasal specimens from rabbits was investigated by using pure cultures of the organism and nasal swabs from infected rabbits. First, the ability of eight transport media to preserve the viabilities of P. multocida strains isolated from rabbits was studied. Cary-Blair medium and Leibovitz medium no. 15 (L-15) were found to be superior to the other six media tested, enabling survival of the organism for more than 14 days at room temperature. Second, the survival of P. multocida in nasal specimens was evaluated on both Cary-Blair medium and L-15. The recovery rate of the organism from these two media was more than 80 to 90% during 4 days of storage and decreased gradually with increasing preservation time. There were no significant differences (P > 0.05) in recovery rates of the organism between Cary-Blair medium and L-15. On the basis of these results, we recommend the use of Cary-Blair medium for the transport of P. multocida in rabbit nasal specimens because of the ease of transport of nasal swabs by mail.

  18. Evaluation of transport media for Pasteurella multocida isolates from rabbit nasal specimens.

    PubMed Central

    Kawamoto, E; Sawada, T; Maruyama, T

    1997-01-01

    A suitable medium for the transport of Pasteurella multocida in nasal specimens from rabbits was investigated by using pure cultures of the organism and nasal swabs from infected rabbits. First, the ability of eight transport media to preserve the viabilities of P. multocida strains isolated from rabbits was studied. Cary-Blair medium and Leibovitz medium no. 15 (L-15) were found to be superior to the other six media tested, enabling survival of the organism for more than 14 days at room temperature. Second, the survival of P. multocida in nasal specimens was evaluated on both Cary-Blair medium and L-15. The recovery rate of the organism from these two media was more than 80 to 90% during 4 days of storage and decreased gradually with increasing preservation time. There were no significant differences (P > 0.05) in recovery rates of the organism between Cary-Blair medium and L-15. On the basis of these results, we recommend the use of Cary-Blair medium for the transport of P. multocida in rabbit nasal specimens because of the ease of transport of nasal swabs by mail. PMID:9230361

  19. Comparative analysis of Pasteurella multocida strains isolated from bovine respiratory infections.

    PubMed

    Sellyei, Boglárka; Rónai, Zsuzsanna; Jánosi, Szilárd; Makrai, László

    2015-12-01

    Bovine respiratory disease (BRD) is the leading cause of significant economic losses in the intensive beef industry worldwide. Beside numerous risk factors Pasteurella multocida, which is regarded as a secondary pathogen, may play a role in the development of the disease. Previous studies of strains from swine pneumonia revealed that there are a few clones associated with clinical disease, suggesting that some strains may be more virulent than others. This linkage may be true in the BRD, however composition of P. multocida populations in the herds are slightly characterized. Thus, we decided to perform phenotypic and genotypic characterisation of strains isolated from calves with respiratory infection at 31 different herds in Hungary. The results demonstrated the presence of two dominant strain types. At the identical taxonomic background (P. multocida subsp. multocida) with slight phenotypic variability they could be separated by trehalose fermentation capacity, α-glucosidase activity and molecular fingerprint patterns of ERIC- and M13-PCR. Independent prevalence and geographical origin of the strain types may refer to their significance in the illness, but their comparison with strains isolated from healthy individuals is taken into consideration.

  20. Comparative Genomics Analysis of Two Different Virulent Bovine Pasteurella multocida Isolates

    PubMed Central

    Pan, Tingting; Li, Tian; Wu, Rui

    2016-01-01

    The Pasteurella multocida capsular type A isolates can cause pneumonia and bovine respiratory disease (BRD). In this study, comparative genomics analysis was carried out to identify the virulence genes in two different virulent P. multocida capsular type A isolates (high virulent PmCQ2 and low virulent PmCQ6). The draft genome sequence of PmCQ2 is 2.32 Mbp and contains 2,002 protein-coding genes, 9 insertion sequence (IS) elements, and 1 prophage region. The draft genome sequence of PmCQ6 is 2.29 Mbp and contains 1,970 protein-coding genes, 2 IS elements, and 3 prophage regions. The genome alignment analysis revealed that the genome similarity between PmCQ2 and PmCQ6 is 99% with high colinearity. To identify the candidate genes responsible for virulence, the PmCQ2 and PmCQ6 were compared together with that of the published genomes of high virulent Pm36950 and PmHN06 and avirulent Pm3480 and Pm70 (capsular type F). Five genes and two insertion sequences are identified in high virulent strains but not in low virulent or avirulent strains. These results indicated that these genes or insertion sequences might be responsible for the virulence of P. multocida, providing prospective candidates for further studies on the pathogenesis and the host-pathogen interactions of P. multocida. PMID:28070502

  1. Antibodies against Pasteurella multocida in snow geese in the western arctic

    USGS Publications Warehouse

    Samuel, M.D.; Shadduck, D.J.; Goldberg, D.R.; Baranyuk, V.; Sileo, L.; Price, J.I.

    1999-01-01

    To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993-1996. Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P. multocida in these populations. Geese with serum antibody levels indicating recent infection with P. multocida were found at both breeding colonies. Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics. Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected. Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease. This pattern of prevalence indicated that enzootic levels of infection with P. multocida occurred at both breeding colonies. When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%).

  2. Virulence Genes and Antimicrobial Resistance Profiles of Pasteurella multocida Strains Isolated from Rabbits in Brazil

    PubMed Central

    Ferreira, Thais Sebastiana Porfida; Felizardo, Maria Roberta; Sena de Gobbi, Débora Dirani; Gomes, Cleise Ribeiro; Nogueira Filsner, Pedro Henrique de Lima; Moreno, Marina; Paixão, Renata; Pereira, Jucélia de Jesus; Micke Moreno, Andrea

    2012-01-01

    Pasteurella multocida is responsible for a wide range of diseases in domestic animals. In rabbits, the agent is related to nasal discharge, pneumonia, otitis media, pyometra, orchitis, abscess, and septicemia. One hundred and forty rabbits with respiratory diseases from four rabbitries in São Paulo State, Brazil were evaluated for the detection of P. multocida in their nasal cavities. A total of twenty-nine animals were positive to P. multocida isolation, and 46 strains were selected and characterized by means of biochemical tests and PCR. P. multocida strains were tested for capsular type, virulence genes, and resistance profile. A total of 45.6% (21/46) of isolates belonged to capsular type A, and 54.34% (25/46) of the isolates were untypeable. None of the strains harboured toxA or pfhA genes. The frequency of the other twenty genes tested was variable, and the data generated was used to build a dendrogram, showing the relatedness of strains, which were clustered according to origin. Resistance revealed to be more common against sulfonamides and cotrimoxazole, followed by erythromycin, penicillin, and amoxicillin. PMID:22919347

  3. Antibodies against Pasteurella multocida in snow geese in the western Arctic.

    PubMed

    Samuel, M D; Shadduck, D J; Goldberg, D R; Baranyuk, V; Sileo, L; Price, J I

    1999-07-01

    To determine if lesser snow geese (Chen caerulescens caerulescens) are a potential reservoir for the Pasteurella multocida bacterium that causes avian cholera, serum samples and/or pharyngeal swabs were collected from > 3,400 adult geese breeding on Wrangel Island (Russia) and Banks Island (Canada) during 1993-1996. Pharyngeal swab sampling rarely (> 0.1%) detected birds that were exposed to P. multocida in these populations. Geese with serum antibody levels indicating recent infection with P. multocida were found at both breeding colonies. Prevalence of seropositive birds was 3.5% at Wrangel Island, an area that has no recorded history of avian cholera epizootics. Prevalence of seropositive birds was 2.8% at Banks Island in 1994, but increased to 8.2% during 1995 and 1996 when an estimated 40,000-60,000 snow geese were infected. Approximately 50% of the infected birds died during the epizootic and a portion of the surviving birds may have become carriers of the disease. This pattern of prevalence indicated that enzootic levels of infection with P. multocida occurred at both breeding colonies. When no avian cholera epizootics occurred (Wrangel Island, Banks Island in 1994), female snow geese (4.7%) had higher antibody prevalence than males (2.0%).

  4. [The effect of local and systemic immunization of suckling pigs on bronchoalveolar clearance and on intrabronchial infections with Pasteurella multocida].

    PubMed

    Müller, G; Heilmann, P

    1990-01-01

    In vivo investigations of bactericidal bronchoalveolar clearance of weaning pigs locally or systemically immunized with Pasteurella antigens exhibited clearly distinctive results, as early as 3 days after the last immunization. They differed from controls by significant increase in the elimination of the homologous Pasteurella multocida A wild strain. That increase in the clearance remains for 21 days and is specific only to the immunized capsular type. There are no relations of this result to the cytological findings of the lavage fluids and to results of the intrabronchial challenge infection. Frequency and severity of pneumonias were reduced by as little as 3 aerogenic immunizations with Pasteurella mutants.

  5. Experimental study of pathogenicity of Pasteurella multocida serogroup F in rabbits.

    PubMed

    Jaglic, Zoran; Jeklova, Edita; Leva, Lenka; Kummer, Vladimir; Kucerova, Zdenka; Faldyna, Martin; Maskova, Jarmila; Nedbalcova, Katerina; Alexa, Pavel

    2008-01-01

    The role of Pasteurella multocida serogroup F in inducing disease in rabbits was investigated in this study. Three groups of 12 Pasteurella-free rabbits each were intranasally (i.n.), subcutaneously (s.c.), and perorally (p.o.) challenged, respectively. Six rabbits of each group were immunosuppressed using dexamethasone. Eight rabbits (four of them immunosuppressed) inoculated i.n. showed symptoms of respiratory distress resulting in respiratory failure and died or were euthanized in the terminal stage of the disease 3-6 days post-infection (p.i.). The main pathological findings were fibrinopurulent pleuropneumonia (immunocompetent rabbits) or diffuse haemorrhagic pneumonia (immunosuppressed rabbits). Septicemic syndrome ending with shock occurred in 11 rabbits (6 of them immunosuppressed) inoculated s.c., which died or were euthanized in the terminal stage of the disease 2-3 days p.i. The most significant pathological findings were extensive cutaneous and subcutaneous lesions. All of the p.o. inoculated rabbits survived the challenge showing no clinical signs of the disease and no macroscopic lesions. The observations in this study indicate that in addition to serogroups A and D of P. multocida, serogroup F also can be highly pathogenic for rabbits and therefore might be a cause of considerable economic loss in commercial rabbit production.

  6. [A case of Pasteurella multocida subsp. multocida septicemia due to cat bites in liver cirrhosis patient].

    PubMed

    Shimizu, T; Hasegawa, K; Mitsuhashi, Y; Kojima, S; Ishikawa, K; Hayashi, N; Sawada, T

    1995-11-01

    A 60-year-old male who had been suffering from liver cirrhosis was admitted to our hospital with high grade fever accompanied by right chest pain. Chest X-rays revealed a moderate amount of pleural fluid suggesting pleuritis. Multocida was isolated from the blood culture as well as the pleural fluid. Antibiotic therapy was initiated according to the drug susceptibility of the isolates. Ten days treatment was effective on the cessation of both septicemia and the clinical symptoms. Since the patient had been bitten several times by his own pet cats, their mouth swabs were taken for pathogenic investigations. Serotypes of the cats' isolates coincided with that of the patient's which consequently indicated the route of infection. P. multocida is a Gram negative coccobacillary organism that resides as normal flora in the oral cavity of animals, including dogs and cats. It has been originally known to be a causative agent for hemorrhagic septicemia in domestic animals. However, recently reports of P. multocida infections in man has been increasing due to the enlargement of pet populations. Although outbreaks of septicemia is rare, it occurs most often in immunologically compromised hosts, including patients with liver cirrhosis as in this case. Therefore, it is important to initiate an urgent antibiotic therapy in such cases. Overall, it is of utmost importance to instruct immunosuppressed patients to avoid excessive exposure to animals including pets.

  7. Inhibition of Pasteurella multocida Adhesion to Rabbit Respiratory Epithelium Using Lectins

    PubMed Central

    Carrillo, Magda Patricia; Martinez, Nhora María; Patiño, María del Pilar; Iregui, Carlos Arturo

    2015-01-01

    This study aimed to evaluate the ability of a panel of lectins to inhibit the ability of Pasteurella multocida to adhere to and affect the rabbit respiratory epithelium. Nasal septa from rabbit fetuses were cultured with various lectins before the addition of P. multocida. The percentage of bacteria adhering to the epithelium was evaluated semiquantitatively by indirect immunoperoxidase (IIP) staining. The goblet cells (GCs) were counted in semithin sections stained with toluidine blue and served as the main morphological criterion to evaluate the inhibitory effect of the lectins. The lectins PNA, WGA, RCA120, and DBA significantly inhibited the adhesion of P. multocida to the ciliated epithelium (P < 0.05) and prevented the pathogen-induced increase in the number of GCs (P < 0.05) compared with those of positive control tissues. In addition, VVA, SJA, UEA I, DSL, SBA, and ECL significantly inhibited the increase in GCs compared with that of the control tissues. The results suggest that less aggressive therapeutic strategies, such as treatment with lectins, may represent alternative approaches to control bacterial respiratory infections. PMID:25810949

  8. In vivo antimicrobial activity of marbofloxacin against Pasteurella multocida in a tissue cage model in calves

    PubMed Central

    Cao, Changfu; Qu, Ying; Sun, Meizhen; Qiu, Zhenzhen; Huang, Xianhui; Huai, Binbin; Lu, Yan; Zeng, Zhenling

    2015-01-01

    Marbofloxacin is a fluoroquinolone specially developed for use in veterinary medicine with broad-spectrum antibacterial activity. The objective of our study was to re-evaluate in vivo antimicrobial activity of marbofloxacin against Pasteurella multocida using subcutaneously implanted tissue cages in calves. Calves were infected by direct injection into tissue cages with P. multocida(type B, serotype 2), then intramuscularly received a range of marbofloxacin doses 24 h after inoculation. The ratio of 24 h area under the concentration-time curve divided by the minimum inhibitory concentration or the mutant prevention concentration (AUC24 h/MIC or AUC24 h/MPC) was the pharmacokinetic-pharmacodynamic (PK/PD) index that best described the effectiveness of marbofloxacin against P. multocida (R2 = 0.8514) by non-linear regression analysis. Marbofloxacin exhibited a good antimicrobial activity in vivo. The levels of AUC24 h/MIC and AUC24 h/MPC that produced 50% (1.5log10 CFU/mL reduction) and 90% (3log10 CFU/mL reduction) of maximum response were 18.60 and 50.65 h, 4.67 and 12.89 h by using sigmoid Emax model WINNONLIN software, respectively. The in vivo PK/PD integrated methods by tissue cage model display the advantage of the evaluation of antimicrobial activity and the optimization of the dosage regimen for antibiotics in the presence of the host defenses, especially in target animal of veterinary interest. PMID:26257726

  9. Studies on the presence and persistence of Pasteurella multocida serovars and genotypes in fowl cholera outbreaks.

    PubMed

    Singh, Reema; Blackall, Patrick J; Remington, Bruce; Turni, Conny

    2013-12-01

    Pasteurella multocida was isolated from poultry on six farms (one free-range duck farm, one free-range turkey farm, one conventional enclosed turkey farm, and three free-range layer farms) suffering fowl cholera outbreaks. In addition, historical isolates from previous outbreaks were available for the conventional turkey farm and the three free-range layer farms. The isolates were serotyped using the Heddleston scheme and genotyped using multi-locus sequencing typing. In the current outbreaks, two of the farms had two different sequence types (STs) of P. multocida in the investigated outbreak (the free-range turkey farm and one of the free-range layer farms). The remaining four farms had one ST within the investigated outbreak. In looking at the historical isolates, two of the four farms had multiple genotypes involved. On the four farms with historical isolates from previous outbreaks, at least one new genotype was present in the investigated outbreak as compared with the historical isolates. On one layer farm, one genotype persisted over a 10-year period. Serotyping revealed the presence of multiple serovars in the current and historical outbreaks, with serovars sometimes changing over time. This study has shown that several STs of P. multocida can be present during some outbreaks of fowl cholera, although other outbreaks involve a single ST. Also, the STs present on a property suffering repeated fowl cholera can both persist and change over time.

  10. Development of a multi-locus sequence typing scheme for avian isolates of Pasteurella multocida.

    PubMed

    Subaaharan, S; Blackall, L L; Blackall, P J

    2010-03-24

    A total of 63 isolates of Pasteurella multocida from Australian poultry, all associated with fowl cholera outbreaks, and three international reference strains, representing the three subspecies within P. multocida were used to develop a multi-locus sequence typing scheme. Primers were designed for conserved regions of seven house-keeping enzymes -adk, est, gdh, mdh, pgi, pmi and zwf - and internal fragments of 570-784 bp were sequenced for all isolates and strains. The number of alleles at the different loci ranged from 11 to 20 and a total of 29 allelic profiles or sequence types were recognised amongst the 66 strains. There was a strong concordance between the MLST data and the existing multi-locus enzyme electrophoresis and ribotyping data. When used to study a sub-set of isolates with a known detailed epidemiological history, the MLST data matched the results given by restriction endonuclease analysis, pulsed-field gel electrophoresis, ribotyping and REP-PCR. The MLST scheme provides a high level of resolution and is an excellent tool for studying the population structure and epidemiology of P. multocida.

  11. A Pasteurella multocida strain affecting nulliparous heifers and calves in different ways.

    PubMed

    Turni, Conny; Dayao, Denise; Aduriz, Gorka; Cortabarria, Nekane; Tejero, Carolina; Ibabe, Jose C; Singh, Reema; Blackall, Pat

    2016-11-15

    Pasteurella multocida isolates from dairy cattle on a farm in Spain were associated with pneumonia of calves (six isolates) and mastitis of heifers (five isolates). The objective was to determine if the P. multocida isolates retrieved from both disease scenarios were the same strain or whether more than one strain was present. The isolates were identified by a species-specific polymerase chain (PCR) assay, serotyped by the Heddleston scheme and then typed by a number of molecular genotyping assays including multi-locus sequence typing (MLST). The 11 isolates were confirmed as P. multocida but failed to react with any of the 16 Heddleston antisera. The PCR targeting the genes associated with the lipopolysaccharide outer core biosynthesis locus assigned all the isolates to L3-the type that contains Heddleston serovars 3 and 4. The MLST analysis showed all isolates belonging to ST 79 within the clonal complex of ST13. Only one of the isolates showed a slight different profile by the repetitive extragenic palindromic PCR. The conclusion was that the same strain was associated with pneumonia in calves and mastitis in heifers.

  12. Experimental pathogenicity and complete genome characterization of a pig origin Pasteurella multocida serogroup F isolate HN07.

    PubMed

    Peng, Zhong; Liang, Wan; Wang, Yuanguo; Liu, Wenjing; Zhang, Hongfeng; Yu, Teng; Zhang, Anding; Chen, Huanchun; Wu, Bin

    2017-01-01

    Pasteurella multocida serotype F isolates are predominately prevalent in avian hosts, but rarely seen in pigs. However, we isolated several strains of P. multocida serotype F from clinical samples of pigs in China. To understand the pathogenicity of these strains, one of the serotype F isolates designated HN07, was used to challenge experimental chickens, as P. multocida of this serotype is predominately prevalent in avian hosts. However, strain HN07 could not resulted in significant clinical signs in experimental chickens even at an infective dose of ∼10(9) CFU, suggesting the isolate was avirulent to chickens and therefore raising the possibility that the porcine serotype F isolate is not transmitted by chickens. We then used HN07 to challenge experimental pigs, as this strain was isolated from pigs. As expected, the strain led to the clinical signs and the pathological lesions in experimental pigs that are similar to the pasteurellosis disease. We then determined the complete genome sequence of the pig origin serogroup F isolate HN07 for the first time. Genome comparison between HN07 and the avian serotype F P. multocida Pm70 identified a novel integrative conjugative element (ICE) ICEpmcn07 which was likely to harbor a series of genes responsible for a putative type IV secretion system (T4SS) in HN07. This is the first time that we determined an ICE carrying a T4SS in P. multocida. Besides, comparative analysis also defined a number of virulence-associated genes in HN07 but absent in Pm70 which may have a contribution to the pathogenicity of the strain. This is the first report of the pathogenicity and genome characterization of a pig origin Pasteurella multocida serogroup F isolate. The pathogenic and genomic definition of the pig origin P. multocida serogroup F in our study would have significance on the pathogenesis and genetic diversity and virulence variability of P. multocida.

  13. [A case of cat-scratch-induced Pasteurella multocida infection presenting with disseminated intravascular coagulation and acute renal failure].

    PubMed

    Fukuchi, Takahiko; Morisawa, Yuji

    2009-09-01

    Domestic animals are the main reservoirs of Pasteurella species for human zoonosis due to bites and scratches. Pasterurella multocida may cause serious soft-tissue infection and, less commonly, sepsis or septic shock, particularly in insufficient initial therapy and an immunocompromised host. We report a case of cat-scratch-induced P. multocida infection, presenting with disseminated intravascular coagulation and acute renal failure. A febrile 83-year-old woman with consciousness disturbance and a subcutaneous left-foot abscess due to a scratch from a pet cat. She was successfully treated with antibiotic piperacillin and clindamycin therapy and aggressive wound drainage.

  14. Efficacy of a novel Pasteurella multocida vaccine against progressive atrophic rhinitis of swine

    USGS Publications Warehouse

    Hsuan, Shih-Ling; Liao, Chih-Ming; Huang, Chienjin; Winton, James R.; Chen, Zeng-Weng; Lee, Wei-Cheng; Liao, Jiunn-Wang; Chen, Ter-Hsin; Chiou, Chwei-Jang; Yeh, Kuang-Sheng; Chien, Maw-Sheng

    2009-01-01

    The efficacy of a novel vaccine composed of three short recombinant subunit Pasteurella multocida toxin (PMT) proteins in combination with a bi-valent P. multocida whole-cell bacterin (rsPMT–PM) was evaluated in field studies for prevention and control of progressive atrophic rhinitis (PAR) of swine at 15 conventional farrow-to-finish farms. Experimental piglets that were immunized twice with the rsPMT–PM vaccine developed detectable titers of neutralizing antibodies (greater than 1:8) that prevented the growth retardation and pathological lesions typically observed following challenge with authentic PMT. A total of 542 sows were vaccinated once or twice prior to parturition and serum neutralizing antibody titers were evaluated. Both single and double vaccination protocols induced neutralizing antibody titers of 1:16 or higher in 62% and 74% of sows, respectively. Notably, neither sows nor piglets at a farm experiencing a severe outbreak of PAR at the time of the vaccination trial had detectable antibody titers, but antibody titers increased significantly to 1:16 or higher in 40% of sows following double vaccination. During the year after vaccination, clinical signs of PAR decreased in fattening pigs and growth performance improved sufficiently to reduce the rearing period until marketing by 2 weeks. Collectively, these results indicate that the rsPMT–PM vaccine could be used to provide protective immunity for controlling the prevalence and severity of PAR among farm-raised swine.

  15. Identification and antimicrobial susceptibility patterns of Pasteurella multocida isolated from chickens and japanese quails in Brazil.

    PubMed

    Rigobelo, Everlon Cid; Blackall, Patrick Joseph; Maluta, Renato Pariz; de Ávila, Fernando Antonio

    2013-01-01

    A study was performed to verify the presence of Pasteurella multocida in eight different poultry groups of 90 birds each. Groups I to IV were chickens (I being > 6 weeks of age with a history of respiratory illness, II > 6 weeks of age and free of respiratory illness, III < 6 weeks of age with respiratory illness and IV being < 6 weeks of age and with no respiratory illness. Groups V to VIII had the matching characteristics of Groups I to V but consisted of Japanese Quails. The P. multocida isolation rate from the groups was as follows; Group I 56/90 (62.3%) Group II 18/90 (20.0%), Group III 12/90 (13.3%), Group IV 3/90 (3.33%), Group V 8/90 (8.88%), Group VI 2/90 (2.22%) Group VII 2/90 (2.22%) and Group VIII 1/90 (1.11%). These isolation rates were not significantly different within the groups of a bird type but the overall chicken isolation rate was significantly higher than the quail isolation rate (p < 0.01). All isolates were examined for their sensitivity to four antimicrobial agents. The results showed only low levels of resistance to the agents tested. The highest level of resistance detected was to cephalothin (5.1% of isolates) followed by amikacin (3.4%).

  16. Identification and antimicrobial susceptibility patterns of Pasteurella multocida isolated from chickens and japanese quails in Brazil

    PubMed Central

    Rigobelo, Everlon Cid; Blackall, Patrick Joseph; Maluta, Renato Pariz; de Ávila, Fernando Antonio

    2013-01-01

    A study was performed to verify the presence of Pasteurella multocida in eight different poultry groups of 90 birds each. Groups I to IV were chickens (I being > 6 weeks of age with a history of respiratory illness, II > 6 weeks of age and free of respiratory illness, III < 6 weeks of age with respiratory illness and IV being < 6 weeks of age and with no respiratory illness. Groups V to VIII had the matching characteristics of Groups I to V but consisted of Japanese Quails. The P. multocida isolation rate from the groups was as follows; Group I 56/90 (62.3%) Group II 18/90 (20.0%), Group III 12/90 (13.3%), Group IV 3/90 (3.33%), Group V 8/90 (8.88%), Group VI 2/90 (2.22%) Group VII 2/90 (2.22%) and Group VIII 1/90 (1.11%). These isolation rates were not significantly different within the groups of a bird type but the overall chicken isolation rate was significantly higher than the quail isolation rate (p < 0.01). All isolates were examined for their sensitivity to four antimicrobial agents. The results showed only low levels of resistance to the agents tested. The highest level of resistance detected was to cephalothin (5.1% of isolates) followed by amikacin (3.4%). PMID:24159299

  17. Identification of avian strains of Pasteurella multocida in India by conventional and PCR assays.

    PubMed

    Shivachandra, S B; Kumar, A A; Gautam, R; Singh, Vijendra P; Saxena, M K; Srivastava, S K

    2006-11-01

    The prevalence of capsular and somatic serotypes were studied among 123 Pasteurella multocida strains isolated from chickens (n=94), ducks (22), quails (4), turkeys (2) and geese (1) from different geographical regions of India. All strains exhibited similar cultural and morphological characteristics. Ninety-two of the isolates belonged to serotype A:1, the most prevalent serotype, with serotypes A:3, A:1,3, D:3 and F:3 having two isolates each. Only one isolate was positive for serotypes A:4 and D:1. Twenty isolates were untyped. A multiplex capsular PCR assay generated amplicons of sizes approximately 460, approximately 1044, approximately 657 and approximately 854 bp in 106 isolates identified as capsular serotype-A, 15 in serotype D and two in serotype F. Capsular types B and E were not detected in any of the avian isolates studied. The present findings suggest that a multiplex capsular PCR assay may be suitable for the rapid initial identification serotypes P. multocida during epidemiological studies of fowl cholera.

  18. Serotypes and DNA fingerprint profiles of Pasteurella multocida isolated from raptors

    USGS Publications Warehouse

    Wilson, M.A.; Duncan, R.M.; Nordholm, G.E.; Berlowski, B.M.

    1995-01-01

    Pasteurella multocida isolates from 21 raptors were examined by DNA fingerprint profile and serotyping methods. Isolates were obtained from noncaptive birds of prey found in 11 states from November 28, 1979, through February 10, 1993. Nine isolates were from bald eagles, and the remaining isolates were from hawks, falcons, and owls. Seven isolates were members of capsule group A, and 14 were nonencapsulated. One isolate was identified as somatic type 3, and another was type 3,4,7; both had unique HhaI DNA fingerprint profiles. Nineteen isolates expressed somatic type 1 antigen; HhaI profiles of all type 1 isolates were identical to each other and to the HhaI profile of the reference somatic type 1, strain X-73. The 19 type 1 isolates were differentiated by sequential digestion of DNA with HpaII; four HpaII fingerprint profiles were obtained. The HpaII profile of one isolate was identical to the HpaII profile of strain X-73. Incidence of P. multocida somatic type 1 in raptors suggests that this type may be prevalent in other wildlife or wildlife environments.

  19. Ultrastructural Comparison of the Nasal Epithelia of Healthy and Naturally Affected Rabbits with Pasteurella multocida A

    PubMed Central

    Esquinas, Paula; Botero, Lucía; Patiño, María del Pilar; Iregui, Carlos

    2013-01-01

    An ultrastructural comparison between the nasal cavities of healthy rabbits and those suffering from two forms of spontaneous infection with Pasteurella multocida was undertaken. Twelve commercially produced rabbits of different ages and respiratory health status were divided into four groups: healthy from 0 to 21 days (G1, n = 2); healthy from 23 to 49 days (G2, n = 2); healthy from 51 to 69 days (G3, n = 2); diseased rabbits with septicemia and the rhinitic form of P. multocida infection (G4, n = 3). The main ultrastructural changes observed were a widening of the interepithelial spaces, increased activity and number of goblet cells, the formation of two types of vacuoles in epithelial cells, the degranulation and migration of heterophils between the epithelial cells, and the association of this migration with some of the other changes. No bacteria were observed adhering to the epithelium, and very few were observed free in the mucus. Scant inter-epithelial spaces were found in healthy rabbits, but they were not as large and numerous as those found in diseased animals. We discuss the origin and meaning of these changes but, we focus on the significance of the inter-epithelial spaces and goblet cells for the defense of the upper respiratory airways against the bacterium and its lipopolysaccharide. PMID:23577280

  20. Characterization of Pasteurella multocida isolates from wetland ecosystems during 1996 to 1999

    USGS Publications Warehouse

    Samuel, M.D.; Shadduck, D.J.; Goldberg, D.R.; Wilson, M.A.; Joly, D.O.; Lehr, M.A.

    2003-01-01

    We cultured 126 Pasteurella multocida isolates, 92 from water and 34 from sediment samples collected from wetlands in the Pacific and Central flyways of the United States between 1996 and 1999. Most (121) of the isolates were P. multocida serotype 1, but serotypes 3, 3/4, 10, and 11 were also found. Many (82) of the isolates were further characterized by DNA fingerprinting procedures and tested in Pekin ducks for virulence. Almost all the serotype 1 isolates we tested caused mortality in Pekin ducks. Serotype 1 isolates varied in virulence, but the most consistent pattern was higher mortality in male ducks than in females. We found no evidence that isolates found in sediment vs. water, between Pacific and Central flyways, or during El Nino years had consistently different virulence. We also found a number of non-serotype 1 isolates that were avirulent in Pekin ducks. Isolates had DNA fingerprint profiles similar to those found in birds that died during avian cholera outbreaks.

  1. Pasteurella multocida toxin: Targeting mast cell secretory granules during kiss-and-run secretion.

    PubMed

    Danielsen, Elisabeth M; Christiansen, Nina; Danielsen, E Michael

    2016-02-01

    Pasteurella multocida toxin (PMT), a virulence factor of the pathogenic Gram-negative bacterium P. multocida, is a 146 kDa protein belonging to the A-B class of toxins. Once inside a target cell, the A domain deamidates the α-subunit of heterotrimeric G-proteins, thereby activating downstream signaling cascades. However, little is known about how PMT selects and enters its cellular targets. We therefore studied PMT binding and uptake in porcine cultured intestinal mucosal explants to identify susceptible cells in the epithelium and underlying lamina propria. In comparison with Vibrio cholera B-subunit, a well-known enterotoxin taken up by receptor-mediated endocytosis, PMT binding to the epithelial brush border was scarce, and no uptake into enterocytes was detected by 2h, implying that none of the glycolipids in the brush border are a functional receptor for PMT. However, in the lamina propria, PMT distinctly accumulated in the secretory granules of mast cells. This also occurred at 4 °C, ruling out endocytosis, but suggestive of uptake via pores that connect the granules to the cell surface. Mast cell granules are known to secrete their contents by a "kiss-and-run" mechanism, and we propose that PMT may exploit this secretory mechanism to gain entry into this particular cell type.

  2. Molecular characterization of Pasteurella multocida isolates obtained from poultry, ruminant, cats and dogs using RAPD and REP-PCR analysis

    PubMed Central

    Shirzad-Aski, Hesamaddin; Tabatabaei, Mohammad

    2016-01-01

    In the present study, Randomly Amplified Polymorphic DNA (RAPD) and Repetitive Extragenic Palindromic sequence-based Polymerase Chain Reaction (REP- PCR) were used to characterize 131 isolates of Pasteurella multocida, originating from different healthy and diseased animal species obtained from several geographical regions of Iran. The RAPD and REP-PCR generated amplified products in the range of 300 to 3400 bp and 200 to 2850 bp, respectively. Among all of the P. multocida isolates, cluster analysis revealed that 63 clusters and nine untypable isolates and 81 clusters and six untypable isolates were produced with RAPD and REP-PCR methods, respectively. The results indicated that the REP-PCR method showed a slightly higher level of discrimination power in differentiating of P. multocida isolates as compared with RAPD. The results showed that a considerable level of genetic diversity exists among P. multocida isolates even in the isolates with the same animal or geographical origins. There was no host- and region-specific pattern. In addition, the isolates obtained from the healthy and diseased animal did not reveal any correlation genotypic profiles, which could be supported by the hypothesis that P. multocida is a strictly opportunistic pathogen. In conclusion, because of a large amount of genetic heterogeneity in the P. multocida isolates, Pasteurellosis may be caused by different clones in the same herd or animal. PMID:28097166

  3. Comparative clinicopathological changes in buffalo and cattle following infection by Pasteurella multocida B:2.

    PubMed

    Annas, S; Zamri-Saad, M; Jesse, F F A; Zunita, Z

    2015-11-01

    Haemorrhagic septicaemia (HS) is an acute, septicaemic disease of cattle and buffalo of Asia and Africa caused by Pasteurella multocida B:2 or E:2. Buffaloes are believed to be more susceptible than cattle. In this study, 9 buffaloes of 8 months old were divided equally into 3 groups (Groups 1, 3, 5). Similarly, 9 cattle of 8 months old were equally divided into 3 groups (Groups 2, 4, 6). Animals of Groups 1 and 2 were inoculated with PBS while Groups 3 and 4 were inoculated subcutaneously with 10(5) cfu/ml of P. multocida B:2. Animals of Groups 5 and 6 were inoculated intranasally with the same inoculum. Both buffaloes and cattle that were inoculated subcutaneously succumbed to the infection at 16 h and 18 h, respectively. Two buffaloes that were inoculated intranasally (Group 5) succumbed at 68 h while the remaining cattle and buffaloes survived the 72-h study period. Endotoxin was detected in the blood of infected cattle (Group 4) and buffaloes (Groups 3 and 5) prior to the detection of P. multocida B:2 in the blood. The endotoxin was detected in the blood of buffaloes of Group 3 and cattle of Group 4 at 0.5 h post-inoculation while buffaloes of Group 5 and cattle of Group 6 at 1.5 h. On the other hand, bacteraemia was detected at 2.5 h in buffaloes of Group 3 and cattle of Group 4 and at 12 h in buffaloes of Group 5 and cattle of Group 6. Affected cattle and buffaloes showed lesions typical of haemorrhagic septicaemia. These included congestion and haemorrhages in the organs of respiratory, gastrointestinal and urinary tracts with evidence of acute inflammatory reactions. The severity of gross and histopathology lesions in cattle and buffalo calves that succumbed to the infection showed insignificant (p > 0.05) difference. However, inoculated buffalo and cattle that survived the infection showed significantly (p < 0.05) less severe gross and histopathological changes than those that succumbed. In general, cattle are more resistant to intranasal infection by P

  4. Localization of the Intracellular Activity Domain of Pasteurella multocida Toxin to the N Terminus

    PubMed Central

    Wilson, Brenda A.; Ponferrada, Virgilio G.; Vallance, Jefferson E.; Ho, Mengfei

    1999-01-01

    We have shown that Pasteurella multocida toxin (PMT) directly causes transient activation of Gqα protein that is coupled to phosphatidylinositol-specific phospholipase Cβ1 in Xenopus oocytes (B. A. Wilson, X. Zhu, M. Ho, and L. Lu, J. Biol. Chem. 272:1268–1275, 1997). We found that antibodies directed against an N-terminal peptide of PMT inhibited the toxin-induced response in Xenopus oocytes, but antibodies against a C-terminal peptide did not. To test whether the intracellular activity domain of PMT is localized to the N terminus, we conducted a deletion mutational analysis of the PMT protein, using the Xenopus oocyte system as a means of screening for toxin activity. Using PCR and conventional cloning techniques, we cloned from a toxinogenic strain of P. multocida the entire toxA gene, encoding the 1,285-amino-acid PMT protein, and expressed the recombinant toxin as a His-tagged fusion protein in Escherichia coli. We subsequently generated a series of N-terminal and C-terminal deletion mutants and expressed the His-tagged PMT fragments in E. coli. These proteins were screened for cytotoxic activity on cultured Vero cells and for intracellular activity in the Xenopus oocyte system. Only the full-length protein without the His tag exhibited activity on Vero cells. The full-length PMT and N-terminal fragments containing the first 500 residues elicited responses in oocytes, but the C-terminal 780 amino acid fragment did not. Our results confirm that the intracellular activity domain of PMT is localized to the N-terminal 500 amino acids of the protein and that the C terminus is required for entry into cells. PMID:9864199

  5. Protective efficacy afforded by live Pasteurella multocida vaccines in chickens is independent of lipopolysaccharide outer core structure.

    PubMed

    Harper, Marina; John, Marietta; Edmunds, Mark; Wright, Amy; Ford, Mark; Turni, Conny; Blackall, P J; Cox, Andrew; Adler, Ben; Boyce, John D

    2016-03-29

    Pasteurella multocida is a major animal pathogen that causes a range of diseases including fowl cholera. P. multocida infections result in considerable losses to layer and breeder flocks in poultry industries worldwide. Both killed whole-cell and live-attenuated vaccines are available; these vaccines vary in their protective efficacy, particularly against heterologous strains. Moreover, until recently there was no knowledge of P. multocida LPS genetics and structure to determine precisely how LPS structure affects the protective capacity of these vaccines. In this study we show that defined lipopolysaccharide (LPS) mutants presented as killed whole-cell vaccines elicited solid protective immunity only against P. multocida challenge strains expressing highly similar or identical LPS structures. This finding indicates that vaccination of commercial flocks with P. multocida killed cell formulations will not protect against strains producing an LPS structure different to that produced by strains included in the vaccine formulation. Conversely, protective immunity conferred by vaccination with live P. multocida strains was found to be largely independent of LPS structure. Birds vaccinated with a range of live mutants belonging to the L1 and L3 LPS genotypes, each expressing a specific truncated LPS structure, were protected against challenge with the parent strain. Moreover, birds vaccinated with any of the five LPS mutants belonging to the L1 LPS genotype were also protected against challenge with an unrelated strain and two of the five groups vaccinated with live LPS mutants belonging to the L3 genotype were protected against challenge with an unrelated strain. In summary, vaccination with live P. multocida aroA mutants producing full-length L1 or L3 LPS or vaccination with live strains producing shortened L1 LPS elicited strong protective immunity against both homologous and heterologous challenge.

  6. Surveillance of antimicrobial resistance in clinical isolates of Pasteurella multocida and Streptococcus suis from Ontario swine.

    PubMed

    Glass-Kaastra, Shiona K; Pearl, David L; Reid-Smith, Richard J; McEwen, Beverly; Slavic, Durda; Fairles, Jim; McEwen, Scott A

    2014-10-01

    Susceptibility results for Pasteurella multocida and Streptococcus suis isolated from swine clinical samples were obtained from January 1998 to October 2010 from the Animal Health Laboratory at the University of Guelph, Guelph, Ontario, and used to describe variation in antimicrobial resistance (AMR) to 4 drugs of importance in the Ontario swine industry: ampicillin, tetracycline, tiamulin, and trimethoprim-sulfamethoxazole. Four temporal data-analysis options were used: visualization of trends in 12-month rolling averages, logistic-regression modeling, temporal-scan statistics, and a scan with the "What's strange about recent events?" (WSARE) algorithm. The AMR trends varied among the antimicrobial drugs for a single pathogen and between pathogens for a single antimicrobial, suggesting that pathogen-specific AMR surveillance may be preferable to indicator data. The 4 methods provided complementary and, at times, redundant results. The most appropriate combination of analysis methods for surveillance using these data included temporal-scan statistics with a visualization method (rolling-average or predicted-probability plots following logistic-regression models). The WSARE algorithm provided interesting results for quality control and has the potential to detect new resistance patterns; however, missing data created problems for displaying the results in a way that would be meaningful to all surveillance stakeholders.

  7. Pasteurella multocida toxin as a transporter of non-cell-permeating proteins.

    PubMed

    Bergmann, Stefan; Jehle, Doris; Schwan, Carsten; Orth, Joachim H C; Aktories, Klaus

    2013-07-01

    The protein toxin Pasteurella multocida toxin (PMT) is the causative agent of atrophic rhinitis in pigs, leading to atrophy of the nasal turbinate bones by affecting osteoblasts and osteoclasts. The mechanism of PMT-induced intoxication is a deamidation of α-subunits of heterotrimeric G proteins, including Gαq, Gα13, and Gαi, thereby causing persistent activation of the G proteins. Here we utilized PMT as a transporter of the non-cell-permeating A domain of diphtheria toxin (DTa). Fusion proteins of PMT and DTa ADP-ribosylated elongation factor 2, the natural target of diphtheria toxin, leading to cell toxicity. PMT-DTa effects were competed by PMT, indicating binding to the same cell surface receptor. Fluorescently labeled PMT-DTa and PMT colocalized with specific markers of early and late endosomes. Bafilomycin A, which inhibits vacuolar H(+)-ATPase, blocked PMT-DTa-induced intoxication of HEK-293 cells. By constructing various PMT-DTa chimeras, we identified a minimal region of PMT necessary for uptake of DTa. The data suggest that PMT is able to transport cargo proteins into eukaryotic cells by utilizing the PMT-specific uptake route.

  8. A systemic Pasteurella multocida toxin aggravates cardiac hypertrophy and fibrosis in mice.

    PubMed

    Weise, Markus; Vettel, Christiane; Spiger, Katharina; Gilsbach, Ralf; Hein, Lutz; Lorenz, Kristina; Wieland, Thomas; Aktories, Klaus; Orth, Joachim H C

    2015-09-01

    Pasteurella multocida toxin (PMT) persistently activates heterotrimeric G proteins of the Gαq/11 , Gα12/13 and Gαi family without interaction with G protein-coupled receptors (GPCRs). We show that PMT acts on heart tissue in vivo and on cardiomyocytes and cardiac fibroblasts in vitro by deamidation of heterotrimeric G proteins. Increased normalized ventricle weights and fibrosis were detected after intraperitoneal administration of PMT in combination with the GPCR agonist phenylephrine. In neonatal rat cardiomyocytes, PMT stimulated the mitogen-activated protein kinase pathway, which is crucial for the development of cellular hypertrophy. The toxin induced phosphorylation of the canonical phosphorylation sites of the extracellular-regulated kinase 1/2 and, additionally, caused phosphorylation of the recently recognized autophosphorylation site, which appears to be important for the development of cellular hypertrophy. Moreover, PMT stimulated the small GTPases Rac1 and RhoA. Both switch proteins are involved in cardiomyocyte hypertrophy. In addition, PMT stimulated RhoA and Rac1 in neonatal rat cardiac fibroblasts. RhoA and Rac1 have been implicated in the regulation of connective tissue growth factor (CTGF) secretion and expression. Accordingly, we show that PMT treatment increased secretion and expression of CTGF in cardiac fibroblasts. Altogether, the data indicate that PMT is an inducer of pathological remodelling of cardiac cells and identifies the toxin as a promising tool for studying heterotrimeric G protein-dependent signalling in cardiac cells.

  9. Selective Membrane Redistribution and Depletion of Gαq-Protein by Pasteurella multocida Toxin.

    PubMed

    Clemons, Nathan C; Luo, Shuhong; Ho, Mengfei; Wilson, Brenda A

    2016-08-01

    Pasteurella multocida toxin (PMT), the major virulence factor responsible for zoonotic atrophic rhinitis, is a protein deamidase that activates the alpha subunit of heterotrimeric G proteins. Initial activation of G alpha-q-coupled phospholipase C-beta-1 signaling by PMT is followed by uncoupling of G alpha-q-dependent signaling, causing downregulation of downstream calcium and mitogenic signaling pathways. Here, we show that PMT decreases endogenous and exogenously expressed G alpha-q protein content in host cell plasma membranes and in detergent resistant membrane (DRM) fractions. This membrane depletion of G alpha-q protein was dependent upon the catalytic activity of PMT. Results indicate that PMT-modified G alpha-q redistributes within the host cell membrane from the DRM fraction into the soluble membrane and cytosolic fractions. In contrast, PMT had no affect on G alpha-s or G beta protein levels, which are not substrate targets of PMT. PMT also had no affect on G alpha-11 levels, even though G alpha-11 can serve as a substrate for deamidation by PMT, suggesting that membrane depletion of PMT-modified G-alpha-q has specificity.

  10. Bordetella

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genus Bordetella includes 8 formally recognized species, of which Bordetella parapertussis, Bordetella bronchiseptica, Bordetella avium, and Bordetella hinzii are of veterinary interest. Bordetella pertussis, the type species, is an obligate human pathogen and the causative agent of whooping co...

  11. Cirrhosis, cellulitis and cats: a 'purrfect' combination for life-threatening spontaneous bacterial peritonitis from Pasteurella multocida.

    PubMed

    Hey, Penelope; Gow, Paul; Torresi, Joseph; Testro, Adam

    2012-11-11

    Pasteurella multocida is a Gram-negative coccobacillus that colonises the upper airways of many animals, in particular, dogs and cats. It acts as an opportunistic infection in humans following an animal bite or scratch and is associated with soft tissue infections, septicaemia and pneumonia, particularly in patients with a compromised immune response, such as patients with liver failure. Spontaneous bacterial peritonitis (SBP) is a serious complication of cirrhosis with a death rate of 10-15%. We report a case of a 47-year-old man with cirrhosis who presented with life-threatening P multocida SBP and bacteraemia secondary to a lick from a cat to a cellulitic leg wound. This case highlights the potential severity of an infection from domestic animals and an otherwise innocuous organism in an immunocompromised host.

  12. Detection of multiple strains of Pasteurella multocida in fowl cholera outbreaks by polymerase chain reaction-based typing.

    PubMed

    Shivachandra, S B; Kumar, A A; Gautam, R; Saxena, M K; Chaudhuri, P; Srivastava, S K

    2005-12-01

    Applicability of molecular methods for the detection and differentiation of Pasteurella multocida strains involved in two separate fowl cholera outbreaks in a single poultry farm was investigated. A total of 12 and 18 strains of P. multocida obtained from two separate outbreaks were subjected to phenotypic and genotypic characterization. Phenotypically, all strains were similar; however, DNA-based techniques by employing polymerase chain reaction (PCR) assays were found to be highly specific and sensitive for rapid detection and differentiation of strains. All 30 strains gave amplicons of approximately 460 bp and approximately 1,044 bp specific for P. multocida and capsular serogroup A in the Multiplex Capsular PCR typing system. Molecular typing techniques such as repetitive extragenic palindromic PCR, enterobacterial repetitive intergenic consensus PCR and single primer PCR differentiated all 30 strains into different profiles. However, similar patterns of genome fragments were observed among all strains following restriction endonuclease analysis using the enzyme HpaII. The current investigation revealed involvement of the same and multiple strains of P. multocida in two outbreaks. The results also indicated that molecular methods of detection and typing are rapid in comparison with conventional methods for epidemiological investigations of fowl cholera outbreaks.

  13. Wetland environmental conditions associated with the risk of avian cholera outbreaks and the abundance of Pasteurella multocida

    USGS Publications Warehouse

    Blanchong, Julie A.; Samuel, Michael D.; Goldberg, Diana R.; Shadduck, Daniel J.; Creekmore, L.H.

    2006-01-01

    Avian cholera is a significant infectious disease affecting waterfowl across North America and occurs worldwide among various avian species. Despite the importance of this disease, little is known about the factors that cause avian cholera outbreaks and what management strategies might be used to reduce disease mortality. Previous studies indicated that wetland water conditions may affect survival and transmission of Pasteurella multocida, the agent that causes avian cholera. These studies hypothesized that water conditions affect the likelihood that avian cholera outbreaks will occur in specific wetlands. To test these predictions, we collected data from avian cholera outbreak and non-outbreak (control) wetlands throughout North America (wintera??spring 1995a??1996 to 1998a??1999) to evaluate whether water conditions were associated with outbreaks. Conditional logistic regression analysis on paired outbreak and non-outbreak wetlands indicated no significant association between water conditions and the risk of avian cholera outbreaks. For wetlands where avian cholera outbreaks occurred, linear regression showed that increased eutrophic nutrient concentrations (Potassium [K], nitrate [NO3], phosphorus [P], and phosphate [PO3]) were positively related to the abundance of P. multocida recovered from water and sediment samples. Wetland protein concentration and an El Ni??o event were also associated with P. multocida abundance. Our results indicate that wetland water conditions are not strongly associated with the risk of avian cholera outbreaks; however, some variables may play a role in the abundance of P. multocida bacteria and might be important in reducing the severity of avian cholera outbreaks.

  14. Purification and characterization of protein H, the major porin of Pasteurella multocida.

    PubMed Central

    Chevalier, G; Duclohier, H; Thomas, D; Shechter, E; Wróblewski, H

    1993-01-01

    Protein H (B. Lugtenberg, R. van Boxtel, D. Evenberg, M. de Jong, P. Storm, and J. Frik, Infect. Immun. 52:175-182, 1986) is the major polypeptide of the outer membrane of Pasteurella multocida, a bacterium pathogenic for humans and animals. We have purified this protein to homogeneity by size exclusion chromatography after selective extraction with surfactants and demonstrated its pore-forming ability after reincorporation into planar lipid bilayers. In these experiments, the current through the pores was a linear function of the applied voltage in the range of -50 to +50 mV. Voltages beyond +/- 50 mV tended to partially close the channels, giving rise to apparent negative resistances. These observations suggest that protein H channels are probably not voltage regulated in vivo. With the patch clamp technique, single-channel conductance fluctuations of 0.33 nS were recorded in 1 M KCl. Electrophoretic and circular dichroism analyses showed that protein H forms homotrimers stable in sodium dodecyl sulfate at room temperature, with a high content of beta-sheet secondary structure. Upon boiling, the trimers were fully dissociated into monomers with an increase of alpha helix and irregular structure, at the expense of beta sheets. The apparent molecular mass of fully denatured monomers ranged between 37 and 41.8 kDa, depending on the electrophoretic system used for analysis. The trimeric arrangement of protein H was confirmed by image analysis of negatively stained, two-dimensional crystal arrays. This morphological study revealed, in agreement with electrophoretical data, a trimeric structure with an overall diameter of 7.7 nm. Each monomer appeared to contain a pore with an average diameter of 1 nm. Quantitative comparisons revealed that the amino acid composition (hydropathy index of -0.40) and the N-terminal sequence (determined over 36 residues) of protein H are similar to those of bacterial general porins, notably porin P2 of Haemophilus influenzae. We conclude

  15. Florfenicol As a Modulator Enhancing Antimicrobial Activity: Example Using Combination with Thiamphenicol against Pasteurella multocida

    PubMed Central

    Wei, Chia-Fong; Shien, Jui-Hung; Chang, Shao-Kuang; Chou, Chi-Chung

    2016-01-01

    Synergistic effects between the same class of antibiotics are rarely reported. Our previous study found synergistic-like interaction between florfenicol (FFC) and thiamphenicol (TAP) against Staphylococcus aureus. Here, the enhanced antimicrobial activity was evaluated in 97 clinical isolates of both Gram-negative and Gram-positive bacteria. Susceptible strains were initially identified by checkerboard microdilution assay (fractional inhibitory concentration index [FICI] ≤ 0.625), followed by confirmation of synergism using the time-kill methodology (≥2 log10 CFU/ml reduction). In all, 43% of Pasteurella multocida tested were susceptible to the enhanced bactericidal effect. In chicken fowl cholera models, FFC and TAP combination at much lower dosage that is correspondent to their MIC deduction provided maximum protection in vivo. Furthermore, synergistic combination of FFC with oxytetracycline (OTC) against Pseudomonas aeruginosa in vitro was also demonstrated. Based on the enhanced uptake of TAP and OTC, FFC presumably elicits enhanced antimicrobial activity in an orderly manner through alteration of bacterial membrane permeability or efflux systems and subsequent increase of intracellular concentration of the antibiotics used in combination. Results of ethidium bromide accumulation assay and RNA-seq showed little evidence for the involvement of efflux pumps in the synergy but further investigation is required. This study suggests the potentiality of a novel combination regimen involving FFC as an initiating modulator effective against both Gram-positive and Gram-negative bacteria depending on the antibiotics that are combined. The observed improvement of bacteriostatic effect to bactericidal, and the extended effectiveness against FFC-resistant bacterial strains warrant further studies. PMID:27065961

  16. Characterization of the membrane-targeting C1 domain in Pasteurella multocida toxin.

    PubMed

    Kamitani, Shigeki; Kitadokoro, Kengo; Miyazawa, Masayuki; Toshima, Hirono; Fukui, Aya; Abe, Hiroyuki; Miyake, Masami; Horiguchi, Yasuhiko

    2010-08-13

    Pasteurella multocida toxin (PMT) is a virulence factor responsible for the pathogenesis of some forms of pasteurellosis. The toxin activates G(q)- and G(12/13)-dependent pathways through the deamidation of a glutamine residue in the alpha-subunit of heterotrimeric GTPases. We recently reported the crystal structure of the C terminus (residues 575-1285) of PMT (C-PMT), which is composed of three domains (C1, C2, and C3), and that the C1 domain is involved in the localization of C-PMT to the plasma membrane, and the C3 domain possesses a cysteine protease-like catalytic triad. In this study, we analyzed the membrane-targeting function of the C1 domain in detail. The C1 domain consists of seven helices of which the first four (residues 590-670), showing structural similarity to the N terminus of Clostridium difficile toxin B, were found to be involved in the recruitment of C-PMT to the plasma membrane. C-PMT lacking these helices (C-PMT DeltaC1(4H)) neither localized to the plasma membrane nor stimulated the G(q/12/13)-dependent signaling pathways. When the membrane-targeting property was complemented by a peptide tag with an N-myristoylation motif, C-PMT DeltaC1(4H) recovered the PMT activity. Direct binding between the C1 domain and liposomes containing phospholipids was evidenced by surface plasmon resonance analyses. These results indicate that the C1 domain of C-PMT functions as a targeting signal for the plasma membrane.

  17. Clinico-pathology, hematology, and biochemistry responses toward Pasteurella multocida Type B: 2 via oral and subcutaneous route of infections

    PubMed Central

    Chung, Eric Lim Teik; Abdullah, Faez Firdaus Jesse; Adamu, Lawan; Marza, Ali Dhiaa; Ibrahim, Hayder Hamzah; Zamri-Saad, Mohd; Haron, Abdul Wahid; Saharee, Abdul Aziz; Lila, Mohd Azmi Mohd; Omar, Abdul Rahman; Bakar, Md Zuki Abu; Norsidin, Mohd Jefri

    2015-01-01

    Background: Pasteurella multocida a Gram-negative bacterium has been identified as the causative agent of many economically important diseases in a wide range of hosts. Hemorrhagic septicemia is a disease caused by P. multocida serotype B:2 and E:2. The organism causes acute, a highly fatal septicemic disease with high morbidity and mortality in cattle and more susceptible in buffaloes. Therefore, the aim of this study was to investigate the clinical signs, blood parameters, post mortem and histopathology changes caused by P. multocida Type B:2 infections initiated through the oral and subcutaneous routes. Methods: Nine buffalo heifers were divided equally into 3 treatment groups. Group 1 was inoculated orally with 10 ml of phosphate buffer saline; Groups 2 and 3 were inoculated with 10 ml of 1012 colony forming unit of P. multocida Type B:2 subcutaneously and orally respectively. Results: There was a significant difference (p<0.05) in temperature between the subcutaneous and the control group. The results revealed significant differences (p<0.05) in erythrocytes, hemoglobin, packed cell volume, leukocytes, monocytes, and A: G ratio between the subcutaneous and the control group. Furthermore, there were significant differences (p<0.05) in leukocytes, band neutrophils, segmented neutrophils, lymphocytes, eosinophils, basophils, thrombocytes, plasma protein, icterus index, gamma glutamyl tranferase and A: G ratio between the oral and the control group. The post mortem lesions of the subcutaneous group buffaloes showed generalized hyperemia, congestion and hemorrhage of the immune organs, gastro-intestinal tract organs and vital organs. The oral group buffaloes showed mild lesions in the lung and liver. Histologically, there were significant differences (p<0.05) in hemorrhage and congestion; necrosis and degeneration; inflammatory cells infiltration; and edema in between the groups. Conclusion: This study was a proof that oral route infection of P. multocida Type B:2

  18. Evaluation of immunopathologic effects of aqueous extract of Echinacea purpurea in mice after experimental challenge with Pasteurella multocida serotype A

    PubMed Central

    Rezaie, A; Gharibi, D; Ghorbanpoor, M; Anbari, S; Pourmahdi Broojeni, M

    2014-01-01

    In order to assess the immunopathological effects of aqueous Echinacea purpurea extract (EPE) on mice experimentally challenged with Pasteurella multocida serotype A, forty female BALB/c mice were randomly divided into four groups. The groups included a control group (received sterile distilled water 2 times/week for 2 weeks, intraperitoneally and then 100 µl sterile saline intranasally), a PMA group (received sterile distilled water as the control group and after 2 weeks, 5.6 × 103 CFU/ml of P. multocida serotype A, intranasally), an EPE+PMA group (received E. purpurea extract intraperitoneally 2 times/week for 2 weeks and then challenged as the PMA group) and an EPE group (received E. purpurea extract as EPE+PMA group and then 100 µl sterile saline intranasally). After 24 and 48 h post challenge, half of the animals in each group were sacrificed and analyzed for bacterial counts in their lungs and livers, TNFα serum levels and histapathological changes. The results showed significant differences in lung bacterial counts between PMA and EPE+PMA groups. TNFα serum level was significantly higher in the PMA group. Histopathological examination revealed infiltration of neutrophils in alveolar septa and hyperemia in the PMA group. In addition, the criteria of bronchopneumonia were partially recovered in the EPE+PMA compared to the PMA group. According to the results, it seems that E. purpurea extract has an immunomodulatory effect and can be used to prevent or control of pneumonia caused by Pasteurella. PMID:27175135

  19. Involvement of the nervous system following experimental infection with Pasteurella multocida B:2 in buffalo (Bubalus bubalis): A clinicopathological study.

    PubMed

    Marza, Ali Dhiaa; Jesse, Faez Firdaus Abdullah; Ahmed, Ihsan Muneer; Chung, Eric Lim Teik; Ibrahim, Hayder Hamzah; Zamri-Saad, Mohd; Omar, Abdul Rahman; Abu Bakar, Md Zuki; Saharee, Abdul Aziz; Haron, Abdul Wahid; Alwan, Mohammed Jwaid; Lila, Mohd Azmi Mohd

    2016-04-01

    Haemorrhagic septicaemia (HS) is an acute, fatal, septicaemic disease of cattle and buffaloes caused by one of two specific serotypes of Pasteurella multocida B:2 and E:2 in Asian and African, respectively. It is well known that HS affect mainly the respiratory and digestive tracts. However, involvement of the nervous system in pathogenesis of HS has been reported in previous studies without details. In this study, nine buffalo calves of 8 months old were distributed into three groups. Animals of Group 1 and 2 were inoculated orally and subcutaneously with 10 ml of 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while animals of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. All calves in Group 1 and Group 3 were euthanised after 504 h (21 day) post-infection, while calves in Group 2 had to euthanise after 12 h post-infection as they develop sever clinical signs of HS. Significant differences were found in Group 2 in the mean scores of clinical signs, gross and histopathological changes which mainly affect different anatomic regions of the nervous system. In addition, successful bacterial isolation of P. multocida B:2 were obtained from different sites of the nervous system. On the other hand, less sever, clinical, gross and histopathological changes were found in Group 1. These results provide for the first time strong evidence of involving of the nervous system in pathogenesis of HS, especially in the peracute stage of the disease.

  20. Phenotypic and genotypic characters of isolates of Pasteurella multocida obtained from back-yard poultry and from two outbreaks of avian cholera in avifauna in Denmark.

    PubMed

    Christensen, J P; Dietz, H H; Bisgaard, M

    1998-01-01

    Two outbreaks of fowl cholera in the avifauna in Denmark, affecting primarily eiders but also cormorants, gulls and oyster-catchers were shown to be caused by the same clone of Pasteurella multocida ssp. multocida by restriction enzyme analysis (REA) and ribotyping, using the enzymes HpaII and HhaI and phenotypic characterization. This observation indicated spread by migratory birds. It was shown that the outbreak clone was closely related to isolates of Pasteurella multocida ssp. multocida obtained from back-yard poultry in Denmark, including chickens, pheasants, turkeys and ducks. The only detectable difference between the outbreak clone and some of these strains concerned the size of one fragment. These results indicate a possible exchange of P. multocida ssp. multocida between populations of wild birds and back-yard poultry. Among the DNA fingerprinting methods used, restriction enzyme analysis offered the highest discrimination among thirty strains obtained from back-yard poultry. The restriction enzymes HpaII and HhaI generated almost the same number of profile types, 17 and 15 respectively, but only HpaII differentiated the outbreak clone from the group of closely related strains isolated from back-yard poultry. Ribotyping, using the same enzymes, resulted in 12 and 10 different profile types, respectively. The outbreak isolates did not harbour any plasmids, while six out of the 30 strains originating from back-yard poultry (20%) carried a cryptic plasmid of approximately 3.4 kb.

  1. Pharmacokinetic/pharmacodynamic integration and modelling of amoxicillin for the calf pathogens Mannheimia haemolytica and Pasteurella multocida.

    PubMed

    Lees, P; Pelligand, L; Illambas, J; Potter, T; Lacroix, M; Rycroft, A; Toutain, P-L

    2015-10-01

    The antimicrobial properties of amoxicillin were determined for the bovine respiratory tract pathogens, Mannheima haemolytica and Pasteurella multocida. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and time-kill curves were established. Pharmacokinetic (PK)/pharmacodynamic (PD) modelling of the time-kill data, based on the sigmoidal Emax equation, generated parameters for three levels of efficacy, namely bacteriostatic, bactericidal (3log10 reduction) and 4log10 reduction in bacterial counts. For these levels, mean AUC(0-24 h) /MIC serum values for M. haemolytica were 29.1, 57.3 and 71.5 h, respectively, and corresponding values for P. multocida were 28.1, 44.9 and 59.5 h. Amoxicillin PK was determined in calf serum, inflamed (exudate) and noninflamed (transudate) tissue cage fluids, after intramuscular administration of a depot formulation at a dosage of 15 mg/kg. Mean residence times were 16.5 (serum), 29.6 (exudate) and 29.0 h (transudate). Based on serum MICs, integration of in vivo PK and in vitro PD data established maximum concentration (Cmax )/MIC ratios of 13.9:1 and 25.2:1, area under concentration-time curve (AUC0-∞ )/MIC ratios of 179 and 325 h and T>MIC of 40.3 and 57.6 h for P. multocida and M. haemolytica, respectively. Monte Carlo simulations for a 90% target attainment rate predicted single dose to achieve bacteriostatic and bactericidal actions over 48 h of 17.7 and 28.3 mg/kg (M. haemolytica) and 17.7 and 34.9 mg/kg (P. multocida).

  2. Pasteurella multocida non-native joint infection after a dog lick: A case report describing a complicated two-stage revision and a comprehensive review of the literature.

    PubMed

    Lam, Philip W; Page, Andrea V

    2015-01-01

    Prosthetic joint infections (PJIs) are commonly caused by pathogens such as Staphylococcus aureus and coagulase-negative staphylococci; however, other microbial etiologies and specific risk factors are increasingly recognized. Pasteurella multocida is a Gram-negative coccobacillus that is part of the normal oral flora in many animals, and is particularly common in dogs and cats. PJIs caused by P multocida have been reported only rarely in the literature and typically occur in the context of an animal bite or scratch. The present article describes a P multocida joint infection that occurred after a dog lick and complicated a two-stage revision arthroplasty. A comprehensive review of the literature regarding P multocida PJIs follows.

  3. Pasteurella multocida non-native joint infection after a dog lick: A case report describing a complicated two-stage revision and a comprehensive review of the literature

    PubMed Central

    Philip W, Lam; Page, Andrea V

    2015-01-01

    Prosthetic joint infections (PJIs) are commonly caused by pathogens such as Staphylococcus aureus and coagulase-negative staphylococci; however, other microbial etiologies and specific risk factors are increasingly recognized. Pasteurella multocida is a Gram-negative coccobacillus that is part of the normal oral flora in many animals, and is particularly common in dogs and cats. PJIs caused by P multocida have been reported only rarely in the literature and typically occur in the context of an animal bite or scratch. The present article describes a P multocida joint infection that occurred after a dog lick and complicated a two-stage revision arthroplasty. A comprehensive review of the literature regarding P multocida PJIs follows. PMID:26361490

  4. Mucoadhesive and pH-sensitive thiolated Eudragit microspheres for oral delivery of Pasteurella multocida antigens containing dermonecrotoxin.

    PubMed

    Islam, Mohammad Ariful; Jiang, Hu-Lin; Quan, Ji-Shan; Arote, Rohidas B; Kang, Mi-Lan; Yoo, Han-Sang; Yun, Cheol-Heui; Choi, Yun-Jaie; Cho, Chong-Su

    2011-05-01

    In this study, cysteine was conjugated to the Eudragit to have mucoadhesive and pH-sensitive properties. Pasteurella multocida dermonecrotoxin (PMT) is a major virulence factor as a causative agent of atrophic rhinitis (AR) in swine and, therefore, inactivated P. multocida was used as a candidate vaccine in the current study. PMT-loaded thiolated Eudragit microspheres (TEMS) prepared using W/O/W emulsion-solvent evaporation method were characterized to assess their efficacy in oral vaccination. PMT-loaded TEMS were observed as spherical shapes with smooth surfaces and average particle sizes were 5.2 +/- 0.55 microm. The loading efficiency of PMT in the TEMS was about 75.3%. A significantly higher percentage of PMT from PMT-loaded TEMS was released at pH 7.4 than at pH 1.5. Murine macrophage stimulated with PMT-loaded TEMS facilitated a gradual secretion of tumor necrosis factor-alpha and nitric oxide as immune stimulatory mediators in a time dependent manner, suggesting that the released PMT from PMT-loaded TEMS had immune stimulating activity of AR vaccine in vitro.

  5. A Rare Case of Glossitis due to Pasteurella multocida after a Cat Scratch

    PubMed Central

    Doan, Thien; Revere, Elizabeth

    2016-01-01

    Pasteurella is one of the zoonotic pathogens that can cause variety of serious infections in animals and humans such as bacteremia, septic shock, endocarditis, meningitis, prosthetic and native valve infections, osteomyelitis, skin and soft tissue infections, abscesses, and even pneumonia with empyema. However, there have been few reports of upper respiratory involvements like tonsillitis and epiglottitis in humans. We present a case of recurrent Pasteurella glossitis after a cat scratch which has not been reported in humans. PMID:27840749

  6. A Rare Case of Glossitis due to Pasteurella multocida after a Cat Scratch.

    PubMed

    Niknam, Negin; Doan, Thien; Revere, Elizabeth

    2016-01-01

    Pasteurella is one of the zoonotic pathogens that can cause variety of serious infections in animals and humans such as bacteremia, septic shock, endocarditis, meningitis, prosthetic and native valve infections, osteomyelitis, skin and soft tissue infections, abscesses, and even pneumonia with empyema. However, there have been few reports of upper respiratory involvements like tonsillitis and epiglottitis in humans. We present a case of recurrent Pasteurella glossitis after a cat scratch which has not been reported in humans.

  7. Cloning of the gene and characterization of the enzymatic properties of the monomeric alkaline phosphatase (PhoX) from Pasteurella multocida strain X-73.

    PubMed

    Wu, Jin-Ru; Shien, Jui-Hung; Shieh, Happy K; Hu, Chung-Chi; Gong, Shuen-Rong; Chen, Ling-Yun; Chang, Poa-Chun

    2007-02-01

    We have identified a new phoX gene encoding the monomeric alkaline phosphatase from Pasteurella multocida X-73. This gene was not found in the published genome sequence of Pasteurella multocida pm70. Characterization of the recombinant PhoX of Pasteurella multocida X-73 showed that it is a monomeric enzyme, activated by Ca(2+) and possibly secreted by the Tat pathway. These features distinguish phosphatases of the PhoX family from those of the PhoA family. All proteins of the PhoX family were found to contain a conserved motif that shares significant sequence homology with the calcium-binding site of a phosphotriesterase known as diisopropylfluorophosphatase. Site-directed mutagenesis revealed that D527 of PhoX might be the ligand bound to the catalytic calcium. This is the first report on identification of homologous sequences between PhoX and the phosphotriesterase and on the potential calcium-binding site of PhoX.

  8. Pasteurella multocida infection of a total knee arthroplasty after a "dog lick".

    PubMed

    Heym, B; Jouve, F; Lemoal, M; Veil-Picard, A; Lortat-Jacob, A; Nicolas-Chanoine, M H

    2006-10-01

    The patient we report here underwent a total knee arthroplasty (TKA) which got infected with P. multocida after her dog had licked a small wound at the third toe of the same foot. Despite a correct treatment comprising synovectomy and cleansing, and an active antibiotic treatment for 3 months, the patient was readmitted for persistent infection of the same knee 2 weeks after the end of the antibiotic treatment. Sampling during surgery allowed for the growth of a P. multocida isolate proven by a molecular method to be identical to the previously isolated strain. This recurrent P. multocida infection was treated by a two-step change of the TKA comprising a 2-month period of antibiotic treatment between the two surgical interventions.

  9. Role of outer membrane protein H (OmpH)- and OmpA-specific monoclonal antibodies from hybridoma tumors in protection of mice against Pasteurella multocida.

    PubMed Central

    Vasfi Marandi, M; Mittal, K R

    1997-01-01

    Two major outer membrane proteins of Pasteurella multocida, designated OmpH and OmpA, were characterized and shown to be related to the families of porin and heat-modifiable proteins, respectively. The backpack hybridoma tumor system in BALB/c mice was used to continuously deliver immunoglobulin G2b (IgG2b) monoclonal antibodies (MAbs) specific for OmpH (MAb MT1) and OmpA (MAb MT4.1). MAbs were detected in serum and peritoneal lavage samples of mice bearing hybridoma tumors by an enzyme-linked immunosorbent assay and an immunoblot assay. Highly significant protection was observed in mice bearing MT1 hybridoma tumors against both intraperitoneal and intranasal challenge infections with homologous nontoxigenic P. multocida strains possessing MAb MT1-reacting epitopes, whereas the mice bearing MT4.1 hybridoma tumors were not protected. The numbers of P. multocida organisms in the lungs of mice bearing MT1 hybridoma tumors were significantly less than those in lungs of mice bearing MT4.1 hybridoma tumors at 48 h postchallenge. These results indicate that the OmpH-specific MAb inhibited proliferation of P. multocida in the lungs. MAb MT1 was unable to kill P. multocida in vitro in the presence of complement. However, an enhanced phagocytosis by polymorphonuclear cells (PMNs) was observed in mice bearing MT1 hybridoma tumors. P. multocida induced a more extensive and rapid influx of PMNs into the peritoneal cavity of mice bearing MT1 hybridoma tumors than of mice bearing MT4.1 hybridoma tumors. The results of this study demonstrate for the first time that IgG MAbs against OmpH of P. multocida are involved in the protection of mice against lethal challenge infection by means of opsonization and inhibition of proliferation of P. multocida as a result of increased influx of PMNs into the infection site. PMID:9353026

  10. Development of a Rapid Multiplex PCR Assay To Genotype Pasteurella multocida Strains by Use of the Lipopolysaccharide Outer Core Biosynthesis Locus

    PubMed Central

    Harper, Marina; John, Marietta; Turni, Conny; Edmunds, Mark; St. Michael, Frank; Adler, Ben; Blackall, P. J.; Cox, Andrew D.

    2014-01-01

    Pasteurella multocida is a Gram-negative bacterial pathogen that is the causative agent of a wide range of diseases in many animal species, including humans. A widely used method for differentiation of P. multocida strains involves the Heddleston serotyping scheme. This scheme was developed in the early 1970s and classifies P. multocida strains into 16 somatic or lipopolysaccharide (LPS) serovars using an agar gel diffusion precipitin test. However, this gel diffusion assay is problematic, with difficulties reported in accuracy, reproducibility, and the sourcing of quality serovar-specific antisera. Using our knowledge of the genetics of LPS biosynthesis in P. multocida, we have developed a multiplex PCR (mPCR) that is able to differentiate strains based on the genetic organization of the LPS outer core biosynthesis loci. The accuracy of the LPS-mPCR was compared with classical Heddleston serotyping using LPS compositional data as the “gold standard.” The LPS-mPCR correctly typed 57 of 58 isolates; Heddleston serotyping was able to correctly and unambiguously type only 20 of the 58 isolates. We conclude that our LPS-mPCR is a highly accurate LPS genotyping method that should replace the Heddleston serotyping scheme for the classification of P. multocida strains. PMID:25428149

  11. Structural basis for substrate specificity and mechanism of N-acetyl-D-neuraminic acid lyase from Pasteurella multocida#

    PubMed Central

    Huynh, Nhung; Aye, Aye; Li, Yanhong; Yu, Hai; Cao, Hongzhi; Tiwari, Vinod Kumar; Shin, Don-Wook; Chen, Xi; Fisher, Andrew J.

    2013-01-01

    N -Acetylneuraminate lyases (NALs) or sialic acid aldolases catalyze the reversible aldol cleavage of N-acetylneuraminic acid (Neu5Ac, the most common form of sialic acid) to form pyruvate and N-acetyl-D-mannosamine (ManNAc). Although equilibrium favors sialic acid cleavage, these enzymes can be used for high-yield chemoenzymatic synthesis of structurally diverse sialic acids in the presence of excess pyruvate. Engineering these enzymes to synthesize structurally modified natural sialic acids and their non-natural derivatives holds promise in creating novel therapeutic agents. Atomic resolution structures of these enzymes will greatly assist in guiding mutagenic and modeling studies to engineer enzymes with altered substrate specificity. We report here the crystal structures of wild-type Pasteurella multocida N-acetylneuraminate lyase and its K164A mutant. Like other bacterial lyases, it assembles into a homotetramer with each monomer folding into a classic (β/α)8 TIM barrel. Two wild-type structures were determined; in the absence of substrates, and trapped in a Schiff base intermediate between Lys164 and pyruvate, respectively. Three structures of the K164A variant were determined: one in the absence of substrates and two binary complexes with N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), respectively. Both sialic acids bind to the active site in the open-chain ketone form of the monosaccharide. The structures reveal that every hydroxyl group of the linear sugars makes hydrogen bond interactions with the enzyme and the residues that determine specificity were identified. Additionally, the structures lend some clues in explaining the natural discrimination of sialic acid substrates between the P. multocida and E. coli NALs. PMID:24152047

  12. A semi-multifunctional sialyltransferase from Bibersteinia trehalosi and its comparison to the Pasteurella multocida ST1 mutants.

    PubMed

    Talafová, Klaudia; Hrabárová, Eva; Nahálka, Jozef

    2015-12-20

    Sialic acids are well known for their crucial roles in many physiological and pathological processes. Improvement in the efficacy of protein drugs, an increase in the anti-inflammatory activity of intravenous immunoglobulin, preparation of infant milk and the diagnosis of diseases are examples of why there is a need for efficient in vitro sialylation. Sialyltransferases are crucial enzymes for the synthesis of sialo-oligosaccharides. Here, we introduce a new α2,3-sialyltransferase from bacteria Bibersteinia trehalosi (BtST1), which is homological to sialyltransferase from Pasteurella multocida (PmST1), Pasteurella dagmatis (PdST1) and Haemophilus ducreyi (Hd0053). BtST1 is active in a wide pH range and shows considerable acceptor flexibility. Very good specific activities have been detected with lactose and LacNAc as acceptors, and these activities were comparable to those of efficient multifunctional PmST1 and higher than PdST1, Hd0053 and also PmST1 M144D which was constructed to decrease the high sialidase activity of PmST1. Testing of PmST1 mutant forms revealed that mutations that included S143 caused only the restriction of sialyltransferase activity, whereas mutations including G142 resulted in the loss of activity with lactose. BtST1 possesses only low sialidase and trans-sialidase activities that are comparable to mutant PmST1 M144D, which are detected only in the presence of CMP. The combination of large acceptor flexibility, high activity for lactose and LacNAc and naturally low sialidase activity make BtST1 an attractive enzyme for biotechnological applications.

  13. Immunogenicity and efficacy of three recombinant subunit Pasteurella multocida toxin vaccines against progressive atrophic rhinitis in pigs

    USGS Publications Warehouse

    Liao, Chih-Ming; Huang, Chienjin; Hsuan, Shih-Ling; Chen, Zeng-Weng; Lee, Wei-Cheng; Liu, Cheng-I; Winton, James R.; Chien, Maw-Sheng

    2006-01-01

    Three short fragments of recombinant subunit Pasteurella multocida toxin (rsPMT) were constructed for evaluation as candidate vaccines against progressive atrophic rhinitis (PAR) of swine. PMT-specific antibody secreting cells and evidence of cellular immunity were detected in rsPMT-immunized pigs following authentic PMT challenge or homologous antigen booster. Piglets immunized with rsPMT fragments containing either the N-terminal or the C-terminal portions of PMT developed high titers of neutralizing antibodies. Pregnant sows immunized with rsPMT had higher levels of maternal antibodies in their colostrum than did those immunized with a conventional PAR-toxoid vaccine. Offspring from rsPMT vaccinated sows had better survival after challenge with a five-fold lethal dose of authentic PMT and had better growth performance after challenge with a sublethal dose of toxin. Our findings indicate these non-toxic rsPMT proteins are attractive candidates for development of a subunit vaccine against PAR in pigs.

  14. Pharmacodynamics of amoxicillin against Mannheimia haemolytica and Pasteurella multocida and pharmacokinetic/pharmacodynamic (PK/PD) correlation in sheep.

    PubMed

    Delis, G A; Koutsoviti-Papadopoulou, M; Siarkou, V I; Kounenis, G; Batzias, G C

    2010-12-01

    Silicone-made tissue cages were implanted in sheep. Blood serum (SBS) and tissue cage fluid (TCF) samples were collected after amoxicillin intravenous and intramuscular administrations, at the dose of 15 mg/kg. Amoxicillin pharmacodynamics were studied in an artificial culture medium, SBS and TCF with use of a Mannheimia haemolytica and a Pasteurella multocida strain. A concentration-independent antimicrobial activity of amoxicillin was confirmed for levels higher than 0.79-1.75×MIC. This result favored the use of the percentage of the 24 h dosing interval during which drug levels remain above MIC as the appropriate pharmacokinetic/pharmacodynamic index. The subsequent correlation revealed that intravenous administration could be considered effective against "deep" infections caused by bacteria with MICs<1 μg/mL or "shallow" infections caused by bacteria with MICs<0.1 μg/mL. Intramuscular administration could be safely considered effective against both "deep" and "shallow" infections when the MICs of the targeted pathogens are lower than 1 μg/mL.

  15. Auranofin Inhibits the Enzyme Activity of Pasteurella multocida Toxin PMT in Human Cells and Protects Cells from Intoxication

    PubMed Central

    Carle, Stefan; Brink, Thorsten; Orth, Joachim H. C.; Aktories, Klaus; Barth, Holger

    2017-01-01

    The AB-type protein toxin from Pasteurella multocida (PMT) contains a functionally important disulfide bond within its catalytic domain, which must be cleaved in the host cell cytosol to render the catalytic domain of PMT into its active conformation. Here, we found that the reductive potential of the cytosol of target cells, and more specifically, the activity of the thioredoxin reductase (TrxR) is crucial for this process. This was demonstrated by the strong inhibitory effect of the pharmacological TrxR inhibitor auranofin, which inhibited the intoxication of target cells with PMT, as determined by analyzing the PMT-catalyzed deamidation of GTP-binding proteins (G-proteins) in the cytosol of cells. The amount of endogenous substrate levels modified by PMT in cells pretreated with auranofin was reduced compared to cells treated with PMT alone. Auranofin had no inhibitory effect on the activity of the catalytic domain of constitutively active PMT in vitro, demonstrating that auranofin did not directly inhibit PMT activity, but interferes with the mode of action of PMT in cells. In conclusion, the results show that TrxR is crucial for the mode of action of PMT in mammalian cells, and that the drug auranofin can serve as an efficient inhibitor, which might be a starting point for novel therapeutic options against toxin-associated diseases. PMID:28098782

  16. Modification of heterotrimeric G-proteins in Swiss 3T3 cells stimulated with Pasteurella multocida toxin.

    PubMed

    Babb, Rebecca C; Homer, Karen A; Robbins, Jon; Lax, Alistair J

    2012-01-01

    Many bacterial toxins covalently modify components of eukaryotic signalling pathways in a highly specific manner, and can be used as powerful tools to decipher the function of their molecular target(s). The Pasteurella multocida toxin (PMT) mediates its cellular effects through the activation of members of three of the four heterotrimeric G-protein families, G(q), G(12) and G(i). PMT has been shown by others to lead to the deamidation of recombinant Gα(i) at Gln-205 to inhibit its intrinsic GTPase activity. We have investigated modification of native Gα subunits mediated by PMT in Swiss 3T3 cells using 2-D gel electrophoresis and antibody detection. An acidic change in the isoelectric point was observed for the Gα subunit of the G(q) and G(i) families following PMT treatment of Swiss 3T3 cells, which is consistent with the deamidation of these Gα subunits. Surprisingly, PMT also induced a similar modification of Gα(11), a member of the G(q) family of G-proteins that is not activated by PMT. Furthermore, an alkaline change in the isoelectric point of Gα(13) was observed following PMT treatment of cells, suggesting differential modification of this Gα subunit by PMT. G(s) was not affected by PMT treatment. Prolonged treatment with PMT led to a reduction in membrane-associated Gα(i), but not Gα(q). We also show that PMT inhibits the GTPase activity of G(q).

  17. Enzymatic actions of Pasteurella multocida toxin detected by monoclonal antibodies recognizing the deamidated α subunit of the heterotrimeric GTPase Gq.

    PubMed

    Kamitani, Shigeki; Ao, Shinpei; Toshima, Hirono; Tachibana, Taro; Hashimoto, Makiko; Kitadokoro, Kengo; Fukui-Miyazaki, Aya; Abe, Hiroyuki; Horiguchi, Yasuhiko

    2011-08-01

    Pasteurella multocida toxin (PMT) is a virulence factor responsible for the pathogenesis of some Pasteurellosis. PMT exerts its toxic effects through the activation of heterotrimeric GTPase (G(q), G(12/13) and G(i))-dependent pathways, by deamidating a glutamine residue in the α subunit of these GTPases. However, the enzymatic characteristics of PMT are yet to be analyzed in detail because the deamidation has only been observed in cell-based assays. In the present study, we developed rat monoclonal antibodies, specifically recognizing the deamidated Gα(q), to detect the actions of PMT by immunological techniques such as western blotting. Using the monoclonal antibodies, we found that the toxin deamidated Gα(q) only under reducing conditions. The C-terminal region of PMT, C-PMT, was more active than the full-length PMT. The C3 domain possessing the enzyme core catalyzed the deamidation in vitro without any other domains. These results not only support previous observations on toxicity, but also provide insights into the enzymatic nature of PMT. In addition, we present several lines of evidence that Gα(11), as well as Gα(q), could be a substrate for PMT.

  18. Auranofin Inhibits the Enzyme Activity of Pasteurella multocida Toxin PMT in Human Cells and Protects Cells from Intoxication.

    PubMed

    Carle, Stefan; Brink, Thorsten; Orth, Joachim H C; Aktories, Klaus; Barth, Holger

    2017-01-13

    The AB-type protein toxin from Pasteurella multocida (PMT) contains a functionally important disulfide bond within its catalytic domain, which must be cleaved in the host cell cytosol to render the catalytic domain of PMT into its active conformation. Here, we found that the reductive potential of the cytosol of target cells, and more specifically, the activity of the thioredoxin reductase (TrxR) is crucial for this process. This was demonstrated by the strong inhibitory effect of the pharmacological TrxR inhibitor auranofin, which inhibited the intoxication of target cells with PMT, as determined by analyzing the PMT-catalyzed deamidation of GTP-binding proteins (G-proteins) in the cytosol of cells. The amount of endogenous substrate levels modified by PMT in cells pretreated with auranofin was reduced compared to cells treated with PMT alone. Auranofin had no inhibitory effect on the activity of the catalytic domain of constitutively active PMT in vitro, demonstrating that auranofin did not directly inhibit PMT activity, but interferes with the mode of action of PMT in cells. In conclusion, the results show that TrxR is crucial for the mode of action of PMT in mammalian cells, and that the drug auranofin can serve as an efficient inhibitor, which might be a starting point for novel therapeutic options against toxin-associated diseases.

  19. Quantitative resistance level (MIC) of Pasteurella multocida isolated from pigs between 2004 and 2006: national resistance monitoring by the BVL.

    PubMed

    Kaspar, Heike; Schröer, Ulrike; Wallmann, Jürgen

    2007-01-01

    The National Resistance Monitoring of the Federal Office of Consumer Protection and Food Safety (BVL) is to determine the prevalence of resistance of bacterial pathogens from animals using a valid database. From 2004 to 2006, a total of 1,472 Pasteurella multocida strains isolated from pigs with acute respiratory tract diseases was submitted to the BVL and examined. Of these, 1,11 (75.5 %) were included in the study and tested using 24 different antimicrobial substances. The results showed that the resistance level is generally low, with the exception of the substances tetracycline, trimethoprim, and the combination trimethoprim/sulfamethoxazole. It also became clear that resistance data need to be evaluated separately for each of the animal production categories, so that a realistic figure of the current resistance level can be presented. This knowledge provides information about the resistance situation in Germany, and helps deduce the necessary management measures that must be taken to minimize resistance to antibiotics. Furthermore, it provides valuable information that can form the basis for empirical therapy, so that the National Resistance Monitoring makes an important contribution to the safety of food derived from animals and consequently aids the improvement of consumer protection.

  20. Application of intact cell-based NFAT-β-lactamase reporter assay for Pasteurella multocida toxin-mediated activation of calcium signaling pathway

    PubMed Central

    Luo, Shuhong; Ho, Mengfei; Wilson, Brenda A.

    2009-01-01

    Pasteurella multocida toxin (PMT) stimulates and subsequently uncouples phospholipase C β1 (PLCβ1) signal transduction through its selective action on the alpha subunit of the Gq protein. Here, we describe the application of an NFAT-β-lactamase reporter assay as a functional readout for PMT-induced activation of the Gq-protein-coupled PLCβ1-IP3-Ca2+ signaling pathway. Use of the NFAT-β-lactamase reporter assay with a cell-permeable fluorogenic substrate provides high sensitivity due to the absence of endogenous β-lactamase activity in mammalian cells. This assay system was optimized for cell density, dose and time exposure of PMT stimulation. It is suited for quantitative characterization of PMT activity in mammalian cells and for use as a high-throughput screening method for PMT deletion and point mutants suitable for vaccine development. This method has application for diagnostic screening of clinical isolates of toxinogenic P. multocida. PMID:18190943

  1. Acute infection of a total knee arthroplasty caused by Pasteurella multocida: a case report and a comprehensive review of the literature in the last 10 years.

    PubMed

    Heydemann, John; Heydemann, Jacob S; Antony, Suresh

    2010-09-01

    Total knee arthroplasty (TKA) infection are most commonly due Staphylococcus aureus followed by coagulase-negative staphylococci, and streptococci, while gram-negative rods are seldom isolated.(1,3,4) In the last 20 years, cases of Pasteurella multocida TKA and total hip arthroplasty (THA) infection resulting from cat and dog bites, scratches, or licks have been published reporting varying presentations and treatment options. Most commonly, P. multocida infected arthroplasties result in local tenderness, cellulitis, and purulent discharge followed by regional adenopathy, and in immunocompromised patients it may progress to septicemia, meningitis, and septic arthritis.(5) Treatment antibiotics include penicillins or 2nd and 3rd generation cephalosporins, and surgical options involve one-stage, or two-stage revision arthroplasties.(6,9,17,19) We report a case of P. multocida TKA infection in a patient who was treated successfully with a 3rd generation cephalosporin, synovectomy and tibial interspacer exchange, along with a review of the literature published in the last 10 years. Our findings show that there is usually a history of exposure to the animal, early appearance of cat bite related infections, and multifactorial decision making for the treatment of P. multocida joint infections.

  2. Immune responses against chimeric DNA and protein vaccines composed of plpEN-OmpH and PlpEC-OmpH from Pasteurella multocida A:3 in mice.

    PubMed

    Okay, Sezer; Ozcengiz, Erkan; Ozcengiz, Gülay

    2012-12-01

    Pasteurella multocida is a pathogenic bacterium causing many diseases that are of significant economic importance to livestock industries. Outer membrane protein H (ompH) gene and two fragments of Pasteurella lipoprotein E (plpE) gene, namely plpEN and plpEC, were cloned from P. multocida A:3. Three DNA vaccine formulations, namely pCMV-ompH, pCMV-plpEN-ompH and pCMV-plpEC-ompH and two protein-based prototype vaccines, alum adjuvanted PlpEN-OmpH and PlpEC-OmpH, were generated. Antibody levels were induced in mice vaccinated with chimeric DNA or protein vaccines. A significant (p < 0.05) increase in serum IFN-g titer was obtained by vaccination with 100 μg of pCMV-ompH, pCMV-plpEC-ompH and PlpEC-OmpH. DNA vaccines did not provide protection upon intraperitoneal challenge with 10 LD50 of live P. multocida A:3. However, 40% protection was conferred by 100 μg of PlpEC-OmpH which was not statistically significant. These results showed that plpEN-ompH and plpEC-ompH chimeric DNA vaccines and alum adjuvanted PlpEN-OmpH or PlpEC-OmpH protein vaccines were immunogenic but not protective against P. multocida A:3 in mice. Prime-boost strategies, i.e. priming with DNA vaccines and boost with protein formulations or different adjuvants can be utilized to obtain significant protection.

  3. A Pasteurella multocida sialyltransferase displaying dual trans-sialidase activities for production of 3'-sialyl and 6'-sialyl glycans.

    PubMed

    Guo, Yao; Jers, Carsten; Meyer, Anne S; Arnous, Anis; Li, Haiying; Kirpekar, Finn; Mikkelsen, Jørn D

    2014-01-20

    This study examined a recombinant Pasteurella multocida sialyltransferase exhibiting dual trans-sialidase activities. The enzyme catalyzed trans-sialylation using either 2-O-(p-nitrophenyl)-α-d-N-acetylneuraminic acid or casein glycomacropeptide (whey protein) as the sialyl donor and lactose as the acceptor, resulting in production of both 3'-sialyllactose and 6'-sialyllactose. This is the first study reporting α-2,6-trans-sialidase activity of this sialyltransferase (EC 2.4.99.1 and 2.4.99.4). A response surface design was used to evaluate the effects of three reaction parameters (pH, temperature, and lactose concentration) on enzymatic production of 3'- and 6'-sialyllactoses using 5% (w/v) casein glycomacropeptide (equivalent to 9mM bound sialic acid) as the donor. The maximum yield of 3'-sialyllactose (2.75±0.35mM) was achieved at a reaction condition with pH 6.4, 40°C, 100mM lactose after 6h; and the largest concentration of 6'-sialyllactose (3.33±0.38mM) was achieved under a condition with pH 5.4, 40°C, 100mM lactose after 8h. 6'-sialyllactose was presumably formed from α-2,3 bound sialic acid in the casein glycomacropeptide as well as from 3'-sialyllactose produced in the reaction. The kcat/Km value for the enzyme using 3'-sialyllactose as the donor for 6'-sialyllactose synthesis at pH 5.4 and 40°C was determined to be 23.22±0.7M(-1)s(-1). Moreover, the enzyme was capable of catalyzing the synthesis of both 3'- and 6'-sialylated galactooligosaccharides, when galactooligosaccharides served as acceptors.

  4. Arf6-Dependent Intracellular Trafficking of Pasteurella multocida Toxin and pH-Dependent Translocation from Late Endosomes

    PubMed Central

    Repella, Tana L.; Ho, Mengfei; Chong, Tracy P. M.; Bannai, Yuka; Wilson, Brenda A.

    2011-01-01

    The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH4Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity. PMID:22053287

  5. Pasteurella multocida toxin (PMT) upregulates CTGF which leads to mTORC1 activation in Swiss 3T3 cells.

    PubMed

    Oubrahim, Hammou; Wong, Allison; Wilson, Brenda A; Chock, P Boon

    2013-05-01

    Pasteurella multocida toxin (PMT) is a mitogenic protein that hijacks cellular signal transduction pathways via deamidation of heterotrimeric G proteins. We previously showed that rPMT activates mTOR signaling via a Gαq/11/PLCβ/PKC mediated pathway, leading in part to cell proliferation and migration. Herein, we show that mTOR and MAPK, but not membrane-associated tyrosine kinases, are activated in serum-starved 3T3 cells by an autocrine/paracrine substance(s) secreted into the conditioned medium following rPMT treatment. Surprisingly, this diffusible factor(s) is capable of activating mTOR and MAPK pathways even in MEF Gαq/11 double knockout cells. Microarray analysis identified connective tissue growth factor (CTGF) mRNA as the most upregulated gene in rPMT-treated serum-starved 3T3 cells relative to untreated cells. These results were further confirmed using RT-PCR and Western blot analyses. In accord with rPMT-induced mTOR activation, upregulation of CTGF protein was observed in WT MEF, but not in Gαq/11 double knockout MEF cells. Although CTGF expression is regulated by TGFβ, rPMT did not activate TGFβ pathway. In addition, MEK inhibitors U0126 or PD98059, but not mTOR specific inhibitors, rapamycin and Torin 1, inhibited rPMT-induced upregulation of CTGF. Importantly, CTGF overexpression in serum-starved 3T3 cells using adenovirus led to phosphorylation of ribosomal protein S6, a downstream target of mTOR. However, despite the ability of CTGF to activate the mTOR pathway, upregulation of CTGF alone could not induce morphological changes as those observed in rPMT-treated cells. Our findings reveal that CTGF plays an important role, but there are additional factors involved in the mitogenic action of PMT.

  6. Arf6-dependent intracellular trafficking of Pasteurella multocida toxin and pH-dependent translocation from late endosomes.

    PubMed

    Repella, Tana L; Ho, Mengfei; Chong, Tracy P M; Bannai, Yuka; Wilson, Brenda A

    2011-03-01

    The potent mitogenic toxin from Pasteurella multocida (PMT) is the major virulence factor associated with a number of epizootic and zoonotic diseases caused by infection with this respiratory pathogen. PMT is a glutamine-specific protein deamidase that acts on its intracellular G-protein targets to increase intracellular calcium, cytoskeletal, and mitogenic signaling. PMT enters cells through receptor-mediated endocytosis and then translocates into the cytosol through a pH-dependent process that is inhibited by NH(4)Cl or bafilomycin A1. However, the detailed mechanisms that govern cellular entry, trafficking, and translocation of PMT remain unclear. Co-localization studies described herein revealed that while PMT shares an initial entry pathway with transferrin (Tfn) and cholera toxin (CT), the trafficking pathways of Tfn, CT, and PMT subsequently diverge, as Tfn is trafficked to recycling endosomes, CT is trafficked retrograde to the ER, and PMT is trafficked to late endosomes. Our studies implicate the small regulatory GTPase Arf6 in the endocytic trafficking of PMT. Translocation of PMT from the endocytic vesicle occurs through a pH-dependent process that is also dependent on both microtubule and actin dynamics, as evidenced by inhibition of PMT activity in our SRE-based reporter assay, with nocodazole and cytochalasin D, respectively, suggesting that membrane translocation and cytotoxicity of PMT is dependent on its transfer to late endosomal compartments. In contrast, disruption of Golgi-ER trafficking with brefeldin A increased PMT activity, suggesting that inhibiting PMT trafficking to non-productive compartments that do not lead to translocation, while promoting formation of an acidic tubulovesicle system more conducive to translocation, enhances PMT translocation and activity.

  7. Pasteurella multocida in backyard chickens in Upper Egypt: incidence with polymerase chain reaction analysis for capsule type, virulence in chicken embryos and antimicrobial resistance.

    PubMed

    Mohamed, Moemen A; Mohamed, Mohamed-Wael A; Ahmed, Ahmed I; Ibrahim, Awad A; Ahmed, Mohamed S

    2012-01-01

    The prevalence of Pasteurella multocida strains among 275 backyard chickens from different regions of Upper Egypt was studied. A total of 21 isolates of P. multocida were recovered in 21 out of 275 chickens tested (7.6%) and were confirmed using phenotypic characterisation. Somatic serotyping of the 21 isolates resulted in 12 isolates being classed as serotype A:1 (57.14%), 4 as serotype A:3 (19.05%) and 5 could not be typed (23.8%). Capsular typing, using multiplex polymerase chain reaction (PCR), demonstrated that 18 strains were capsular type A (85.7%), and 3 were type D (14.3%). The present findings suggest that a multiplex capsular PCR could be valuable for the rapid identification of P. multocida in cases of fowl cholera infection. A total of 5 isolates of P. multocida were selected to study their pathogenicity in embryonated chicken eggs instead of conducting a study in mature chickens. The results showed a variation in pathogenicity between the strains tested, namely: serotype A:1 strains caused 80% mortality, in contrast to 20% mortality by type D strains. Pathological findings included severe congestion of the entire embryo, haemorrhaging of the skin, feather follicles and toe, and ecchymotic haemorrhages on the liver of the inoculated embryos. The observations in this study indicate that P. multocida serogroup A could be highly pathogenic for mature chickens and therefore might be a cause of considerable economic losses in commercial production. A total of 10 isolates were subjected to antimicrobial susceptibility to determine the minimal inhibitory concentration of 7 antimicrobials. All isolates were susceptible to ciprofloxacin, florfenicol, streptomycin and sulphamethoxazol with trimethoprim and with varying degrees of sensitivity to the other agents.

  8. Pasteurella multocida toxin (PMT) activates RhoGTPases, induces actin polymerization and inhibits migration of human dendritic cells, but does not influence macropinocytosis.

    PubMed

    Blöcker, Dagmar; Berod, Luciana; Fluhr, Joachim W; Orth, Joachim; Idzko, Marco; Aktories, Klaus; Norgauer, Johannes

    2006-03-01

    Dendritic cells (DCs) are considered as one of the principal initiators of immune responses. In their immature state, they migrate into peripheral tissue in order to uptake antigen and to patrol for danger signals. Upon maturation, they acquire the ability to migrate to the lymph nodes and present the captured antigens to T cells in order to direct the development of specific immune responses. There is evidence that microbial compounds interfere with proper functions of DCs in order to block innate and specific immunity. Here we characterized the influence of Pasteurella multocida toxin (PMT) on monocyte-derived DCs. Using pull-down assays with recombinant rhotekin or p21-activated kinase, we demonstrated the activation of RhoGTPases by PMT in DCs. Moreover, PMT induced changes in DC morphology and actin polymerization, impaired chemotaxin-induced actin re-organization and inhibited their migration response. However, macropinocytosis was not influenced by PMT. In summary, these data indicate that PMT inhibits proper function of the motility machinery in DCs, which might limit the development of adaptive immune surveillance during infection with Pasteurella multocida.

  9. Virulence genotyping of Pasteurella multocida isolated from multiple hosts from India.

    PubMed

    Sarangi, Laxmi Narayan; Priyadarshini, Adyasha; Kumar, Santosh; Thomas, Prasad; Gupta, Santosh Kumar; Nagaleekar, Viswas Konasagara; Singh, Vijendra Pal

    2014-01-01

    In this study, 108 P. multocida isolates recovered from various host animals such as cattle, buffalo, swine, poultry (chicken, duck, and emu) and rabbits were screened for carriage of 8 virulence associated genes. The results revealed some unique information on the prevalence of virulence associated genes among Indian isolates. With the exception of toxA gene, all other virulence associated genes were found to be regularly distributed among host species. Association study between capsule type and virulence genes suggested that pfhA, nanB, and nanH genes were regularly distributed among all serotypes with the exception of CapD, whereas toxA gene was found to be positively associated with CapD and CapA. The frequency of hgbA and nanH genes among swine isolates of Indian origin was found to be less in comparison to its equivalents around the globe. Interestingly, very high prevalence of tbpA gene was observed among poultry, swine, and rabbit isolates. Likewise, very high prevalence of pfhA gene (95.3%) was observed among Indian isolates, irrespective of host species origin.

  10. Serotyping of foot and mouth disease virus and Pasteurella multocida from Indian gaurs (Bos gaurus), concurrently infected with foot and mouth disease and haemorrhagic septicaemia.

    PubMed

    Chandranaik, Basavegowdanadoddi Marinaik; Hegde, Raveendra; Shivashankar, Beechagondahalli Papanna; Giridhar, Papanna; Muniyellappa, Handenahally Kaverappa; Kalge, Rajeshwar; Sumathi, Benamanahalli Raju; Nithinprabhu, Kumble; Chandrashekara, Narasimhaiah; Manjunatha, Venkataramanappa; Jaisingh, Nirupama; Mayanna, Asha; Chandrakala, Gowda Kallenahalli; Kanaka, Sermaraja; Venkatesha, Mudalagiri Dasappagupta

    2015-06-01

    We report the serotyping of foot-and-mouth disease virus (FMDV) and Pasteurella multocida from Indian gaurs which were concurrently infected with foot-and-mouth disease (FMD) and haemorrhagic septicaemia. Bannerghatta biological park (BBP), a national park located in the outskirts of Bengaluru city, Karnataka, India, is bordered by several villages. These villages witnessed massive outbreaks of FMD which spread rapidly to the herbivores at BBP. Post-mortem was conducted on carcasses of two Indian gaurs that died with symptoms of FMD. The salient gross findings included extensive vesicular lesions on the tongue, gums, cheeks, upper palate and hooves. Haemorrhagic tracheitis and ecchymotic haemorrhages on the heart were characteristic. The vesicular lesions of oral cavity were positive for 'O' type of FMD virus by sandwich enzyme-linked immuno sorbent assay (ELISA). The heart blood and spleen samples yielded growth of pure cultures of P. multocida. The isolates were typed as P. multocida type B using KTSP61 and KTT72 primers yielding specific amplicons of 620 bp. The phylogenetic analysis of the isolates was carried by sequencing of 1.4-Kbp nucleotides on the 16S ribosomal RNA (rRNA) gene of the isolates.

  11. Pasteurella multocida Heddleston Serovar 3 and 4 Strains Share a Common Lipopolysaccharide Biosynthesis Locus but Display both Inter- and Intrastrain Lipopolysaccharide Heterogeneity

    PubMed Central

    Harper, Marina; St. Michael, Frank; John, Marietta; Vinogradov, Evgeny; Steen, Jennifer A.; van Dorsten, Lieke; Steen, Jason A.; Turni, Conny; Blackall, Patrick J.; Adler, Ben; Cox, Andrew D.

    2013-01-01

    Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and chronic forms. The major outer membrane component lipopolysaccharide (LPS) is both an important virulence factor and a major immunogen. Our previous studies determined the LPS structures expressed by different P. multocida strains and revealed that a number of strains belonging to different serovars contain the same LPS biosynthesis locus but express different LPS structures due to mutations within glycosyltransferase genes. In this study, we report the full LPS structure of the serovar 4 type strain, P1662, and reveal that it shares the same LPS outer core biosynthesis locus, L3, with the serovar 3 strains P1059 and Pm70. Using directed mutagenesis, the role of each glycosyltransferase gene in LPS outer core assembly was determined. LPS structural analysis of 23 Australian field isolates that contain the L3 locus revealed that at least six different LPS outer core structures can be produced as a result of mutations within the LPS glycosyltransferase genes. Moreover, some field isolates produce multiple but related LPS glycoforms simultaneously, and three LPS outer core structures are remarkably similar to the globo series of vertebrate glycosphingolipids. Our in-depth analysis showing the genetics and full range of P. multocida lipopolysaccharide structures will facilitate the improvement of typing systems and the prediction of the protective efficacy of vaccines. PMID:23974032

  12. Pasteurella multocida Heddleston serovar 3 and 4 strains share a common lipopolysaccharide biosynthesis locus but display both inter- and intrastrain lipopolysaccharide heterogeneity.

    PubMed

    Harper, Marina; St Michael, Frank; John, Marietta; Vinogradov, Evgeny; Steen, Jennifer A; van Dorsten, Lieke; Steen, Jason A; Turni, Conny; Blackall, Patrick J; Adler, Ben; Cox, Andrew D; Boyce, John D

    2013-11-01

    Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and chronic forms. The major outer membrane component lipopolysaccharide (LPS) is both an important virulence factor and a major immunogen. Our previous studies determined the LPS structures expressed by different P. multocida strains and revealed that a number of strains belonging to different serovars contain the same LPS biosynthesis locus but express different LPS structures due to mutations within glycosyltransferase genes. In this study, we report the full LPS structure of the serovar 4 type strain, P1662, and reveal that it shares the same LPS outer core biosynthesis locus, L3, with the serovar 3 strains P1059 and Pm70. Using directed mutagenesis, the role of each glycosyltransferase gene in LPS outer core assembly was determined. LPS structural analysis of 23 Australian field isolates that contain the L3 locus revealed that at least six different LPS outer core structures can be produced as a result of mutations within the LPS glycosyltransferase genes. Moreover, some field isolates produce multiple but related LPS glycoforms simultaneously, and three LPS outer core structures are remarkably similar to the globo series of vertebrate glycosphingolipids. Our in-depth analysis showing the genetics and full range of P. multocida lipopolysaccharide structures will facilitate the improvement of typing systems and the prediction of the protective efficacy of vaccines.

  13. Cross protection against fowl cholera disease with the use of recombinant Pasteurella multocida FHAB2 peptides vaccine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It has been demonstrated that fhaB2 (filamentous hemagglutinin) is an important virulence factor for P. multocida in development of fowl cholera disease and that recombinant FHAB2 peptides derived from P. multocida, Pm-1059, protect turkeys against Pm-1059 challenge. To test the hypothesis that rFHA...

  14. Antimicrobial susceptibility of Escherichia coli F4, Pasteurella multocida, and Streptococcus suis isolates from a diagnostic veterinary laboratory and recommendations for a surveillance system.

    PubMed

    Glass-Kaastra, Shiona K; Pearl, David L; Reid-Smith, Richard J; McEwen, Beverly; Slavic, Durda; McEwen, Scott A; Fairles, Jim

    2014-04-01

    Antimicrobial susceptibility data on Escherichia coli F4, Pasteurella multocida, and Streptococcus suis isolates from Ontario swine (January 1998 to October 2010) were acquired from a comprehensive diagnostic veterinary laboratory in Ontario, Canada. In relation to the possible development of a surveillance system for antimicrobial resistance, data were assessed for ease of management, completeness, consistency, and applicability for temporal and spatial statistical analyses. Limited farm location data precluded spatial analyses and missing demographic data limited their use as predictors within multivariable statistical models. Changes in the standard panel of antimicrobials used for susceptibility testing reduced the number of antimicrobials available for temporal analyses. Data consistency and quality could improve over time in this and similar diagnostic laboratory settings by encouraging complete reporting with sample submission and by modifying database systems to limit free-text data entry. These changes could make more statistical methods available for disease surveillance and cluster detection.

  15. Clinico-pathology, hematology and biochemistry responses in buffaloes towards Pasteurella multocida type B: 2 immunogen lypopolysaccharide via oral and intravenous routes of infection.

    PubMed

    Chung, Eric Lim Teik; Abdullah, Faez Firdaus Jesse; Ibrahim, Hayder Hamzah; Marza, Ali Dhiaa; Zamri-Saad, Mohd; Haron, Abdul Wahid; Lila, Mohd Azmi Mohd; Norsidin, Mohd Jefri

    2016-02-01

    Haemorrhagic septicaemia is a disease caused by Pasteurella multocida serotype B: 2 and E: 2. The organism causes acute, highly fatal septicaemic disease with high morbidity and mortality in cattle and more susceptible in buffaloes. Lipopolysaccharide can be found on the outer cell wall of the organism. Lipopolysaccharide is released during multiplication which leads to inflammatory reaction. It represents the endotoxin of P. multocida type B: 2 and responsible for toxicity in haemorrhagic septicaemia which plays an important role in the pathogenesis of the disease. Therefore, the aim of this study was to investigate the clinical signs, blood parameters, gross post mortem lesions and histopathology changes caused by P. multocida type B:2 immunogen lipopolysaccharide infections initiated through intravenous and oral routes of infection. 9 buffalo heifers were divided equally into 3 treatment groups. Group 1 was inoculated orally with 10 ml of phosphate buffer saline (PBS); Group 2 and 3 were inoculated with 10 ml of lipopolysaccharide broth intravenously and orally respectively. For the clinical signs, there were significant differences (p < 0.05) in temperature between the control, intravenous and oral group. In hematology and biochemistry findings, there were significant differences (p < 0.05) in erythrocytes, haemoglobin, PCV, MCV, lymphocytes, monocytes, eosinophils, GGT and albumin between the control, intravenous and oral group. However, there were no significant differences (p > 0.05) in the MCHC, leukocytes, band neutrophils, basophils, thrombocytes, plasma protein, icterus index, total protein, globulin and A:G ratio between intravenous and oral group. For Group 2 buffaloes, there were gross lesions in the lung, trachea, heart, liver, spleen, and kidney. In contrast, lesions were only observed in the lung, trachea and liver of Group 3 buffaloes. There were significant differences (p < 0.05) in hemorrhage and congestion; necrosis and degeneration; and

  16. Assessment of Pasteurella multocida A Lipopolysaccharide, as an Adhesin in an In Vitro Model of Rabbit Respiratory Epithelium

    PubMed Central

    Romero, Stefany; Esquinas, Paula; Patiño, Pilar; Martínez, Nhora

    2017-01-01

    The role of the P. multocida lipopolysaccharide (LPS) as a putative adhesin during the early stages of infection with this bacterium in the respiratory epithelium of rabbits was investigated. By light microscopy and double enzyme labeling of nasal septa tissues, the amount of bacteria attached to the respiratory epithelium and the amount of LPS present in goblet cells at different experimental times were estimated. Transmission electron microscopy (TEM) and LPS labeling with colloidal gold particles were also used to determine the exact location of LPS in the cells. Septa that were challenged with LPS of P. multocida and 30 minutes later with P. multocida showed more adherent bacteria and more severe lesions than the other treatments. Free LPS was observed in the lumen of the nasal septum, forming bilamellar structures and adhering to the cilia, microvilli, cytoplasmic membrane, and cytoplasm of epithelial ciliated and goblet cells. The above findings suggest that P. multocida LPS plays an important role in the process of bacterial adhesion and that it has the ability of being internalized into host cells. PMID:28251016

  17. Pasteurella multocida isolated from wild birds of North America: a serotype and DNA fingerprint study of isolates from 1978 to 1993

    USGS Publications Warehouse

    Wilson, M.A.; Duncan, R.M.; Nordholm, G.E.; Berlowski, B.M.

    1995-01-01

    Serotype and DNA fingerprint methods were used to study Pasteurella multocida isolated from 320 wild birds of North America. Isolates were collected during 1978-93. The HhaI profiles of 314 isolates matched the HhaI profile of somatic reference type 1, strain X-73; somatic type 1 antigen was expressed by 310 isolates, and the serotype of four isolates was undetected. Differentiation of the 314 isolates was observed by digestion of DNA with HpaII. None of the HpaII profiles matched the HpaII profile of X-73 (designated HhaI 001/HpaII 001). Three HpaII profiles were recognized among the somatic type 1 isolates: HpaII 002 (n = 18), HpaII 003 (n = 122), and HpaII 004 (n = 174). Profile HpaII 002 was found among isolates collected during 1979-83. Profile HpaII 003 was identified from isolates collected during 1979-89, with the exception of two isolates in 1992. The HpaII 004 profile was identified from isolates collected during 1983-93. Of the six remaining isolates, four expressed somatic type 4 and had HhaI profiles identical to the somatic type 4 reference strain P-1662 profile (designated HhaI 004); these isolates were differentiated by digestion of DNA with HpaII. One isolate was identified as serotype F:11, and another was serotype A:3,4. In the present study, 314 of 316 (99.4%) isolates from wild birds in the Central, Mississippi, and Pacific flyways during 1978-93, were P. multocida somatic type 1.

  18. Differences in Virulence Between Bovine-Derived Clinical Isolates of Pasteurella multocida Serotype A from the UK and the USA in a Model of Bovine Pneumonic Pasteurellosis.

    PubMed

    Dagleish, M P; Bayne, C W; Moon, G G; Finlayson, J; Sales, J; Williams, J; Hodgson, J C

    2016-07-01

    The time of onset and subsequent degree and progression of clinical signs, bacterial colonization and tissue pathology during experimental disease induced by intratracheal inoculation of either a UK or USA isolate of Pasteurella multocida serotype A recovered from clinical cases of bovine pneumonia were determined. Calves aged 8 weeks were challenged with 300 ml phosphate buffered saline (PBS) alone (group 1, n = 3, negative control) or containing 7.1 × 10(8) colony forming units (cfu) of UK isolate (group 2, n = 8) or 5.8 × 10(8) cfu of USA isolate (group 3, n = 8). Bronchoalveolar lavage (BAL) at 0, 1 and 4 days post challenge (dpc) and at the time of necropsy examination (7-8 dpc) showed no significant differences between groups 2 and 3 in bacterial numbers recovered. No P. multocida were recovered from group 1 animals. No clinical disease was present in group 1 calves and in group 3 was limited to scour in 1 calf at 1 dpc. All calves in group 2 had reduced food intake at 4-5 dpc, five had periods of dullness, three a mild nasal discharge at 1 dpc, four had mild to substantial respiratory stridor and one was killed at 6 dpc for humane reasons. Rectal temperatures remained about 39°C in group 1 calves, but increased in P. multocida-challenged calves to 40-41°C within 8-12 h of challenge. Significantly (P = 0.01) greater percentages of lung surface area were consolidated in group 2 (mean ± SD, 21 ± 10.1) compared with group 3 (7 ± 8.6) calves. Significantly more extensive and severe histological lesions were present in the lung lobes (P = 0.006) and lymph nodes (P = 0.02) of group 2 compared with group 3 calves. Pleurisy was present in group 2 calves only and no pathology was present in group 1. Pulsed-field gel electrophoresis (PFGE) produced 11 (group 2, UK isolate) or 10 (group 3, USA isolate) bands with differences in banding patterns. Results overall showed that two isolates, distinct geographically and genetically (by PFGE

  19. Decreasing the sialidase activity of multifunctional Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) by site-directed mutagenesis.

    PubMed

    Sugiarto, Go; Lau, Kam; Li, Yanhong; Khedri, Zahra; Yu, Hai; Le, Diem-Thuy; Chen, Xi

    2011-11-01

    Pasteurella multocida α2-3-sialyltransferase 1 (PmST1) is a multifunctional enzyme which has α2-6-sialyltransferase, α2-3-sialidase, and α2-3-trans-sialidase activities in addition to its major α2-3-sialyltransferase activity. The presence of the α2-3-sialidase activity of PmST1 complicates its application in enzymatic synthesis of α2-3-linked sialosides as the product formed can be hydrolyzed by the enzyme. Herein we show that the α2-3-sialidase activity of PmST1 can be significantly decreased by protein crystal structure-based site-directed mutagenesis. A PmST1 double mutant E271F/R313Y showed a significantly (6333-fold) decreased sialidase activity without affecting its α2-3-sialyltransferase activity. The double mutant E271F/R313Y, therefore, is a superior enzyme for enzymatic synthesis of α2-3-linked sialosides.

  20. Impact of early versus later fluoroquinolone treatment on the clinical; microbiological and resistance outcomes in a mouse-lung model of Pasteurella multocida infection.

    PubMed

    Ferran, Aude A; Toutain, Pierre-Louis; Bousquet-Mélou, Alain

    2011-03-24

    The early curative uses of antimicrobial drugs such as fluoroquinolones before the onset of symptoms in veterinary medicine may be regarded as irrational antibiotic consumption. However, it should be stressed that in early curative antimicrobial treatment as in metaphylaxis, the bacterial burden at the infection site is often very low, and so the rapid eradication of the bacterial population could result. We investigated the impact of early versus later curative administrations of 1 or 40 mg/kg of marbofloxacin on the survival of mice, the eradication of the targeted pathogen and the selection of resistant bacteria in a mouse lung infection with Pasteurella multocida. In this model, for a given marbofloxacin dose, the clinical and bacteriological outcomes were better, and the selection of resistance less frequent, for the early rather than for the late treatment. Moreover, the early administration of 1mg/kg led to better clinical and similar bacteriological (eradication and selection of resistance) outcomes than the late administration of 40 mg/kg marbofloxacin. Our results suggest that the optimal doses for the animals' cure could be lower when administered early during the time course of the infection than when administered after the disease outbreak. As the main argument against early treatments such as metaphylaxis is the possible enhancement of resistance at the gut level, further studies should assess if lower doses of antibiotic administered to all the animals of a herd could have less impact on the commensal digestive flora than higher doses only administered to animals showing clinical symptoms.

  1. Feeding of waste milk to Holstein calves affects antimicrobial resistance of Escherichia coli and Pasteurella multocida isolated from fecal and nasal swabs.

    PubMed

    Maynou, G; Bach, A; Terré, M

    2017-04-01

    The use of milk containing antimicrobial residues in calf feeding programs has been shown to select for resistant fecal Escherichia coli in dairy calves. However, information is scarce about the effects of feeding calves waste milk (WM) on the prevalence of multidrug-resistant bacteria. The objective of this study was to determine the antimicrobial resistance patterns of fecal E. coli and nasal Pasteurella multocida isolates from calves fed either milk replacer (MR) or WM in 8 commercial dairy farms (4 farms per feeding program). Fecal and nasal swabs were collected from 20 ± 5 dairy calves at 42 ± 3.2 d of age, and from 10 of these at approximately 1 yr of age in each study farm to isolate the targeted bacteria. Furthermore, resistance of E. coli isolates from calf-environment and from 5 calves at birth and their dams was also evaluated in each study farm. Resistances were tested against the following antimicrobial agents: amoxicillin-clavulanic acid, ceftiofur, colistin, doxycycline (DO), enrofloxacin (ENR), erythromycin, florfenicol, imipenem, and streptomycin. A greater number of fecal E. coli resistant to ENR, florfenicol, and streptomycin and more multidrug-resistant E. coli phenotypes were isolated in feces of calves fed WM than in those fed MR. However, the prevalence of fecal-resistant E. coli was also influenced by calf age, as it increased from birth to 6 wk of age for ENR and DO and decreased from 6 wk to 1 yr of age for DO regardless of the feeding program. From nasal samples, an increase in the prevalence of colistin-resistant P. multocida was observed in calves fed WM compared with those fed MR. The resistance patterns of E. coli isolates from calves and their dams tended to differ, whereas similar resistance profiles among E. coli isolates from farm environment and calves were observed. The findings of this study suggest that feeding calves WM fosters the presence of resistant bacteria in the lower gut and respiratory tracts of dairy calves.

  2. The RNA-Binding Chaperone Hfq Is an Important Global Regulator of Gene Expression in Pasteurella multocida and Plays a Crucial Role in Production of a Number of Virulence Factors, Including Hyaluronic Acid Capsule

    PubMed Central

    Mégroz, Marianne; Kleifeld, Oded; Wright, Amy; Powell, David; Harrison, Paul; Adler, Ben; Harper, Marina

    2016-01-01

    The Gram-negative bacterium Pasteurella multocida is the causative agent of a number of economically important animal diseases, including avian fowl cholera. Numerous P. multocida virulence factors have been identified, including capsule, lipopolysaccharide (LPS), and filamentous hemagglutinin, but little is known about how the expression of these virulence factors is regulated. Hfq is an RNA-binding protein that facilitates riboregulation via interaction with small noncoding RNA (sRNA) molecules and their mRNA targets. Here, we show that a P. multocida hfq mutant produces significantly less hyaluronic acid capsule during all growth phases and displays reduced in vivo fitness. Transcriptional and proteomic analyses of the hfq mutant during mid-exponential-phase growth revealed altered transcript levels for 128 genes and altered protein levels for 78 proteins. Further proteomic analyses of the hfq mutant during the early exponential growth phase identified 106 proteins that were produced at altered levels. Both the transcript and protein levels for genes/proteins involved in capsule biosynthesis were reduced in the hfq mutant, as were the levels of the filamentous hemagglutinin protein PfhB2 and its secretion partner LspB2. In contrast, there were increased expression levels of three LPS biosynthesis genes, encoding proteins involved in phosphocholine and phosphoethanolamine addition to LPS, suggesting that these genes are negatively regulated by Hfq-dependent mechanisms. Taken together, these data provide the first evidence that Hfq plays a crucial role in regulating the global expression of P. multocida genes, including the regulation of key P. multocida virulence factors, capsule, LPS, and filamentous hemagglutinin. PMID:26883595

  3. Pasteurella multocida toxin activates the inositol triphosphate signaling pathway in Xenopus oocytes via G(q)alpha-coupled phospholipase C-beta1.

    PubMed

    Wilson, B A; Zhu, X; Ho, M; Lu, L

    1997-01-10

    Pasteurella multocida toxin (PMT) has been hypothesized to cause activation of a GTP-binding protein (G-protein)-coupled phosphatidylinositol-specific phospholipase C (PLC) in intact cells. We used voltage-clamped Xenopus oocytes to test for direct PMT-mediated stimulation of PLC by monitoring the endogenous Ca2+-dependent C1- current. Injection of PMT induced an inward, two-component Cl- current, similar to that evoked by injection of IP3 through intracellular Ca2+ mobilization and Ca2+ influx through voltage-gated Ca2+ channels. These PMT-induced currents were blocked by specific inhibitors of Ca2+ and Cl- channels, removal of extracellular Ca2+, or chelation of intracellular Ca2+. Specific antibodies directed against an N-terminal, but not a C-terminal, peptide of PMT inhibited the toxin-induced currents, implicating that the N terminus of PMT is important for toxin activity. Injection with specific antibodies against PLCbeta1, PLCbeta2, PLCbeta3, or PLCgamma1 identified PLCbeta1 as the primary mediator of the PMT-induced Cl- currents. Injection with guanosine 5'-O-(2-(thio)diphosphate), antibodies to the common GTP-binding region of G-protein alpha subunits, or antibodies to different regions of G-protein beta subunits established the involvement of a G-protein alpha subunit in PMT-activation of PLCbeta1. Injection with specific antibodies against the alpha-subunits of G(q/11), G(s/olf), G(i/o/t/z), or G(i-1/i-2/i-3) isoforms confirmed the involvement of Gq/11alpha. Preinjection of oocytes with pertussis toxin enhanced the PMT response. Overexpression of G(q)alpha in oocytes could enhance the PMT response by 30-fold to more than 300-fold, whereas introduction of antisense G(q)alpha cRNA reduced the response by 7-fold. The effects of various specific antibodies on the PMT response were reproduced in oocytes overexpressing G(q)alpha.

  4. One-Step Multiplex RT-qPCR Assay for the Detection of Peste des petits ruminants virus, Capripoxvirus, Pasteurella multocida and Mycoplasma capricolum subspecies (ssp.) capripneumoniae

    PubMed Central

    Lamien, Charles Euloge; Spergser, Joachim; Lelenta, Mamadou; Wade, Abel; Gelaye, Esayas; Loitsch, Angelika; Minoungou, Germaine; Thiaucourt, Francois; Diallo, Adama

    2016-01-01

    Respiratory infections, although showing common clinical symptoms like pneumonia, are caused by bacterial, viral or parasitic agents. These are often reported in sheep and goats populations and cause huge economic losses to the animal owners in developing countries. Detection of these diseases is routinely done using ELISA or microbiological methods which are being reinforced or replaced by molecular based detection methods including multiplex assays, where detection of different pathogens is carried out in a single reaction. In the present study, a one-step multiplex RT-qPCR assay was developed for simultaneous detection of Capripoxvirus (CaPV), Peste de petits ruminants virus (PPRV), Pasteurella multocida (PM) and Mycoplasma capricolum ssp. capripneumonia (Mccp) in pathological samples collected from small ruminants with respiratory disease symptoms. The test performed efficiently without any cross-amplification. The multiplex PCR efficiency was 98.31%, 95.48%, 102.77% and 91.46% whereas the singleplex efficiency was 93.43%, 98.82%, 102.55% and 92.0% for CaPV, PPRV, PM and Mccp, respectively. The correlation coefficient was greater than 0.99 for all the targets in both multiplex and singleplex. Based on cycle threshold values, intra and inter assay variability, ranged between the limits of 2%–4%, except for lower concentrations of Mccp. The detection limits at 95% confidence interval (CI) were 12, 163, 13 and 23 copies/reaction for CaPV, PPRV, PM and Mccp, respectively. The multiplex assay was able to detect CaPVs from all genotypes, PPRV from the four lineages, PM and Mccp without amplifying the other subspecies of mycoplasmas. The discriminating power of the assay was proven by accurate detection of the targeted pathogen (s) by screening 58 viral and bacterial isolates representing all four targeted pathogens. Furthermore, by screening 81 pathological samples collected from small ruminants showing respiratory disease symptoms, CaPV was detected in 17 samples

  5. Characterization of the lipopolysaccharide from Pasteurella multocida Heddleston serovar 9: identification of a proposed bi-functional dTDP-3-acetamido-3,6-dideoxy-α-D-glucose biosynthesis enzyme.

    PubMed

    Harper, Marina; St Michael, Frank; Vinogradov, Evgeny; John, Marietta; Boyce, John D; Adler, Ben; Cox, Andrew D

    2012-03-01

    Pasteurella multocida strains are classified into 16 different lipopolysaccharide (LPS) serovars using the Heddleston serotyping scheme. Ongoing studies in our laboratories on the LPS aim to determine the core oligosaccharide (OS) structures expressed by each of the Heddleston type strains and identify the genes and transferases required for the biosynthesis of the serovar-specific OSs. In this study, we have determined the core OS of the LPS expressed by the Heddleston serovar 9 type strain, P2095. Structural information was established by a combination of monosaccharide and methylation analyses, nuclear magnetic resonance spectroscopy and mass spectrometry revealing the following structure: . The serovar 9 OS contains an inner core that is conserved among P. multocida strains with an elaborate outer core extension containing rhamnose (Rha), a D-glycero-D-manno isomer of heptose, and the unusual deoxyamino sugar, 3-acetamido-3,6-dideoxy-α-D-glucose (Qui3NAc). Genetic analyses of the LPS outer core biosynthesis locus revealed that in addition to the glycosyltransferases predicted to transfer the sugars to the nascent LPS molecule, the locus also contained the complete set of genes required for the biosynthesis of the nucleotide sugar donors dTDP-Rha and dTDP-Qui3NAc. One of the genes identified as part of the dTDP-Qui3NAc biosynthesis pathway, qdtD, encodes a proposed bi-functional enzyme with N-terminal amino acid identity to dTDP-4-oxo-6-deoxy-D-glucose-3,4-oxoisomerase and C-terminal amino acid identity to dTDP-3-oxo-6-deoxy-α-D-glucose transacetylase.

  6. Mammalian target of rapamycin complex 1 (mTORC1) plays a role in Pasteurella multocida toxin (PMT)-induced protein synthesis and proliferation in Swiss 3T3 cells.

    PubMed

    Oubrahim, Hammou; Wong, Allison; Wilson, Brenda A; Chock, P Boon

    2013-01-25

    Pasteurella multocida toxin (PMT) is a potent mitogen known to activate several signaling pathways via deamidation of a conserved glutamine residue in the α subunit of heterotrimeric G-proteins. However, the detailed mechanism behind mitogenic properties of PMT is unknown. Herein, we show that PMT induces protein synthesis, cell migration, and proliferation in serum-starved Swiss 3T3 cells. Concomitantly PMT induces phosphorylation of ribosomal S6 kinase (S6K1) and its substrate, ribosomal S6 protein (rpS6), in quiescent 3T3 cells. The extent of the phosphorylation is time and PMT concentration dependent, and is inhibited by rapamycin and Torin1, the two specific inhibitors of the mammalian target of rapamycin complex 1 (mTORC1). Interestingly, PMT-mediated mTOR signaling activation was observed in MEF WT but not in Gα(q/11) knock-out cells. These observations are consistent with the data indicating that PMT-induced mTORC1 activation proceeds via the deamidation of Gα(q/11), which leads to the activation of PLCβ to generate diacylglycerol and inositol trisphosphate, two known activators of the PKC pathway. Exogenously added diacylglycerol or phorbol 12-myristate 13-acetate, known activators of PKC, leads to rpS6 phosphorylation in a rapamycin-dependent manner. Furthermore, PMT-induced rpS6 phosphorylation is inhibited by PKC inhibitor, Gö6976. Although PMT induces epidermal growth factor receptor activation, it exerts no effect on PMT-induced rpS6 phosphorylation. Together, our findings reveal for the first time that PMT activates mTORC1 through the Gα(q/11)/PLCβ/PKC pathway. The fact that PMT-induced protein synthesis and cell migration is partially inhibited by rapamycin indicates that these processes are in part mediated by the mTORC1 pathway.

  7. 21 CFR 558.630 - Tylosin and sulfamethazine.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... pneumonias caused by bacterial pathogens (Pasteurella multocida and/or Corynebacterium pyogenes); for... Bordetella bronchiseptica rhinitis; prevention of swine dysentery (vibrionic); control of swine pneumonias... hyodysenteriae; and control of swine pneumonias caused by bacterial pathogens (Pasteurella multocida...

  8. Characterization of Pasteurella species isolated from lungs of calves with pneumonia.

    PubMed Central

    Madsen, E B; Bisgaard, M; Mutters, R; Pedersen, K B

    1985-01-01

    During routine bacteriological examination of pneumonic calf lungs it was experienced that many Pasteurella multocida-like isolates had a fermentation pattern different from what is generally accepted for P. multocida sensu stricto. Forty-one out of 50 strains selected for further investigation were phenotypically related and formed a group of indole-, mannitol-and sorbitol-negative P. multocida-like strains, which was tentatively designated taxon 13. Deoxyribonucleic acid/deoxyribonucleic acid hybridizations including both ornithine positive and ornithine negative strains of taxon 13 allowed the classification of the former as P. multocida biovar 6 and the latter as V factor independent strains of Haemophilus avium. PMID:3986681

  9. Bordetella avium

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetellosis is an acute, highly contagious disease of the upper respiratory tract of young turkeys (4-8 wk of age). The disease is caused by a gram-negative, nonfermentative bacterium, Bordetella avium. Members of the genus Bordetella are well known for their ability to colonize and damage ciliate...

  10. Characterization of TolC Efflux Pump Proteins from Pasteurella multocida▿ †

    PubMed Central

    Hatfaludi, Tamas; Al-Hasani, Keith; Dunstone, Michelle; Boyce, John; Adler, Ben

    2008-01-01

    Two TolC homologs, PM0527 and PM1980, were identified for Pasteurella multocida. A pm0527 mutant displayed increased susceptibility to a range of chemicals, including rifampin (512-fold) and acridine orange (128-fold). A pm1980 mutant showed increased susceptibility to rifampin, ceftazidime, and vancomycin. PMID:18725450

  11. A numerical taxonomic study of Actinobacillus, Pasteurella and Yersinia.

    PubMed

    Sneath, P H; Stevens, M

    1985-10-01

    A numerical taxonomic study of strains of Actinobacillus, Pasteurella and Yersinia, with some allied bacteria, showed 23 reasonably distinct groups. These fell into three major areas. Area A contained species of Actinobacillus and Pasteurella: A. suis, A. equuli, A. lignieresii, P. haemolytica biovar A, P. haemolytica biovar T, P. multocida, A. actinomycetemcomitans, 'P. bettii', 'A. seminis', P. ureae and P. aerogenes. Also included in A was a composite group of Pasteurella pneumotropica and P. gallinarum, together with unnamed groups referred to as 'BLG', 'Mair', 'Ross' and 'aer-2'. Area B contained species of Yersinia: Y. enterocolitica, Y. pseudotuberculosis, Y. pestis and a group 'ent-b' similar to Y. enterocolitica. Area C contained non-fermenting strains: Y. philomiragia, Moraxella anatipestifer and a miscellaneous group 'past-b'. There were also a small number of unnamed single strains.

  12. Septic shock from Pasturella multocida following a cat bite: case report and review of literature.

    PubMed

    Adler, Adam C; Cestero, Cesar; Brown, Robert B

    2011-01-01

    Pasteurella multocida is a gram-negative organism categorized morphologically as a coccobacillus. P. multocida is a natural inhabitant found in the nasopharynx and oropharynx of numerous animal hosts, but serves as an opportunistic pathogen in humans. Pasturella multocida has multiple subspecies; when identified as the cause of infection they are broadly termed pasturelloses. Infections involving P. multocida are typically reported to occur in immune-compromised patients. Few cases in the literature identify pasturellosis as the causative agent of septic shock, especially in cirrhotic patients. Our patient's underlying cirrhosis and past splenectomy place her in the higher risk category for developing invasive Pasturella infection. We report a patient who presented with septic shock that was initially thought to be related to a urinary tract infection. It was later revealed that the patient's condition was caused by a recent cat bite leading to Pasturella bacteremia compounded by hepatic cirrhosis and previous splenectomy.

  13. Bordetella pertussis.

    PubMed

    Nieves, Delma J; Heininger, Ulrich

    2016-06-01

    Pertussis is a highly infectious vaccine-preventable cough illness that continues to be a significant source of morbidity and mortality around the world. The majority of human illness is caused by Bordetella pertussis, and some is caused by Bordetella parapertussis. Bordetella is a Gram-negative, pleomorphic, aerobic coccobacillus. In the past several years, even countries with high immunization rates in early childhood have experienced rises in pertussis cases. Reasons for the resurgence of reported pertussis may include molecular changes in the organism and increased awareness and diagnostic capabilities, as well as lessened vaccine efficacy and waning immunity. The most morbidity and mortality with pertussis infection is seen in infants too young to benefit from immunization. Severe infection requiring hospitalization, including in an intensive care setting, is mostly seen in those under 3 months of age. As a result, research and public health actions have been aimed at better understanding and reducing the spread of Bordetella pertussis. Studies comparing the cost benefit of cocooning strategies versus immunization of pregnant women have been favorable towards immunizing pregnant women. This strategy is expected to prevent a larger number of pertussis cases, hospitalizations, and deaths in infants <1 year old while also being cost-effective. Studies have demonstrated that the source of infection in infants usually is a family member. Efforts to immunize children and adults, in particular pregnant women, need to remain strong.

  14. Antimicrobial Susceptibility of Bordetella bronchiseptica Isolates from Swine and Companion Animals and Detection of Resistance Genes

    PubMed Central

    Prüller, Sandra; Rensch, Ulrike; Meemken, Diana; Kaspar, Heike; Kopp, Peter A.; Klein, Günter; Kehrenberg, Corinna

    2015-01-01

    Bordetella bronchiseptica causes infections of the respiratory tract in swine and other mammals and is a precursor for secondary infections with Pasteurella multocida. Treatment of B. bronchiseptica infections is conducted primarily with antimicrobial agents. Therefore it is essential to get an overview of the susceptibility status of these bacteria. The aim of this study was to comparatively analyse broth microdilution susceptibility testing according to CLSI recommendations with an incubation time of 16 to 20 hours and a longer incubation time of 24 hours, as recently proposed to obtain more homogenous MICs. Susceptibility testing against a panel of 22 antimicrobial agents and two fixed combinations was performed with 107 porcine isolates from different farms and regions in Germany and 43 isolates obtained from companion animals in Germany and other European countries. Isolates with increased MICs were investigated by PCR assays for the presence of resistance genes. For ampicillin, all 107 porcine isolates were classified as resistant, whereas only a single isolate was resistant to florfenicol. All isolates obtained from companion animals showed elevated MICs for β-lactam antibiotics and demonstrated an overall low susceptibility to cephalosporines. Extension of the incubation time resulted in 1–2 dilution steps higher MIC50 values of porcine isolates for seven antimicrobial agents tested, while isolates from companion animals exhibited twofold higher MIC50/90 values only for tetracycline and cefotaxime. For three antimicrobial agents, lower MIC50 and MIC90 values were detected for both, porcine and companion animal isolates. Among the 150 isolates tested, the resistance genes blaBOR-1 (n = 147), blaOXA-2, (n = 4), strA and strB (n = 17), sul1 (n = 10), sul2 (n = 73), dfrA7 (n = 3) and tet(A) (n = 8) were detected and a plasmid localisation was identified for several of the resistance genes. PMID:26275219

  15. Antimicrobial resistance and virulence gene profiles in P. multocida strains isolated from cats

    PubMed Central

    Ferreira, Thais Sebastiana Porfida; Felizardo, Maria Roberta; de Gobbi, Debora Dirani Sena; Moreno, Marina; Moreno, Andrea Micke

    2015-01-01

    Cats are often described as carriers of Pasteurella multocida in their oral microbiota. This agent is thought to cause pneumonia, conjunctivitis, rhinitis, gingivostomatitis, abscess and osteonecrosis in cats. Human infection with P. multocida has been described in several cases affecting cat owners or after cat bites. In Brazil, the cat population is approximately 21 million animals and is increasing, but there are no studies of the presence of P. multocida in the feline population or of human cases of infection associated with cats. In this study, one hundred and ninety-one healthy cats from owners and shelters in São Paulo State, Brazil, were evaluated for the presence of P. multocida in their oral cavities. Twenty animals were positive for P. multocida , and forty-one strains were selected and characterized by means of biochemical tests and PCR. The P. multocida strains were tested for capsular type, virulence genes and resistance profile. A total of 75.6% (31/41) of isolates belonged to capsular type A, and 24.4% (10/41) of the isolates were untypeable. None of the strains harboured toxA, tbpA or pfhA genes. The frequencies of the other genes tested were variable, and the data generated were used to build a dendrogram showing the relatedness of strains, which were clustered according to origin. The most common resistance profile observed was against sulfizoxazole and trimethoprim-sulphamethoxazole. PMID:26221117

  16. Antimicrobial resistance and virulence gene profiles in P. multocida strains isolated from cats.

    PubMed

    Ferreira, Thais Sebastiana Porfida; Felizardo, Maria Roberta; de Gobbi, Debora Dirani Sena; Moreno, Marina; Moreno, Andrea Micke

    2015-03-01

    Cats are often described as carriers of Pasteurella multocida in their oral microbiota. This agent is thought to cause pneumonia, conjunctivitis, rhinitis, gingivostomatitis, abscess and osteonecrosis in cats. Human infection with P. multocida has been described in several cases affecting cat owners or after cat bites. In Brazil, the cat population is approximately 21 million animals and is increasing, but there are no studies of the presence of P. multocida in the feline population or of human cases of infection associated with cats. In this study, one hundred and ninety-one healthy cats from owners and shelters in São Paulo State, Brazil, were evaluated for the presence of P. multocida in their oral cavities. Twenty animals were positive for P. multocida , and forty-one strains were selected and characterized by means of biochemical tests and PCR. The P. multocida strains were tested for capsular type, virulence genes and resistance profile. A total of 75.6% (31/41) of isolates belonged to capsular type A, and 24.4% (10/41) of the isolates were untypeable. None of the strains harboured toxA, tbpA or pfhA genes. The frequencies of the other genes tested were variable, and the data generated were used to build a dendrogram showing the relatedness of strains, which were clustered according to origin. The most common resistance profile observed was against sulfizoxazole and trimethoprim-sulphamethoxazole.

  17. A retrospective six-year national survey of P. multocida infections in Israel.

    PubMed

    Nseir, William; Giladi, M; Moroz, I; Moses, A E; Benenson, S; Finkelstein, R; Dan, M; Chazan, B; Bishara, J; Ben-Dror, G; Hassin, D; Peled, N; Rahav, G; Grupper, M; Potasman, I

    2009-01-01

    Pasteurella multocida is the commonest organism infecting pet bites. Anecdotal reports tend to overemphasize dramatic outcomes. We aimed to study a large database of P. multocida infections. This retrospective survey of P. multocida infections in Israeli hospitals refers to the y 2000-2005. Clinical microbiologists were contacted by email and asked to perform a back-search of their hospital's records for isolates of P. multocida. The charts of patients growing P. multocida were abstracted into a structured questionnaire. 77 cases were identified in 12 hospitals, yielding an annual incidence of 0.19/100,000. The mean age was 49.2+/-26.5 y and the mortality rate was 2.6%. Those who died were >65 y of age, had diabetes mellitus or cirrhosis and were bacteraemic. One-third of the cases occurred in people aged > or =65 y. Cats caused most of these infections (54%). Surgery for debridement was common (53.7%), but no-one required amputation; a second- and third-look operation was necessary for these patients. Bacteraemia was found in 32.5% of patients and was significantly more common among those aged >60 y (p =0.044). Hospitalized patients with P. multocida have a favourable prognosis, apart from elderly and bacteraemic patients with comorbidities. Surgery and reoperations may be required in about half of the patients.

  18. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... product from each serial shall be tested for potency using the mouse test provided in this paragraph. A mouse dose shall be 1/20 of the least dose recommended on the label for other animals which shall not be less than 2 ml. (1) The ability of the bacterin being tested (Unknown) to protect mice shall...

  19. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... product from each serial shall be tested for potency using the mouse test provided in this paragraph. A mouse dose shall be 1/20 of the least dose recommended on the label for other animals which shall not be less than 2 ml. (1) The ability of the bacterin being tested (Unknown) to protect mice shall...

  20. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... product from each serial shall be tested for potency using the mouse test provided in this paragraph. A mouse dose shall be 1/20 of the least dose recommended on the label for other animals which shall not be less than 2 ml. (1) The ability of the bacterin being tested (Unknown) to protect mice shall...

  1. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... product from each serial shall be tested for potency using the mouse test provided in this paragraph. A mouse dose shall be 1/20 of the least dose recommended on the label for other animals which shall not be less than 2 ml. (1) The ability of the bacterin being tested (Unknown) to protect mice shall...

  2. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... product from each serial shall be tested for potency using the mouse test provided in this paragraph. A mouse dose shall be 1/20 of the least dose recommended on the label for other animals which shall not be less than 2 ml. (1) The ability of the bacterin being tested (Unknown) to protect mice shall...

  3. Pets and Pasteurella Infections

    MedlinePlus

    ... Ear Nose & Throat Emotional Problems Eyes Fever From Insects or Animals Genitals and Urinary Tract Glands & Growth ... Preventable Diseases Healthy Children > Health Issues > Conditions > From Insects or Animals > Pets and Pasteurella Infections Health Issues ...

  4. Bordetella pertussis transmission

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella pertussis and Bordetella bronchiseptica are Gram negative bacterial respiratory pathogens. B. pertussis is the causative agent of whooping cough and is considered a human-adapted variant of B. bronchiseptica. B. pertussis and B. bronchiseptica share mechanisms of pathogenesis and are gene...

  5. Bordetella pertussis transmission.

    PubMed

    Trainor, Elizabeth A; Nicholson, Tracy L; Merkel, Tod J

    2015-11-01

    Bordetella pertussis and B. bronchiseptica are Gram-negative bacterial respiratory pathogens. Bordetella pertussis is the causative agent of whooping cough and is considered a human-adapted variant of B. bronchiseptica. Bordetella pertussis and B. bronchiseptica share mechanisms of pathogenesis and are genetically closely related. However, despite the close genetic relatedness, these Bordetella species differ in several classic fundamental aspects of bacterial pathogens such as host range, pathologies and persistence. The development of the baboon model for the study of B. pertussis transmission, along with the development of the swine and mouse model for the study of B. bronchiseptica, has enabled the investigation of different aspects of transmission including the route, attack rate, role of bacterial and host factors, and the impact of vaccination on transmission. This review will focus on B. pertussis transmission and how animal models of B. pertussis transmission and transmission models using the closely related B. bronchiseptica have increased our understanding of B. pertussis transmission.

  6. Actinobacillus rossii sp. nov., Actinobacillus seminis sp. nov., nom. rev., Pasteurella bettii sp. nov., Pasteurella lymphangitidis sp. nov., Pasteurella mairi sp. nov., and Pasteurella trehalosi sp. nov.

    PubMed

    Sneath, P H; Stevens, M

    1990-04-01

    Evidence from numerical taxonomic analysis and DNA-DNA hybridization supports the proposal of new species in the genera Actinobacillus and Pasteurella. The following new species are proposed: Actinobacillus rossii sp. nov., from the vaginas of postparturient sows; Actinobacillus seminis sp. nov., nom. rev., associated with epididymitis of sheep; Pasteurella bettii sp. nov., associated with human Bartholin gland abscess and finger infections; Pasteurella lymphangitidis sp. nov. (the BLG group), which causes bovine lymphangitis; Pasteurella mairi sp. nov., which causes abortion in sows; and Pasteurella trehalosi sp. nov., formerly biovar T of Pasteurella haemolytica, which causes septicemia in older lambs.

  7. Discovery of a novel species of Bordetella

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella species are Gram-negative coccobacilli. There are currently nine described species that constitute the genus Bordetella. Historically, this genus is subdivided into two groups of species: the “classical” and “non-classical” Bordetella. The classical Bordetella are the most studied group r...

  8. Evaluation of the Specificity of BP3385 for Bordetella pertussis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BP3385 has been proposed as a diagnostic PCR target for discriminating between Bordetella pertussis and other Bordetella species that also infect humans. Our results demonstrate this gene is also present in some strains of Bordetella hinzii and Bordetella bronchiseptica....

  9. Classification of Pasteurella species B as Pasteurella oralis sp. nov.

    PubMed

    Christensen, Henrik; Bertelsen, Mads F; Bojesen, Anders Miki; Bisgaard, Magne

    2012-06-01

    Pasteurella species B has so far only been reported from the oral cavity of dogs, cats and a ferret. In the present study, information from 15 recent isolates from different sources, including African hedgehogs (Atelerix albiventris), banded mongoose (Mungos mungo), Moholi bushbabies (Galago moholi) and pneumonia of a cat, were compared to five strains investigated previously from bite wounds in humans inflicted by a cat and dog and from gingiva of a cat. rpoB gene sequence comparison showed that 17 isolates, including the reference strain (CCUG 19794(T)), had identical sequences, whereas two were closely related and demonstrated 97.9 and 99.6 % similarity to strain CCUG 19794(T), respectively; the type strain of Pasteurella stomatis was the most closely related strain, with 92.3 % similarity. This is within the mean range (76-100 %) of rpoB gene sequence similarity between species of the same genus within the family Pasteurellaceae. 16S rRNA gene sequencing of four strains selected based on rpoB sequence comparison showed at least 99.7 % similarity between strains of Pasteurella species B, with 96.2 % similarity to the type strain of the closest related species (Pasteurella canis), indicating that Pasteurella species B should have separate species status. Separate species status was also documented when recN sequence comparisons were converted to a genome similarity of 93.7 % within Pasteurella species B and 59.0 % to the type strain of the closest related species (P. canis). Based on analysis of the phylogenetic and phenotypic data, and since most isolates originate from the oral cavities of a diverse group of animals, it is suggested that these bacteria be classified as Pasteurella oralis sp. nov.; the type strain is P683(T) ( = CCUG 19794(T) = CCM 7950(T) = strain 23193(T) = MCCM 00102(T)), obtained from a cat. Previous reports of the type strain have shown ubiquinone-8, demethylmenaquinone-8 and menaquinone-8 as the major quinones. Polyamines in the type

  10. Invasive Bordetella holmesii infections.

    PubMed

    Fishbain, Joel T; Riederer, Kathleen; Sawaf, Hadi; Mody, Rupal

    2015-02-01

    Bordetella holmesii is a rare cause of invasive human disease. The fastidious and unusual nature of this organism makes routine isolation and identification challenging. We report two cases of B. holmesii bacteremia that were rapidly identified by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS) when standard techniques failed to provide speciation. There are no current standards for susceptibility testing or treatment recommendations. The rare occurrence and challenges in identifying this pathogen led us to perform a comprehensive review of the epidemiology, clinical presentations, and treatment options for this potentially invasive pathogen.

  11. 9 CFR 113.116 - Pasteurella Multocida Bacterin, Avian Isolate, Type 4.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... turkeys during the prechallenge period of the potency test provided in paragraph (c) of this section shall... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  12. 9 CFR 113.118 - Pasteurella Multocida Bacterin, Avian Isolate, Type 3.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated turkeys... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  13. 9 CFR 113.116 - Pasteurella Multocida Bacterin, Avian Isolate, Type 4.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... turkeys during the prechallenge period of the potency test provided in paragraph (c) of this section shall... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  14. 9 CFR 113.118 - Pasteurella Multocida Bacterin, Avian Isolate, Type 3.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated turkeys... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  15. 9 CFR 113.116 - Pasteurella Multocida Bacterin, Avian Isolate, Type 4.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... turkeys during the prechallenge period of the potency test provided in paragraph (c) of this section shall... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  16. 9 CFR 113.118 - Pasteurella Multocida Bacterin, Avian Isolate, Type 3.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated turkeys... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  17. 9 CFR 113.116 - Pasteurella Multocida Bacterin, Avian Isolate, Type 4.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... turkeys during the prechallenge period of the potency test provided in paragraph (c) of this section shall... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  18. 9 CFR 113.118 - Pasteurella Multocida Bacterin, Avian Isolate, Type 3.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated turkeys... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  19. 9 CFR 113.116 - Pasteurella Multocida Bacterin, Avian Isolate, Type 4.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... turkeys during the prechallenge period of the potency test provided in paragraph (c) of this section shall... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  20. 9 CFR 113.118 - Pasteurella Multocida Bacterin, Avian Isolate, Type 3.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated turkeys... turkey, test results shall be determined by observing the remaining 20 turkeys. The test is inconclusive... more turkeys, but the serial is unsatisfactory if the test is not repeated. (c) Potency test. Bulk...

  1. 9 CFR 113.117 - Pasteurella Multocida Bacterin, Avian Isolate, Type 1.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated... week prechallenge period. (2) Unvaccinated controls. Each of not more than 11 chickens shall be held as controls. (3) Challenge. Not less than 14 days after the second injection, each of 20 vaccinates, and...

  2. 9 CFR 113.117 - Pasteurella Multocida Bacterin, Avian Isolate, Type 1.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated... week prechallenge period. (2) Unvaccinated controls. Each of not more than 11 chickens shall be held as controls. (3) Challenge. Not less than 14 days after the second injection, each of 20 vaccinates, and...

  3. 9 CFR 113.117 - Pasteurella Multocida Bacterin, Avian Isolate, Type 1.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated... week prechallenge period. (2) Unvaccinated controls. Each of not more than 11 chickens shall be held as controls. (3) Challenge. Not less than 14 days after the second injection, each of 20 vaccinates, and...

  4. 9 CFR 113.117 - Pasteurella Multocida Bacterin, Avian Isolate, Type 1.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated... week prechallenge period. (2) Unvaccinated controls. Each of not more than 11 chickens shall be held as controls. (3) Challenge. Not less than 14 days after the second injection, each of 20 vaccinates, and...

  5. 9 CFR 113.117 - Pasteurella Multocida Bacterin, Avian Isolate, Type 1.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... viable bacteria and fungi as provided in § 113.26. (b) Safety test. Observation of the vaccinated... week prechallenge period. (2) Unvaccinated controls. Each of not more than 11 chickens shall be held as controls. (3) Challenge. Not less than 14 days after the second injection, each of 20 vaccinates, and...

  6. 9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Haemolytica Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.68 Pasteurella Haemolytica Vaccine, Bovine. Pasteurella Haemolytica Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  7. 9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 9 Animals and Animal Products 1 2013-01-01 2013-01-01 false Pasteurella Haemolytica Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.68 Pasteurella Haemolytica Vaccine, Bovine. Pasteurella Haemolytica Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  8. 9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 9 Animals and Animal Products 1 2014-01-01 2014-01-01 false Pasteurella Haemolytica Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.68 Pasteurella Haemolytica Vaccine, Bovine. Pasteurella Haemolytica Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  9. 9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 9 Animals and Animal Products 1 2012-01-01 2012-01-01 false Pasteurella Haemolytica Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.68 Pasteurella Haemolytica Vaccine, Bovine. Pasteurella Haemolytica Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  10. 9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 9 Animals and Animal Products 1 2011-01-01 2011-01-01 false Pasteurella Haemolytica Vaccine... REQUIREMENTS Live Bacterial Vaccines § 113.68 Pasteurella Haemolytica Vaccine, Bovine. Pasteurella Haemolytica Vaccine, Bovine, shall be prepared as a desiccated live culture bacterial vaccine of an avirulent...

  11. Environmental Origin of the Genus Bordetella

    PubMed Central

    Hamidou Soumana, Illiassou; Linz, Bodo; Harvill, Eric T.

    2017-01-01

    Members of the genus Bordetella include human and animal pathogens that cause a variety of respiratory infections, including whooping cough in humans. Despite the long known ability to switch between a within-animal and an extra-host lifestyle under laboratory growth conditions, no extra-host niches of pathogenic Bordetella species have been defined. To better understand the distribution of Bordetella species in the environment, we probed the NCBI nucleotide database with the 16S ribosomal RNA (16S rRNA) gene sequences from pathogenic Bordetella species. Bacteria of the genus Bordetella were frequently found in soil, water, sediment, and plants. Phylogenetic analyses of their 16S rRNA gene sequences showed that Bordetella recovered from environmental samples are evolutionarily ancestral to animal-associated species. Sequences from environmental samples had a significantly higher genetic diversity, were located closer to the root of the phylogenetic tree and were present in all 10 identified sequence clades, while only four sequence clades possessed animal-associated species. The pathogenic bordetellae appear to have evolved from ancestors in soil and/or water. We show that, despite being animal-adapted pathogens, Bordetella bronchiseptica, and Bordetella hinzii have preserved the ability to grow and proliferate in soil. Our data implicate soil as a probable environmental origin of Bordetella species, including the animal-pathogenic lineages. Soil may further constitute an environmental niche, allowing for persistence and dissemination of the bacterial pathogens. Spread of pathogenic bordetellae from an environmental reservoir such as soil may potentially explain their wide distribution as well as frequent disease outbreaks that start without an obvious infectious source. PMID:28174558

  12. Cloning of a serotype-specific antigen from Pasteurella haemolytica A1.

    PubMed Central

    Gonzalez-Rayos, C; Lo, R Y; Shewen, P E; Beveridge, T J

    1986-01-01

    Recombinant plasmids coding for a soluble (or surface) antigen of Pasteurella haemolytica A1 were identified. Two plasmids, both containing the same 5.4 kilobase pairs of insert DNA, were recovered independently by screening a clone band of P. haemolytica A1 genomic DNA in Escherichia coli for the expression of P. haemolytica A1 soluble antigens (R. Y. C. Lo and L.A. Cameron, Can. J. Biochem. Cell Biol. 64:73-76, 1986). E. coli cells carrying the plasmids were found to be agglutinated by an antiserum raised against the P. haemolytica A1 soluble antigens. Analysis of the E. coli clones by electron microscopy revealed patches of amorphous material on the surface of the cells which were not present on the controls. Further characterization with protein A-colloidal gold labeled both these patches and the outer membranes of these cloned cells pretreated with the specific antiserum. These results indicated that the cloned antigen was expressed on the surface of the E. coli cells. The cloned antigen was found to be specific for serotype 1 when tested by slide agglutination against a collection of P. haemolytica typing antisera. Southern blot hybridization, using the cloned DNA as a probe, labeled the genomic DNA from P. haemolytica serotype 1 as well as the cross-agglutinating serotypes 2 and 7, but not DNA from the non-cross-agglutinating serotypes 3 and 4 and Pasteurella multocida. These results demonstrated that serotype specificity could be attributed to the particular antigenic determinants in the genome of the organism. Images PMID:3527985

  13. Ovine pulmonary surfactant induces killing of Pasteurella haemolytica, Escherichia coli, and Klebsiella pneumoniae by normal serum.

    PubMed Central

    Brogden, K A

    1992-01-01

    Pulmonary surfactant has been shown to play an increasingly important role in bacterial clearance at the alveolar surface in the lung. This study describes a bactericidal mechanism in which ovine pulmonary surfactant induces killing of Pasteurella haemolytica by normal serum. To demonstrate killing, six bacterial species were incubated first with pulmonary surfactant for 60 min at 37 degrees C and then with serum for an additional 60 min at 37 degrees C. P. haemolytica type A1 strains 82-25 and L101, a P. haemolytica type 2 strain, Escherichia coli, and Klebsiella pneumoniae were susceptible and Pasteurella multocida, Serratia marcescens, and Pseudomonas aeruginosa were not susceptible to killing by ovine pulmonary surfactant and normal serum. No bacteria incubated with bovine pulmonary surfactant were killed by normal serum. Although the species origin of pulmonary surfactant was selective, the species origin of serum was not. P. haemolytica incubated with ovine pulmonary surfactant was killed by fetal calf serum, gnotobiotic calf serum, pooled normal sheep serum, pooled normal rabbit serum, and pooled guinea pig serum. Ultrastructurally, killed P. haemolytica suspensions contained dead cells and cells distorted with vacuoles between the cytoplasmic membrane and the cytoplasm. The mechanism of killing did not correlate with concentrations of complement or lysozyme or titers of residual antibody in either the pulmonary surfactant or the serum, and killing was reduced by preincubation of surfactant with P. haemolytica lipopolysaccharide. Preliminary characterization of both surfactant and serum implicate a low-molecular-weight proteinaceous component in the surfactant and serum albumin in the serum. This mechanism may help clear certain gram-negative bacteria from the lungs of sheep as a part of the pulmonary innate defense system. Images PMID:1452351

  14. 21 CFR 558.630 - Tylosin and sulfamethazine.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... (vibrionic); control of swine pneumonias caused by bacterial pathogens (Pasteurella multocida and/or... pneumonias caused by bacterial pathogens (Pasteurella multocida and/or Corynebacterium pyogenes). (iii) For... hyodysenteriae; and control of swine pneumonias caused by bacterial pathogens (Pasteurella multocida...

  15. 21 CFR 558.630 - Tylosin and sulfamethazine.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... (vibrionic); control of swine pneumonias caused by bacterial pathogens (Pasteurella multocida and/or... pneumonias caused by bacterial pathogens (Pasteurella multocida and/or Corynebacterium pyogenes). (iii) For... hyodysenteriae; and control of swine pneumonias caused by bacterial pathogens (Pasteurella multocida...

  16. Differentiation among closely related organisms of the Actinobacillus-Haemophilus-Pasteurella group by means of lysozyme and EDTA.

    PubMed Central

    Olsen, I; Brondz, I

    1985-01-01

    Bacteriolysis in Tris-maleate buffer (0.005 M, pH 7.2) supplemented with EDTA (0.01 M) and hen egg white lysozyme (HEWL, 1.0 microgram/ml) was set up to assist differentiation between the taxonomically closely related Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus. A. actinomycetemcomitans was more sensitive to lysis in this system than H. aphrophilus. The standard method for bacteriolysis separated the 10 tested strains of A. actinomycetemcomitans into two groups (I and II) based on their lysis patterns, whereas the 7 strains of H. aphrophilus examined were homogeneous. In group I of A. actinomycetemcomitans, EDTA displayed a considerable lytic effect, which was not increased by supplementation with HEWL. In group II, the lytic effect of EDTA was much less, but HEWL had a considerable supplementary lytic effect. When the turbidity of A. actinomycetemcomitans (ATCC 29522) or H. aphrophilus (ATCC 33389) suspended in Tris buffer was monitored at close pH intervals (0.2) from pH 5.2 to 9.2, maximal lysis of ATCC 29522 occurred with EDTA at pH 8.0 and with EDTA-HEWL at pH 7.6, while ATCC 33389 lysed with EDTA at pH 9.0 and with EDTA-HEWL at pH 9.2. When other members of the family Pasteurellaceae (Haemophilus influenzae type b, Haemophilus paraphrophilus, Pasteurella multocida, Pasteurella haemolytica, and Pasteurella ureae) were included for comparison, the group I strains of A. actinomycetemcomitans were the most rapidly lysed by EDTA. H. paraphrophilus was the least sensitive of the gram-negative strains tested, but not as resistant as Micrococcus luteus (control). M. luteus was the organism most sensitive to lysozyme, followed by P. ureae and the group II strains of A. actinomycetemcomitans, while the group I strains of A. actinomycetemcomitans, H. paraphrophilus, and P. haemolytica were the least sensitive organisms. Images PMID:3935663

  17. Airborne transmission of Bordetella pertussis.

    PubMed

    Warfel, Jason M; Beren, Joel; Merkel, Tod J

    2012-09-15

    Pertussis is a contagious, acute respiratory illness caused by the bacterial pathogen Bordetella pertussis. Although it is widely believed that transmission of B. pertussis occurs via aerosolized respiratory droplets, no controlled study has ever documented airborne transmission of pertussis. We set out to determine if airborne transmission occurs between infected and naive animals, utilizing the baboon model of pertussis. Our results showed that 100% of exposed naive animals became infected even when physical contact was prevented, demonstrating that pertussis transmission occurs via aerosolized respiratory droplets.

  18. [Isolation of Bordetella trematum from bacteremia].

    PubMed

    Halim, Ilham; Ihbibane, Fatima; Belabbes, Houria; Zerouali, Khalid; El Mdaghri, Naima

    2014-01-01

    The species Bordetella trematum was first described in 1996. Currently only eleven cases were published. We describe the first case of Bordetella trematum issued from bacteremia with a patient who has severe burns in Morocco. The identification was not possible by conventional microbiological methods where the resort to 16S ARNr sequencing. The use of molecular methods, including sequencing of the 16S ARNr, is currently an essential complementary tool to identify microbiological pathogens.

  19. 21 CFR 520.2218 - Sulfamerazine, sulfamethazine, and sulfaquinoxaline powder.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine,...

  20. 21 CFR 520.2218 - Sulfamerazine, sulfamethazine, and sulfaquinoxaline powder.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine,...

  1. 21 CFR 520.2218 - Sulfamerazine, sulfamethazine, and sulfaquinoxaline powder.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine,...

  2. 21 CFR 520.2218 - Sulfamerazine, sulfamethazine, and sulfaquinoxaline powder.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine,...

  3. 21 CFR 520.2218 - Sulfamerazine, sulfamethazine, and sulfaquinoxaline powder.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine,...

  4. Bordetella holmesii: an under-recognised Bordetella species.

    PubMed

    Pittet, Laure F; Emonet, Stéphane; Schrenzel, Jacques; Siegrist, Claire-Anne; Posfay-Barbe, Klara M

    2014-06-01

    Bordetella holmesii, first described in 1995, is believed to cause both invasive infections (bacteraemia, meningitis, endocarditis, pericarditis, pneumonia, and arthritis) and pertussis-like symptoms. Infection with B holmesii is frequently misidentified as being with B pertussis, the cause of whooping cough, because routine diagnostic tests for pertussis are not species-specific. In this Review, we summarise knowledge about B holmesii diagnosis and treatment, and assess research needs. Although no fatal cases of B holmesii have been reported, associated invasive infections can cause substantial morbidities, even in previously healthy individuals. Antimicrobial treatment can be problematic because B holmesii's susceptibility to macrolides (used empirically to treat B pertussis) and third-generation cephalosporins (often used to treat invasive infections) is lower than would be expected. B holmesii's adaptation to human beings is continuing, and virulence might increase, causing the need for better diagnostic assays and epidemiological surveillance.

  5. Development of a PCR assay for identification of Bordetella hinzii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella hinzii infects primarily poultry and immunocompromised humans. Although initially thought to be nonpathogenic in poultry, it was recently shown that some strains cause disease in turkey poults indistinguishable from the clinical presentation of turkey coryza caused by Bordetella avium. ...

  6. A Quantitative Flurometric Assay for the Measurement of Antibody to Pasteurella haemolytica in Cattle

    PubMed Central

    Confer, A.W.; Fox, J.C.; Newman, P.R.; Lawson, G.W.; Corstvet, R.E.

    1983-01-01

    A rapid, simple fluorometric method is described for measuring antibody to Pasteurella haemolytica in sera of cattle. Various antigen preparations were compared for the test including live, formalin-killed and phenol-killed P. haemolytica. A preparation composed of formalin-killed organisms from a 22 hour culture gave consistent results and was used in the studies. The test was reproduciable with percent coefficients of variation for fluorescent signal unit values on ten or more replicate samples ranging from 5.7 to 28.0. Sera from calves vaccinated by aerosol exposure to live P. haemolytica had up to a five-fold increase in antibody titer as measured by the flurometric method test during a 21 day period. Fluorometric method titers were comparable to those obtained by the indirect bacterial agglutination test. There was no seroconversion to P. haemolytica in calves vaccinated by aerosol exposure of P. multocida. The major advantages of the fluorometric method test over conventional methods are that the assay does not require serial dilutions of serum samples and thus limits time and effort to determine antibody titers. PMID:6339016

  7. Pasteurella haemolytica complicated respiratory infections in sheep and goats.

    PubMed

    Brogden, K A; Lehmkuhl, H D; Cutlip, R C

    1998-01-01

    Respiratory infections which commonly occur in sheep and goats often result from adverse physical and physiological stress combined with viral and bacterial infections. Inevitably, Pasteurella haemolytica pneumonia occurs as a result of these interactions. In this review, we present recent advances in research on the complex etiology of pneumonia involving P. haemolytica. Initially stress, induced by factors such as heat, overcrowding, exposure to inclement weather, poor ventilation, handling and transport is a major predisposing factor. Respiratory viruses including parainfluenza 3 (PI-3) virus, adenovirus type 6 and respiratory syncytial virus (RSV), and to a lesser extent bovine adenovirus type 2, ovine adenovirus types 1 and 5, and reovirus type 1 cause respiratory infections and pneumonia. More importantly these viruses also dramatically increase the susceptibility of sheep and goats to secondary P. haemolytica infection. Primary infection of the lower respiratory tract, with Mycoplasma ovipneumoniae and Bordetella parapertussis can increase the susceptibility of sheep and goats to secondary P. haemolytica infection. It is possible that initial infections with viral or primary bacterial agents break down the antimicrobial barrier consisting of beta defensins and anionic peptides found in epithelial cells, resident and inflammatory cells, and serous and mucous secretions of the respiratory tract. Loss of barrier integrity may release P. haemolytica from its usual commensal status. Once in the lung, P. haemolytica becomes opportunistic. To grow and colonize, P. haemolytica uses extracellular products like O-sialoglycoprotein endopeptidase, neuraminidase and RTX leukotoxin, as well as cell-associated products such as capsular polysaccharide, lipopolysaccharide, outer membrane proteins, proteins involved in iron acquisition and a periplasmic superoxide dismutase. In lambs and kids, pneumonic pasteurellosis can be acute, characterized by fever, listlessness, poor

  8. Growth Phase dependent gene regulation in Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetellae are Gram negative bacterial respiratory pathogens. Bordetella pertussis, the causative agent of whooping cough, is a human-restricted variant of Bordetella bronchiseptica, which infects a broad range of mammals causing chronic and often asymptomatic infections. Growth phase dependent gen...

  9. Whooping cough in Pakistan: Bordetella pertussis vs Bordetella parapertussis in 2005-2009.

    PubMed

    Bokhari, Habib; Said, Fahad; Syed, Muhammad A; Mughal, Amjad; Kazi, Yasmeen F; Heuvelman, Kees; Mooi, Frits R

    2011-10-01

    Pertussis, or whooping cough, is an acute respiratory disease mainly affecting infants and children and is caused by Bordetella pertussis and Bordetella parapertussis. The aim of this study was to investigate the share of Bordetella species from potential whooping cough cases during 2005-2009. Eight hundred and two samples from suspected pertussis cases were collected, mainly from 2 provinces of Pakistan. Bacterial culture, identification, DNA extraction and routinely used polymerase chain reaction (PCR) methods using IS1001, IS1002 and IS481 were used to identify the Bordetella species. The results were unexpected, because all of the isolates collected from the different cities were identified as B. parapertussis (7.4%); B. pertussis was not isolated from any sample. However, PCR results indicated the presence of a small percentage (0.6%) of B. pertussis among the total cases studied. This study suggests that vaccines to protect against both B. pertussis and B. parapertussis should be considered.

  10. Structure of Bordetella pertussis peptidoglycan

    SciTech Connect

    Folkening, W.J.; Nogami, W.; Martin, S.A.; Rosenthal, R.S.

    1987-09-01

    Bordetella pertussis Tohama phases I and III were grown to the late-exponential phase in liquid medium containing (/sup 3/H)diaminopimelic acid and treated by a hot (96/sup 0/C) sodium dodecyl sulfate extraction procedure. Washed sodium dodecyl sulfate-insoluble residue from phases I and III consisted of complexes containing protein (ca. 40%) and peptidoglycan (60/sup 6/). Subsequent treatment with proteinase K yielded purified peptidoglycan which contained N-acetylglucosamine, N-acetylmuramic acid, alanine, glutamic acid, and diaminopimelic acid in molar ratios of 1:1:2:1:1 and <2% protein. Radiochemical analyses indicated that /sup 3/H added in diaminopimelic acid was present in peptidoglycan-protein complexes and purified peptidoglycan as diaminopimelic acid exclusively and that pertussis peptidoglycan was not O acetylated, consistent with it being degraded completely by hen egg white lysozyme. Muramidase-derived disaccharide peptide monomers and peptide-cross-linked dimers and higher oligomers were isolated by molecular-sieve chromatography; from the distribution of these peptidoglycan fragments, the extent of peptide cross-linking of both phase I and III peptidoglycan was calculated to be ca. 48%. Unambiguous determination of the structure of muramidase-derived pepidoglycan fragments by fast atom bombardment-mass spectrometry and tandem mass spectrometry indicated that the pertussis peptidoglycan monomer fraction was surprisingly homogeneous, consisting of >95% N-acetylglucosaminyl-N-acetylmuramyl-alanyl-glutamyl-diaminopimelyl-alanine.

  11. Virulence and Genome Evolution of Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The genetic changes that cause some isolates to be more pathogenic than others are generally not well understood. Bordetella bronchiseptica is a prime model to study the underlying factor(s) that cause some strains to be more pathogenic than others, as strains of this clonal species cause different ...

  12. Diagnosis of whooping cough in Switzerland: differentiating Bordetella pertussis from Bordetella holmesii by polymerase chain reaction.

    PubMed

    Pittet, Laure F; Emonet, Stéphane; François, Patrice; Bonetti, Eve-Julie; Schrenzel, Jacques; Hug, Melanie; Altwegg, Martin; Siegrist, Claire-Anne; Posfay-Barbe, Klara M

    2014-01-01

    Bordetella holmesii, an emerging pathogen, can be misidentified as Bordetella pertussis by routine polymerase chain reaction (PCR). In some reports, up to 29% of the patients diagnosed with pertussis have in fact B. holmesii infection and invasive, non-respiratory B. holmesii infections have been reported worldwide. This misdiagnosis undermines the knowledge of pertussis' epidemiology, and may lead to misconceptions on pertussis vaccine's efficacy. Recently, the number of whooping cough cases has increased significantly in several countries. The aim of this retrospective study was to determine whether B. holmesii was contributing to the increase in laboratory-confirmed cases of B. pertussis in Switzerland. A multiplex species-specific quantitative PCR assay was performed on 196 nasopharyngeal samples from Swiss patients with PCR-confirmed Bordetella infection (median age: 6 years-old, minimum 21 days-old, maximum 86 years-old), formerly diagnosed as Bordetella pertussis (IS481+). No B. holmesii (IS481+, IS1001-, hIS1001+) was identified. We discuss whether laboratories should implement specific PCR to recognize different Bordetella species. We conclude that in Switzerland B. holmesii seems to be circulating less than in neighboring countries and that specific diagnostic procedures are not necessary routinely. However, as the epidemiological situation may change rapidly, periodic reevaluation is suggested.

  13. Diagnosis of Whooping Cough in Switzerland: Differentiating Bordetella pertussis from Bordetella holmesii by Polymerase Chain Reaction

    PubMed Central

    Pittet, Laure F.; Emonet, Stéphane; François, Patrice; Bonetti, Eve-Julie; Schrenzel, Jacques; Hug, Melanie; Altwegg, Martin; Siegrist, Claire-Anne; Posfay-Barbe, Klara M.

    2014-01-01

    Bordetella holmesii, an emerging pathogen, can be misidentified as Bordetella pertussis by routine polymerase chain reaction (PCR). In some reports, up to 29% of the patients diagnosed with pertussis have in fact B. holmesii infection and invasive, non-respiratory B. holmesii infections have been reported worldwide. This misdiagnosis undermines the knowledge of pertussis' epidemiology, and may lead to misconceptions on pertussis vaccine's efficacy. Recently, the number of whooping cough cases has increased significantly in several countries. The aim of this retrospective study was to determine whether B. holmesii was contributing to the increase in laboratory-confirmed cases of B. pertussis in Switzerland. A multiplex species-specific quantitative PCR assay was performed on 196 nasopharyngeal samples from Swiss patients with PCR-confirmed Bordetella infection (median age: 6 years-old, minimum 21 days-old, maximum 86 years-old), formerly diagnosed as Bordetella pertussis (IS481+). No B. holmesii (IS481+, IS1001−, hIS1001+) was identified. We discuss whether laboratories should implement specific PCR to recognize different Bordetella species. We conclude that in Switzerland B. holmesii seems to be circulating less than in neighboring countries and that specific diagnostic procedures are not necessary routinely. However, as the epidemiological situation may change rapidly, periodic reevaluation is suggested. PMID:24586447

  14. Expression of pertussis toxin in Bordetella bronchiseptica and Bordetella parapertussis carrying recombinant plasmids.

    PubMed Central

    Lee, C K; Roberts, A; Perrin, S

    1989-01-01

    Pertussis toxin is produced only by strains of Bordetella pertussis. Cloned genes encoding pertussis toxin from B. pertussis were transferred into Bordetella bronchiseptica and Bordetella parapertussis by conjugation. These transconjugants expressed pertussis toxin at levels comparable to those expressed by B. pertussis. The toxin made by these strains was biologically active in the Chinese hamster cell clumping assay, contained all five subunits, and was mostly periplasmic. Toxin expression appeared to be modulated in the same way as are the vir-regulated genes of B. pertussis. Introduction of these plasmids into B. pertussis failed to produce hypertoxigenic strains. Instead, these transconjugants underwent plasmid loss, gene deletions, or conversion to the avirulent phase. Images PMID:2707851

  15. Complete Genome Sequences of Four Different Bordetella sp. Isolates Causing Human Respiratory Infections

    PubMed Central

    Peng, Yanhui; Loparev, Vladimir; Batra, Dhwani; Bowden, Katherine E.; Cassiday, Pamela K.; Davis, Jamie K.; Johnson, Taccara; Juieng, Phalasy; Miner, Christine E.; Rowe, Lori; Sheth, Mili; Tondella, M. Lucia; Williams, Margaret M.

    2016-01-01

    Species of the genus Bordetella associate with various animal hosts, frequently causing respiratory disease. Bordetella pertussis is the primary agent of whooping cough and other Bordetella species can cause similar cough illness. Here, we report four complete genome sequences from isolates of different Bordetella species recovered from human respiratory infections. PMID:27795250

  16. Significant finding of Bordetella holmesii DNA in nasopharyngeal samples from French patients with suspected pertussis.

    PubMed

    Njamkepo, Elisabeth; Bonacorsi, Stéphane; Debruyne, Monique; Gibaud, Sophie Anne; Guillot, Sophie; Guiso, Nicole

    2011-12-01

    Pertussis is routinely diagnosed with real-time PCR based on insertion sequence IS481, which is not specific for Bordetella pertussis. We conducted a retrospective study using real-time PCRs specific for Bordetella pertussis and for Bordetella holmesii on 177 samples positive for IS481 PCR. Bordetella holmesii DNA was detected in 20.3% samples collected from adolescents and adults.

  17. Complete Bordetella avium, Bordetella hinzii and Bordetella trematum lipid A structures and genomic sequence analyses of the loci involved in their modifications.

    PubMed

    Novikov, Alexey; Shah, Nita R; AlBitar-Nehme, Sami; Basheer, Soorej M; Trento, Ilaria; Tirsoaga, Alina; Moksa, Michelle; Hirst, Martin; Perry, Malcolm B; Hamidi, Asmaa El; Fernandez, Rachel C; Caroff, Martine

    2014-08-01

    Endotoxin is recognized as one of the virulence factors of the Bordetella avium bird pathogen, and characterization of its structure and corresponding genomic features are important for an understanding of its role in pathogenicity and for an improved general knowledge of Bordetella spp virulence factors. The structure of the biologically active part of B. avium LPS, lipid A, is described and compared to those of another bird pathogen, opportunistic in humans, Bordetella hinzii, and to that of Bordetella trematum, a human pathogen. Sequence analyses showed that the three strains have homologues of acyl-chain modifying enzymes PagL, PagP and LpxO, of the 1-phosphatase LpxE, in addition to LgmA, LgmB and LgmC, which are required for the glucosamine modification. MALDI mass spectrometry identified a high amount of glucosamine substituting the phosphate groups of B. avium lipid A; this modification was absent from B. hinzii and B. trematum. The acylation patterns of the three lipid As were similar, but they differed from those of Bordetella pertussis and Bordetella parapertussis. They were also found to be close to the lipid A structure of Bordetella bronchiseptica, a mammalian pathogen, only differing from the latter by the degree of hydroxylation of the branched fatty acid.

  18. A PCR assay for identification of Bordetella hinzii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella hinzii infects primarily poultry and immunocompromised humans. Although initially thought to be nonpathogenic in poultry, it was recently shown that some strains cause disease in turkey poults indistinguishable from the clinical presentation of turkey coryza caused by Bordetella avium. B....

  19. Bordetella bronchiseptica and fatal pneumonia of dogs and cats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less well-studied. The objectives of this study were to identify the frequency of Bordetella bronchiseptica infection in fatal cases of bronchopneumonia in dogs and cats and to compare the diag...

  20. The effect of Pasteurella haemolytica and the leukotoxin of Pasteurella haemolytica on bovine lung explants.

    PubMed Central

    Wilkie, I W; Fallding, M H; Shewen, P E; Yager, J A

    1990-01-01

    Bovine lung explants were used in a study designed to compare the pathogenic effects of Pasteurella haemolytica type 1, a nonpathogenic organism Neisseria subflava, or the crude leukotoxin of P. haemolytica on alveolar macrophages and lung parenchymal cells. Concentrated, purified peripheral blood neutrophil suspensions were added with the bacteria to some explants. Duplicate pairs of cultures from each treatment group were fixed at regular intervals up to 24 hours after seeding and morphological changes were assessed by light and electron microscopy. Pasteurella haemolytica caused deterioration of alveolar macrophages within one hour but did not affect parenchymal cells for more than 12 hours. Neisseria subflava did not affect alveolar macrophages initially, but caused an accelerated deterioration after four hours. After 24 hours, bacterial overgrowth caused similar deterioration of all cells in explants seeded with either bacterium. Alveolar macrophages phagocytosed large numbers of N. subflava but rarely ingested P. haemolytica. Added neutrophils did not have any discernible effect on any of the explants and did not potentiate bacterial effects. Addition of crude leukotoxin of P. haemolytica to the culture medium significantly accelerated alveolar macrophage deterioration without apparent effect on parenchymal cell survival. These results support the hypothesis that the severe tissue destruction of fulminant pneumonic pasteurellosis is not a direct result of bacterial infection. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. Fig. 8. PMID:2306666

  1. Pasteurella caballi, a new species from equine clinical specimens.

    PubMed Central

    Schlater, L K; Brenner, D J; Steigerwalt, A G; Moss, C W; Lambert, M A; Packer, R A

    1989-01-01

    The name Pasteurella caballi is proposed for a group of organisms represented by 29 strains isolated from respiratory and other infections in horses. P. caballi strains are gram-negative, oxidase-positive, nonmotile, fermentative rods with the key characteristics of the genus Pasteurella. These strains differed from other Pasteurella species in that all were aerogenic and catalase negative, and some strains produced acid from myo-inositol and L-rhamnose. The levels of DNA relatedness of 28 P. caballi strains with labeled DNA from the proposed type strain averaged 91 and 85% (hydroxyapatite method at 55 and 70 degrees C). P. caballi was 13 to 53% related to strains representing 22 other species of the family Pasteurellaceae. The guanine-plus-cytosine content of the DNA of four strains was 41 to 42 mol%. The type strain is 83851 (=ATCC 49197). PMID:2584369

  2. Isolation of Pasteurella haemolytica leukotoxin mutants.

    PubMed Central

    Chidambaram, M; Sharma, B; Petras, S F; Reese, C P; Froshauer, S; Weinstock, G M

    1995-01-01

    Two mutants of Pasteurella haemolytica A1 that do not produce leukotoxin were isolated. Following mutagenesis, colonies were screened with antiserum by a filter assay for absence of the secreted leukotoxin. The two mutants both appeared to produce normal amounts of other antigens, as judged by reactivity with polyclonal serum from an animal with pasteurellosis, and were not altered in beta-hemolytic activity as seen on blood agar plates. There was no evidence of either cell-associated or secreted leukotoxin protein when Western blots (immunoblots) were carried out with the polyclonal serum or with a monoclonal antibody directed against the leukotoxin. Southern blots revealed that both mutants show the wild-type restriction pattern at the leukotoxin locus, although the strain with the lktA2 mutation showed differences in other regions of the chromosome on analysis by pulsed-field gel electrophoresis. The strain with the lktA2 mutation grew more slowly than did the wild-type strain, while the strain with the lktA1 mutation was indistinguishable from the wild-type strain in its growth properties. The strain with the lktA1 mutation should be valuable in determining the role of the leukotoxin in virulence as well as in identifying other virulence factors of P. haemolytica. PMID:7868223

  3. A transport medium for specimens containing Pasteurella pestis.

    PubMed

    Cavanaugh, D C; Vivona, S; Do-Van-Quy; Gibson, F L; Deuber, G L; Rust, J H

    1967-01-01

    A medium, originally designed by Stuart and co-workers and later modified by Cary & Blair, for the maintenance and transport, without multiplication, of pathogenic bacteria contained in bacteriological specimens was tested in the laboratory and in the field in Viet-Nam to determine its effectiveness in preserving specimens known to contain Pasteurella pestis.The results indicate that this medium should be useful in diagnostic plague studies in areas where transport facilities are inadequate. Properly collected clinical specimens, sent to a central laboratory by any means and under any climatic conditions likely to be encountered in the hot tropics, should yield viable Pasteurella pestis for at least 30 days.

  4. Bacteriocin Produced by Bordetella pertussis1

    PubMed Central

    Litkenhous, Claudia; Liu, P. V.

    1967-01-01

    Of the 24 strains of Bordetella pertussis examined, 2 produced bacteriocins that inhibited the growth of all but 2 other strains of this species. The two strains producing the bacteriocin and the two resistant strains were rough, whereas all susceptible strains were smooth. The bacteriocin was not active on the B. parapertussis or B. bronchiseptica strains tested. These bacteriocins appeared to be protein in nature, since they were heat-labile and partially inactivated by trypsin. They were antigenic but the neutralizing antibodies did not precipitate the antigens. Absorption of the antiserum with homologous cell suspensions removed the agglutinating, but not the neutralizing, antibody. Images PMID:4290577

  5. Bordetella pertussis pathogenesis: current and future challenges

    PubMed Central

    Melvin, Jeffrey A.; Scheller, Erich V.; Miller, Jeff F.; Cotter, Peggy A.

    2014-01-01

    Pertussis, or whooping cough, has recently reemerged as a major public health threat despite high levels of vaccination against the etiological agent, Bordetella pertussis. In this Review, we describe the pathogenesis of this disease, with a focus on recent mechanistic insights into virulence factor function. We also discuss the changing epidemiology of pertussis and the challenges of vaccine development. Despite decades of research, many aspects of B. pertussis physiology and pathogenesis remain poorly understood. We highlight knowledge gaps that must be addressed to develop improved vaccines and therapeutic strategies. PMID:24608338

  6. Resemblance and divergence: the "new" members of the genus Bordetella.

    PubMed

    Gross, Roy; Keidel, Kristina; Schmitt, Karin

    2010-08-01

    Bordetella pertussis, the etiological agent of whooping cough, belongs to the bacterial pathogens first described in the so-called golden era of microbiology more than 100 years ago. In the course of the following decades, several other closely related pathogens were described which are nowadays classified in the genus Bordetella together with B. pertussis. These are the human and animal pathogens B. parapertussis, B. bronchiseptica and B. avium which are of high medical or veterinary interest, and which, together with B. pertussis, are referred to as the "classical" Bordetella species. Only in the past 15 years, several additional species were classified in the genus, frequently isolated from patients with underlying disease, animals or from the environment. Very little is known about most of these bacteria. In the present review, the current knowledge about these "new" Bordetella species is briefly summarized.

  7. Bordetella holmesii, an emerging cause of septic arthritis.

    PubMed

    Abouanaser, Salaheddin F; Srigley, Jocelyn A; Nguyen, Tram; Dale, Suzanne E; Johnstone, Jennie; Wilcox, Lindsay; Jamieson, Frances; Rawte, Prasad; Pernica, Jeffrey M

    2013-04-01

    Bordetella holmesii is a well-described pathogen in asplenic and immunocompromised patients. Here we report the first two published cases of septic arthritis caused by B. holmesii documented in apparently immunocompetent patients and unaccompanied by bacteremia.

  8. Bordetella holmesii: initial genomic analysis of an emerging opportunist.

    PubMed

    Planet, Paul J; Narechania, Apurva; Hymes, Saul R; Gagliardo, Christina; Huard, Richard C; Whittier, Susan; Della-Latta, Phyllis; Ratner, Adam J

    2013-03-01

    Bordetella holmesii is an emerging opportunistic pathogen that causes respiratory disease in healthy individuals and invasive infections among patients lacking splenic function. We used 16S rRNA gene analysis to confirm B. holmesii as the cause of bacteremia in a child with sickle cell disease. Semiconductor-based draft genome sequencing provided insight into B. holmesii phylogeny and potential virulence mechanisms and also identified a toluene-4-monoxygenase locus unique among bordetellae.

  9. Genetic diversity and relationships in populations of Bordetella spp.

    PubMed Central

    Musser, J M; Hewlett, E L; Peppler, M S; Selander, R K

    1986-01-01

    Genetic diversity in 60 strains of three nominal Bordetella species recovered from humans and other mammalian hosts was assessed by analyzing electrophoretically demonstrable allelic variation at structural genes encoding 15 enzymes. Eleven of the loci were polymorphic, and 14 distinctive electrophoretic types, representing multilocus genotypes, were identified. The population structure of Bordetella spp. is clonal, and genetic diversity is relatively limited compared with most other pathogenic bacteria and is insufficient to justify recognition of three species. All isolates of Bordetella parapertussis were of one electrophoretic type, which was closely similar to 9 of the 10 electrophoretic types represented by isolates of Bordetella bronchiseptica. Bordetella pertussis 18-323, which is used in mouse potency tests of vaccines, is more similar genetically to isolates of B. bronchiseptica and B. parapertussis than to other isolates currently assigned to the species B. pertussis. Apart from strain 18-323, the isolates of B. pertussis represented only two closely related clones, and all isolates of B. pertussis from North America (except strain 18-323) were genotypically identical. Strain Dejong, which has been classified as B. bronchiseptica, was strongly differentiated from all of the other Bordetella isolates examined. Images PMID:3957867

  10. Differential in vitro expression of the brkA gene in Bordetella pertussis and Bordetella parapertussis clinical isolates.

    PubMed

    Stefanelli, Paola; Sanguinetti, Maurizio; Fazio, Cecilia; Posteraro, Brunella; Fadda, Giovanni; Mastrantonio, Paola

    2006-09-01

    In this study, we set up a real-time reverse transcriptase PCR assay to measure the relative amounts of brkA transcripts in 50 Bordetella isolates. The results suggested that brkA expression is strain dependent and its level may play a role in determining the serum resistance or susceptibility phenotype. Pertussis immunocompetent sera were unable to kill Bordetella parapertussis via complement deposition.

  11. Reclassification of Pasteurella gallinarum, [Haemophilus] paragallinarum, Pasteurella avium and Pasteurella volantium as Avibacterium gallinarum gen. nov., comb. nov., Avibacterium paragallinarum comb. nov., Avibacterium avium comb. nov. and Avibacterium volantium comb. nov.

    PubMed

    Blackall, Patrick J; Christensen, Henrik; Beckenham, Tim; Blackall, Linda L; Bisgaard, Magne

    2005-01-01

    This paper describes a phenotypic and genotypic investigation of the taxonomy of [Haemophilus] paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium, a major subcluster within the avian 16S rRNA cluster 18 of the family Pasteurellaceae. An extended phenotypic characterization was performed of the type strain of [Haemophilus] paragallinarum, which is NAD-dependent, and eight NAD-independent strains of [Haemophilus] paragallinarum. Complete 16S rRNA gene sequences were obtained for one NAD-independent and four NAD-dependent [Haemophilus] paragallinarum strains. These five sequences along with existing 16S rRNA gene sequences for 11 other taxa within avian 16S rRNA cluster 18 as well as seven other taxa from the Pasteurellaceae were subjected to phylogenetic analysis. The analysis demonstrated that [Haemophilus] paragallinarum, Pasteurella gallinarum, Pasteurella avium and Pasteurella volantium formed a monophyletic group with a minimum of 96.8 % sequence similarity. This group can also be separated by phenotypic testing from all other recognized and named taxa within the Pasteurellaceae. As both genotypic and phenotypic testing support the separate and distinct nature of this subcluster, the transfer is proposed of Pasteurella gallinarum, [Haemophilus] paragallinarum, Pasteurella avium and Pasteurella volantium to a new genus Avibacterium as Avibacterium gallinarum gen. nov., comb. nov., Avibacterium paragallinarum comb. nov., Avibacterium avium comb. nov. and Avibacterium volantium comb. nov. The type strains are NCTC 1118T (Avibacterium gallinarum), NCTC 11296T (Avibacterium paragallinarum), NCTC 11297T (Avibacterium avium) and NCTC 3438T (Avibacterium volantium). Key characteristics that separate these four species are catalase activity (absent only in Avibacterium paragallinarum) and production of acid from galactose (negative only in Avibacterium paragallinarum), maltose (negative only in Avibacterium avium) and mannitol (negative

  12. Human infections associated with Bordetella bronchiseptica.

    PubMed Central

    Woolfrey, B F; Moody, J A

    1991-01-01

    This study examines the potential of Bordetella bronchiseptica to act as a human pathogen. After encountering two patients from whom B. bronchiseptica was isolated, we searched the literature and found 23 reports in which a human infection was reported in association with B. bronchiseptica. As a basis for evaluating these cases, we summarize the literature about the current microbiological status of B. bronchiseptica, the pathology and pathogenic mechanisms associated with the microorganism, and the likelihood of it acting as a commensal or colonizer. From this review we conclude that B. bronchiseptica has been rarely isolated from humans despite their considerable exposure to animal sources. Evidence suggests that B. bronchiseptica may be rarely encountered as a commensal or colonizer of the respiratory tract of humans and rarely in association with infection. When found as a probable pathogen, most infections have been respiratory tract in origin and have occurred in severely compromised hosts. PMID:1889042

  13. A simplified sequence-based identification scheme for Bordetella reveals several putative novel species.

    PubMed

    Spilker, Theodore; Leber, Amy L; Marcon, Mario J; Newton, Duane W; Darrah, Rebecca; Vandamme, Peter; Lipuma, John J

    2014-02-01

    The differentiation of Bordetella species, particularly those causing human infection, is problematic. We found that sequence analysis of an internal fragment of nrdA allowed differentiation of the currently named Bordetella species. Analysis of 107 "Bordetella" isolates recovered almost exclusively from human respiratory tract specimens identified several putative novel species.

  14. Role of a Putative Polysaccharide Locus in Bordetella Biofilm Development▿

    PubMed Central

    Parise, Gina; Mishra, Meenu; Itoh, Yoshikane; Romeo, Tony; Deora, Rajendar

    2007-01-01

    Bordetellae are gram-negative bacteria that colonize the respiratory tracts of animals and humans. We and others have recently shown that these bacteria are capable of living as sessile communities known as biofilms on a number of abiotic surfaces. During the biofilm mode of existence, bacteria produce one or more extracellular polymeric substances that function, in part, to hold the cells together and to a surface. There is little information on either the constituents of the biofilm matrix or the genetic basis of biofilm development by Bordetella spp. By utilizing immunoblot assays and by enzymatic hydrolysis using dispersin B (DspB), a glycosyl hydrolase that specifically cleaves the polysaccharide poly-β-1,6-N-acetyl-d-glucosamine (poly-β-1,6-GlcNAc), we provide evidence for the production of poly-β-1,6-GlcNAc by various Bordetella species (Bordetella bronchiseptica, B. pertussis, and B. parapertussis) and its role in their biofilm development. We have investigated the role of a Bordetella locus, here designated bpsABCD, in biofilm formation. The bps (Bordetella polysaccharide) locus is homologous to several bacterial loci that are required for the production of poly-β-1,6-GlcNAc and have been implicated in bacterial biofilm formation. By utilizing multiple microscopic techniques to analyze biofilm formation under both static and hydrodynamic conditions, we demonstrate that the bps locus, although not essential at the initial stages of biofilm formation, contributes to the stability and the maintenance of the complex architecture of Bordetella biofilms. PMID:17114249

  15. VIRULENCE AND CITRULLINE UREIDASE ACTIVITY OF PASTEURELLA TULARENSIS12

    PubMed Central

    Marchette, Nyven J.; Nicholes, Paul S.

    1961-01-01

    Marchette, Nyven J. (University of Utah, Salt Lake City), and Paul S. Nicholes. Virulence and citrulline ureidase activity of Pasteurella tularensis. J. Bacteriol. 82:26–32. 1961.—The presence of a citrulline ureidase system in Pasteurella tularensis strains of high virulence, and its absence in avirulent strains and strains of low virulence was confirmed. The presence of this system, however, was shown to be not directly related to virulence. The only wild strain of P. tularensis tested that lacked a citrulline ureidase system was isolated from a rodent. All the strains, isolated from rabbits, rabbit ticks, a human being, and a horse, that were tested possessed this system. The existence of two North American varieties of P. tularensis was postulated on the basis of virulence and citrulline ureidase activity. PMID:13766500

  16. Virulence and citrulline ureidase activity of Pasteurella tularensis.

    PubMed

    MARCHETTE, N J; NICHOLES, P S

    1961-07-01

    Marchette, Nyven J. (University of Utah, Salt Lake City), and Paul S. Nicholes. Virulence and citrulline ureidase activity of Pasteurella tularensis. J. Bacteriol. 82:26-32. 1961.-The presence of a citrulline ureidase system in Pasteurella tularensis strains of high virulence, and its absence in avirulent strains and strains of low virulence was confirmed. The presence of this system, however, was shown to be not directly related to virulence. The only wild strain of P. tularensis tested that lacked a citrulline ureidase system was isolated from a rodent. All the strains, isolated from rabbits, rabbit ticks, a human being, and a horse, that were tested possessed this system. The existence of two North American varieties of P. tularensis was postulated on the basis of virulence and citrulline ureidase activity.

  17. Systemic infection by Pasteurella canis biotype 1 in newborn puppies.

    PubMed

    de la Puente Redondo, V A; Gutiérrez Martín, C B; García del Blanco, N; Antolín Ayala, M I; Alonso Alonso, P; Rodríguez Ferri, E F

    2000-01-01

    Pasteurella canis biotype 1, usually associated with the oral cavity of dogs and cats, or with human wound infections following dog bites, was isolated from newborn puppies with a fatal systemic infection. The identity of P. canis was confirmed by arbitrarily primed polymerase chain reaction and the organism was susceptible to all the penicillins, cephalosporins, tetracyclines and fluoroquinolones tested and to most of the aminoglycosides tested. This represents the first report of systemic pasteurellosis caused by P. canis in dogs.

  18. [Simultaneous screening method for Bordetella species by conventional PCR assay].

    PubMed

    Akiyama, Yumi; Saito, Etsuko; Enomoto, Miki; Tsuji, Hidetaka; Chikahira, Masatsugu; Yoshida, Masashi

    2013-11-01

    A simultaneous screening method using conventional PCR was developed for the detection and discrimination of Bordetella pertussis, Bordetella parapertussis, and Bordetella holmesii. A formulated multiprex method employing 4 kinds of paired primers on amplification of 4 corresponding different insertion sequences (IS481, IS1001, IS1002 and hIS1001) enabled rapid screening and identification. The detection limits of each DNA extracted from 3 kinds of Bordetella species were 5fg/microL for each. Obscure existences of B. pertussis and B. holmesii at low levels were confirmed with the LAMP method. This multiplex assay was applied to the clinical specimens obtained from patients with pertussis-like symptoms at sentinel clinics under the epidemiological surveillance of infectious diseases of Hyogo prefecture in FY2012. Among 42 nasopharyngeal swabs, B. pertussis was detected from 12 samples including 8 samples collected at outbreak in nursery school. The use of this method for the surveillance of infectious agents enabled us to search for 3 kinds of Bordetella species at once with low costs.

  19. Genetic Basis for Lipopolysaccharide O-Antigen Biosynthesis in Bordetellae

    PubMed Central

    Preston, Andrew; Allen, Andrew G.; Cadisch, Joanna; Thomas, Richard; Stevens, Kim; Churcher, Carol M.; Badcock, K. L.; Parkhill, Julian; Barrell, Bart; Maskell, Duncan J.

    1999-01-01

    Bordetella bronchiseptica and Bordetella parapertussis express a surface polysaccharide, attached to a lipopolysaccharide, which has been called O antigen. This structure is absent from Bordetella pertussis. We report the identification of a large genetic locus in B. bronchiseptica and B. parapertussis that is required for O-antigen biosynthesis. The locus is replaced by an insertion sequence in B. pertussis, explaining the lack of O-antigen biosynthesis in this species. The DNA sequence of the B. bronchiseptica locus has been determined and the presence of 21 open reading frames has been revealed. We have ascribed putative functions to many of these open reading frames based on database searches. Mutations in the locus in B. bronchiseptica and B. parapertussis prevent O-antigen biosynthesis and provide tools for the study of the role of O antigen in infections caused by these bacteria. PMID:10417135

  20. Adenylate cyclase toxin (ACT) from Bordetella hinzii: characterization and differences from ACT of Bordetella pertussis.

    PubMed

    Donato, Gina M; Hsia, Hung-Lun J; Green, Candace S; Hewlett, Erik L

    2005-11-01

    Bordetella hinzii is a commensal respiratory microorganism in poultry but is increasingly being recognized as an opportunistic pathogen in immunocompromised humans. Although associated with a variety of disease states, practically nothing is known about the mechanisms employed by this bacterium. In this study, we show by DNA sequencing and reverse transcription-PCR that both commensal and clinical strains of B. hinzii possess and transcriptionally express cyaA, the gene encoding adenylate cyclase toxin (ACT) in other pathogenic Bordetella species. By Western blotting, we also found that B. hinzii produces full-length ACT protein in quantities that are comparable to those made by B. pertussis. In contrast to B. pertussis ACT, however, ACT from B. hinzii is less extractable from whole bacteria, nonhemolytic, has a 50-fold reduction in adenylate cyclase activity, and is unable to elevate cyclic AMP levels in host macrophages (nontoxic). The decrease in enzymatic activity is attributable, at least in part, to a decreased binding affinity of B. hinzii ACT for calmodulin, the eukaryotic activator of B. pertussis ACT. In addition, we demonstrate that the lack of intoxication by B. hinzii ACT may be due to the absence of expression of cyaC, the gene encoding the accessory protein required for the acylation of B. pertussis ACT. These results demonstrate the expression of ACT by B. hinzii and represent the first characterization of a potential virulence factor of this organism.

  1. Towards Improved Accuracy of Bordetella pertussis Nucleic Acid Amplification Tests

    PubMed Central

    2012-01-01

    In many clinical microbiology laboratories, nucleic acid amplification tests such as PCR have become the routine methods for the diagnosis of pertussis. While PCR has greatly increased the ability of laboratories to detect Bordetella pertussis infections, it has also been associated with false-positive results that can, given the tendency of B. pertussis to cause outbreaks, result in unnecessary and costly control measures. The species specificity of Bordetella gene targets and their number of copies per genome greatly impact the performance characteristics of nucleic acid amplification tests for B. pertussis. It is crucial that laboratorians recognize these characteristics, to limit false-positive test results and prevent pseudo-outbreaks. PMID:22442315

  2. Aerosol infection of mice with Bordetella pertussis.

    PubMed Central

    Sato, Y; Izumiya, K; Sato, H; Cowell, J L; Manclark, C R

    1980-01-01

    Aerosol inhalation of Bordetella pertussis Tohama phase I resulted in a reproducible and uniform infection of mice (strain DDY or ICR). Mice in groups of 10 exposed for 30 min to aerosols generated from bacterial suspensions of 10(9) and 10(10) organisms per ml resulted in mean bacterial counts of 2.3 (+/- 0.3) X 10(4) and 1.0 (+/- 0.3) X 10(5) colony-forming units, respectively, in the lung of each animal. Subsequent studies using a 30-min aerosol inoculation of ICR mice with 2 X 10(9) bacterial cells per ml showed: (i) B. pertussis cells reached a maximum of about 10(7) colony-forming units per lung 14 days after inhalation. (ii) Deaths (10 to 100%, depending on mouse age) occurred 10 to 14 days after exposure. (iii) The lung weight and the leukocyte count increased from basal values of 100 mg and 10(4) leukocytes per mm3 to a plateau of 950 mg and 1.95 X 10(5) leukocytes per mm3, respectively, 14 days after challenge. (iv) There was a significantly reduced rate of body weight gain by infected mice compared to noninfected mice. (v) With mortality as the criterion for disease, susceptibility varied with the age of mice as follows: 10 days old greater than 18 greater than 28 greater than 49. (vi) Bacteria were associated with ciliated respiratory epithelial cells by scanning electron microscopy. Images Fig. 4 PMID:6249758

  3. Bordetella pertussis fimbriae (Fim): relevance for vaccines.

    PubMed

    Gorringe, Andrew R; Vaughan, Thomas E

    2014-10-01

    Bordetella pertussis produces two serologically distinct fimbriae, Fim2 and Fim3. Expression of these antigens is governed by the BvgA/S system and by the length of a poly(C) tract in the promoter of each gene. Fim2 and Fim3 are important antigens for whole cell pertussis vaccines as clinical trials have shown an association of anti-fimbriae antibody-mediated agglutination and protection. The current five component acellular pertussis vaccine contains co-purified Fim2/3 and provided good efficacy in clinical trials with the anti-Fim antibody response correlating with protection when pre and post exposure antibody levels were analysed. The predominant serotype of B. pertussis isolates has changed over time in most countries but it is not understood whether this is vaccine-driven or whether serotype is linked to the prevailing predominant genotype. Recent studies have shown that both Fim2 and Fim3 are expressed during infection and that Fim2 is more immunogenic than Fim3 in the acellular vaccine.

  4. Bordetella species in children with cystic fibrosis: what do we know? The role in acute exacerbations and chronic course.

    PubMed

    Bos, A C; Beemsterboer, P; Wolfs, T F W; Versteegh, F G A; Arets, H G M

    2011-09-01

    Despite vaccination, pertussis is still endemic in the Netherlands. A literature search was performed to verify what is known about the role of Bordetella species in children with cystic fibrosis, with regard to the incidence of Bordetella infections, the involvement in pulmonary exacerbations and the influence on chronic course. Little is known about the frequency of Bordetella infections and the involvement of Bordetella species both in relation to the chronic course of cystic fibrosis and to pulmonary exacerbations. Since it is difficult to detect Bordetella species in cultures and few sputum cultures investigated have been obtained during an exacerbation, it is likely that the frequency of Bordetella species in CF patients is underestimated. Identification of Bordetella species in these patients may have serious consequences for the treatment of exacerbations in CF. Future research investigating the role of Bordetella species in cystic fibrosis should use specific techniques to detect Bordetella in cultures.

  5. Misidentification of Bordetella bronchiseptica as Bordetella pertussis using a newly described real-time PCR targeting the pertactin gene.

    PubMed

    Register, Karen B; Nicholson, Tracy L

    2007-12-01

    Recently, a real-time PCR (RT-PCR) assay based on sequence from the gene for pertactin was proposed for identification of Bordetella pertussis. Here, it is reported that the B. pertussis pertactin gene sequence for the region that encompasses the RT-PCR probe and primers is nearly identical to that of many Bordetella bronchiseptica strains of human and avian origin. Additionally, it is demonstrated that such strains are erroneously identified as B. pertussis using the RT-PCR assay. These data suggest that the use of the assay without confirmatory testing may result in erroneous identification of a significant proportion of human isolates of B. bronchiseptica as B. pertussis.

  6. Septic Arthritis and Osteomyelitis Due to Bordetella petrii

    PubMed Central

    Bankowski, Matthew J.; Pien, Francis D.

    2014-01-01

    A case of Bordetella petrii septic arthritis and osteomyelitis in an elbow resulted from a dirt bike accident in Hawaii. Two months of intravenous antibiotics and repeated surgeries were required to cure this infection. Our case, and literature review, suggests that extended-spectrum penicillins, tetracycline, and trimethoprim-sulfamethoxazole are good treatment options. PMID:25540393

  7. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  8. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  9. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  10. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  11. 21 CFR 866.3065 - Bordetella spp. serological reagents.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Bordetella spp. serological reagents. 866.3065 Section 866.3065 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065...

  12. Significant Decrease in Pertactin-Deficient Bordetella pertussis Isolates, Japan

    PubMed Central

    Miyaji, Yusuke; Otsuka, Nao; Arakawa, Yoshichika; Shibayama, Keigo; Kamachi, Kazunari

    2017-01-01

    Prevalence of pertactin-lacking Bordetella pertussis isolates has been observed worldwide. In Japan, however, we found that the frequency of pertactin-deficient isolates in 2014–2016 (8%) was significantly lower than the frequency in 2005–2007 (41%), 2008–2010 (35%), and 2011–2013 (25%). This reduction was closely associated with changes in genotypes. PMID:28322702

  13. Bordetella pseudohinzii spp. nov. infects C57Bl6 mice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clinical studies rely heavily on mouse models of infection. Precise identification and control of contaminating pathogens that circulate in mouse colonies is an important task. Over the past decade, there have been several reports documenting the isolation of Bordetella spp. from purported pathog...

  14. Comparison of Ribotyping and MLST for Genotyping Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica is a widespread bacterial pathogen that infects a variety of domesticated and wild animals. Multilocus sequence typing (MLST) and PvuII ribotyping have proven useful to distinguish among strains of B. bronchiseptica. Both are highly discriminatory and have been used to in...

  15. Strain-specific virulence of Bordetella hinzii in turkeys

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella hinzii is commonly acquired from the respiratory tract of diseased poultry but regarded as nonpathogenic in avian hosts. Recently, it was recognized that some previously used isolates were misidentified at the time of their acquisition as B. avium, B. avium-like or Alcaligenes faecalis ty...

  16. Identification of a CO2 responsive regulon in Bordetella

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sensing the environment allows pathogenic bacteria to coordinately regulate gene expression to maximize survival within or outside of a host. Here we show that Bordetella species regulate virulence factor expression in response to carbon dioxide levels that mimic in vivo conditions. We found strains...

  17. Transmission of Bordetella holmesii during pertussis outbreak, Japan.

    PubMed

    Kamiya, Hajime; Otsuka, Nao; Ando, Yuka; Odaira, Fumito; Yoshino, Shuji; Kawano, Kimiko; Takahashi, Hirokazu; Nishida, Toshihide; Hidaka, Yoshio; Toyoizumi-Ajisaka, Hiromi; Shibayama, Keigo; Kamachi, Kazunari; Sunagawa, Tomimasa; Taniguchi, Kiyosu; Okabe, Nobuhiko

    2012-07-01

    We describe the epidemiology of a pertussis outbreak in Japan in 2010-2011 and Bordetella holmesii transmission. Six patients were infected; 4 patients were students and a teacher at the same junior high school. Epidemiologic links were found between 5 patients. B. holmesii may have been transmitted from person to person.

  18. Identification of a CO2 responsive regulon in Bordetella.

    PubMed

    Hester, Sara E; Lui, Minghsun; Nicholson, Tracy; Nowacki, Daryl; Harvill, Eric T

    2012-01-01

    Sensing the environment allows pathogenic bacteria to coordinately regulate gene expression to maximize survival within or outside of a host. Here we show that Bordetella species regulate virulence factor expression in response to carbon dioxide levels that mimic in vivo conditions within the respiratory tract. We found strains of Bordetella bronchiseptica that did not produce adenylate cyclase toxin (ACT) when grown in liquid or solid media with ambient air aeration, but produced ACT and additional antigens when grown in air supplemented to 5% CO(2). Transcriptome analysis and quantitative real time-PCR analysis revealed that strain 761, as well as strain RB50, increased transcription of genes encoding ACT, filamentous hemagglutinin (FHA), pertactin, fimbriae and the type III secretion system in 5% CO(2) conditions, relative to ambient air. Furthermore, transcription of cyaA and fhaB in response to 5% CO(2) was increased even in the absence of BvgS. In vitro analysis also revealed increases in cytotoxicity and adherence when strains were grown in 5% CO(2). The human pathogens B. pertussis and B. parapertussis also increased transcription of several virulence factors when grown in 5% CO(2), indicating that this response is conserved among the classical bordetellae. Together, our data indicate that Bordetella species can sense and respond to physiologically relevant changes in CO(2) concentrations by regulating virulence factors important for colonization, persistence and evasion of the host immune response.

  19. Bordetella pertussis Strain Lacking Pertactin and Pertussis Toxin.

    PubMed

    Williams, Margaret M; Sen, Kathryn; Weigand, Michael R; Skoff, Tami H; Cunningham, Victoria A; Halse, Tanya A; Tondella, M Lucia

    2016-02-01

    A Bordetella pertussis strain lacking 2 acellular vaccine immunogens, pertussis toxin and pertactin, was isolated from an unvaccinated infant in New York State in 2013. Comparison with a French strain that was pertussis toxin-deficient, pertactin wild-type showed that the strains carry the same 28-kb deletion in similar genomes.

  20. Septic arthritis and osteomyelitis due to Bordetella petrii.

    PubMed

    Nogi, Masayuki; Bankowski, Matthew J; Pien, Francis D

    2015-03-01

    A case of Bordetella petrii septic arthritis and osteomyelitis in an elbow resulted from a dirt bike accident in Hawaii. Two months of intravenous antibiotics and repeated surgeries were required to cure this infection. Our case, and literature review, suggests that extended-spectrum penicillins, tetracycline, and trimethoprim-sulfamethoxazole are good treatment options.

  1. Bordetella bronchiseptica pneumonia in a patient with AIDS.

    PubMed Central

    de la Fuente, J; Albo, C; Rodríguez, A; Sopeña, B; Martínez, C

    1994-01-01

    Bordetella bronchiseptica is recognised as a respiratory tract pathogen in many mammalian species, but has rarely been implicated in human infection. A case is reported of pneumonia caused by B bronchiseptica in a patient suffering from acquired immunodeficiency syndrome (AIDS). Images PMID:8066571

  2. Opportunistic Pulmonary Bordetella hinzii Infection after Avian Exposure

    PubMed Central

    Dupin, Clarisse; Bénézit, François; Goret, Julien; Piau, Caroline; Jouneau, Stéphane; Guillot, Sophie; Mégraud, Francis; Kayal, Samer; Desrues, Benoit; Le Coustumier, Alain; Guiso, Nicole

    2015-01-01

    We report 2 cases of pulmonary Bordetella hinzii infection in immunodeficient patients. One of these rare cases demonstrated the potential transmission of the bacteria from an avian reservoir through occupational exposure and its persistence in humans. We establish bacteriologic management of these infections and suggest therapeutic options if needed. PMID:26584467

  3. Two Distinct Episodes Of Whooping Cough Caused By Consecutive Bordetella Pertussis And Bordetella Parapertussis Infections In A Fully Immunized Healthy Boy.

    PubMed

    Heininger, Ulrich; Schlassa, Detlef

    2016-11-01

    We describe a 5-year-old, fully immunized boy with polymerase chain reaction-proven consecutive Bordetella pertussis and Bordetella parapertussis infections causing typical whooping cough at the age of 2 and 5 years, respectively. Neither pertussis immunization nor disease provides reliable immunity against further episodes of whooping cough.

  4. Development of real-time PCR assay for differential detection of Bordetella bronchiseptica and Bordetella parapertussis.

    PubMed

    Tizolova, Anette; Brun, Delphine; Guiso, Nicole; Guillot, Sophie

    2014-04-01

    Bordetella parapertussis is a causative agent of whooping cough in humans, and B. bronchiseptica is causing wide variety of respiratory infections in mammals, including humans. Specific diagnostic tests are not currently available. Our first objective was to develop a real-time PCR test for the specific detection of B. bronchiseptica based on the previously described end-point PCR, targeting an intergenomic sequence of the fla gene locus, but it has not been reached. However, there is cross-reactivity between B. parapertussis and B. bronchiseptica. Therefore, the targeted region of several clinical isolates of both species was sequenced, and alignment of the sequences allowed the development of a 2-step real-time PCR assay. The first PCR assay detected the DNA of all clinical isolates of both B. bronchiseptica and B. parapertussis tested. The second PCR assay detected only the DNA of B. parapertussis clinical isolates, thereby allowing discrimination between B. parapertussis and B. bronchiseptica.

  5. Serological titers to bovine herpesvirus 1, bovine viral diarrhea virus, parainfluenza 3 virus, bovine respiratory syncytial virus and Pasteurella haemolytica in feedlot calves with respiratory disease: associations with bacteriological and pulmonary cytological variables.

    PubMed Central

    Allen, J W; Viel, L; Bateman, K G; Nagy, E; Røsendal, S; Shewen, P E

    1992-01-01

    Acute and convalescent serum samples were taken from 59 calves with signs of respiratory disease (cases) and 60 clinically normal animals (controls) during their first month in the feedlot. Sera were analyzed for antibodies to bovine parainfluenza 3 (PI3) virus by hemagglutination inhibition, to bovine viral diarrhea (BVD) virus, bovine respiratory syncytial (BRS) virus and bovine herpesvirus 1 (BHV1) by virus neutralization, and to Pasteurella haemolytica by indirect agglutination (PhIA) and cytotoxin neutralization (PhCN) tests. There was minimal evidence of serological activity to BHV1. Serological activity to the other agents occurred commonly and the prevalence of acute titers and their mean values was similar in case and control groups. Mean convalescent PI3 and P. haemolytica (PhIA) titers were higher in controls than cases (p < 0.01) but, otherwise, convalescent titers did not differ between groups. The incidence of seroconversion was similar in both groups for all agents except for PI3 virus which was more frequent in controls than cases (p < 0.0001). There was a positive association between PhIA and CN seroconversion and isolation of P. haemolytica from bronchoalveolar lavage (BAL) fluid (p < 0.1). The measure of agreement (kappa) between seroconversion with the P. haemolytica PhIA and PhCN tests was 0.51. Bacteriological and cytological evaluations of the respiratory tract were made using BAL. No associations were evident between serological titers and pulmonary cytology. A multivariate logistic analysis was used to evaluate associations between disease status and serological, bacteriological and cytological data. Cases were positively associated with the presence of neutrophils and Pasteurella multocida in BAL fluid and negatively associated with PI3 virus and PhIA seroconversion. PMID:1335831

  6. Misidentification of Bordetella bronchiseptica as Bordetella pertussis using a Newly Described RT-PCR Targeting the Pertactin Gene

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recently a real-time PCR (RT-PCT) assay based on sequence from the gene for pertactin was proposed for identification of Bordetella pertussis. Here we report that the B. pertussis pertactin gene sequence for the region encompassing the RT-PCR probe and primers is nearly identical to that of many B....

  7. Molecular Pathogenesis, Epidemiology, and Clinical Manifestations of Respiratory Infections Due to Bordetella pertussis and Other Bordetella Subspecies

    PubMed Central

    Mattoo, Seema; Cherry, James D.

    2005-01-01

    Bordetella respiratory infections are common in people (B. pertussis) and in animals (B. bronchiseptica). During the last two decades, much has been learned about the virulence determinants, pathogenesis, and immunity of Bordetella. Clinically, the full spectrum of disease due to B. pertussis infection is now understood, and infections in adolescents and adults are recognized as the reservoir for cyclic outbreaks of disease. DTaP vaccines, which are less reactogenic than DTP vaccines, are now in general use in many developed countries, and it is expected that the expansion of their use to adolescents and adults will have a significant impact on reducing pertussis and perhaps decrease the circulation of B. pertussis. Future studies should seek to determine the cause of the unique cough which is associated with Bordetella respiratory infections. It is also hoped that data gathered from molecular Bordetella research will lead to a new generation of DTaP vaccines which provide greater efficacy than is provided by today's vaccines. PMID:15831828

  8. Molecular Analysis of Tetracycline Resistance in Pasteurella aerogenes

    PubMed Central

    Kehrenberg, Corinna; Schwarz, Stefan

    2001-01-01

    Tetracycline-resistant Pasteurella aerogenes isolates obtained from the intestinal tract of swine were investigated for their tet genes by PCR analysis and hybridization experiments. In contrast to Pasteurella isolates from the respiratory tract, tet(H) genes were detected in the chromosomal DNA of only 2 of the 24 isolates, one of which also carried two copies of a tet(B) gene. All other P. aerogenes isolates carried tet(B) genes, which are the predominant tet genes among Enterobacteriaceae. A single isolate harbored a tet(B) gene as part of a truncated Tn10 element on the 4.8-kb plasmid pPAT2. Comparative analysis of the pPAT2 sequence suggested that the Tn10 relic on plasmid pPAT2 is the result of several illegitimate recombination events. The remaining 21 P. aerogenes isolates carried one or two copies of the tet(B) gene in their chromosomal DNA. In the majority of the cases, these tet(B) genes were associated with copies of Tn10 as confirmed by their SfuI and BamHI hybridization patterns. No correlation between the number of tet gene copies and the MICs of tetracycline, doxycyline and minocycline was observed. PMID:11557485

  9. Genetic Variation of Bordetella pertussis in Austria.

    PubMed

    Wagner, Birgit; Melzer, Helen; Freymüller, Georg; Stumvoll, Sabine; Rendi-Wagner, Pamela; Paulke-Korinek, Maria; Repa, Andreas; Mooi, Frits R; Kollaritsch, Herwig; Mittermayer, Helmut; Kessler, Harald H; Stanek, Gerold; Steinborn, Ralf; Duchêne, Michael; Wiedermann, Ursula

    2015-01-01

    In Austria, vaccination coverage against Bordetella pertussis infections during infancy is estimated at around 90%. Within the last years, however, the number of pertussis cases has increased steadily, not only in children but also in adolescents and adults, indicating both insufficient herd immunity and vaccine coverage. Waning immunity in the host and/or adaptation of the bacterium to the immunised hosts could contribute to the observed re-emergence of pertussis. In this study we therefore addressed the genetic variability in B. pertussis strains from several Austrian cities. Between the years 2002 and 2008, 110 samples were collected from Vienna (n = 32), Linz (n = 63) and Graz (n = 15) by nasopharyngeal swabs. DNA was extracted from the swabs, and bacterial sequence polymorphisms were examined by MLVA (multiple-locus variable number of tandem repeat analysis) (n = 77), by PCR amplification and conventional Sanger sequencing of the polymorphic regions of the prn (pertactin) gene (n = 110), and by amplification refractory mutation system quantitative PCR (ARMS-qPCR) (n = 110) to directly address polymorphisms in the genes encoding two pertussis toxin subunits (ptxA and ptxB), a fimbrial adhesin (fimD), tracheal colonisation factor (tcfA), and the virulence sensor protein (bvgS). Finally, the ptxP promoter region was screened by ARMS-qPCR for the presence of the ptxP3 allele, which has been associated with elevated production of pertussis toxin. The MLVA analysis revealed the highest level of polymorphisms with an absence of MLVA Type 29, which is found outside Austria. Only Prn subtypes Prn1/7, Prn2 and Prn3 were found with a predominance of the non-vaccine type Prn2. The analysis of the ptxA, ptxB, fimD, tcfA and bvgS polymorphisms showed a genotype mixed between the vaccine strain Tohama I and a clinical isolate from 2006 (L517). The major part of the samples (93%) displayed the ptxP3 allele. The consequences for the vaccination strategy are discussed.

  10. Genetic Variation of Bordetella pertussis in Austria

    PubMed Central

    Wagner, Birgit; Melzer, Helen; Freymüller, Georg; Stumvoll, Sabine; Rendi-Wagner, Pamela; Paulke-Korinek, Maria; Repa, Andreas; Mooi, Frits R.; Kollaritsch, Herwig; Kessler, Harald H.; Stanek, Gerold; Steinborn, Ralf; Duchêne, Michael; Wiedermann, Ursula

    2015-01-01

    In Austria, vaccination coverage against Bordetella pertussis infections during infancy is estimated at around 90%. Within the last years, however, the number of pertussis cases has increased steadily, not only in children but also in adolescents and adults, indicating both insufficient herd immunity and vaccine coverage. Waning immunity in the host and/or adaptation of the bacterium to the immunised hosts could contribute to the observed re-emergence of pertussis. In this study we therefore addressed the genetic variability in B. pertussis strains from several Austrian cities. Between the years 2002 and 2008, 110 samples were collected from Vienna (n = 32), Linz (n = 63) and Graz (n = 15) by nasopharyngeal swabs. DNA was extracted from the swabs, and bacterial sequence polymorphisms were examined by MLVA (multiple-locus variable number of tandem repeat analysis) (n = 77), by PCR amplification and conventional Sanger sequencing of the polymorphic regions of the prn (pertactin) gene (n = 110), and by amplification refractory mutation system quantitative PCR (ARMS-qPCR) (n = 110) to directly address polymorphisms in the genes encoding two pertussis toxin subunits (ptxA and ptxB), a fimbrial adhesin (fimD), tracheal colonisation factor (tcfA), and the virulence sensor protein (bvgS). Finally, the ptxP promoter region was screened by ARMS-qPCR for the presence of the ptxP3 allele, which has been associated with elevated production of pertussis toxin. The MLVA analysis revealed the highest level of polymorphisms with an absence of MLVA Type 29, which is found outside Austria. Only Prn subtypes Prn1/7, Prn2 and Prn3 were found with a predominance of the non-vaccine type Prn2. The analysis of the ptxA, ptxB, fimD, tcfA and bvgS polymorphisms showed a genotype mixed between the vaccine strain Tohama I and a clinical isolate from 2006 (L517). The major part of the samples (93%) displayed the ptxP3 allele. The consequences for the vaccination strategy are discussed. PMID

  11. Evaluation of Four Commercial Real-Time PCR Assays for Detection of Bordetella spp. in Nasopharyngeal Aspirates ▿

    PubMed Central

    Lanotte, Philippe; Plouzeau, Chloé; Burucoa, Christophe; Grélaud, Carole; Guillot, Sophie; Guiso, Nicole; Garnier, Fabien

    2011-01-01

    We evaluated the performances of 4 commercial real-time PCR kits for Bordetella pertussis IS481 sequence detection in nasopharyngeal aspirates by comparison with an in-house real-time PCR assay. Among them, the Simplexa Bordetella pertussis/parapertussis assay (Focus Diagnostics), the SmartCycler Bordetella pertussis/parapertussis assay (Cepheid), and Bordetella R-gene (Argene) present sensitivities over 90%. One kit proved unsuitable for routine clinical use. PMID:21918018

  12. Development of a PCR for identification of Bordetella hinzii.

    PubMed

    Register, Karen B

    2013-06-01

    Bordetella hinzii infects primarily poultry and immunocompromised humans. It is closely related to the etiologic agent of turkey coryza, Bordetella avium. Distinguishing between B. avium and B. hinzii is difficult, and there is no method for identification of B. hinzii suitable for use by diagnostic laboratories. This report details the development of a B. hinzii-specific PCR targeting the ompA gene. Assay sensitivity is 100% based on analysis of 48 B. hinzii isolates from diverse geographic locations representing all known ribotypes. Evaluation of 71 isolates of B. avium and 20 other bacterial isolates from poultry, comprising gram-negative and gram-positive commensals and pathogens of nine genera, demonstrated an assay specificity of 100%. The ompA PCR is a rapid, reliable, and accurate method for identification of B. hinzii and provides a valuable new tool for veterinary diagnostic laboratories investigating poultry respiratory disease outbreaks.

  13. Epidemiology of whooping cough & typing of Bordetella pertussis.

    PubMed

    Hegerle, Nicolas; Guiso, Nicole

    2013-11-01

    Bordetella pertussis is a Gram-negative human-restricted bacterium that evolved from the broad-range mammalian pathogen, Bordetella bronchiseptica. It causes whooping cough or pertussis in humans, which is the most prevalent vaccine-preventable disease worldwide. The introduction of the pertussis whole-cell vaccination for young children, followed by the introduction of the pertussis acellular vaccination (along with booster vaccination) for older age groups, has affected the bacterial population and epidemiology of the disease. B. pertussis is relatively monomorphic worldwide, but nevertheless, different countries are facing different epidemiological evolutions of the disease. Although it is tempting to link vaccine-driven phenotypic and genotypic evolution of the bacterium to epidemiology, many other factors should be considered and surveillance needs to continue, in addition to studies investigating the impact of current clinical isolates on vaccine efficacy.

  14. PURIFICATION OF HISTAMINE SENSITIZING FACTOR OF BORDETELLA PERTUSSIS.

    DTIC Science & Technology

    Histamine Sensitizing Factor ( HSF ) of Bordetella pertussis was studied. To improve the cultivation method for HSF preparation, growth conditions of...maintenance and surface bulk culture was devised. Intracellular localization of HSF and lethal toxin were studied. HSF was mainly localized in the cell...wall prepared by Mickle disintegration, and toxin was found in protoplasm Ribosomes prepared from sonicate contained considerable amount of HSF and

  15. The Bordetella bhu Locus Is Required for Heme Iron Utilization

    PubMed Central

    Vanderpool, Carin K.; Armstrong, Sandra K.

    2001-01-01

    Bordetella pertussis and Bordetella bronchiseptica are capable of obtaining iron from hemin and hemoglobin. Genes encoding a putative bacterial heme iron acquisition system (bhu, for Bordetella heme utilization) were identified in a B. pertussis genomic sequence database, and the corresponding DNA was isolated from a virulent strain of B. pertussis. A B. pertussis bhuR mutant, predicted to lack the heme outer membrane receptor, was generated by allelic exchange. In contrast to the wild-type strain, bhuR mutant PM5 was incapable of acquiring iron from hemin and hemoglobin; genetic complementation of PM5 with the cloned bhuRSTUV genes restored heme utilization to wild-type levels. In parallel studies, B. bronchiseptica bhu sequences were also identified and a B. bronchiseptica bhuR mutant was constructed and confirmed to be defective in heme iron acquisition. The wild-type B. bronchiseptica parent strain grown under low-iron conditions produced the presumptive BhuR protein, which was absent in the bhuR mutant. Furthermore, production of BhuR by iron-starved B. bronchiseptica was markedly enhanced by culture in hemin-supplemented medium, suggesting that these organisms sense and respond to heme in the environment. Analysis of the genetic region upstream of the bhu cluster identified open reading frames predicted to encode homologs of the Escherichia coli ferric citrate uptake regulators FecI and FecR. These putative Bordetella regulators may mediate heme-responsive positive transcriptional control of the bhu genes. PMID:11418569

  16. Bordetella biofilms: a lifestyle leading to persistent infections.

    PubMed

    Cattelan, Natalia; Dubey, Purnima; Arnal, Laura; Yantorno, Osvaldo M; Deora, Rajendar

    2016-02-01

    Bordetella bronchiseptica and B. pertussis are Gram-negative bacteria that cause respiratory diseases in animals and humans. The current incidence of whooping cough or pertussis caused by B. pertussis has reached levels not observed since the 1950s. Although pertussis is traditionally known as an acute childhood disease, it has recently resurged in vaccinated adolescents and adults. These individuals often become silent carriers, facilitating bacterial circulation and transmission. Similarly, vaccinated and non-vaccinated animals continue to be carriers of B. bronchiseptica and shed bacteria resulting in disease outbreaks. The persistence mechanisms of these bacteria remain poorly characterized. It has been proposed that adoption of a biofilm lifestyle allows persistent colonization of the mammalian respiratory tract. The history of Bordetella biofilm research is only a decade long and there is no single review article that has exclusively focused on this area. We systematically discuss the role of Bordetella factors in biofilm development in vitro and in the mouse respiratory tract. We further outline the implications of biofilms to bacterial persistence and transmission in humans and for the design of new acellular pertussis vaccines.

  17. Bordetella bronchiseptica responses to physiological reactive nitrogen and oxygen stresses

    PubMed Central

    Omsland, Anders; Miranda, Katrina M.; Friedman, Richard L.; Boitano, Scott

    2008-01-01

    Bordetella bronchiseptica can establish prolonged airway infection consistent with a highly developed ability to evade mammalian host immune responses. Upon initial interaction with the host upper respiratory tract mucosa, B. bronchiseptica are subjected to antimicrobial reactive nitrogen species (RNS) and reactive oxygen species (ROS), effector molecules of the innate immune system. However, the responses of B. bronchiseptica to redox species at physiologically relevant concentrations (nM-µM) have not been investigated. Using predicted physiological concentrations of nitric oxide (NO), superoxide (O2.−) and hydrogen peroxide (H2O2) on low numbers of colony forming units (CFU) of B. bronchiseptica, all redox active species displayed dose-dependent antimicrobial activity. Susceptibility to individual redox active species was significantly increased upon introduction of a second species at sub-antimicrobial concentrations. An increased bacteriostatic activity of NO was observed relative to H2O2. The understanding of Bordetella responses to physiologically relevant levels of exogenous RNS and ROS will aid in defining the role of endogenous production of these molecules in host innate immunity against Bordetella and other respiratory pathogens. PMID:18462394

  18. Bordetella biofilms: a lifestyle leading to persistent infections

    PubMed Central

    Cattelan, Natalia; Dubey, Purnima; Arnal, Laura; Yantorno, Osvaldo M.; Deora, Rajendar

    2015-01-01

    Bordetella bronchiseptica and B. pertussis are Gram-negative bacteria that cause respiratory diseases in animals and humans. The current incidence of whooping cough or pertussis caused by B. pertussis has reached levels not observed since the 1950s. Although pertussis is traditionally known as an acute childhood disease, it has recently resurged in vaccinated adolescents and adults. These individuals often become silent carriers, facilitating bacterial circulation and transmission. Similarly, vaccinated and non-vaccinated animals continue to be carriers of B. bronchiseptica and shed bacteria resulting in disease outbreaks. The persistence mechanisms of these bacteria remain poorly characterized. It has been proposed that adoption of a biofilm lifestyle allows persistent colonization of the mammalian respiratory tract. The history of Bordetella biofilm research is only a decade long and there is no single review article that has exclusively focused on this area. We systematically discuss the role of Bordetella factors in biofilm development in vitro and in the mouse respiratory tract. We further outline the implications of biofilms to bacterial persistence and transmission in humans and for the design of new acellular pertussis vaccines. PMID:26586694

  19. Bordetella bronchialis sp. nov., Bordetella flabilis sp. nov. and Bordetella sputigena sp. nov., isolated from human respiratory specimens, and reclassification of Achromobacter sediminum Zhang et al. 2014 as Verticia sediminum gen. nov., comb. nov.

    PubMed

    Vandamme, Peter A; Peeters, Charlotte; Cnockaert, Margo; Inganäs, Elisabeth; Falsen, Enevold; Moore, Edward R B; Nunes, Olga C; Manaia, Célia M; Spilker, Theodore; LiPuma, John J

    2015-10-01

    The phenotypic and genotypic characteristics of four Bordetella hinzii-like strains from human respiratory specimens and representing nrdA gene sequence based genogroups 3, 14 and 15 were examined. In a 16S rRNA gene sequence based phylogenetic tree, the four strains consistently formed a single coherent lineage but their assignment to the genus Bordetella was equivocal. The respiratory quinone, polar lipid and fatty acid profiles generally conformed to those of species of the genus Bordetella and were characterized by the presence of ubiquinone 8, of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol and several aminolipids, and of high percentages of C16 : 0, cyclo-C17 : 0 and summed feature 2, as major chemotaxonomic marker molecules, respectively. The DNA G+C content was about 66 mol%, which corresponded with that of the high-percentage DNA G+C content genera of the family Alcaligenaceae including the genus Bordetella. DNA–DNA hybridization experiments revealed the presence of three distinct genomospecies and thus confirmed phenotypic differences as revealed by means of extensive biochemical characterization. We therefore propose to formally classify Bordetella genogroups 3, 14 and 15 as Bordetella bronchialis sp. nov. (type strain LMG 28640T = AU3182T = CCUG 56828T), Bordetella sputigena sp. nov. (type strain LMG 28641T = CCUG 56478T) and Bordetella flabilis sp. nov. (type strain LMG 28642T = AU10664T = CCUG 56827T). In addition, we propose to reclassify Achromobacter sediminum into the novel genus Verticia, as Verticia sediminum, gen. nov., comb. nov., on the basis of its unique phylogenetic position, its marine origin and its distinctive phenotypic, fatty acid and polar lipid profile.

  20. 21 CFR 520.2261b - Sulfamethazine powder.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... (Pasteurella multocida), and pullorum disease (Salmonella pullorum). (iii) Limitations. Add the required dose... (bacterial scours) (E. coli), and bacterial pneumonia (Pasteurella spp.). (iii) Limitations. Add the required... and bovine respiratory disease complex (shipping fever complex) (Pasteurella spp.),...

  1. BpsR functions as a dual activator and repressor of Bordetella gene expression

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bacteria are Gram-negative respiratory pathogens of animals, birds, and humans. A hallmark feature of some Bordetella species is their ability to efficiently survive in the respiratory tract even after vaccination. Our overall hypothesis is that establishment of colonization and persisten...

  2. Cilia-associated bacteria in fatal Bordetella bronchiseptica pneumonia of dogs and cats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less well-studied. The objectives of this study were to identify the frequency of Bordetella bronchiseptica infection in fatal cases of bronchopneumonia in dogs and cats and to compare the diag...

  3. Evaluation of 3 analyte-specific reagents for detection of Bordetella pertussis and Bordetella parapertussis in clinical specimens.

    PubMed

    Hassan, Ferdaus; Hays, Lindsay; Bell, Jeremiah; Selvarangan, Rangaraj

    2014-11-01

    The performance of 3 analyte-specific reagents (ASRs), Elitech Biosciences, EraGen Biosciences, and Focus Diagnostic, was evaluated for detection of Bordetella pertussis (BP) and Bordetella parapertussis (BPP) in nasopharyngeal swab specimens. A total of 104 frozen, leftover clinical specimens obtained from pediatric patients during 2011-2012 were included in this study. Performance was compared to the Bordetella real-time polymerase chain reaction (PCR) laboratory-developed test (LDT). The positive percent agreement for detection of BP by Elitech was 96% (95% confidence interval [CI]: 85.14-99.30); EraGen and Focus was 98% (95% CI: 87.99-99.89) in comparison to LDT PCR assay. The negative percent agreement of Elitech, EraGen, and Focus in comparison to LDT was 96% (95% CI: 85.14-99.30), 92% (95% CI: 79.89-97.41), and 96% (95% CI: 85.14-99.30), respectively. Limit of detection (LOD) for BP was 0.1 CFU/reaction by both Focus and EraGen and 1.0 CFU/reaction by Elitech. However, LOD for BPP was lower by EraGen (0.1 CFU/reaction) compared to Focus (1.0 CFU/reaction) and Elitech (1.0 CFU/reaction). These results demonstrate that all 3 ASRs tested are comparable and reliable for routine clinical diagnosis of pertussis and parapertussis.

  4. Prevalence of Bordetella holmesii and Bordetella bronchiseptica in respiratory tract samples from Belgian patients with pertussis-like symptoms by sensitive culture method and mass spectrometry.

    PubMed

    Van den Bossche, D; De Bel, A; De Smet, D; Heylen, O; Vekens, E; Vandoorslaer, K; Soetens, O; Piérard, D

    2013-01-01

    Insertion sequences IS481 and IS1001 are targets for molecular detection of respectively Bordetella pertussis and Bordetella parapertussis. There is a raising concern about specificity of these targets due to sequence similarity with Bordetella holmesii and Bordetella bronchiseptica. The likelihood of false (para)pertussis diagnoses should be correlated with the prevalence of these organisms in the respiratory tract (RT). From October 2010 until September 2011, 2,207 RT samples were submitted to the Belgian reference laboratory for pertussis diagnosis. End-point IS481/IS 1001 PCR and culture were performed for B. pertussis and B. parapertussis. We developed a sensitive culture method followed by screening with matrix-assisted laser desorption/ionisation- time of flight mass spectrometry (MALDI-TOF MS) to look for B. holmesii and B. bronchiseptica in our samples,. Only one B. bronchiseptica and no B. holmesii were detected in RT samples from Belgian patients with pertussis-like symptoms.

  5. Identification and Cloning of waaF (rfaF) from Bordetella pertussis and Use To Generate Mutants of Bordetella spp. with Deep Rough Lipopolysaccharide

    PubMed Central

    Allen, Andrew G.; Isobe, Tomoko; Maskell, Duncan J.

    1998-01-01

    A DNA locus from Bordetella pertussis capable of reconstituting lipopolysaccharide (LPS) O-antigen biosynthesis in Salmonella typhimurium SL3789 (rfaF511) has been isolated, by using selection with the antibiotic novobiocin. DNA within the locus encodes a protein with amino acid sequence similarity to heptosyltransferase II, encoded by waaF (previously rfaF) in other gram-negative bacteria. Mutation of this gene in B. pertussis, Bordetella parapertussis, and Bordetella bronchiseptica by allelic exchange generated bacteria with deep rough LPS phenotypes consistent with the proposed function of the gene as an inner core heptosyltransferase. These are the first LPS mutants generated in B. parapertussis and B. bronchiseptica and the first deep rough mutants of any of the bordetellae. PMID:9422589

  6. Effects of atmospheric ammonia on young pigs experimentally infected with Bordetella bronchiseptica

    SciTech Connect

    Drummond, J.G.; Curtis, S.E.; Meyer, R.C.; Simon, J.; Norton, H.W.

    1981-06-01

    Effects of atmospheric ammonia on performance and respiratory tract health of young pigs experimentally infected with Bordetella bronchiseptica were studied. Treatments were: (1) control, (2) Bordetella inoculation (approx 10(9) bacteria/naris) alone, (3) Bordetella inoculation plus exposure to atmospheric ammonia at 34.7 mg/m3 (50 ppm), and (4) Bordetella inoculation plus exposure to atmospheric ammonia at 69.4 mg/m3 (100 ppm). Pigs weighted 8.01 kg (av) at start of treatment. Body weight and feed disappearance were measured weekly. After 4 weeks, all pigs were killed and examined grossly, and appropriate specimens were obtained for histopathologic examination. Regression models were fitted to growth, feed disappearance, and gain-to-feed data. The growth model indicated that Bordetella-inoculated pigs gained 26% less body weight than did controls, regardless of atmospheric ammonia concentration. Bordetella inoculation, regardless of ammonia exposure, reduced feed disappearance 12% below the control rate. Treatment difference was not noted in gain/feed data. Shrunken turbinates were observed in Bordetella-inoculated pigs. Shrinkage also appeared to be related directly to ammonia concentration. Rhinitis was confirmed histopathologically, and its severity was related with atmospheric ammonia concentration, but no difference was seen in the osseous core of the turbinates.

  7. Transcriptional Analysis of the Bordetella Alcaligin Siderophore Biosynthesis Operon

    PubMed Central

    Kang, Ho Young; Armstrong, Sandra K.

    1998-01-01

    The alc gene cluster of Bordetella pertussis includes three genes, alcA, alcB, and alcC, which are involved in alcaligin siderophore biosynthesis in response to iron starvation. The production of AlcA, AlcB, and AlcC in Bordetella cells and the transcriptional organization of alcA, alcB, and alcC were investigated by using a set of three alc′-′lacZ gene fusion constructs that were contiguous with the known promoter upstream of alcA and extended to fusion junctions within each alc cistron. All three alc′-′lacZ fusions exhibited iron-repressible reporter gene expression which was abolished by deletion of the 105-bp alcA promoter-operator region. In an immunoblot analysis using a monoclonal antibody specific for β-galactosidase, the AlcA-LacZ, AlcB-LacZ, and AlcC-LacZ hybrid proteins were detected in Bordetella cells grown under iron-depleted conditions. A B. pertussis mutant in which the 105-bp alcA promoter-operator region was deleted by allelic exchange was unable to produce detectable levels of siderophore. Hybridization analysis using gene-specific probes showed that alc-specific transcript levels in the mutant were negligible compared with those of the wild-type parent. These results confirm that alcA, alcB, and alcC are cotranscribed from an iron-regulated control region immediately upstream of alcA. Transcript analysis using hybridization probes representing regions downstream of alcC demonstrated that alc transcription extends approximately 3.6 kb further downstream from the alcC coding region, suggesting the cotranscription of additional, uncharacterized alcaligin system genes. PMID:9473039

  8. Bordetella pertussis, B. parapertussis, vaccines and cycles of whooping cough.

    PubMed

    Bouchez, Valérie; Guiso, Nicole

    2015-10-01

    Whooping cough is a vaccine-preventable disease due to Bordetella pertussis and B. parapertussis. This highly contagious respiratory disease occurs through epidemic cycles every 3-5 years and vaccination did not change this frequency. Models suggest that the cyclic increase of susceptibles is linked to demographic differences and different vaccine coverage. However, differences in surveillance of the disease as well as adaptation of the agents of the disease to their human hosts and to vaccine pressure might also play an important role. These parameters are discussed in this review.

  9. Immunological characterization of the lipooligosaccharide B band of Bordetella pertussis.

    PubMed

    Martin, D; Peppler, M S; Brodeur, B R

    1992-07-01

    Two structurally and immunologically different components of Bordetella pertussis endotoxin can be visualized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining: a major A band and a faster-migrating minor B band. Certain mutant strains of B. pertussis express only the B band, while the wild-type strains produce both lipooligosaccharides (LOS). Two monoclonal antibodies (MAbs) directed against the minor LOS B band were generated, allowing the study of this surface molecule on different strains of Bordetella. These two MAbs, designated BL-8 and BL-9, reacted strongly with phenol-water-purified LOS obtained from a B. pertussis LOS B mutant strain. Sodium periodate treatment of the purified LOS prevented binding of the MAbs, indicating the carbohydrate nature of the epitope(s). Western immunoblotting experiments revealed that the epitope(s) recognized by these MAbs is conserved on all B. pertussis and Bordetella bronchiseptica Vir- (avirulent) variant strains tested but is not present on Bordetella parapertussis and B. bronchiseptica Vir+ (virulent) wild-type strains. Further studies showed that although present in the lipopolysaccharide B band expressed by Vir- strains, the epitope(s) recognized by the MAbs is not accessible on the surface of intact B. bronchiseptica cells. For B. pertussis, the density and accessibility of this epitope(s) are dependent on the virulence-associated or LOS phenotype expressed by the strain. Our data demonstrate that the expression and accessibility of the epitope(s) are significantly greater on the LOS B variant strains and LOS AB Vir- strains compared with fresh B. pertussis clinical isolates. For these latter strains, which are Vir+, this epitope(s) was barely detectable on the surface of intact bacteria, despite Western blot analyses that revealed specific reactions between the MAbs and the LOS B band. The two LOS B-specific MAbs had no bacteriolytic activity against a LOS AB wild-type strain, while the

  10. Dual infection with Bordetella pertussis and Mycoplasma pneumoniae in three infants: case reports.

    PubMed

    Zouari, A; Touati, A; Smaoui, H; Brun, D; Kasdaghli, K; Menif, K; Ben Jaballah, N; Ben Hassen, E; Guiso, N; Kechrid, A

    2012-04-01

    Studying pertussis-like respiratory infections, we report the cases of three infants with evidence of both Bordetella pertussis and Mycoplasma pneumoniae. Bordetella infection was identified by the real-time polymerase chain reaction (RT-PCR) of nasopharyngeal specimens. Neither B. pertussis nor B. parapertussis were recovered on the culture of nasopharyngeal aspirates (NPAs) from any subjects. M. pneumoniae etiology was diagnosed by culture and RT-PCR. The evolution was fatal for all of the subjects. We conclude that, among patients with Bordetella infection, co-infection with another respiratory pathogen is often probable, and these mixed infections might cause a more severe form of illness, sometimes leading to death.

  11. Antigenic and virulence properties of Pasteurella haemolytica leukotoxin mutants.

    PubMed Central

    Petras, S F; Chidambaram, M; Illyes, E F; Froshauer, S; Weinstock, G M; Reese, C P

    1995-01-01

    Antigenic properties of two mutants of Pasteurella haemolytica, strains 59B0071 and 59B0072, that do not produce detectable leukotoxin were investigated. Western blot (immunoblot) analysis with a number of polyclonal sera from animals recovering from pasteurellosis revealed that both mutants secreted a variety of antigens that were also present in cultures of several wild-type strains. These antigens ranged from about 100 to 15 kDa. Mutant strain 59B0071 was found to be totally deficient in leukotoxin, as judged not only by Western blotting but also by cytotoxicity assays with bovine lymphoma (BL-3) cells or bovine polymorphonuclear cells as targets. The mutant strain 59B0071 had normal levels of a secreted sialylglycoprotease, however. When strains were tested for virulence in goat and cattle challenge experiments, a reduction in mortality and lung lesions was observed with the mutant 59B0071 in comparison with results obtained with wild-type strains. These results are consistent with an important role for leukotoxin in P. haemolytica virulence and suggest that leukotoxin-negative mutants may be useful tools in the investigation of other virulence properties involved in P. haemolytica infections. PMID:7868224

  12. Pasteurella haemolytica antigens associated with resistance to pneumonic pasteurellosis.

    PubMed Central

    Mosier, D A; Simons, K R; Confer, A W; Panciera, R J; Clinkenbeard, K D

    1989-01-01

    Antigens associated with whole Pasteurella haemolytica biotype A serotype 1, a capsular carbohydrate-protein extract of the organism, and P. haemolytica leukotoxin were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Antigens of the electrophoresed preparations were detected by Western blotting (immunoblotting) with sera from cattle which were either nonvaccinated or vaccinated with live or killed P. haemolytica vaccines and had variable degrees of resistance to experimental pneumonic pasteurellosis. Distinct, easily recognizable antigens of these preparations were identified, and the antibody responses to these antigens were quantified by densitometry. To determine their importance to disease resistance, we then compared antibody responses with experimental lesion scores. Antibody reactivity to surface antigens which were significantly correlated with resistance and present in two or more of the preparations were detected at 86, 66, 51, 49, 34, 31, and 16 kilodaltons (kDa). Of these, antibody responses to antigens at 86, 49, and 31 kDa appeared most important based on their concentration and significance levels. Antibody reactivity to leukotoxin antigens which were significantly correlated with resistance and common with important surface antigens were detected at 86, 66, and 49 kDa. Antibody responses to unique leukotoxin antigens which were significantly correlated with resistance were present at 92 and 58 kDa. Images PMID:2917783

  13. Characterisation of Pasteurella dagmatis-like isolates recovered from the feline oral cavity.

    PubMed

    Sellyei, Boglárka; Wehmann, Eniko; Makrai, László; Magyar, Tibor

    2010-10-26

    Three Pasteurella dagmatis-like strains recovered from the feline oral cavity were analysed by traditional biochemical tests, the Biolog Microstation™ ID System, and 16S rRNA and sodA gene sequence analysis. The molecular biological methods revealed that these strains differ from P. dagmatis, forming a new genomospecies in the genus Pasteurella sensu stricto. Furthermore, sequence analysis and multiple alignments of 16S rRNA and the sodA gene established that the P. pneumotropica NCTC10827 and the P. dagmatis-like strains described here possess high genetic similarity.

  14. Pasteurella haemolytica bacteriophage: identification, partial characterization, and relationship of temperate bacteriophages from isolates of Pasteurella haemolytica (biotype A, serotype 1).

    PubMed

    Richards, A B; Renshaw, H W; Sneed, L W

    1985-05-01

    Pasteurella haemolytica (biotype A, serotype 1) isolates (n = 15) from the upper respiratory tract of clinically normal cattle, as well as from lung lesions from cases of fatal bovine pasteurellosis, were examined for the presence of bacteriophage after irradiation with UV light. Treatment of all P haemolytica isolates with UV irradiation resulted in lysis of bacteria due to the induction of vegetative development of bacteriophages. The extent of growth inhibition and bacterial lysis in irradiated cultures was UV dose-dependent. Bacterial cultures exposed to UV light for 20 s reached peak culture density between 60 and 70 minutes after irradiation; thereafter, culture density declined rapidly, so that by 120 minutes, it was approximately 60% of the original value. When examined ultrastructurally, lytic cultures from each isolate revealed bacteriophages with an overall length of approximately 200 nm and that appeared to have a head with icosahedral symmetry and a contractile tail. Cell-free filtrate from each noninduced bacterial isolate was inoculated onto the other bacterial isolates in a cross-culture sensitivity assay for the presence of phages lytic for the host bacterial isolates. Zones of lysis (plaques) did not develop when bacterial lawns grown from the different isolates were inoculated with filtrates from the heterologous isolates.

  15. Resident microbiota affect Bordetella pertussis infectious dose and host specificity.

    PubMed

    Weyrich, Laura S; Feaga, Heather A; Park, Jihye; Muse, Sarah J; Safi, Chetan Y; Rolin, Olivier Y; Young, Sarah E; Harvill, Eric T

    2014-03-01

    Before contacting host tissues, invading pathogens directly or indirectly interact with host microbiota, but the effects of such interactions on the initial stages of infection are poorly understood. Bordetella pertussis is highly infectious among humans but requires large doses to colonize rodents, unlike a closely related zoonotic pathogen, Bordetella bronchiseptica, raising important questions about the contributions of bacterial competition to initial colonization and host selection. We observed that <100 colony-forming units (CFU) of B. bronchiseptica efficiently infected mice and displaced culturable host microbiota, whereas 10 000 CFU of B. pertussis were required to colonize murine nasal cavities and did not displace host microorganisms. Bacteria isolated from murine nasal cavities but not those from the human lower respiratory tract limited B. pertussis growth in vitro, indicating that interspecies competition may limit B. pertussis colonization of mice. Further, a broad-spectrum antibiotic treatment delivered before B. pertussis inoculation reduced the infectious dose to <100 CFU, and reintroduction of single Staphylococcus or Klebsiella species was sufficient to inhibit B. pertussis colonization of antibiotic-treated mice. Together, these results reveal that resident microorganisms can prevent B. pertussis colonization and influence host specificity, and they provide rationale for manipulating microbiomes to create more-accurate animal models of infectious diseases.

  16. Virulence of Bordetella bronchiseptica: role of adenylate cyclase-hemolysin.

    PubMed Central

    Gueirard, P; Guiso, N

    1993-01-01

    Bordetella bronchiseptica is a pathogen of laboratory, domestic, and wild animals and sometimes of humans. In the present study some characteristics of the virulence of B. bronchiseptica isolates of different origin were studied. All isolates had similar phenotypes, similar bacteriological characters, and synthesized adenylate cyclase-hemolysin, filamentous hemagglutinin and pertactin but not pertussis toxin. These isolates, however, differed in their ability to express dermonecrotic toxin and to cause a lethal infection, but no correlation was found with the human or animal origin of the isolates. The fact that the most virulent isolate did not express dermonecrotic toxin suggests that this toxin does not play an important role in the virulence of the bacteria in the murine model. After infection with virulent B. bronchiseptica a very early synthesis and a persistence of anti-adenylate cyclase-hemolysin and anti-filamentous hemagglutinin antibodies were observed in the sera of infected mice, suggesting a persistence of the bacteria or of its antigens. B. bronchiseptica adenylate cyclase-hemolysin was purified and was shown to be a major protective antigen against B. bronchiseptica infection. Furthermore, we showed that its immunological and protective properties were different from that of B. pertussis adenylate cyclase-hemolysin, confirming that Bordetella species are immunologically different. Images PMID:8406794

  17. Novel environmental species isolated from the plaster wall surface of mural paintings in the Takamatsuzuka tumulus: Bordetella muralis sp. nov., Bordetella tumulicola sp. nov. and Bordetella tumbae sp. nov.

    PubMed

    Tazato, Nozomi; Handa, Yutaka; Nishijima, Miyuki; Kigawa, Rika; Sano, Chie; Sugiyama, Junta

    2015-12-01

    Ten strains of Gram-stain-negative, non-spore-forming, non-motile coccobacilli were isolated from the plaster wall surface of 1300-year-old mural paintings inside the stone chamber of the Takamatsuzuka tumulus in Asuka village (Asuka-mura), Nara Prefecture, Japan. Based on 16S rRNA gene sequence analysis of the isolates, they belonged to the proteobacterial genus Bordetella (class Betaproteobacteria) and could be separated into three groups representing novel lineages within the genus Bordetella. Three isolates were selected, one from each group, and identified carefully using a polyphasic approach. The isolates were characterized by the presence of Q-8 as their major ubiquinone system and C16 : 0 (30.0-41.8 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c; 10.1-27.0 %) and C17 : 0 cyclo (10.8-23.8 %) as the predominant fatty acids. The major hydroxy fatty acids were C12 : 0 2-OH and C14 : 0 2-OH. The DNA G+C content was 59.6-60.0 mol%. DNA-DNA hybridization tests confirmed that the isolates represented three separate novel species, for which the names Bordetella muralis sp. nov. (type strain T6220-3-2bT = JCM 30931T = NCIMB 15006T), Bordetella tumulicola sp. nov. (type strain T6517-1-4bT = JCM 30935T = NCIMB 15007T) and Bordetella tumbae sp. nov. (type strain T6713-1-3bT = JCM 30934T = NCIMB 15008T) are proposed. These results support previous evidence that members of the genus Bordetella exist in the environment and may be ubiquitous in soil and/or water.

  18. Complete Genome Sequences of Bordetella pertussis Vaccine Reference Strains 134 and 10536

    PubMed Central

    Peng, Yanhui; Loparev, Vladimir; Batra, Dhwani; Burroughs, Mark; Johnson, Taccara; Juieng, Phalasy; Rowe, Lori; Tondella, M. Lucia; Williams, Margaret M.

    2016-01-01

    Vaccine formulations and vaccination programs against whooping cough (pertussis) vary worldwide. Here, we report the complete genome sequences of two divergent Bordetella pertussis reference strains used in the production of pertussis vaccines. PMID:27635001

  19. Contribution of the Bordetella Bps Polysaccharide to Biofilm Formation and Respiratory Disease in Swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica is pervasive in swine populations and plays multiple roles in respiratory disease. Additionally, B. bronchiseptica is capable of establishing long-term or chronic infections in swine. Bacterial biofilms are increasingly recognized as important contributors to chronic bacter...

  20. Simple and specific detection of Bordetella holmesii by using a loop-mediated isothermal amplification assay.

    PubMed

    Otsuka, Nao; Yoshino, Shuji; Kawano, Kimiko; Toyoizumi-Ajisaka, Hiromi; Shibayama, Keigo; Kamachi, Kazunari

    2012-07-01

    A loop-mediated isothermal amplification (LAMP) assay for simple detection of Bordetella holmesii was developed. This assay discriminates between B. holmesii and other Bordetella species and successfully detect B. holmesii DNA in nasopharyngeal swab samples from subjects with suspected pertussis. The LAMP assay results were in complete agreement with the results of previously published real-time PCR assay, indicating that the former is a powerful tool for the accurate diagnosis and surveillance of B. holmesii.

  1. A specific real-time PCR assay for the detection of Bordetella pertussis.

    PubMed

    Vincart, Benoit; De Mendonça, Ricardo; Rottiers, Sylvianne; Vermeulen, Françoise; Struelens, Marc J; Denis, Olivier

    2007-07-01

    A novel real-time PCR (RT-PCR) assay was developed for detection of Bordetella pertussis in respiratory specimens by targeting the pertactin gene. In vitro evaluation with reference strains and quality control samples showed analytical sensitivity equivalent to and specificity superior to those of PCR assays which target the IS481 element. The pertactin-based RT-PCR assay offers better discrimination between B. pertussis and other Bordetella species than previously described assays.

  2. Secretion and expression of the Pasteurella haemolytica Leukotoxin.

    PubMed Central

    Highlander, S K; Engler, M J; Weinstock, G M

    1990-01-01

    The Pasteurella haemolytica leukotoxin gene cluster (lktCABD) is homologous to the Escherichia coli hemolysin locus (hlyCABD). Since the cloned leukotoxin (LktA) is not secreted from E. coli cells, a heteroplasmid complementation system was developed that permits secretion of the leukotoxin from cells expressing the hemolysin transport proteins HlyB and HlyD. We observed that the secreted leukotoxin protein had weak hemolytic activity when activated by either the HlyC or LktC proteins and that LktC expressed in E. coli could confer weak hemolytic activity upon hemolysin. Thus, it appears that the accessory proteins of the leukotoxin and hemolysin gene clusters are functionally similar, although their expression in E. coli is not equivalent. Northern (RNA) blot analysis of the P. haemolytica leukotoxin gene cluster revealed a major 3.5-kilobase transcript that includes the lktC and lktA genes. The start site for this transcript mapped to a cytosine residue 30 nucleotides upstream from the putative start of lktC; a similar initiation site was observed in E. coli, although adjacent cytosine and adenine residues were also utilized. The 3.5-kilobase transcript terminated near the rho-independent terminator structure between lktA and lktB, but transcription may continue, via antitermination or de novo transcription initiation, into the downstream lktB and lktD genes. We propose that the lack of LktB and LktD function in E. coli is a result, at least in part, of poor lktBD transcription and suggest that a P. haemolytica-specific regulator is required for optimal expression of the leukotoxin genes. Images PMID:2185213

  3. Bordetella adenylate cyclase toxin: entry of bacterial adenylate cyclase into mammalian cells.

    PubMed

    Confer, D L; Slungaard, A S; Graf, E; Panter, S S; Eaton, J W

    1984-01-01

    We have identified an adenylate cyclase toxin in urea extracts and culture supernatant fluids of Bordetella pertussis (2). The ability of this toxin and the lack of a strong correlation between its activity and adenylate cyclase activity found in urea extracts suggest that it is an oligomer of readily dissociable subunits. The mechanism by which Bordetella adenylate cyclase toxin interacts with target cells is unknown, but polyvalent cations are necessary. Neutrophils exposed to the toxin acquire a 39,000 Mr protein that can also be photoaffinity labeled with 32P-ATP. We anticipate that this protein will prove to be a catalytic component of Bordetella adenylate cyclase toxin. Susceptible cells exposed to Bordetella adenylate cyclase toxin are functionally aberrant. In phagocytes, decreased bactericidal capacity may be important in the pathogenesis of human whooping cough and other Bordetella infections occurring in domestic animals. The effects of the toxin on neoplastic cells may offer new insights into the factors controlling their growth and differentiation. Bordetella adenylate cyclase toxin is a unique bacterial product. Further purification and characterization of this toxin will add to our understanding of cell-protein interactions and pathogen-host relationships.

  4. Draft Genome Sequence of the Rodent Opportunistic Pathogen Pasteurella pneumotropica ATCC 35149T.

    PubMed

    Sasaki, Hiraku; Ishikawa, Hiroki; Asano, Ryoki; Ueshiba, Hidehiro; Matsumoto, Tetsuya; Boot, Ron; Kawamoto, Eiichi

    2014-08-07

    Pasteurella pneumotropica is an opportunistic pathogen in rodents that is commonly isolated from upper respiratory tracts in laboratory rodents. Here, we report the draft genome sequence of the P. pneumotropica type strain ATCC 35149, which was first isolated and characterized as biotype Jawetz.

  5. Pasteurella species peritoneal dialysis-associated peritonitis: Household pets as a risk factor

    PubMed Central

    Poliquin, Philippe Guillaume; Lagacé-Wiens, Philippe; Verrelli, Mauro; Allen, David W; Embil, John M

    2015-01-01

    BACKGROUND: Pasteurella species are Gram-negative coccobacilli that are a part of the normal oropharyngeal flora of numerous domestic animals. They have been recognized as a rare but significant cause of peritonitis in patients undergoing peritoneal dialysis (PD). A consensus about management strategies for PD-associated peritonitis caused by Pasteurella species currently does not exist. METHODS: The microbiological database serving the Manitoba Renal Program was searched from 1997 to 2013 for cases of Pasteurella species PD-associated peritonitis, and charts were reviewed. PubMed was searched for case reports and data were abstracted. RESULTS: Seven new local cases and 30 previously reported cases were analyzed. This infection is clinically similar to other forms of PD peritonitis, with household pet exposure appearing to be the strongest risk factor. Cats are the most commonly implicated pet. Direct contact between the pet and the equipment was commonly reported (25 of 37 patients) but was not necessary for infection to develop. The mean duration of treatment was 15 days. Complication rates were low, with only 11% of patients requiring PD catheter removal. There was no mortality reported. CONCLUSION: Pasteurella species are a rare cause of PD-associated peritonitis that can be successfully treated with a two-week course of intraperitoneal antibiotics with a high likelihood of catheter salvage. PMID:25798157

  6. Bordetella petrii infection with long-lasting persistence in human.

    PubMed

    Le Coustumier, Alain; Njamkepo, Elisabeth; Cattoir, Vincent; Guillot, Sophie; Guiso, Nicole

    2011-04-01

    We report the repeated isolation of Bordetella petrii in the sputum of a 79-year-old female patient with diffuse bronchiectasis and persistence of the bacterium for >1 year. The patient was first hospitalized due to dyspnea, which developed into severe cough with purulent sputum that yielded B. petrii on culture. After this first episode, the patient was hospitalized an additional 4 times with bronchorrhea symptoms. The isolates collected were analyzed by using biochemical, genotypic, and proteomic tools. Expression of specific proteins was analyzed by using serum samples from the patient. The B. petrii isolates were compared with other B. petrii isolates collected from humans or the environment and with isolates of B. pertussis, B. parapertussis, B. bronchiseptica, and B. holmesii, obtained from human respiratory tract infections. Our observations indicate that B. petrii can persist in persons with chronic pulmonary obstructive disease as has been previously demonstrated for B. bronchiseptica.

  7. Rapid increase in pertactin-deficient Bordetella pertussis isolates, Australia.

    PubMed

    Lam, Connie; Octavia, Sophie; Ricafort, Lawrence; Sintchenko, Vitali; Gilbert, Gwendolyn L; Wood, Nicholas; McIntyre, Peter; Marshall, Helen; Guiso, Nicole; Keil, Anthony D; Lawrence, Andrew; Robson, Jenny; Hogg, Geoff; Lan, Ruiting

    2014-04-01

    Acellular vaccines against Bordetella pertussis were introduced in Australia in 1997. By 2000, these vaccines had replaced whole-cell vaccines. During 2008-2012, a large outbreak of pertussis occurred. During this period, 30% (96/320) of B. pertussis isolates did not express the vaccine antigen pertactin (Prn). Multiple mechanisms of Prn inactivation were documented, including IS481 and IS1002 disruptions, a variation within a homopolymeric tract, and deletion of the prn gene. The mechanism of lack of expression of Prn in 16 (17%) isolates could not be determined at the sequence level. These findings suggest that B. pertussis not expressing Prn arose independently multiple times since 2008, rather than by expansion of a single Prn-negative clone. All but 1 isolate had ptxA1, prn2, and ptxP3, the alleles representative of currently circulating strains in Australia. This pattern is consistent with continuing evolution of B. pertussis in response to vaccine selection pressure.

  8. Inhaled Bordetella pertussis vaccine decreases airway responsiveness in guinea pigs.

    PubMed

    Vargas, M H; Bazán-Perkins, B; Segura, P; Campos, M G; Selman, M; Montaño, L M

    1995-01-01

    Bordetella pertussis (BP) has been used as adjuvant for experimental animal immunization, but its effects on airway responsiveness are uncertain. Three groups of guinea pigs were used: animals with a single exposure to inhaled BP vaccine (strain 134, total dose 1.24 x 10(12) germs), animals submitted to a sensitization procedure through inhalation of ovalbumin plus BP, and healthy control animals. Four weeks after inhalation of BP or after the beginning of sensitization, dose- or concentration-response curves to histamine were constructed in vivo and in vitro (tracheal and parenchymal preparations). We found that BP alone produced lower responses to histamine than control guinea pigs in vivo (insufflation pressure, p = 0.0003) and in tracheal tissues (p = 0.04), but not in parenchymal preparations. Sensitization did not modify the responsiveness compared with their respective controls. These results suggest that some BP component(s), probably pertussis toxin, causes a long lasting airway hyporesponsiveness in guinea pigs.

  9. Prevalence of Bordetella pertussis and Bordetella parapertussis infections in Tunisian hospitalized infants: results of a 4-year prospective study.

    PubMed

    Zouari, Asma; Smaoui, Hanen; Brun, Delphine; Njamkepo, Elisabeth; Sghaier, Soufien; Zouari, Emna; Félix, Renaud; Menif, Khaled; Ben Jaballah, Najla; Guiso, Nicole; Kechrid, Amel

    2012-04-01

    The prevalence of pertussis in Tunisia remains undetermined essentially because of the unavailability of a basic laboratory diagnostic service. Specific diagnostic tools were applied for the first time in a Tunisian prospective study in order to get a first estimation of the prevalence of Bordetella pertussis/parapertussis infections and to evaluate their use to determine the epidemiologic characteristics of these infections in Tunisian infants. Between 2007 and 2011, a total of 626 samples from 599 infants aged <1 year with and without pertussoid cough were investigated for the presence of B. pertussis/parapertussis using culture and real-time polymerase chain reaction (PCR). The real-time PCR (RT-PCR) targets include IS481 commonly found in B. pertussis, B. bronchiseptica, and B. holmesii; IS1001 specific of B. parapertussis, in combination with the pertussis toxin promoter region gene (ptx) of B. pertussis; and the recA gene specific of B. holmesii. When possible, patients' household contacts provided nasopharyngeal aspirates (NPAs) for RT-PCR detection of B. pertussis/parapertussis or single-serum samples for anti-PT IgG quantification. All except 1 NPAs were negative by conventional culture, whereas PCR gave positive signals for 126 specimens (21%): B. pertussis, B. parapertussis, and Bordetella spp. were detected in 82%, 6%, and 4% of the samples, respectively. The simultaneous presence of B. pertussis and B. parapertussis was noted in 8% of the cases. Pertussis was reported throughout the year with a peak during the summer of the year 2009. The prevalence of Bordetella infection was 20% between 2007 and 2011. Most of these cases corresponded to patients younger than 6 months who received <3 doses of pertussis vaccine. Among the household contacts enrolled in the study, mothers seemed to be the likely source of infection. This study showed that pertussis is still prevalent in Tunisia and that the disease remains a public health problem affecting not only

  10. Interspecies variations in Bordetella catecholamine receptor gene regulation and function.

    PubMed

    Brickman, Timothy J; Suhadolc, Ryan J; Armstrong, Sandra K

    2015-12-01

    Bordetella bronchiseptica can use catecholamines to obtain iron from transferrin and lactoferrin via uptake pathways involving the BfrA, BfrD, and BfrE outer membrane receptor proteins, and although Bordetella pertussis has the bfrD and bfrE genes, the role of these genes in iron uptake has not been demonstrated. In this study, the bfrD and bfrE genes of B. pertussis were shown to be functional in B. bronchiseptica, but neither B. bronchiseptica bfrD nor bfrE imparted catecholamine utilization to B. pertussis. Gene fusion analyses found that expression of B. bronchiseptica bfrA was increased during iron starvation, as is common for iron receptor genes, but that expression of the bfrD and bfrE genes of both species was decreased during iron limitation. As shown previously for B. pertussis, bfrD expression in B. bronchiseptica was also dependent on the BvgAS virulence regulatory system; however, in contrast to the case in B. pertussis, the known modulators nicotinic acid and sulfate, which silence Bvg-activated genes, did not silence expression of bfrD in B. bronchiseptica. Further studies using a B. bronchiseptica bvgAS mutant expressing the B. pertussis bvgAS genes revealed that the interspecies differences in bfrD modulation are partly due to BvgAS differences. Mouse respiratory infection experiments determined that catecholamine utilization contributes to the in vivo fitness of B. bronchiseptica and B. pertussis. Additional evidence of the in vivo importance of the B. pertussis receptors was obtained from serologic studies demonstrating pertussis patient serum reactivity with the B. pertussis BfrD and BfrE proteins.

  11. Interspecies Variations in Bordetella Catecholamine Receptor Gene Regulation and Function

    PubMed Central

    Brickman, Timothy J.; Suhadolc, Ryan J.

    2015-01-01

    Bordetella bronchiseptica can use catecholamines to obtain iron from transferrin and lactoferrin via uptake pathways involving the BfrA, BfrD, and BfrE outer membrane receptor proteins, and although Bordetella pertussis has the bfrD and bfrE genes, the role of these genes in iron uptake has not been demonstrated. In this study, the bfrD and bfrE genes of B. pertussis were shown to be functional in B. bronchiseptica, but neither B. bronchiseptica bfrD nor bfrE imparted catecholamine utilization to B. pertussis. Gene fusion analyses found that expression of B. bronchiseptica bfrA was increased during iron starvation, as is common for iron receptor genes, but that expression of the bfrD and bfrE genes of both species was decreased during iron limitation. As shown previously for B. pertussis, bfrD expression in B. bronchiseptica was also dependent on the BvgAS virulence regulatory system; however, in contrast to the case in B. pertussis, the known modulators nicotinic acid and sulfate, which silence Bvg-activated genes, did not silence expression of bfrD in B. bronchiseptica. Further studies using a B. bronchiseptica bvgAS mutant expressing the B. pertussis bvgAS genes revealed that the interspecies differences in bfrD modulation are partly due to BvgAS differences. Mouse respiratory infection experiments determined that catecholamine utilization contributes to the in vivo fitness of B. bronchiseptica and B. pertussis. Additional evidence of the in vivo importance of the B. pertussis receptors was obtained from serologic studies demonstrating pertussis patient serum reactivity with the B. pertussis BfrD and BfrE proteins. PMID:26371128

  12. BpsR Modulates Bordetella Biofilm Formation by Negatively Regulating the Expression of the Bps Polysaccharide

    PubMed Central

    Conover, Matt S.; Redfern, Crystal J.; Ganguly, Tridib; Sukumar, Neelima; Sloan, Gina; Mishra, Meenu

    2012-01-01

    Bordetella bacteria are Gram-negative respiratory pathogens of animals, birds, and humans. A hallmark feature of some Bordetella species is their ability to efficiently survive in the respiratory tract even after vaccination. Bordetella bronchiseptica and Bordetella pertussis form biofilms on abiotic surfaces and in the mouse respiratory tract. The Bps exopolysaccharide is one of the critical determinants for biofilm formation and the survival of Bordetella in the murine respiratory tract. In order to gain a better understanding of regulation of biofilm formation, we sought to study the mechanism by which Bps expression is controlled in Bordetella. Expression of bpsABCD (bpsA-D) is elevated in biofilms compared with levels in planktonically grown cells. We found that bpsA-D is expressed independently of BvgAS. Subsequently, we identified an open reading frame (ORF), BB1771 (designated here bpsR), that is located upstream of and in the opposite orientation to the bpsA-D locus. BpsR is homologous to the MarR family of transcriptional regulators. Measurement of bpsA and bpsD transcripts and the Bps polysaccharide levels from the wild-type and the ΔbpsR strains suggested that BpsR functions as a repressor. Consistent with enhanced production of Bps, the bpsR mutant displayed considerably more structured biofilms. We mapped the bpsA-D promoter region and showed that purified BpsR protein specifically bound to the bpsA-D promoter. Our results provide mechanistic insights into the regulatory strategy employed by Bordetella for control of the production of the Bps polysaccharide and biofilm formation. PMID:22056934

  13. BpsR modulates Bordetella biofilm formation by negatively regulating the expression of the Bps polysaccharide.

    PubMed

    Conover, Matt S; Redfern, Crystal J; Ganguly, Tridib; Sukumar, Neelima; Sloan, Gina; Mishra, Meenu; Deora, Rajendar

    2012-01-01

    Bordetella bacteria are Gram-negative respiratory pathogens of animals, birds, and humans. A hallmark feature of some Bordetella species is their ability to efficiently survive in the respiratory tract even after vaccination. Bordetella bronchiseptica and Bordetella pertussis form biofilms on abiotic surfaces and in the mouse respiratory tract. The Bps exopolysaccharide is one of the critical determinants for biofilm formation and the survival of Bordetella in the murine respiratory tract. In order to gain a better understanding of regulation of biofilm formation, we sought to study the mechanism by which Bps expression is controlled in Bordetella. Expression of bpsABCD (bpsA-D) is elevated in biofilms compared with levels in planktonically grown cells. We found that bpsA-D is expressed independently of BvgAS. Subsequently, we identified an open reading frame (ORF), BB1771 (designated here bpsR), that is located upstream of and in the opposite orientation to the bpsA-D locus. BpsR is homologous to the MarR family of transcriptional regulators. Measurement of bpsA and bpsD transcripts and the Bps polysaccharide levels from the wild-type and the ΔbpsR strains suggested that BpsR functions as a repressor. Consistent with enhanced production of Bps, the bpsR mutant displayed considerably more structured biofilms. We mapped the bpsA-D promoter region and showed that purified BpsR protein specifically bound to the bpsA-D promoter. Our results provide mechanistic insights into the regulatory strategy employed by Bordetella for control of the production of the Bps polysaccharide and biofilm formation.

  14. Harmonization of Bordetella pertussis real-time PCR diagnostics in the United States in 2012.

    PubMed

    Williams, Margaret M; Taylor, Thomas H; Warshauer, David M; Martin, Monte D; Valley, Ann M; Tondella, M Lucia

    2015-01-01

    Real-time PCR (rt-PCR) is an important diagnostic tool for the identification of Bordetella pertussis, Bordetella holmesii, and Bordetella parapertussis. Most U.S. public health laboratories (USPHLs) target IS481, present in 218 to 238 copies in the B. pertussis genome and 32 to 65 copies in B. holmesii. The CDC developed a multitarget PCR assay to differentiate B. pertussis, B. holmesii, and B. parapertussis and provided protocols and training to 19 USPHLs. The 2012 performance exercise (PE) assessed the capability of USPHLs to detect these three Bordetella species in clinical samples. Laboratories were recruited by the Wisconsin State Proficiency Testing program through the Association of Public Health Laboratories, in partnership with the CDC. Spring and fall PE panels contained 12 samples each of viable Bordetella and non-Bordetella species in saline. Fifty and 53 USPHLs participated in the spring and fall PEs, respectively, using a variety of nucleic acid extraction methods, PCR platforms, and assays. Ninety-six percent and 94% of laboratories targeted IS481 in spring and fall, respectively, in either singleplex or multiplex assays. In spring and fall, respectively, 72% and 79% of USPHLs differentiated B. pertussis and B. holmesii and 68% and 72% identified B. parapertussis. IS481 cycle threshold (CT) values for B. pertussis samples had coefficients of variation (CV) ranging from 10% to 28%. Of the USPHLs that differentiated B. pertussis and B. holmesii, sensitivity was 96% and specificity was 95% for the combined panels. The 2012 PE demonstrated increased harmonization of rt-PCR Bordetella diagnostic protocols in USPHLs compared to that of the previous survey.

  15. Active and Passive Immunizations with Bordetella Colonization Factor A Protect Mice against Respiratory Challenge with Bordetella bronchiseptica▿

    PubMed Central

    Sukumar, Neelima; Love, Cheraton F.; Conover, Matt S.; Kock, Nancy D.; Dubey, Purnima; Deora, Rajendar

    2009-01-01

    Bordetella colonization factor A (BcfA) is an outer membrane immunogenic protein, which is critical for efficient colonization of the murine respiratory tract. These properties of BcfA prompted us to examine its utility in inducing a protective immune response against Bordetella bronchiseptica in a mouse model of intranasal infection. Mice vaccinated with BcfA demonstrated reduced pathology in the lungs and harbored lower bacterial burdens in the respiratory tract. Immunization with BcfA led to the generation of BcfA-specific antibodies in both the sera and lungs, and passive immunization led to the reduction of B. bronchiseptica in the tracheas and lungs. These results suggest that protection after immunization with BcfA is mediated in part by antibodies against BcfA. To further investigate the mechanism of BcfA-induced immune clearance, we examined the role of neutrophils and macrophages. Our results demonstrate that neutrophils are critical for anti-BcfA antibody-mediated clearance and that opsonization with anti-BcfA serum enhances phagocytosis of B. bronchiseptica by murine macrophages. We show that immunization with BcfA results in the production of gamma interferon and subclasses of immunoglobulin G antibodies that are consistent with the induction of a Th1-type immune response. In combination, our findings suggest that the mechanism of BcfA-mediated immunity involves humoral and cellular responses. Expression of BcfA is conserved among multiple clinical isolates of B. bronchiseptica. Our results demonstrate the striking protective efficacy of BcfA-mediated immunization, thereby highlighting its utility as a potential vaccine candidate. These results also provide a model for the development of cell-free vaccines against B. bronchiseptica. PMID:19064638

  16. Structure activity characterization of Bordetella petrii lipid A, from environment to human isolates.

    PubMed

    Basheer, Soorej M; Bouchez, Valerie; Novikov, Alexey; Augusto, Luis A; Guiso, Nicole; Caroff, Martine

    2016-01-01

    Bordetella petrii, a facultative anaerobic species, is the only known member of the Bordetella genus with environmental origin. However it was also recently isolated from humans. The structures of the B. petrii lipid A moieties of the endotoxins were characterized here for the first time for an environmental strain and compared to that of human isolates. Characterization was achieved using chemical analyses, gas chromatography-mass spectrometry, and Matrix Assisted Laser Desorption Ionisation mass spectrometry. The analyses revealed that the different lipid A structures contain a common bisphosphorylated β-(1→6)-linked d-glucosamine disaccharide with hydroxytetradecanoic acid in amide as well at the C-3' in ester linkages. Similar to Bordetella pertussis and Bordetella bronchiseptica lipids A, the hydroxytetradecanoic acid at the C-2' position was substituted by tetradecanoic acid. Unlike B. pertussis, the hydroxytetradecanoic acid at the C-2 position was substituted with either 12:0 or 14:0 and/or their 2-OH forms. Depending on the environmental or human origin the structures differed in the length and degree of fatty acid acylation and impacted the IL-6 and TNF-α inflammatory responses tested. In one isolate we showed the presence at the C-3 position of the short-chain 10:0(3-OH), which according to our previous analyses is more characteristic of the human pathogens in the genus like B. pertussis and Bordetella parapertussis.

  17. A Pasteurella sp associated with respiratory disease in captive desert tortoises.

    PubMed

    Snipes, K P; Biberstein, E L; Fowler, M E

    1980-11-01

    Bacteria isolated from captive healthy desert tortoises were compared with bacteria from captive tortoises with respiratory illness and with bacteria from free-ranging tortoises from the Mojave Desert. Major differences were not observed among these groups when bacteria from the mouth, nares, trachea, lungs, and cloaca were compared. Frequently encountered organisms in all 3 groups included: coagulase-negative, catalase-positive, gram-positive cocci; Corynebacterium sp; members of Enterobacteriaceae, including Proteus spp; and a bacterium apparently belonging to the genus Pasteurella. The Pasteurella sp was consistently found to be associated with respiratory lesions in captive tortoises with signs of respiratory disease but was also found to be part of the gastrointestinal and nasal flora of healthy tortoises. It was hypothesized that respiratory disease in captive desert tortoises involves a commensal bacterium with the potential to be an opportunistic pathogen when the tortoise is stressed by a captive environment.

  18. Renal abscess caused by a Providencia stuartii isolate biochemically misidentified as Pasteurella.

    PubMed

    Chamberland, Robin R; McElvania TeKippe, Erin; Burnham, Carey-Ann D; Kennedy, Donald J

    2013-08-01

    Providencia stuartii is associated with urinary tract infection (UTI) in catheterized patients. Here we report an abscess containing P. stuartii in a patient with a history of UTI, renal stones, and stent placement. This organism was identified by matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry and 16S rRNA gene sequencing following biochemical identification as Pasteurella.

  19. Bronchitis caused by Bordetella holmesii in a child with asthma misdiagnosed as mycoplasmal infection.

    PubMed

    Katsukawa, Chihiro; Kushibiki, Chieko; Nishito, Atsumi; Nishida, Rikou; Kuwabara, Norimitsu; Kawahara, Ryuji; Otsuka, Nao; Miyaji, Yusuke; Toyoizumi-Ajisaka, Hiromi; Kamachi, Kazunari

    2013-06-01

    We report a case of a bronchitis caused by Bordetella holmesii in a 2-year-old girl with asthma. The patient had a moderate fever and productive cough, and her condition was initially diagnosed as mycoplasmal bronchitis on the basis of her clinical symptoms and rapid serodiagnosis of mycoplasmal infection. She was treated with a bronchodilator and clarithromycin, which resulted in complete recovery. However, after the initial diagnosis, nucleic acid amplification tests of her sputum showed the absence of both Mycoplasma pneumoniae and Bordetella pertussis infections. Sputum culture showed the presence of a slow-growing, gram-negative bacillus in pure culture on Bordetella agar plates; the bacillus was later identified as B. holmesii. B. holmesii infection is rare in immunocompetent children; however, the organism is a true pathogen that can cause bronchitis in young children with asthma.

  20. Bordetella holmesii bacteremia cases in the United States, April 2010-January 2011.

    PubMed

    Tartof, Sara Y; Gounder, Prabhu; Weiss, Don; Lee, Lillian; Cassiday, Pamela K; Clark, Thomas A; Briere, Elizabeth C

    2014-01-01

    We describe the first report of temporally related cases of Bordetella holmesii bacteremia. Demographic and clinical data were collected through chart abstraction and case-patient interviews. Twenty-two cases were identified from 6 states. Symptom onset dates ranged from April 2010 to January 2011. Median age of patients was 17.1 years and 64% had functional or anatomic asplenia. Pulsed-field gel electrophoresis profiles of a sample of isolates were identical. These cases occurred during a peak in pertussis outbreaks with documented cases of B. holmesii/Bordetella pertussis respiratory coinfection; whether there is a link between B. holmesii respiratory and bloodstream infection is unknown.

  1. Separation and purification of the hemagglutinins from Bordetella pertussis.

    PubMed Central

    Sato, Y; Cowell, J L; Sato, H; Burstyn, D G; Manclark, C R

    1983-01-01

    The role of the filamentous hemagglutinin (FHA) and the lymphocytosis-promoting factor hemagglutinin (LPF) in pertussis pathogenesis and immunity is the subject of active investigation. To be certain of their role as protective antigens, the hemagglutinins must be pure and free of each other. This report describes procedures to separate and purify FHA and LPF from the culture supernatant of stationary cultures of Bordetella pertussis Tohama, using hydroxylapatite, haptoglobin-Sepharose, and Sepharose CL-6B filtration chromatography. Purified FHA contained less than 0.002% active LPF assayed by histamine-sensitizing activity, and both hemagglutinins contained less than 0.01% of each other based on antigenic activity measured by an enzyme-linked immunosorbent assay, using affinity chromatography-purified antibody to each hemagglutinin. LPF and FHA were also shown to be antigenically distinct by immunodiffusion and were judged to be highly purified proteins by polyacrylamide gel electrophoresis. In addition, the purification procedures yielded milligram amounts of each hemagglutinin with very good recovery of starting activities. Images PMID:6305841

  2. Clinical, laboratorial and radiographic predictors of Bordetella pertussis infection☆

    PubMed Central

    Bellettini, Camila Vieira; de Oliveira, Andressa Welter; Tusset, Cintia; Baethgen, Ludmila Fiorenzano; Amantéa, Sérgio Luís; Motta, Fabrizio; Gasparotto, Aline; Andreolla, Huander Felipe; Pasqualotto, Alessandro C.

    2014-01-01

    OBJECTIVE: To identify clinical, laboratorial and radiographic predictors for Bordetella pertussis infection. METHODS: This was a retrospective study, which analyzed medical records of all patients submitted to a molecular dignosis (qPCR) for B. pertussis from September 2011 to January 2013. Clinical and laboratorial data were reviewed, including information about age, sex, signs/symptoms, length of hospitalization, blood cell counts, imaging findings, coinfection with other respiratory pathogens and clinical outcome. RESULTS: 222 cases were revised. Of these, 72.5% had proven pertussis, and 60.9% were under 1 year old. In patients aging up to six months, independent predictors for B. pertussis infection were (OR 8.0, CI 95% 1.8-36.3; p=0.007) and lymphocyte count >104/µL (OR 10.0, CI 95% 1.8-54.5; p=0.008). No independent predictors of B. pertussis infection could be determined for patients older than six months. Co-infection was found in 21.4% of patients, of which 72.7% were up to six months of age. Adenovirus was the most common agent (40.9%). In these patients, we were not able to identify any clinical features to detect patients presenting with a respiratory co-infection, even though longer hospital stay was observed in patients with co-infections (12 vs. 6 days; p=0.009). CONCLUSIONS: Cyanosis and lymphocytosis are independent predictors for pertussis in children up to 6 months old. PMID:25510991

  3. Review of the neutrophil response to Bordetella pertussis infection.

    PubMed

    Eby, Joshua C; Hoffman, Casandra L; Gonyar, Laura A; Hewlett, Erik L

    2015-12-01

    The nature and timing of the neutrophil response to infection with Bordetella pertussis is influenced by multiple virulence factors expressed by the bacterium. After inoculation of the host airway, the recruitment of neutrophils signaled by B. pertussis lipooligosaccharide (LOS) is suppressed by pertussis toxin (PTX). Over the next week, the combined activities of PTX, LOS and adenylate cyclase toxin (ACT) result in production of cytokines that generate an IL-17 response, promoting neutrophil recruitment which peaks at 10-14 days after inoculation in mice. Arriving at the site of infection, neutrophils encounter the powerful local inhibitory activity of ACT, in conjunction with filamentous hemagglutinin. With the help of antibodies, neutrophils contribute to clearance of B. pertussis, but only after 28-35 days in a naïve mouse. Studies of the lasting, antigen-specific IL-17 response to infection in mice and baboons has led to progress in vaccine development and understanding of pathogenesis. Questions remain about the mediators that coordinate neutrophil recruitment and the mechanisms by which neutrophils overcome B. pertussis virulence factors.

  4. Prosthetic valve endocarditis caused by Bordetella holmesii, an Acinetobacter lookalike.

    PubMed

    Jonckheere, Stijn; De Baere, Thierry; Schroeyers, Pascal; Soetens, Oriane; De Bel, Annelies; Surmont, Ignace

    2012-06-01

    We report a case of fulminant endocarditis on a prosthetic homograft aortic valve caused by Bordetella holmesii, which was successfully managed by surgical valve replacement and antibiotic treatment. B. holmesii, a strictly aerobic, small, Gram-negative coccobacillus, has been implicated as an infrequent cause of a pertussis-like syndrome and other respiratory illnesses. However, B. holmesii is also a rare cause of septicaemia and infective endocarditis, mostly in immunocompromised patients. To our knowledge, this is the first report of B. holmesii endocarditis on a prosthetic aortic valve. Routine laboratory testing initially misidentified the strain as Acinetobacter sp. Correct identification was achieved by 16S rRNA gene and outer-membrane protein A (ompA) gene sequencing. Interestingly, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry also produced an accurate species-level identification. Subsequent susceptibility testing and review of the literature revealed ceftazidime, cefepime, carbapenems, aminoglycosides, fluoroquinolones, piperacillin/tazobactam, tigecycline and colistin as possible candidates to treat infections caused by B. holmesii.

  5. The multifaceted RisA regulon of Bordetella pertussis

    PubMed Central

    Coutte, Loïc; Huot, Ludovic; Antoine, Rudy; Slupek, Stephanie; Merkel, Tod J.; Chen, Qing; Stibitz, Scott; Hot, David; Locht, Camille

    2016-01-01

    The whooping cough agent Bordetella pertussis regulates the production of its virulence factors by the BvgA/S system. Phosphorylated BvgA activates the virulence-activated genes (vags) and represses the expression of the virulence-repressed genes (vrgs) via the activation of the bvgR gene. In modulating conditions, with MgSO4, the BvgA/S system is inactive, and the vrgs are expressed. Here, we show that the expression of almost all vrgs depends on RisA, another transcriptional regulator. We also show that some vags are surprisingly no longer modulated by MgSO4 in the risA− background. RisA also regulates the expression of other genes, including chemotaxis and flagellar operons, iron-regulated genes, and genes of unknown function, which may or may not be controlled by BvgA/S. We identified RisK as the likely cognate RisA kinase and found that it is important for expression of most, but not all RisA-regulated genes. This was confirmed using the phosphoablative RisAD60N and the phosphomimetic RisAD60E analogues. Thus the RisA regulon adds a new layer of complexity to B. pertussis virulence gene regulation. PMID:27620673

  6. New Data on Vaccine Antigen Deficient Bordetella pertussis Isolates

    PubMed Central

    Bouchez, Valérie; Hegerle, Nicolas; Strati, Francesco; Njamkepo, Elisabeth; Guiso, Nicole

    2015-01-01

    Evolution of Bordetella pertussis is driven by natural and vaccine pressures. Isolates circulating in regions with high vaccination coverage present multiple allelic and antigenic variations as compared to isolates collected before introduction of vaccination. Furthermore, during the last epidemics reported in regions using pertussis acellular vaccines, isolates deficient for vaccine antigens, such as pertactin (PRN), were reported to reach high proportions of circulating isolates. More sporadic filamentous hemagglutinin (FHA) or pertussis toxin (PT) deficient isolates were also collected. The whole genome of some recent French isolates, deficient or non-deficient in vaccine antigens, were analyzed. Transcription profiles of the expression of the main virulence factors were also compared. The invasive phenotype in an in vitro human tracheal epithelial (HTE) cell model of infection was evaluated. Our genomic analysis focused on SNPs related to virulence genes known to be more likely to present allelic polymorphism. Transcriptomic data indicated that isolates circulating since the introduction of pertussis vaccines present lower transcription levels of the main virulence genes than the isolates of the pre-vaccine era. Furthermore, isolates not producing FHA present significantly higher expression levels of the entire set of genes tested. Finally, we observed that recent isolates are more invasive in HTE cells when compared to the reference strain, but no multiplication occurs within cells. PMID:26389958

  7. Bordetella pertussis: new concepts in pathogenesis and treatment

    PubMed Central

    Carbonetti, Nicholas H.

    2016-01-01

    Purpose of review The purpose of this review is to summarize and discuss recent findings and selected topics of interest in Bordetella pertussis virulence and pathogenesis and treatment of pertussis. It is not intended to cover issues on immune responses to B. pertussis infection or problems with currently used pertussis vaccines. Recent findings Studies on the activities of various B. pertussis virulence factors include the immunomodulatory activities of filamentous hemagglutinin, fimbriae, and adenylate cyclase toxin. Recently emerging B. pertussis strains show evidence of genetic selection for vaccine escape mutants, with changes in vaccine antigen-expressing genes, some of which may have increased the virulence of this pathogen. Severe and fatal pertussis in young infants continues to be a problem, with several studies highlighting predictors of fatality, including the extreme leukocytosis associated with this infection. Treatments for pertussis are extremely limited, though early antibiotic intervention may be beneficial. Neutralizing pertussis toxin activity may be an effective strategy, as well as targeting two host proteins, pendrin and sphingosine-1-phosphate receptors, as novel potential therapeutic interventions. Summary Pertussis is reemerging as a major public health problem and continued basic research is revealing information on bacterial virulence and disease pathogenesis, as well as potential novel strategies for vaccination and targets for therapeutic intervention. PMID:26906206

  8. Review of the neutrophil response to Bordetella pertussis infection

    PubMed Central

    Eby, Joshua C.; Hoffman, Casandra L.; Gonyar, Laura A.; Hewlett, Erik L.

    2015-01-01

    The nature and timing of the neutrophil response to infection with Bordetella pertussis is influenced by multiple virulence factors expressed by the bacterium. After inoculation of the host airway, the recruitment of neutrophils signaled by B. pertussis lipooligosaccharide (LOS) is suppressed by pertussis toxin (PTX). Over the next week, the combined activities of PTX, LOS and adenylate cyclase toxin (ACT) result in production of cytokines that generate an IL-17 response, promoting neutrophil recruitment which peaks at 10–14 days after inoculation in mice. Arriving at the site of infection, neutrophils encounter the powerful local inhibitory activity of ACT, in conjunction with filamentous hemagglutinin. With the help of antibodies, neutrophils contribute to clearance of B. pertussis, but only after 28–35 days in a naïve mouse. Studies of the lasting, antigen-specific IL-17 response to infection in mice and baboons has led to progress in vaccine development and understanding of pathogenesis. Questions remain about the mediators that coordinate neutrophil recruitment and the mechanisms by which neutrophils overcome B. pertussis virulence factors. PMID:26432818

  9. Antibody responses to Bordetella bronchiseptica in vaccinated and infected dogs

    PubMed Central

    Ellis, John; Rhodes, Carrie; Lacoste, Stacey; Krakowka, Steven

    2014-01-01

    Bordetella bronchiseptica (Bb) whole cell bacterins have been replaced with acelluar vaccines. We evaluated the response to the acellular Bb vaccines in Bb-seropositive commingled laboratory beagles and client-owned dogs with various lifestyles and vaccination histories. A single parenteral dose of the acellular Bb vaccine resulted in consistent anamnestic IgG, and to a lesser, but notable extent, IgA, Bb-reactive antibody responses in the seropositive beagles. Associated with the increase in antibodies measured by enzyme-linked immunosorbent assay (ELISA) was an increase in the complement (C)-dependent IgG antibody mediated bactericidal effect on Bb in vitro. Antibody responses in client-owned dogs were more variable and were dependent upon the vaccination history and serological evidence of previous Bb exposure. Antibodies from vaccinated dogs recognized several Bb proteins, notably P68 (pertactin) and P220 (fimbrial hemagglutinin), the response to which has been shown to be disease-sparing in Bp infections. These antibody responses were similar to those in experimentally infected dogs and in dogs that had received a widely used whole cell bacterin. PMID:25183893

  10. Phylogenomic and molecular demarcation of the core members of the polyphyletic pasteurellaceae genera actinobacillus, haemophilus, and pasteurella.

    PubMed

    Naushad, Sohail; Adeolu, Mobolaji; Goel, Nisha; Khadka, Bijendra; Al-Dahwi, Aqeel; Gupta, Radhey S

    2015-01-01

    The genera Actinobacillus, Haemophilus, and Pasteurella exhibit extensive polyphyletic branching in phylogenetic trees and do not represent coherent clusters of species. In this study, we have utilized molecular signatures identified through comparative genomic analyses in conjunction with genome based and multilocus sequence based phylogenetic analyses to clarify the phylogenetic and taxonomic boundary of these genera. We have identified large clusters of Actinobacillus, Haemophilus, and Pasteurella species which represent the "sensu stricto" members of these genera. We have identified 3, 7, and 6 conserved signature indels (CSIs), which are specifically shared by sensu stricto members of Actinobacillus, Haemophilus, and Pasteurella, respectively. We have also identified two different sets of CSIs that are unique characteristics of the pathogen containing genera Aggregatibacter and Mannheimia, respectively. It is now possible to demarcate the genera Actinobacillus sensu stricto, Haemophilus sensu stricto, and Pasteurella sensu stricto on the basis of discrete molecular signatures. The other members of the genera Actinobacillus, Haemophilus, and Pasteurella that do not fall within the "sensu stricto" clades and do not contain these molecular signatures should be reclassified as other genera. The CSIs identified here also provide useful diagnostic targets for the identification of current and novel members of the indicated genera.

  11. Phylogenomic and Molecular Demarcation of the Core Members of the Polyphyletic Pasteurellaceae Genera Actinobacillus, Haemophilus, and Pasteurella

    PubMed Central

    Naushad, Sohail; Adeolu, Mobolaji; Goel, Nisha; Khadka, Bijendra; Al-Dahwi, Aqeel; Gupta, Radhey S.

    2015-01-01

    The genera Actinobacillus, Haemophilus, and Pasteurella exhibit extensive polyphyletic branching in phylogenetic trees and do not represent coherent clusters of species. In this study, we have utilized molecular signatures identified through comparative genomic analyses in conjunction with genome based and multilocus sequence based phylogenetic analyses to clarify the phylogenetic and taxonomic boundary of these genera. We have identified large clusters of Actinobacillus, Haemophilus, and Pasteurella species which represent the “sensu stricto” members of these genera. We have identified 3, 7, and 6 conserved signature indels (CSIs), which are specifically shared by sensu stricto members of Actinobacillus, Haemophilus, and Pasteurella, respectively. We have also identified two different sets of CSIs that are unique characteristics of the pathogen containing genera Aggregatibacter and Mannheimia, respectively. It is now possible to demarcate the genera Actinobacillus sensu stricto, Haemophilus sensu stricto, and Pasteurella sensu stricto on the basis of discrete molecular signatures. The other members of the genera Actinobacillus, Haemophilus, and Pasteurella that do not fall within the “sensu stricto” clades and do not contain these molecular signatures should be reclassified as other genera. The CSIs identified here also provide useful diagnostic targets for the identification of current and novel members of the indicated genera. PMID:25821780

  12. Differential regulation of type III secretion and virulence genes in Bordetella pertussis and Bordetella bronchiseptica by a secreted anti-σ factor

    PubMed Central

    Ahuja, Umesh; Shokeen, Bhumika; Cheng, Ning; Cho, Yeonjoo; Blum, Charles; Coppola, Giovanni; Miller, Jeff F.

    2016-01-01

    The BvgAS phosphorelay regulates ∼10% of the annotated genomes of Bordetella pertussis and Bordetella bronchiseptica and controls their infectious cycles. The hierarchical organization of the regulatory network allows the integration of contextual signals to control all or specific subsets of BvgAS-regulated genes. Here, we characterize a regulatory node involving a type III secretion system (T3SS)-exported protein, BtrA, and demonstrate its role in determining fundamental differences in T3SS phenotypes among Bordetella species. We show that BtrA binds and antagonizes BtrS, a BvgAS-regulated extracytoplasmic function (ECF) sigma factor, to couple the secretory activity of the T3SS apparatus to gene expression. In B. bronchiseptica, a remarkable spectrum of expression states can be resolved by manipulating btrA, encompassing over 80 BtrA-activated loci that include genes encoding toxins, adhesins, and other cell surface proteins, and over 200 BtrA-repressed genes that encode T3SS apparatus components, secretion substrates, the BteA effector, and numerous additional factors. In B. pertussis, BtrA retains activity as a BtrS antagonist and exerts tight negative control over T3SS genes. Most importantly, deletion of btrA in B. pertussis revealed T3SS-mediated, BteA-dependent cytotoxicity, which had previously eluded detection. This effect was observed in laboratory strains and in clinical isolates from a recent California pertussis epidemic. We propose that the BtrA-BtrS regulatory node determines subspecies-specific differences in T3SS expression among Bordetella species and that B. pertussis is capable of expressing a full range of T3SS-dependent phenotypes in the presence of appropriate contextual cues. PMID:26884180

  13. Differential regulation of type III secretion and virulence genes in Bordetella pertussis and Bordetella bronchiseptica by a secreted anti-σ factor.

    PubMed

    Ahuja, Umesh; Shokeen, Bhumika; Cheng, Ning; Cho, Yeonjoo; Blum, Charles; Coppola, Giovanni; Miller, Jeff F

    2016-03-01

    The BvgAS phosphorelay regulates ∼10% of the annotated genomes of Bordetella pertussis and Bordetella bronchiseptica and controls their infectious cycles. The hierarchical organization of the regulatory network allows the integration of contextual signals to control all or specific subsets of BvgAS-regulated genes. Here, we characterize a regulatory node involving a type III secretion system (T3SS)-exported protein, BtrA, and demonstrate its role in determining fundamental differences in T3SS phenotypes among Bordetella species. We show that BtrA binds and antagonizes BtrS, a BvgAS-regulated extracytoplasmic function (ECF) sigma factor, to couple the secretory activity of the T3SS apparatus to gene expression. In B. bronchiseptica, a remarkable spectrum of expression states can be resolved by manipulating btrA, encompassing over 80 BtrA-activated loci that include genes encoding toxins, adhesins, and other cell surface proteins, and over 200 BtrA-repressed genes that encode T3SS apparatus components, secretion substrates, the BteA effector, and numerous additional factors. In B. pertussis, BtrA retains activity as a BtrS antagonist and exerts tight negative control over T3SS genes. Most importantly, deletion of btrA in B. pertussis revealed T3SS-mediated, BteA-dependent cytotoxicity, which had previously eluded detection. This effect was observed in laboratory strains and in clinical isolates from a recent California pertussis epidemic. We propose that the BtrA-BtrS regulatory node determines subspecies-specific differences in T3SS expression among Bordetella species and that B. pertussis is capable of expressing a full range of T3SS-dependent phenotypes in the presence of appropriate contextual cues.

  14. 21 CFR 522.2640 - Tylosin.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... (shipping fever, pneumonia) usually associated with Pasteurella multocida and Arcanobacterium pyogenes; foot... Mycoplasma hyosynoviae; swine pneumonia caused by Pasteurella spp.; swine erysipelas caused by Erysipelothrix..., tonsillitis, and pneumonia caused by Staphylococci spp., hemolytic Streptococci spp., and...

  15. Iron and pH-responsive FtrABCD ferrous iron utilization system of Bordetella species

    PubMed Central

    Brickman, Timothy J.; Armstrong, Sandra K.

    2012-01-01

    Summary A putative operon encoding an uncharacterized ferrous iron transport (FtrABCD) system was previously identified in cDNA microarray studies. In growth studies using buffered medium at pH values ranging from pH 6.0 to 7.6, Bordetella pertussis and Bordetella bronchiseptica FtrABCD system mutants showed dramatic reductions in growth yields under iron-restricted conditions at pH 6.0, but had no growth defects at pH 7.6. Supplementation of culture medium with 2 mM ascorbate reductant was inhibitory to alcaligin siderophore-dependent growth at pH 7.6, but had a neglible effect on FtrABCD system-dependent iron assimilation at pH 6.0 consistent with its predicted specificity for ferrous iron. Unlike Bordetella siderophore-dependent and haem iron transport systems, and in agreement with its hypothesized role in transport of inorganic iron from periplasm to cytoplasm, FtrABCD system function did not require the TonB energy transduction complex. Gene fusion analysis revealed that ftrABCD promoter activity was maximal under iron-restricted growth conditions at acidic pH. The pH of human airway surface fluids ranges from pH 5.5 to 7.9, and the FtrABCD system may supply ferrous iron necessary for Bordetella growth in acidic host microenvironments in which siderophores are ineffective for iron retrieval. PMID:22924881

  16. Contribution of Bordetella bronchiseptica Filamentous Hemagglutinin and Pertactin to Respiratory Disease in Swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Respiratory disease in pigs is the most important health concern for swine producers today. Bordetella bronchiseptica is widely prevalent in swine populations and has multiple roles in respiratory disease. It is the primary etiologic agent of atrophic rhinitis, bronchopneumonia in young pigs, and ha...

  17. Contribution of Bordetella bronchiseptica Filamentous Hemagglutinin and Pertactin to Respiratory Disease in Swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Respiratory disease in pigs is the most important health concern for swine producers today. Bordetella bronchiseptica is pervasive in swine populations and plays multiple roles in respiratory disease. It is a primary etiologic agent of atrophic rhinitis, bronchopneumonia in young pigs, and has been ...

  18. Replacement of Adenylate Cyclase Toxin in a Prominent Lineage of Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica is a gram negative respiratory pathogen that infects a wide-range of hosts and causes a diverse spectrum of disease that may be due in part to the set of virulence factors utilized by different strains. In a murine model of infection, we found that virulence, as measured by...

  19. Comparison of Ribotyping and sequence-based typing for discriminating among isolates of Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: Our goal was to compare the discriminatory power of PvuII ribotyping and MLST using a single set of diverse Bordetella bronchiseptica isolates and to determine whether subtyping based on repeat region sequences of the pertactin gene (prn) provides additional resolution. Methods and Results: ...

  20. Aerosolized vaccine as an unexpected source of false-positive Bordetella pertussis PCR results.

    PubMed

    Salimnia, Hossein; Lephart, Paul R; Asmar, Basim I; Prebelich, Dawn; Paulson, Erin; Fairfax, Marilynn R

    2012-02-01

    When 13 of 13 nasal wash specimens from a single pediatrician's office tested positive for low quantities of Bordetella pertussis DNA, we suspected prelaboratory contamination. Investigation revealed that Pentacel and Adacel vaccines contain high copy numbers of B. pertussis DNA, which can be aerosolized, causing false-positive B. pertussis PCR results.

  1. Bordetella bronchiseptica in a paediatric cystic fibrosis patient: possible transmission from a household cat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica was isolated from the sputum of a cystic fibrosis patient recently exposed to a kitten with an acute respiratory disease. Genetic characterization of the isolate and comparison with other isolates of human or feline origin strongly implicate the kitten as the source of infe...

  2. Identification and taxonomic characterization of Bordetella pseudohinzii spp. nov. isolated from laboratory-raised mice

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella species of the class Betaproteobacteria have historically been subdivided into the "classical” group, represented by the respiratory pathogens B. bronchiseptica, B. pertussis and B. parapertussis, and six less extensively studied species. Among the latter "non-classical" group is B. hinz...

  3. The characterisation of Bordetella/Alcaligenes-like organisms and their effects on turkey poults and chicks.

    PubMed

    Varley, J

    1986-01-01

    Eight isolates of the Bordetella or Alcaligenes-like organisms associated with turkey rhino-tracheitis were examined. Five of these isolates had been recovered from the United Kingdom and three were foreign isolates. Four of the UK isolates came from flocks with mild respiratory disease. The fifth isolate came from birds with no respiratory signs and this appears to be the first report of the recovery of Bordetella/Alcaligenes from apparently normal turkeys. The field isolates and type strains Alcaligenes faecalis NCTC 415 and Bordetella bronchiseptica NCTC 452 were characterised by biochemical tests, but these did not include any electrophoresis or nucleic acid studies. Cluster analysis using the group average method and the similarly coefficient of Sokal and Sneath indicated that all the strains were distinct from Alcaligenes faecalis but were quite closely related to Bordetella bronchiseptica. Each field isolate was used to infect separate groups of day-old turkey poults and chicks, and each group contained birds which were experimentally infected and others which were in-contact. Observations were made over a 32-day period. In turkey poults, some of the isolates induced severe respiratory disease and mortality, and others very little or none. The UK isolates were less pathogenic than the foreign isolates. It was not possible to correlate the pathogenicity of the isolates for turkey poults with their biochemical characteristics. Chicks infected with two of the eight isolates showed slight respiratory signs, but there was no significant mortality.

  4. Evaluation of a commercial loop-mediated isothermal amplification assay for diagnosis of Bordetella pertussis infection.

    PubMed

    Kamachi, Kazunari; Moriuchi, Takumi; Hiramatsu, Yukihiro; Otsuka, Nao; Shibayama, Keigo

    2017-02-01

    We evaluated a commercial loop-mediated isothermal amplification (LAMP) assay kit for Bordetella pertussis detection. The LAMP primers were designed to target the ptxP1 allele of the pertussis toxin promoter, but the assay could detect B. pertussis ptxP3 and ptxP8 strains in addition to ptxP1 strains, with high analytical sensitivity.

  5. Complete Genome Sequences of Four Bordetella pertussis Vaccine Reference Strains from Serum Institute of India

    PubMed Central

    Peng, Yanhui; Loparev, Vladimir; Johnson, Taccara; Juieng, Phalasy; Gairola, Sunil; Kumar, Rakesh; Shaligram, Umesh; Gowrishankar, Ramnath; Moura, Hercules; Rees, Jon; Schieltz, David M.; Williamson, Yulanda; Woolfitt, Adrian; Barr, John; Tondella, M. Lucia; Williams, Margaret M.

    2016-01-01

    Serum Institute of India is among the world’s largest vaccine producers. Here, we report the complete genome sequences for four Bordetella pertussis strains used by Serum Institute of India in the production of whole-cell pertussis vaccines. PMID:28007855

  6. Insertion sequences shared by Bordetella species and implications for the biological diagnosis of pertussis syndrome.

    PubMed

    Tizolova, A; Guiso, N; Guillot, S

    2013-01-01

    The molecular diagnosis of pertussis and parapertussis syndromes is based on the detection of insertion sequences (IS) 481 and 1001, respectively. However, these IS are also detected in the genomes of various Bordetella species, such that they are not specific for either B. pertussis or B. parapertussis. Therefore, we screened the genome of recently circulating isolates of Bordetella species to compare the prevalence of IS481, IS1001 and, also IS1002 with previously published data and to sequence all IS detected. We also investigated whether the numbers of IS481 and IS1001 copies vary in recently circulating isolates of the different Bordetella species. We used the polymerase chain reaction (PCR) method for screening the genome of circulating isolates and to prepare the fragments for sequencing. We used Southern blotting and quantitative real-time PCR for quantification of the numbers of IS. We found no significant diversity in the sequences of the IS harboured in the genomes of the Bordetella isolates screened, except for a 71-nucleotide deletion from IS1002 in B. bronchiseptica. The IS copy numbers in the genome of recently circulating isolates were similar to those in reference strains. Our results confirm that biological diagnosis targeting the IS481 and IS1001 elements are not specific and detect the species B. pertussis, B. holmesii and B. bronchiseptica (IS481), and B. parapertussis and B. bronchiseptica (IS1001).

  7. Bordetella trematum bacteremia in an infant: a cause to look for.

    PubMed

    Saksena, R; Manchanda, V; Mittal, M

    2015-01-01

    Bordetella trematum spp. nov. has been isolated from wounds, ear infections and diabetic ulcers. We report a case of a 7-month-old infant with fever, vomiting and abnormal body movements with bacteremia caused by this novel species. The infant responded to fluoroquinolone and macrolide combination therapy.

  8. Draft genome sequence of the Bordetella bronchiseptica swine isolate KM22

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica swine isolate KM22 has been used in experimental infections of swine as a model of clinical B. bronchiseptica infections within swine herds and to study host-to-host transmission. Here we report the draft genome sequence of KM22....

  9. Draft genome sequences of six Bordetella hinzii isolates acquired from Avian and Mammalian hosts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella hinzii is a Gram-negative bacterium known to infect poultry, humans, rabbits and rodents. It is an opportunistic pathogen in immunocompromised humans and some strains cause mild to moderate respiratory disease in turkeys. Little is known as to the degree of genetic diversity within the ...

  10. Evidence for horizontal gene transfer of two antigenically distinct O antigens in Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Antigenic variation is one mechanism pathogens use to avoid immune-mediated competition between closely related strains. Here, we show that two Bordetella bronchiseptica strains, RB50 and 1289, express two antigenically distinct O-antigen serotypes (O1 and O2 respectively). When 18 additional B. b...

  11. Septic arthritis caused by Bordetella holmesii in an adolescent with chronic haemolytic anaemia.

    PubMed

    Moissenet, Didier; Leverger, Guy; Mérens, Audrey; Bonacorsi, Stéphane; Guiso, Nicole; Vu-Thien, Hoang

    2011-11-01

    We describe a case of septic arthritis caused by Bordetella holmesii in a 15-year-old boy with chronic haemolytic anaemia. B. holmesii was identified by 16S rRNA gene sequence analysis. The patient outcome was favourable. To our knowledge, this is the first case of B. holmesii septic arthritis in an asplenic patient.

  12. Bordetella holmesii meningitis in a 12-year-old anorectic girl.

    PubMed

    Van Balen, Tessa; Nieman, An-Emmie; Hermans, Mirjam H A; Schneeberger, Peter M; de Vries, Esther

    2012-04-01

    We describe a 12-year-old anorectic girl with Bordetella holmesii meningitis, the techniques used for its identification, and minimum inhibitory concentrations of antibiotics for 7 B. Holmesii strains collected in the Netherlands during the past 12 years. B. holmesii meningitis has not been previously reported.

  13. Novel, host-restricted genotypes of Bordetella bronchiseptica associated with phocine respiratory tract isolates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica is a widespread respiratory pathogen in a variety of wild and domesticated animals. During a succession of phocine morbillivirus outbreaks occurring over the past 25 years, it was identified as a frequent secondary invader, often believed to be the cause of death. Prior a...

  14. First report of infectious pericarditis due to Bordetella holmesii in an adult patient with malignant lymphoma.

    PubMed

    Nei, Takahito; Hyodo, Hideya; Sonobe, Kazunari; Dan, Kazuo; Saito, Ryoichi

    2012-05-01

    Bordetella holmesii is a fastidious Gram-negative rod first identified in 1995. Though rare, it is isolated mainly in immunocompromised and asplenic hosts and is associated with bacteremia, pertussis-like respiratory tract infection, and endocarditis. Herein, we describe a unique B. holmesii infectious pericarditis patient with malignant lymphoma.

  15. Iron and pH-responsive FtrABCD ferrous iron utilization system of Bordetella species.

    PubMed

    Brickman, Timothy J; Armstrong, Sandra K

    2012-11-01

    A putative operon encoding an uncharacterized ferrous iron transport (FtrABCD) system was previously identified in cDNA microarray studies. In growth studies using buffered medium at pH values ranging from pH 6.0 to 7.6, Bordetella pertussis and Bordetella bronchiseptica FtrABCD system mutants showed dramatic reductions in growth yields under iron-restricted conditions at pH 6.0, but had no growth defects at pH 7.6. Supplementation of culture medium with 2 mM ascorbate reductant was inhibitory to alcaligin siderophore-dependent growth at pH 7.6, but had a neglible effect on FtrABCD system-dependent iron assimilation at pH 6.0 consistent with its predicted specificity for ferrous iron. Unlike Bordetella siderophore-dependent and haem iron transport systems, and in agreement with its hypothesized role in transport of inorganic iron from periplasm to cytoplasm, FtrABCD system function did not require the TonB energy transduction complex. Gene fusion analysis revealed that ftrABCD promoter activity was maximal under iron-restricted growth conditions at acidic pH. The pH of human airway surface fluids ranges from pH 5.5 to 7.9, and the FtrABCD system may supply ferrous iron necessary for Bordetella growth in acidic host microenvironments in which siderophores are ineffective for iron retrieval.

  16. Novel multitarget real-time PCR assay for rapid detection of Bordetella species in clinical specimens.

    PubMed

    Tatti, Kathleen M; Sparks, Kansas N; Boney, Kathryn O; Tondella, Maria Lucia

    2011-12-01

    A novel multitarget real-time PCR (RT-PCR) assay for the rapid identification of Bordetella pertussis, B. parapertussis, and B. holmesii was developed using multicopy insertion sequences (ISs) in combination with the pertussis toxin subunit S1 (ptxS1) singleplex assay. The RT-PCR targets for the multiplex assay include IS481, commonly found in B. pertussis and B. holmesii; IS1001 of B. parapertussis; and the IS1001-like sequence of B. holmesii. Overall, 402 Bordetella species and 66 non-Bordetella species isolates were tested in the multitarget assay. Cross-reactivity was found only with 5 B. bronchiseptica isolates, which were positive with IS1001 of B. parapertussis. The lower limit of detection (LLOD) of the multiplex assay was similar to the LLOD of each target in an individual assay format, which was approximately 1 genomic equivalent per reaction for all targets. A total of 197 human clinical specimens obtained during cough-illness outbreak investigations were used to evaluate the multitarget RT-PCR assay. The multiplex assay results from 87 clinical specimens were compared to the individual RT-PCR assay and culture results. The multitarget assay is useful as a diagnostic tool to confirm B. pertussis infections and to rapidly identify other Bordetella species. In conclusion, the use of this multitarget RT-PCR approach increases specificity, while it decreases the amount of time, reagents, and specimen necessary for RT-PCRs used for accurate diagnosis of pertussis-like illness.

  17. Early Induction of Cytokines in Pigs Coinfected with Swine Influenza Virus and Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Respiratory disease is one of the most important health issues for the swine industry, and coinfection with two or more pathogens is a common occurrence. Bordetella bronchiseptica and swine influenza virus (SIV) are important and common respiratory pathogens of pigs. A study examining the effect o...

  18. Early Induction of Cytokines in Pigs Coinfected with Swine Influenza Virus and Bordetella bronchiseptica

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Respiratory disease is one of the most important health issues for the swine industry, and coinfection with two or more pathogens is a common occurrence. Bordetella bronchiseptica and swine influenza virus (SIV) are important and common respiratory pathogens of pigs. The effect of coinfection of S...

  19. Bordetella bronchiseptica associated with pulmonary disease in mountain voles (Microtus montanus)

    USGS Publications Warehouse

    Jensen, W.I.; Duncan, R.M.

    1980-01-01

    Bordetella bronchiseptica was isolated from the lungs of all of six mountain voles (Microtus montanus) found dead or dying of pulmonary infection near the Bear River Research Station in northern Utah in January, 1973. The possibility of concomitant viral or mycoplasmal infection was not ruled out.

  20. Identification of Bordetella bronchseptica in fatal pneumonia of dogs and cats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infection with Bordetella bronchiseptica is a common cause of tracheobronchitis and upper respiratory disease in dogs and cats, but it can also lead to fatal pneumonia. Identification of this pathogen is important due the risk of transmission to other animals, availability of vaccines and potential...

  1. 21 CFR 522.2630 - Tulathromycin.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ..., Pasteurella multocida, Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in..., and Mycoplasma hyopneumoniae; and for the control of SRD associated with A. pleuropneumoniae,...

  2. 21 CFR 520.90e - Ampicillin trihydrate soluble powder.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...) Indications for use. Oral treatment of porcine colibacillosis (Escherichia coli) and salmonellosis (Salmonella... Pasteurella multocida, Staphylococcus spp., Streptococcus spp., and Salmonella spp. (3) Limitations. For...

  3. 21 CFR 520.90e - Ampicillin trihydrate soluble powder.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...) Indications for use. Oral treatment of porcine colibacillosis (Escherichia coli) and salmonellosis (Salmonella... Pasteurella multocida, Staphylococcus spp., Streptococcus spp., and Salmonella spp. (3) Limitations. For...

  4. 21 CFR 520.314 - Cefadroxil.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Staphylococcus aureus. For the treatment of genitourinary tract infections (cystitis) due to susceptible strains... strains of Pasteurella multocida, S. aureus, Staphylococcus epidermidis, and Streptococcus spp....

  5. 21 CFR 520.314 - Cefadroxil.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Staphylococcus aureus. For the treatment of genitourinary tract infections (cystitis) due to susceptible strains... strains of Pasteurella multocida, S. aureus, Staphylococcus epidermidis, and Streptococcus spp....

  6. 21 CFR 520.314 - Cefadroxil.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Staphylococcus aureus. For the treatment of genitourinary tract infections (cystitis) due to susceptible strains... strains of Pasteurella multocida, S. aureus, Staphylococcus epidermidis, and Streptococcus spp....

  7. 21 CFR 520.314 - Cefadroxil.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Staphylococcus aureus. For the treatment of genitourinary tract infections (cystitis) due to susceptible strains... strains of Pasteurella multocida, S. aureus, Staphylococcus epidermidis, and Streptococcus spp....

  8. 21 CFR 520.314 - Cefadroxil.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Staphylococcus aureus. For the treatment of genitourinary tract infections (cystitis) due to susceptible strains... strains of Pasteurella multocida, S. aureus, Staphylococcus epidermidis, and Streptococcus spp....

  9. 21 CFR 520.90e - Ampicillin trihydrate soluble powder.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...) Indications for use. Oral treatment of porcine colibacillosis (Escherichia coli) and salmonellosis (Salmonella... Pasteurella multocida, Staphylococcus spp., Streptococcus spp., and Salmonella spp. (3) Limitations. For...

  10. 21 CFR 520.90e - Ampicillin trihydrate soluble powder.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...) Indications for use. Oral treatment of porcine colibacillosis (Escherichia coli) and salmonellosis (Salmonella... Pasteurella multocida, Staphylococcus spp., Streptococcus spp., and Salmonella spp. (3) Limitations. For...

  11. Acid tolerance response of Bordetella bronchiseptica in avirulent phase.

    PubMed

    Fingermann, M; Hozbor, D

    2015-12-01

    Bordetella bronchiseptica is a Gram-negative bacterium responsible for respiratory diseases in many mammalian hosts, including humans. This pathogen has been shown as able to persist inside the host cells, even in the phagosomes that are acidified to pH 4.5-5.0 after bacterial infection. Here we evaluated the resistance of B. bronchiseptica to survive under acidic conditions. In particular we analyzed the bacterial capacity to develop the mechanism known as acid tolerance response (ATR). Our studies were mainly focused on the avirulent phase of the bacteria since this phenotypic phase was reported to be more resistant to environmental stress conditions than the virulent phase. Results from B. bronchiseptica in virulent phase were also included for comparison purposes. In fact, for B. bronchiseptica 9.73 bacteria in virulent phase we observed that the viability of bacteria does not decrease significantly when grown at pH as low as 4.5, but it is affected when the pH of the medium was equal to or less than 4.0. After acid-adaptation at pH 5.5 for several hours, the survival rate of B. bronchiseptica 9.73 at lethal pH 4.0 for 6h was increased. Interestingly, the avirulent phase mediated by the two-component BvgAS system conferred further resistance to lethal acid challenge and a marked increase in the magnitude of the expressed ATR. The ATR for this avirulent phase seems to be associated with changes in LPS and surface protein profiles. 2D-gel electrophoresis revealed at least 25 polypeptides differentially expressed, 17 of which were only expressed or over-expressed under acid conditions. Using MALDI-TOF mass spectrometry, 10 of these differentially expressed polypeptides were identified.

  12. Utilization of multiple real-time PCR assays for the diagnosis of Bordetella spp. in clinical specimens.

    PubMed

    Tatti, Kathleen M; Tondella, Maria Lucia

    2013-01-01

    Bordetella pertussis causes an upper respiratory infection in infants, adolescents, and adults. Diagnosis of pertussis, a vaccine-preventable disease, can be difficult, but recent implementation of real-time PCR assays in laboratories has hastened the ability of clinicians to make an accurate diagnosis. In this paper we describe the method of nasopharyngeal specimen collection, extraction of DNA, and real-time PCR assays that will allow the detection and identification of Bordetella spp. in clinical specimens.

  13. 21 CFR 522.2630 - Tulathromycin.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk of.... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for...

  14. 21 CFR 522.2630 - Tulathromycin.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk of.... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for...

  15. 21 CFR 522.2630 - Tulathromycin.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk of.... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for...

  16. 21 CFR 522.2630 - Tulathromycin.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk of.... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for...

  17. SELECTIVE INHIBITION BY TRYPTOPHAN ANALOGUES OF MURINE TOXIN SYNTHESIS IN PASTEURELLA PESTIS

    PubMed Central

    Montie, Thomas C.; Ajl, Samuel J.

    1964-01-01

    Montie, Thomas C. (Albert Einstein Medical Center, Philadelphia, Pa.), and Samuel J. Ajl. Selective inhibition by tryptophan analogues of murine toxin synthesis in Pasteurella pestis. J. Bacteriol. 88:1467–1475. 1964.—Washed-cell suspensions of Pasteurella pestis, avirulent strain “Tjiwidej,” exhibited a preferential inhibition of toxin synthesis relative to total protein formation, when grown in the presence of various tryptophan analogues. Growth was partially inhibited in the presence of methyl analogues. High concentrations of 5-fluorotryptophan induced slight growth-inhibitory effects. However, toxin production was more sensitive to these levels of the analogue. Growth inhibition appeared not to relate to toxin inhibition. Inhibition of toxin synthesis by analogues was reversed by l-tryptophan and indole. Shikimic acid but not anthranilic acid antagonized the action of 4-methyltryptophan on selective toxin synthesis. The formation of tryptophanless protein accounted for continued protein synthesis in tryptophan-depleted cells. Protein resolved by acrylamide gel electrophoresis from crude cell extracts exhibited two toxic protein bands. The synthesis of one toxin-protein band, the less-mobile of the two, appeared to be associated with the membrane fraction of the cell, and was selectively blocked in cells grown in the presence of tryptophan analogues. Cellular tryptophan levels may determine the quantity and quality of proteins made. Images PMID:14234807

  18. Development of a multiplex real-time PCR assay for the detection of Bordetella pertussis and Bordetella parapertussis in a single tube reaction.

    PubMed

    Arbefeville, Sophie; Levi, Michael H; Ferrieri, Patricia

    2014-02-01

    Pertussis is an infectious respiratory disease caused by the fastidious bacterium Bordetella pertussis, which may infect unvaccinated, previously vaccinated children, and adults in whom immunity has waned. Infants are at a particular risk for severe disease and complications. Bordetella parapertussis may cause a similar illness, however the symptoms are less severe and of shorter duration. Pertussis is a highly contagious disease and early diagnosis is essential. Studies have shown that PCR is 2-4 times more likely than culture to detect Bordetella pertussis. We developed a multiplex, real-time PCR assay using analyte-specific reagent (ASR) primers and probes dispensed in a convenient lyophilized bead format that targeted the multi-copy insertion sequences IS481 and IS1001 of B. pertussis and B. parapertussis, respectively. These specific ASRs were used in conjunction with Cepheid Smartmix. Included in the ASRs is a competitive internal control to evaluate the performance of the PCR reaction. After DNA extraction, amplification and detection were done on the Smart Cycler System, which performs integrated amplification and detection automatically in a single step. Specificity of the assay was confirmed using multiple distinct bacterial strains. Sensitivity of the assay and extraction efficiency were evaluated on DNA isolated from pure bacterial cultures and on spiked respiratory specimens. We also spiked different swab types and transport media to evaluate for interfering substances. To assess accuracy, we studied different patient specimen types received from two outside laboratories that used similar or different methods to detect B. pertussis and B. parapertussis. The sensitivity and the specificity of the assay for B. pertussis were 90% and 96%, respectively, and for B. parapertussis 71% (only 7 positive specimens were available for testing) and 100%, respectively. Our assay was found to be a valid method for the simultaneous detection of B. pertussis and B

  19. Identification of AlcR, an AraC-Type Regulator of Alcaligin Siderophore Synthesis in Bordetella bronchiseptica and Bordetella pertussis

    PubMed Central

    Pradel, Elizabeth; Guiso, Nicole; Locht, Camille

    1998-01-01

    A Fur titration assay was used to isolate DNA fragments bearing putative Fur binding sites (FBS) from a partial Bordetella bronchiseptica genomic DNA library. A recombinant plasmid bearing a 3.5-kb DNA insert was further studied. Successive deletions in the cloned fragment enabled us to map a putative FBS at about 2 kb from one end. Sequence analysis revealed the presence of an FBS upstream from a new gene encoding an AraC-type transcriptional regulator. The deduced protein displays similarity to PchR, an activator of pyochelin siderophore and ferripyochelin receptor synthesis in Pseudomonas aeruginosa. Homologous genes in Bordetella pertussis and Bordetella parapertussis were PCR amplified, and sequence comparisons indicated a very high conservation in the three species. The B. pertussis and B. bronchiseptica chromosomal genes were inactivated by allelic exchange. Under low-iron growth conditions, the mutants did not secrete the alcaligin siderophore and lacked AlcC, an alcaligin biosynthetic enzyme. Alcaligin production was restored after transformation with a plasmid bearing the wild-type gene. On the basis of its role in regulation of alcaligin biosynthesis, the new gene was designated alcR. Additional sequence determination showed that alcR is located about 2 kb downstream from the alcABC operon and is transcribed in the same orientation. Two tightly linked open reading frames, alcD and alcE, were identified between alcC and alcR. AlcE is a putative iron-sulfur protein; AlcD shows no homology with the proteins in the database. The production of major virulence factors and colonization in the mouse respiratory infection model are AlcR independent. PMID:9473041

  20. Identification and Characterization of alcR, a Gene Encoding an AraC-Like Regulator of Alcaligin Siderophore Biosynthesis and Transport in Bordetella pertussis and Bordetella bronchiseptica

    PubMed Central

    Beaumont, Fiona C.; Kang, Ho Young; Brickman, Timothy J.; Armstrong, Sandra K.

    1998-01-01

    A Bordetella bronchiseptica iron transport mutant was isolated following an enrichment procedure based on streptonigrin resistance. The mutant displayed a growth defect on iron-restricted medium containing ferric alcaligin as the sole iron source. In addition to the apparent inability to acquire iron from the siderophore, the mutant failed to produce alcaligin as well as two known iron-regulated proteins, one of which is the AlcC alcaligin biosynthesis protein. A 1.6-kb KpnI-PstI Bordetella pertussis DNA fragment mapping downstream of the alcaligin biosynthesis genes alcABC restored both siderophore biosynthesis and expression of the iron-regulated proteins to the mutant. Nucleotide sequencing of this complementing 1.6-kb region identified an open reading frame predicted to encode a protein with strong similarity to members of the AraC family of transcriptional regulators, for which we propose the gene designation alcR. Primer extension analysis localized an iron-regulated transcription initiation site upstream of the alcR open reading frame and adjacent to sequences homologous to the consensus Fur repressor binding site. The AlcR protein was produced by using an Escherichia coli expression system and visualized in electrophoretic gels. In-frame alcR deletion mutants of B. pertussis and B. bronchiseptica were constructed, and the defined mutants exhibited the alcR mutant phenotype, characterized by the inability to produce and transport alcaligin and express the two iron-repressed proteins. The cloned alcR gene provided in trans restored these siderophore system activities to the mutants. Together, these results indicate that AlcR is involved in the regulation of Bordetella alcaligin biosynthesis and transport genes and is required for their full expression. PMID:9473040

  1. Bordetella holmesii infection: current knowledge and a vision for future research.

    PubMed

    Pittet, Laure F; Posfay-Barbe, Klara M

    2015-08-01

    Bordetella holmesii is a recently recognized Gram-negative bacterium causing both pertussis-like respiratory symptoms and invasive infections, such as bacteremia, pneumonia, meningitis, arthritis, pericarditis and endocarditis. Few data are available on its epidemiological characteristics, mostly related to respiratory infections. However, these are frequently misdiagnosed as a Bordetella pertussis infection as most diagnostic tests routinely used are not species-specific, thus biasing the epidemiological studies of both strains, as well as the efficacy studies on pertussis vaccination. There is no accepted agreement on treatment and it remains unknown if antimicrobial prophylaxis is indicated in certain clinical settings. We review here the current knowledge on B. holmesii and the need for further research.

  2. Bordetella pertussis and pertactin-deficient clinical isolates: lessons for pertussis vaccines.

    PubMed

    Hegerle, Nicolas; Guiso, Nicole

    2014-09-01

    Bordetella pertussis causes whooping cough in humans, a highly transmissible respiratory disease life threatening for unvaccinated infants. Vaccination strategies were thus introduced worldwide with great success in developed countries reaching high vaccine coverage with efficacious vaccines. In the late 20th/early 21st century, acellular pertussis vaccines replaced whole cell pertussis vaccines but B. pertussis still circulates and evolves in humans, its only known reservoir. The latest transformation of this pathogen, and of its close relative Bordetella parapertussis, is the loss of pertactin production, a virulence factor included in different acellular pertussis vaccines. The real impact of this evolution on acellular pertussis vaccines efficacy and effectiveness should be assessed through standardized surveillance and isolation of B. pertussis and B. parapertussis worldwide.

  3. The potential role of subclinical Bordetella Pertussis colonization in the etiology of multiple sclerosis.

    PubMed

    Rubin, Keith; Glazer, Steven

    2016-04-01

    It is established that (1) subclinical Bordetella pertussis colonization of the nasopharynx persists in highly vaccinated populations, and (2) B. pertussis toxin is a potent adjuvant that, when co-administered with neural antigens, induces neuropathology in experimental autoimmune encephalomyelitis, the principle animal model of multiple sclerosis. Building on these observations with supporting epidemiologic and biologic evidence, we propose that, contrary to conventional wisdom that subclinical pertussis infections are innocuous to hosts, B. pertussis colonization is an important cause of multiple sclerosis.

  4. Genome Sequences of 28 Bordetella pertussis U.S. Outbreak Strains Dating from 2010 to 2012

    PubMed Central

    Harvill, Eric T.; Goodfield, Laura L.; Ivanov, Yury; Meyer, Jessica A.; Newth, Christopher; Cassiday, Pamela; Tondella, Maria Lucia; Liao, Patty; Zimmerman, Jerry; Meert, Kathleen; Wessel, David; Berger, John; Dean, J. Michael; Holubkov, Richard; Burr, Jeri; Liu, Teresa; Brinkac, Lauren; Kim, Maria

    2013-01-01

    Despite the availability of highly effective vaccines, Bordetella pertussis incidence has been rapidly rising in highly vaccinated populations. Recent outbreaks have received media attention, feeding concerns about the emergence of dangerous new strains with increased virulence or that escape vaccine-induced immunity. To accelerate the study of this reemerging pathogen, we sequenced the genomes of 28 B. pertussis strains isolated during outbreaks from 2010 through 2012, making both strains and sequence data available to the scientific community. PMID:24356839

  5. A vir-repressed gene of Bordetella pertussis is required for virulence.

    PubMed Central

    Beattie, D T; Shahin, R; Mekalanos, J J

    1992-01-01

    Coordinate regulation of gene expression in Bordetella pertussis is controlled by the products of the vir locus, BvgA and BvgS. In the presence of modulating signals such as MgSO4 and nicotinic acid, expression of vir-activated genes (vag) is reduced, while expression of vir-repressed genes (vrg) is maximal. We have cloned one of these vir-repressed genes, vrg-6, in Escherichia coli. DNA sequencing has shown that vrg-6 is contained on a single EcoRI restriction endonuclease fragment and is predicted to code for a protein of 105 amino acids with a molecular weight of 11,441. The predicted protein product appears to have two domains, one consisting of seven hydrophobic proline-rich pentameric repeats and the other consisting of five alkaline trimeric repeats. Southern blot analysis has revealed vrg-6-homologous sequences in the chromosomes of Bordetella bronchiseptica and Bordetella parapertussis, but, unlike Bordetella pertussis, these species do not express vrg-6-homologous RNA when grown under modulating conditions. In order to assess the role of vrg gene products in B. pertussis pathogenesis, two 18323 derivatives which harbor TnphoA insertions in vrg genes were analyzed in a mouse model of respiratory infection. Strain SK6, which carries a vrg-6::TnphoA mutation, failed to induce lymphocytosis and was significantly less able to colonize lungs and trachea than its parent strain 18323 or than SK18, which harbors a TnphoA fusion in the vrg-18 locus. This is the first evidence that a vir-repressed gene may play an important role in the virulence of B. pertussis and the pathogenesis of whooping cough. Images PMID:1730491

  6. Host Specificity of Ovine Bordetella parapertussis and the Role of Complement.

    PubMed

    Hester, Sara E; Goodfield, Laura L; Park, Jihye; Feaga, Heather A; Ivanov, Yury V; Bendor, Liron; Taylor, Dawn L; Harvill, Eric T

    2015-01-01

    The classical bordetellae are comprised of three subspecies that differ from broad to very limited host specificity. Although several lineages appear to have specialized to particular host species, most retain the ability to colonize and grow in mice, providing a powerful common experimental model to study their differences. One of the subspecies, Bordetella parapertussis, is composed of two distinct clades that have specialized to different hosts: one to humans (Bpphu), and the other to sheep (Bppov). While Bpphu and the other classical bordetellae can efficiently colonize mice, Bppov strains are severely defective in their ability to colonize the murine respiratory tract. Bppov genomic analysis did not reveal the loss of adherence genes, but substantial mutations and deletions of multiple genes involved in the production of O-antigen, which is required to prevent complement deposition on B. bronchiseptica and Bpphu strains. Bppov lacks O-antigen and, like O-antigen mutants of other bordetellae, is highly sensitive to murine complement-mediated killing in vitro. Based on these results, we hypothesized that Bppov failed to colonize mice because of its sensitivity to murine complement. Consistent with this, the Bppov defect in the colonization of wild type mice was not observed in mice lacking the central complement component C3. Furthermore, Bppov strains were highly susceptible to killing by murine complement, but not by sheep complement. These data demonstrate that the failure of Bppov to colonize mice is due to sensitivity to murine, but not sheep, complement, providing a mechanistic example of how specialization that accompanies expansion in one host can limit host range.

  7. Characterization of the Filamentous Hemagglutinin-Like Protein FhaS in Bordetella bronchiseptica

    PubMed Central

    Julio, Steven M.; Cotter, Peggy A.

    2005-01-01

    Filamentous hemagglutinin (FHA) is a large (>200 kDa), rod-shaped protein expressed by bordetellae that is both surface-associated and secreted. FHA mediates bacterial adherence to epithelial cells and macrophages in vitro and is absolutely required for tracheal colonization in vivo. The recently sequenced Bordetella bronchiseptica genome revealed the presence of a gene, fhaS, that is nearly identical to fhaB, the FHA structural gene. We show that although fhaS expression requires the BvgAS virulence control system, it is maximal only under a subset of conditions in which BvgAS is active, suggesting an additional level of regulation. We also show that, like FHA, FhaS undergoes a C-terminal proteolytic processing event and is both surface-associated and secreted and that export across the outer membrane requires the channel-forming protein FhaC. Unlike FHA, however, FhaS was unable to mediate adherence of B. bronchiseptica to epithelial cell lines in vitro and was not required for respiratory tract colonization in vivo. In a coinfection experiment, a ΔfhaS strain was out-competed by wild-type B. bronchiseptica, indicating that fhaS is expressed in vivo and that FhaS contributes to bacterial fitness in a manner revealed when the mutant must compete with wild-type bacteria. These data suggest that FHA and FhaS perform distinct functions during the Bordetella infectious cycle. A survey of various Bordetella strains revealed two distinct fhaS alleles that segregate according to pathogen host range and that B. parapertussishu most likely acquired its fhaS allele from B. pertussis horizontally, suggesting fhaS may contribute to host-species specificity. PMID:16041011

  8. Bordetella bronchiseptica in a Pediatric Cystic Fibrosis Patient: Possible Transmission from a Household Cat

    PubMed Central

    Register, Karen B.; Sukumar, Neelima; Palavecino, Elizabeth L.; Rubin, Bruce K.; Deora, Rajendar

    2011-01-01

    Summary Bordetella bronchiseptica is a zoonotic respiratory pathogen commonly found in domesticated farm and companion animals, including dogs and cats. Here we report isolation of B. bronchiseptica from a sputum sample of a cystic fibrosis patient recently exposed to a kitten with an acute respiratory illness. Genetic characterization of the isolate and comparison with other isolates of human or feline origin strongly suggest the kitten was the source of infection. PMID:22212633

  9. Piracy of adhesins: attachment of superinfecting pathogens to respiratory cilia by secreted adhesins of Bordetella pertussis.

    PubMed

    Tuomanen, E

    1986-12-01

    Two proteins secreted by Bordetella pertussis are known to mediate adherence of these bacteria to mammalian respiratory cilia. When either ciliated cells or other pathogenic bacteria were pretreated with these adhesins, Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus acquired the ability to adhere to cilia in vitro and in vivo. Such piracy of adhesins may contribute to superinfection in mucosal diseases such as whooping cough.

  10. A Case of Lower Respiratory Tract Infection with Canine-associated Pasteurella canis in a Patient with Chronic Obstructive Pulmonary Disease

    PubMed Central

    Acharya, Preetam R.; Biranthabail, Dhanashree; Rangnekar, Aseem; Shiragavi, Sachin

    2015-01-01

    This is the report of lower respiratory tract infection with Pasteurella canis in a chronic obstructive pulmonary disease (COPD) patient with history of casual exposure to cats. Pasteurella species are part of the oral and gastrointestinal flora in the canine animals. These organisms are usually implicated in wound infection following animal bites, but can also be associated with a variety of infections including respiratory tract infections. PMID:26435948

  11. Type Six Secretion System of Bordetella bronchiseptica and Adaptive Immune Components Limit Intracellular Survival During Infection

    PubMed Central

    Bendor, Liron; Weyrich, Laura S.; Linz, Bodo; Rolin, Olivier Y.; Taylor, Dawn L.; Goodfield, Laura L.; Smallridge, William E.; Kennett, Mary J.; Harvill, Eric T.

    2015-01-01

    The Type Six Secretion System (T6SS) is required for Bordetella bronchiseptica cytotoxicity, cytokine modulation, infection, and persistence. However, one-third of recently sequenced Bordetella bronchiseptica strains of the predominantly human-associated Complex IV have lost their T6SS through gene deletion or degradation. Since most human B. bronchiseptica infections occur in immunocompromised patients, we determine here whether loss of Type Six Secretion is beneficial to B. bronchiseptica during infection of immunocompromised mice. Infection of mice lacking adaptive immunity (Rag1-/- mice) with a T6SS-deficient mutant results in a hypervirulent phenotype that is characterized by high numbers of intracellular bacteria in systemic organs. In contrast, wild-type B. bronchiseptica kill their eukaryotic cellular hosts via a T6SS-dependent mechanism that prevents survival in systemic organs. High numbers of intracellular bacteria recovered from immunodeficient mice but only low numbers from wild-type mice demonstrates that B. bronchiseptica survival in an intracellular niche is limited by B and T cell responses. Understanding the nature of intracellular survival during infection, and its effects on the generation and function of the host immune response, are important to contain and control the spread of Bordetella-caused disease. PMID:26485303

  12. Identification of a 68-kilodalton protective protein antigen from Bordetella bronchiseptica.

    PubMed Central

    Montaraz, J A; Novotny, P; Ivanyi, J

    1985-01-01

    A 68-kilodalton (kd) outer membrane protein antigen of Bordetella bronchiseptica has been identified by using monoclonal antibodies that recognized two nonoverlapping determinants. Antibody BB05 also reacted with homologous proteins from Bordetella pertussis and Bordetella parapertussis but not with another 12 organisms from various bacterial genera. Passive injection of BB05 antibody protected mice from aerosol infection with B. bronchiseptica as shown by reduced mortality and reduced pathology of turbinate bones. The 68-kd B. bronchiseptica antigen was purified by BB05-based affinity chromatography and evaluated for its potency to immunize mice actively against either intraperitoneal or aerosol challenge with B. bronchiseptica. Immunization with the 68-kd antigen in incomplete Freund adjuvant significantly reduced the levels of mortality in intraperitoneally challenged mice. In the aerosol infection model, injection of the 68-kd antigen with complete or incomplete Freund adjuvant or saponin reduced the bacterial counts in the lungs of infected mice. These results suggest that the 68-kd protein may represent a potential "protective" antigen of B. bronchiseptica. Images PMID:3972452

  13. Role of feedback and network architecture in controlling virulence gene expression in Bordetella.

    PubMed

    Prajapat, Mahendra Kumar; Saini, Supreet

    2013-11-01

    Bordetella is a Gram-negative bacterium responsible for causing whooping cough in a broad range of host organisms. For successful infection, Bordetella controls expression of four distinct classes of genes (referred to as class 1, 2, 3, and 4 genes) at distinct times in the infection cycle. This control is executed by a single two-component system, BvgAS. Interestingly, the transmembrane component of the two-component system, BvgS, consists of three phospho-transfer domains leading to phosphorylation of the response regulator, BvgA. Phosphorylated BvgA then controls expression of virulence genes and also controls bvgAS transcription. In this work, we perform simulations to characterize the role of the network architecture in governing gene expression in Bordetella. Our results show that the wild-type network is locally optimal for controlling the timing of expression of the different classes of genes involved in infection. In addition, the interplay between environmental signals and positive feedback aids the bacterium identify precise conditions for and control expression of virulence genes.

  14. Prior exposure to Bordetella species as an exclusion criterion in the baboon model of pertussis

    PubMed Central

    NGUYEN, Annalee W.; WAGNER, Ellen K.; POSADA, Luciano; LIU, Xinlei; CONNELLY, Sheila; PAPIN, James F.; WOLF, Roman F.; KALEKO, Michael; MAYNARD, Jennifer A.

    2016-01-01

    The baboon model of Bordetella pertussis infection is the newest and most clinically accurate model of the human disease to date. However, among the 15 experimentally infected baboons in this study, a subset of baboons did not exhibit the expected high bacterial colonization levels or increase in white blood cell count. Moreover, cultures of nasopharyngeal wash samples from several baboons suggested B. bronchiseptica coinfection. Analysis of serum antibodies recognizing filamentous hemagglutinin, pertussis toxin and B. pertussis lipo-oligosaccharide indicated that several baboons had likely been previously exposed to Bordetella species and that prior exposure correlated with partial protection from B. pertussis infection. Notably, all animals with a baseline Fha titer of 5 IU/ml or below exhibited symptoms typical of the model, suggesting this value can be used as inclusion criteria for animals prior to study enrollment. While B. pertussis infection is endemic to human populations and B. bronchiseptica is common in wild small mammals, this study illustrates that baboons can readily harbor both organisms. Awareness of Bordetella species that share antigens capable of generating protective immune responses and tracking of prior exposure to those species is required for successful use of the baboon model of pertussis. PMID:27666464

  15. Occurence of Bordetella bronchiseptica in domestic cats with upper respiratory tract infections.

    PubMed

    Garbal, M; Adaszek, Ł; Łyp, P; Frymus, J; Winiarczyk, M; Winiarczyk, S

    2016-01-01

    Bordetella bronchiseptica is a widespread Gram-negative pathogen occurring in different mammal species. It is known to play a role in the etiology of infectious atrophic rhinitis of swine, canine kennel cough, respiratory syndromes of cats, rabbits and guinea pigs, and sporadic human cases have also been reported. The aim of this article is to present the occurrence of infections caused by these bacteria in domestic cats with respiratory symptoms, as well as to conduct a molecular analysis of the flaA gene B. bronchiseptica for the purpose of ascertaining whether cats become infected with one or more bacteria strains. B. bronchiseptica was isolated from the respiratory system of 16 out of 35 domestic cats with symptoms of respiratory tract infections. Polymorphism analysis of polymerase chain reaction products of B. bronchiseptica flaA was performed to reveal the possible differences in nucleotide sequences of the flagellin gene. The phylogenetic analysis of nucleotide sequences obtained during PCR indicated that the isolates of bacteria from our own studies are characterised by 100% homology of the analysed fragment of the flaA gene, which suggests maintenance of a single genotype of these microorganisms in the cat population. Moreover, the bacteria revealed full homology with reference strain B. bronchiseptica ATCC 4617, and 99.4% homology with strain B. parapertussis ATCC 15311. This indicates that the PCR optimised for the Bordetella spp. flaA gene, combined with sequencing of amplicons obtained in PCR, is an effective diagnostic method allowing differentiation of Bordetella spp. type microorganisms.

  16. Identification and taxonomic characterization of Bordetella pseudohinzii sp. nov. isolated from laboratory-raised mice

    PubMed Central

    Register, Karen B.; Newman, Jeffrey D.; Taylor, Dawn L.; Boschert, Kenneth R.; Le Guyon, Soazig; Wilson, Emily F.; Brinkac, Lauren M.; Sanka, Ravi; Greco, Suellen C.; Klender, Paula M.; Losada, Liliana

    2016-01-01

    Bordetella hinzii is known to cause respiratory disease in poultry and has been associated with a variety of infections in immunocompromised humans. In addition, there are several reports of B. hinzii infections in laboratory-raised mice. Here we sequenced and analysed the complete genome sequences of multiple B. hinzii-like isolates, obtained from vendor-supplied C57BL/6 mice in animal research facilities on different continents, and we determined their taxonomic relationship to other Bordetella species. The whole-genome based and 16S rRNA gene based phylogenies each identified two separate clades in B. hinzii, one was composed of strains isolated from poultry, humans and a rabbit whereas the other clade was restricted to isolates from mice. Distinctly different estimated DNA–DNA hybridization values, average nucleotide identity scores, gene content, metabolic profiles and host specificity all provide compelling evidence for delineation of the two species, B. hinzii – from poultry, humans and rabbit – and Bordetella pseudohinzii sp. nov. type strain 8-296-03T (=NRRL B-59942T=NCTC 13808T) that infect mice. PMID:27707434

  17. Type Six Secretion System of Bordetella bronchiseptica and Adaptive Immune Components Limit Intracellular Survival During Infection.

    PubMed

    Bendor, Liron; Weyrich, Laura S; Linz, Bodo; Rolin, Olivier Y; Taylor, Dawn L; Goodfield, Laura L; Smallridge, William E; Kennett, Mary J; Harvill, Eric T

    2015-01-01

    The Type Six Secretion System (T6SS) is required for Bordetella bronchiseptica cytotoxicity, cytokine modulation, infection, and persistence. However, one-third of recently sequenced Bordetella bronchiseptica strains of the predominantly human-associated Complex IV have lost their T6SS through gene deletion or degradation. Since most human B. bronchiseptica infections occur in immunocompromised patients, we determine here whether loss of Type Six Secretion is beneficial to B. bronchiseptica during infection of immunocompromised mice. Infection of mice lacking adaptive immunity (Rag1-/- mice) with a T6SS-deficient mutant results in a hypervirulent phenotype that is characterized by high numbers of intracellular bacteria in systemic organs. In contrast, wild-type B. bronchiseptica kill their eukaryotic cellular hosts via a T6SS-dependent mechanism that prevents survival in systemic organs. High numbers of intracellular bacteria recovered from immunodeficient mice but only low numbers from wild-type mice demonstrates that B. bronchiseptica survival in an intracellular niche is limited by B and T cell responses. Understanding the nature of intracellular survival during infection, and its effects on the generation and function of the host immune response, are important to contain and control the spread of Bordetella-caused disease.

  18. Biodegradation of endosulfan isomers and its metabolite endosulfate by two biosurfactant producing bacterial strains of Bordetella petrii.

    PubMed

    Odukkathil, Greeshma; Vasudevan, Namasivayam

    2015-01-01

    The main objective of the investigation was to study the biodegradation of endosulfan isomers and its major metabolite endosulfate by two biosurfactant producing bacterial strains of Bordetella petrii. The significance of the study is to evaluate the capability of biosurfactant producing bacterial strains in enhancing the bioavailability of endosulfan. Sixty bacterial strains were isolated from the endosulfan degrading bacterial consortium and were screened for endosulfan degradation and biosurfactant production. Among those, two strains Bordetella petrii I GV 34 (Gene bank Accession No KJ02262) and Bordetella petrii II GV 36 (Gene bank Accession No KJ022625) were capable of degrading endosulfan with simultaneous biosurfactant production. Bordetella petrii I degraded 89% of α and 84% of β isomers of endosulfan whereas Bordetella petrii II degraded 82% of both the isomers. Both the strains were able to reduce the surface tension up to 19.6% and 21.4% with a minimum observed surface tension of 45 Dynes/cm and 44 Dynes/cm, respectively. The study revealed that the strains have the potential to enhance the degradation endosulfan residues in contaminated sites and water by biosurfactant production.

  19. Identification and characterization of BipA, a Bordetella Bvg-intermediate phase protein.

    PubMed

    Stockbauer, K E; Fuchslocher, B; Miller, J F; Cotter, P A

    2001-01-01

    The Bordetella BvgAS sensory transduction system has traditionally been viewed as controlling a transition between two distinct phenotypic phases: the Bvg(+) or virulent phase and the Bvg(-) or avirulent phase. Recently, we identified a phenotypic phase of Bordetella bronchiseptica that displays reduced virulence in a rat model of respiratory infection concomitant with increased ability to survive nutrient deprivation. Characterization of this phase, designated Bvg-intermediate (Bvg(i)), indicated the presence of antigens that are maximally, if not exclusively, expressed in this phase and therefore suggested the existence of a previously unidentified class of Bvg-regulated genes. We now report the identification and characterization of a Bvg(i) phase protein, BipA (Bvg-intermediate phase protein A), and its structural gene, bipA. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicates that bipA is expressed maximally under Bvgi phase conditions and thus represents the first identified Bvgi phase gene. bipA encodes a 1578-amino-acid protein that shares amino acid sequence similarity at its N-terminus with the proposed outer membrane localization domains of intimin (Int) of enteropathogenic and enterohaemorrhagic Escherichia coli and invasin (Inv) of Yersinia spp. Although not apparent at the amino acid level, BipA is also similar to Int and Inv in that the proposed membrane-spanning domain is followed by several 90-amino-acid repeats and a distinct C-terminal domain. Localization studies using an antibody directed against the C-terminus of BipA indicated that its C-terminus is exposed on the bacterial cell surface. Western blot analysis with this same antibody indicated that BipA homologues are expressed in Bvg(i) phase Bordetella pertussis and Bordetella parapertussis. Comparison of a Delta bipA strain with wild-type B. bronchiseptica indicated that BipA is not required for Bvg(i) phase-specific aggregative adherence to rat lung epithelial cells

  20. Simple visual assay for determination of Pasteurella haemolytica cytotoxin neutralizing antibody titers in cattle sera.

    PubMed Central

    Gentry, M J; Confer, A W; Kreps, J A

    1985-01-01

    A simple visual assay is described for determining the capacity of bovine serum to neutralize the cytotoxin produced by Pasteurella haemolytica serotype 1. The test was reproducible from day to day with different target cell populations and cytotoxin preparations. Cytotoxin neutralization titers obtained by the visual assay were comparable to those determined by the trypan blue exclusion and 51Cr-release methods. The visual assay was used to measure neutralization titers of bovine sera obtained from vaccination experiments and fractions of purified serum obtained by gel filtration. The major advantages of the visual assay over other assays are that it is rapid, inexpensive, and does not use radioisotopes. It also does not require specialized equipment, making it adaptable to most laboratories. Images PMID:3905853

  1. Iron acquisition in Pasteurella haemolytica: expression and identification of a bovine-specific transferrin receptor.

    PubMed Central

    Ogunnariwo, J A; Schryvers, A B

    1990-01-01

    Seven type 1 field isolates of Pasteurella haemolytica were screened for their ability to use different transferrins as a source of iron for growth. All seven strains were capable of using bovine but not human, porcine, avian, or equine transferrin. A screening assay failed to detect siderophore production in any of the strains tested. Iron-deficient cells from these strains expressed a binding activity, specific for bovine transferrin, that was regulated by the level of iron in the medium. Inhibition of expression by translation and transcription inhibitors suggested that iron regulation was occurring at the gene level. Affinity isolation of receptor proteins from all seven strains with biotinylated bovine transferrin identified a 100-kilodalton iron-regulated outer membrane protein as the bovine transferrin receptor. Iron-regulated outer membrane proteins of 71 and 77 kilodaltons were isolated along with the 100-kilodalton protein when less stringent washing procedures were employed in the affinity isolation procedure. Images PMID:2365453

  2. Udder orf infection and its role in ovine clinical mastitis caused by Pasteurella haemolytica.

    PubMed

    Burriel, A R

    1997-04-01

    During an experimental study of ovine subclinical mastitis caused by coagulase-negative staphylococci, an outbreak of contagious ecthyma occurred among ewes unvaccinated against parapox virus. The same group of ewes developed a high rate (43.7%) of clinical mastitis caused by Pasteurella haemolytica. The rate of clinical mastitis among ewes vaccinated against parapox virus was very low (3.7%) suggesting that the presence of orf in the unvaccinated ewes was contributing to the high rate of clinical mastitis. An examination of the iron, sodium, potassium and albumin concentration of milk collected from 16 unvaccinated and nine randomly selected vaccinated ewes before experimental infection with coagulase-negative staphylococci or their uninfected control mammary glands indicated significant differences in the iron (p < 0.0001) and sodium (p = 0.01) concentration. Increased iron concentration in the milk may have assisted in the development of udder infection caused by P. haemolytica as iron is easily utilised by this bacterium.

  3. Efficacy of recombinant leukotoxin in protection against pneumonic challenge with live Pasteurella haemolytica A1.

    PubMed Central

    Conlon, J A; Shewen, P E; Lo, R Y

    1991-01-01

    The recombinant leukotoxin (rLKT) of the bacterium Pasteurella haemolytica A1 was examined for its ability to protect cattle from experimental challenge with logarithmic-phase P. haemolytica. Six different vaccines were utilized in the experiment: P. haemolytica culture supernatant, P. haemolytica culture supernatant enriched with rLKT, rLKT alone, P. haemolytica culture supernatant enriched with Escherichia coli supernatant not containing LKT, E. coli supernatant alone, and phosphate-buffered saline. rLKT alone showed no protective capacity against development of clinical signs of respiratory disease or against development of postmortem lung lesions after experimental challenge. It was, however, shown to enhance the efficacy of the culture supernatant vaccine and decrease clinical signs and pneumonic lesions. The complexity of protective immunity in this disease is emphasized in this study, and, although LKT is an important virulence factor of the organism, an immune response to LKT alone does not protect animals against disease. PMID:1987075

  4. Tracheal versus pulmonary deposition and clearance of inhaled Pasteurella haemolytica or Staphylococcus aureus in mice.

    PubMed Central

    Rodríguez, L M; López, A; Merino-Moncada, M; Martínez-Burnes, J; Mondragón, I

    1985-01-01

    The aim of this investigation was to do a comparative study on the deposition and clearance of inhaled bacteria between the lungs and tracheae of mice exposed to aerosols of bacteria. Two hundred and eighty-eight mice were divided into four groups (n = 72) and exposed to aerosols of Pasteurella haemolytica or Staphylococcus aureus in four replicates. The numbers of bacteria were determined in the trachea and lungs of mice sacrificed 0, 2, 4, 8, 12, 24, 48 and 72 hours postexposure. Results indicated that bacterial deposition was greater in lungs than in tracheae. No significant (p greater than 0.05) difference was observed between P. haemolytica and S. aureus clearance rates. Although bacteria were rapidly eliminated from the whole respiratory tract, bacterial clearance was significantly (p less than 0.002) faster in tracheae than lungs. A significant (p less than 0.05) replicate effect was also observed. PMID:4041977

  5. The pulmonary clearance of Pasteurella haemolytica in calves infected with bovine virus diarrhea or Mycoplasma bovis.

    PubMed Central

    Lopez, A; Maxie, M G; Savan, M; Ruhnke, H L; Thomson, R G; Barnum, D A; Geissinger, H D

    1982-01-01

    Based on current literature which commonly associates bovine virus diarrhea virus and Mycoplasma bovis with "pneumonic pasteurellosis," an investigation was conducted into the effect of these two pathogens on the capacity of bovine lung to clear inhaled Pasteurella haemolytica. There was no significant effect (p less than 0.05) of either bovine virus diarrhea virus or M. bovis on the mean clearance rate of P. haemolytica, nor did the time interval of three, five or seven days between the first inoculation and exposure to P. haemolytica and adversely affect the lung clearance rates. However, it was found that the left lungs and a higher bacterial retention (p less than 0.05) than the right lungs. PMID:7127194

  6. Toll-like receptor 4-positive macrophages protect mice from Pasteurella pneumotropica-induced pneumonia

    NASA Technical Reports Server (NTRS)

    Hart, Marcia L.; Mosier, Derek A.; Chapes, Stephen K.

    2003-01-01

    This study investigates Toll-like receptor 4 (TLR4)-positive macrophages in early recognition and clearance of pulmonary bacteria. TLR4 is a trans-membrane receptor that is the primary recognition molecule for lipopolysaccharide of gram-negative bacteria. The TLR4(Lps-del) mouse strains C57BL10/ScN (B10) and STOCK Abb(tm1) TLR4(Lps-del) Slc11a1(s)(B10 x C2D) are susceptible to pulmonary infections and develop pneumonia when naturally or experimentally infected by the opportunistic bacterium Pasteurella pneumotropica. Since these mice have the TLR4(Lps-del) genotype, we hypothesized that reconstitution of mice with TLR4-positive macrophages would provide resistance to this bacterium. A cultured macrophage cell line (C2D macrophages) and bone marrow cells from C2D mice were adoptively transferred to B10 and B10 x C2D mice by intraperitoneal injection. C2D macrophages increased B10 and B10 x C2D mouse resistance to P. pneumotropica. In C2D-recipient mice there was earlier transcription of tumor necrosis factor alpha and chemokines JE and macrophage inflammatory protein 2 (MIP-2) in the lungs of B10 and B10 x C2D mice, and there was earlier transcription of KC and MIP-1alpha in B10 x C2D mice. In addition, the course of inflammation following experimental Pasteurella challenge was altered in C2D recipients. C2D macrophages also protected B10 x C2D mice, which lack CD4(+) T cells. These data indicate that macrophages are critical for pulmonary immunity and can provide host resistance to P. pneumotropica. This study indicates that TLR4-positive macrophages are important for early recognition and clearance of pulmonary bacterial infections.

  7. Detection of genes expressed in Bordetella bronchiseptica colonizing rat trachea by in vivo expressed-tag immunoprecipitation method.

    PubMed

    Abe, Hiroyuki; Kamitani, Shigeki; Fukui-Miyazaki, Aya; Shinzawa, Naoaki; Nakamura, Keiji; Horiguchi, Yasuhiko

    2015-05-01

    Analyses of bacterial genes expressed in response to the host environment provide clues to understanding the host-pathogen interactions that lead to the establishment of infection. In this study, a novel method named In Vivo Expressed-Tag ImmunoPrecipitation (IVET-PI) was developed for detecting genes expressed in bacteria that are recovered in a small numbers from host tissues. IVET-IP was designed to overcome some drawbacks of previous similar methods. We applied IVET-IP to Bordetella bronchiseptica colonizing rat trachea and identified 173 genes that were expressed in the bacteria over the entire course of an infection. These gene products included two transcriptional factors that are involved in the expression of filamentous hemagglutinin, adenylate cyclase toxin, and major virulence factors for the bordetellae. We consider that this method might provide novel insight into the course of Bordetella infection.

  8. Molecular evolution and host adaptation of Bordetella spp.: phylogenetic analysis using multilocus enzyme electrophoresis and typing with three insertion sequences.

    PubMed Central

    van der Zee, A; Mooi, F; Van Embden, J; Musser, J

    1997-01-01

    A total of 188 Bordetella strains were characterized by the electrophoretic mobilities of 15 metabolic enzymes and the distribution and variation in positions and copy numbers of three insertion sequences (IS). The presence or absence of IS elements within certain lineages was congruent with estimates of overall genetic relationships as revealed by multilocus enzyme electrophoresis. Bordetella pertussis and ovine B. parapertussis each formed separate clusters, while human B. parapertussis was most closely related to IS1001-containing B. bronchiseptica isolates. The results of the analysis provide support for the hypothesis that the population structure of Bordetella is predominantly clonal, with relatively little effective horizontal gene flow. Only a few examples of putative recombinational exchange of an IS element were detected. Based on the results of this study, we tried to reconstruct the evolutionary history of different host-adapted lineages. PMID:9352907

  9. Cloning and nucleotide sequence of the aroA gene of Bordetella pertussis.

    PubMed Central

    Maskell, D J; Morrissey, P; Dougan, G

    1988-01-01

    The aroA locus of Bordetella pertussis, encoding 5-enolpyruvylshikimate 3-phosphate synthase, has been cloned into Escherichia coli by using a cosmid vector. The gene is expressed in E. coli and complemented an E. coli aroA mutant. The nucleotide sequence of the B. pertussis aroA gene was determined and contains an open reading frame encoding 442 amino acids, with a calculated molecular weight for 5-enolpyruvylshikimate 3-phosphate synthase of 46,688. The amino acid sequence derived from the nucleotide sequence shows homology with the published amino acid sequences of aroA gene products of other microorganisms. PMID:2897356

  10. In Vitro Activity of Solithromycin against Bordetella pertussis, an Emerging Respiratory Pathogen

    PubMed Central

    Vicino, David; Fernandes, Prabhavathi

    2016-01-01

    There has been an increase in the number of pertussis cases reported since the introduction of the acellular pertussis vaccine. While children that present with pertussis have a characteristic whooping cough, adults can simply have a persistent, nonspecific cough and remain undiagnosed. Macrolide antibiotics, such as azithromycin, are the currently recommended treatment for pertussis. Solithromycin is a new macrolide and the first fluoroketolide with broad activity against a wide spectrum of bacterial pathogens and has completed clinical development for community-acquired bacterial pneumonia. This study reports the potent in vitro activity of solithromycin against a collection of recent isolates of Bordetella pertussis. PMID:27620481

  11. Respiratory disease associated with Bordetella bronchiseptica in a Hoffmann's two-toed sloth (Choloepus hoffmanni).

    PubMed

    Hammond, Elizabeth E; Sosa, Daniel; Beckerman, Robert; Aguilar, Roberto F

    2009-06-01

    A 2-yr-old female captive-born Hoffmann's two-toed sloth (Choloepus hoffmanni) presented with respiratory disease. A severe inspiratory dyspnea with nasal congestion was observed with open-mouthed breathing and bilateral mucopurulent nasal exudate. Despite initial treatment with broad-spectrum antimicrobial therapy and anti-inflammatory and supportive care, the dyspnea persisted. The animal was anesthetized for bronchoscopy to obtain a deep tracheal sample. Based on culture of Bordetella bronchiseptica and sensitivity, a combination of systemic enrofloxacin, dexamethasone, and coupage with nebulization of saline, gentamicin, and albuterol as well as supportive care resulted in full recovery after 6 weeks of treatment.

  12. Bordetella trematum in chronic ulcers: report on two cases and review of the literature.

    PubMed

    Almagro-Molto, M; Eder, W; Schubert, S

    2015-08-01

    We report on two new cases Bordetella trematum in chronic ulcers. Literature shows that this species is infrequently isolated from clinical samples. Only a few cases have been reported so far. B. trematum may be underdiagnosed because of its problematic identification. Today, MALDI-TOF MS allows for an accurate discrimination of these bacteria. Overall, B. trematum was typically associated with severe skin and soft tissue infections, particularly in diabetic patients. The intrinsic resistance to some antibiotics commonly used to treat these infections makes the isolation of B. trematum significant.

  13. Bordetella trematum sepsis with shock in a diabetic patient with rapidly developing soft tissue infection.

    PubMed

    Majewski, Lorrance L; Nogi, Masayuki; Bankowski, Matthew J; Chung, Heath H

    2016-09-01

    Bordetella is a gram-negative, glucose non-fermenting bacillus, consisting of many host-associated species. B. trematum has previously been identified in wound infections, but rarely known to be a source of bacteremia. Currently, 16S rRNA sequencing represents the reference standard method by which identification is made. Herein, we present a case of fatal B. trematum bacteremia with septic shock. The presumed primary site of the infection was a rapidly developing left leg deep soft tissue infection without necrotizing fasciitis. B. trematum should now be considered as a significant pathogen in sepsis.

  14. Filamentous hemagglutinin of Bordetella pertussis: a key adhesin with immunomodulatory properties?

    PubMed

    Villarino Romero, Rodrigo; Osicka, Radim; Sebo, Peter

    2014-01-01

    The filamentous hemagglutinin of pathogenic Bordetellae is a prototype of a large two-partner-system-secreted and β-structure-rich bacterial adhesin. It exhibits several binding activities that may facilitate bacterial adherence to airway mucosa and host phagocytes in the initial phases of infection. Despite three decades of research on filamentous hemagglutinin, there remain many questions on its structure-function relationships, integrin interactions and possible immunomodulatory signaling capacity. Here we review the state of knowledge on this important virulence factor and acellular pertussis vaccine component. Specific emphasis is placed on outstanding questions that are yet to be answered.

  15. Kitten-transmitted Bordetella bronchiseptica infection in a patient receiving temozolomide for glioblastoma.

    PubMed

    Redelman-Sidi, Gil; Grommes, Christian; Papanicolaou, Genovefa

    2011-04-01

    Bordetella bronchiseptica is a gram negative coccobacillus that can be transmitted from domestic animals and cause severe infections in immunocompromised patients. A 56-year-old man with a left parietal glioblastoma was treated with resection, radiation and concomitant and adjuvant temozolomide chemotherapy. He received bevacizumab for progression, and dose dense metronomic temozolomide was added for additional progression. He developed chronic cough and was diagnosed with B. bronchiseptica infection. This is the first reported case of B. bronchiseptica infection in a patient receiving temozolomide. The infection was likely acquired from an infected kitten. Patients receiving temozolomide should be counseled on the risks of acquiring zoonotic infections, including B. bronchiseptica, from their pets.

  16. Antimicrobial resistance in bacteria associated with porcine respiratory disease in Australia.

    PubMed

    Dayao, Denise Ann E; Gibson, Justine S; Blackall, Patrick J; Turni, Conny

    2014-06-25

    The porcine respiratory disease complex greatly affects the health and production of pigs. While antimicrobial agents are used to treat the respiratory infections caused by bacterial pathogens, there is no current information on antimicrobial resistance in Australian pig respiratory bacterial isolates. The aim of this study was to determine the antimicrobial resistance profiles, by determining the minimum inhibitory concentration of nine antimicrobial agents for 71 Actinobacillus pleuropneumoniae, 51 Pasteurella multocida and 18 Bordetella bronchiseptica cultured from Australian pigs. The majority of A. pleuropneumoniae isolates were resistant to erythromycin (89%) and tetracycline (75%). Resistance to ampicillin (8.5%), penicillin (8.5%) and tilmicosin (25%) was also identified. The P. multocida isolates exhibited resistance to co-trimoxazole (2%), florfenicol (2%), ampicillin (4%), penicillin (4%), erythromycin (14%) and tetracycline (28%). While all the B. bronchiseptica isolates showed resistance to beta-lactams (ampicillin, ceftiofur and penicillin), some were resistant to erythromycin (94%), florfenicol (6%), tilmicosin (22%) and tetracycline (39%). The incidence of multiple drug resistance (MDR) varied across the species - in B. bronchiseptica, 27.8% of resistant isolates showed MDR, while 9.1% of the resistant isolates in A. pleuropneumoniae, and 4.8% in P. multocida showed MDR. This study illustrated that Australian pig strains of bacterial respiratory pathogens exhibited low levels of resistance to antimicrobial agents commonly used in the pig industry.

  17. Septicemic pasteurellosis in farmed elk (Cervus canadensis) in Alberta.

    PubMed

    Malhi, Pritpal S; Ngeleka, Musangu; Woodbury, Murray R

    2016-09-01

    Septicemic pasteurellosis is a bacterial disease of domestic and wild animals including bison, elk, and pronghorn antelope caused by Pasteurella multocida. Here we report 2 cases of septicemic pasteurellosis in farmed elk. Pasteurella multocida serogroup B was isolated from multiple tissues in both animals. Gene sequencing (16S ribosomal RNA) and BLAST query confirmed that the sequence is 99% to 100% homologous to the P. multocida sequences in the database.

  18. Septicemic pasteurellosis in farmed elk (Cervus canadensis) in Alberta

    PubMed Central

    Malhi, Pritpal S.; Ngeleka, Musangu; Woodbury, Murray R.

    2016-01-01

    Septicemic pasteurellosis is a bacterial disease of domestic and wild animals including bison, elk, and pronghorn antelope caused by Pasteurella multocida. Here we report 2 cases of septicemic pasteurellosis in farmed elk. Pasteurella multocida serogroup B was isolated from multiple tissues in both animals. Gene sequencing (16S ribosomal RNA) and BLAST query confirmed that the sequence is 99% to 100% homologous to the P. multocida sequences in the database. PMID:27587888

  19. PK and PK/PD of doxycycline in drinking water after therapeutic use in pigs.

    PubMed

    Prats, C; El Korchi, G; Giralt, M; Cristòfol, C; Peña, J; Zorrilla, I; Saborit, J; Pérez, B

    2005-12-01

    A commercial doxycycline formulation was administered in drinking water to 12 pigs at the recommended dose of 10 mg/kg daily for 5 days. The mean plasma concentration at steady-state was 1.37 +/- 1.21 microg/mL, which was reached at 68 +/- 27.2 h postadministration. Absorption and elimination half-life values were 7.20 +/- 2.42 and 7.01 +/- 2.10 h, respectively. Most plasma concentrations during dosing were higher than the minimum inhibitory concentrations (MICs) described for the main porcine bacterial pathogens of the respiratory tract (Pasteurella multocida, Actinobacillus pleuropneumoniae, Bordetella bronchiseptica and Mycoplasma hyopneumoniae). It is concluded that when pigs were treated with doxycycline in drinking water at the recommended rate, therapeutically effective concentrations were achieved throughout the treatment period, supporting the clinical use of this tetracycline in the control of respiratory infections. However, inter-animal differences were marked.

  20. Acquisition and loss of virulence-associated factors during genome evolution and speciation in three clades of Bordetella species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacteria in the genus Bordetella include nine species that are important pathogens. B. pertussis causes whooping cough, a serious and sometimes fatal disease in infants and in elderly people. Some strains of B. parapertussis also cause whooping cough-like disease in children while others cause pn...

  1. Phenotypic modulation of the Bvg+ phase is not required for pathogenesis and transmission of Bordetella bronchiseptica in swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The majority of virulence gene expression in Bordetella is regulated by a two-component sensory transduction system encoded by the bvg locus. In response to environmental cues, the BvgAS regulatory system controls expression of a spectrum of phenotypic phases transitioning between a virulent (Bvg+) ...

  2. Swine influenza virus co-infection with Bordetella bronchiseptica enhances bacterial colonization and host immune responses exacerbating pulmonary lesions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Swine influenza virus (SIV) is one of the most important disease causing agents for the U.S. swine industry, not only as a primary pathogen but as a predisposing agent to secondary bacterial infection. Bordetella bronchiseptica (Bb) is often isolated from swine and has been shown to contribute to th...

  3. The Bordetella bronchiseptica type III secretion system is required for persistence and disease severity but not transmission in swine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica is pervasive in swine populations and plays multiple roles in respiratory disease. Most studies addressing virulence factors of B. bronchiseptica utilize isolates derived from hosts other than pigs in conjunction with rodent infection models. Based on previous in vivo mouse...

  4. Draft genome sequences of 53 genetically distinct isolates of Bordetella bronchiseptica representing 11 terrestrial and aquatic hosts

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bordetella bronchiseptica infects a variety of mammalian and avian hosts. Here we report the genome sequences of 53 genetically distinct isolates, acquired from a broad range of terrestrial and aquatic animals. These data will greatly facilitate ongoing efforts to better understand evolution, host...

  5. Bordetella evades the host immune system by inducing IL-10 through a type III effector, BopN

    PubMed Central

    Nagamatsu, Kanna; Kuwae, Asaomi; Konaka, Tadashi; Nagai, Shigenori; Yoshida, Sei; Eguchi, Masahiro; Watanabe, Mineo; Mimuro, Hitomi; Koyasu, Shigeo

    2009-01-01

    The inflammatory response is one of several host alert mechanisms that recruit neutrophils from the circulation to the area of infection. We demonstrate that Bordetella, a bacterial pathogen, exploits an antiinflammatory cytokine, interleukin-10 (IL-10), to evade the host immune system. We identified a Bordetella effector, BopN, that is translocated into the host cell via the type III secretion system, where it induces enhanced production of IL-10. Interestingly, the BopN effector translocates itself into the nucleus and is involved in the down-regulation of mitogen-activated protein kinases. Using pharmacological blockade, we demonstrated that BopN-induced IL-10 production is mediated, at least in part, by its ability to block the extracellular signal-regulated kinase pathway. We also showed that BopN blocks nuclear translocation of nuclear factor κB p65 (NF-κBp65) but, in contrast, promotes nuclear translocation of NF-κBp50. A BopN-deficient strain was unable to induce IL-10 production in mice, resulting in the elimination of bacteria via neutrophil infiltration into the pulmonary alveoli. Furthermore, IL-10–deficient mice effectively eliminated wild-type as well as BopN mutant bacteria. Thus, Bordetella exploits BopN as a stealth strategy to shut off the host inflammatory reaction. These results explain the ability of Bordetella species to avoid induction of the inflammatory response. PMID:20008527

  6. Bordetella bronchiseptica Pneumonia in an Infant and Genetic Comparison of Clinical Isolates with Veterinary Kennel Cough Vaccines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An infant with recurrent episodes of respiratory failure was diagnosed with pertussis based on immunofluorescence testing, but culture revealed macrolide-resistant Bordetella bronchiseptica. Genetic analysis demonstrated that the child was not infected with a kennel cough vaccine strain, although th...

  7. Persistent Bordetella bronchiseptica pneumonia in an immunocompetent infant and genetic comparison of clinical isolates with kennel cough vaccine strains.

    PubMed

    Rath, Barbara A; Register, Karen B; Wall, Jeffrey; Sokol, Dawn M; Van Dyke, Russell B

    2008-03-15

    An infant who experienced recurrent episodes of respiratory failure received a diagnosis of pertussis on the basis of immunofluorescence testing, but culture revealed macrolide-resistant Bordetella bronchiseptica. Genetic analysis demonstrated that the child was not infected with a kennel cough vaccine strain, although the family's dog had recently been vaccinated. The infection cleared with imipenem therapy.

  8. Bordetella avium causes induction of apoptosis and nitric oxide synthase in turkey tracheal explant cultures.

    PubMed

    Miyamoto, David M; Ruff, Kristin; Beach, Nathan M; Stockwell, Stephanie B; Dorsey-Oresto, Angella; Masters, Isaac; Temple, Louise M

    2011-09-01

    Bordetellosis is an upper respiratory disease of turkeys caused by Bordetella avium in which the bacteria attach specifically to ciliated respiratory epithelial cells. Little is known about the mechanisms of pathogenesis of this disease, which has a negative impact in the commercial turkey industry. In this study, we produced a novel explant organ culture system that was able to successfully reproduce pathogenesis of B. avium in vitro, using tracheal tissue derived from 26 day-old turkey embryos. Treatment of the explants with whole cells of B. avium virulent strain 197N and culture supernatant, but not lipopolysaccharide (LPS) or tracheal cytotoxin (TCT), specifically induced apoptosis in ciliated cells, as shown by annexin V and TUNEL staining. LPS and TCT are known virulence factors of Bordetella pertussis, the causative agent of whooping cough. Treatment with whole cells of B. avium and LPS specifically induced NO response in ciliated cells, shown by uNOS staining and diaphorase activity. The explant system is being used as a model to elucidate specific molecules responsible for the symptoms of bordetellosis.

  9. The vaccine potential of Bordetella pertussis biofilm-derived membrane proteins

    PubMed Central

    de Gouw, Daan; Serra, Diego O; de Jonge, Marien I; Hermans, Peter WM; Wessels, Hans JCT; Zomer, Aldert; Yantorno, Osvaldo M; Diavatopoulos, Dimitri A; Mooi, Frits R

    2014-01-01

    Pertussis is an infectious respiratory disease of humans caused by the gram-negative pathogen Bordetella pertussis. The use of acellular pertussis vaccines (aPs) which induce immunity of relative short duration and the emergence of vaccine-adapted strains are thought to have contributed to the recent resurgence of pertussis in industrialized countries despite high vaccination coverage. Current pertussis vaccines consist of antigens derived from planktonic bacterial cultures. However, recent studies have shown that biofilm formation represents an important aspect of B. pertussis infection, and antigens expressed during this stage may therefore be potential targets for vaccination. Here we provide evidence that vaccination of mice with B. pertussis biofilm-derived membrane proteins protects against infection. Subsequent proteomic analysis of the protein content of biofilm and planktonic cultures yielded 11 proteins which were ≥three-fold more abundant in biofilms, of which Bordetella intermediate protein A (BipA) was the most abundant, surface-exposed protein. As proof of concept, mice were vaccinated with recombinantly produced BipA. Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria. Finally, we confirmed that bipA is expressed during respiratory tract infection of mice, and that anti-BipA antibodies are present in the serum of convalescent whooping cough patients. Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines. PMID:26038752

  10. Pneumonia in lambs inoculated with Bordetella parapertussis: clinical and pathological studies.

    PubMed

    Chen, W; Alley, M R; Manktelow, B W

    1988-09-01

    Eight one-week-old, colostrum-deprived lambs were inoculated intratracheally with ovine isolates of Bordetella parapertussis. After inoculation, the lambs showed slight depression and anorexia. The total circulating leucocyte counts in these animals rose gradually to a peak five days post-inoculation. Neutrophil counts also increased and were highest at Day 3. Lesions grossly and histologically similar to those of naturally-occurring ovine chronic non-progressive pneumonia were seen in the majority of infected animals. Grossly, they consisted of many small areas of collapse and dull-red consolidation. Histologically, B. parapertussis caused mild acute tracheobronchitis, severe alveolar collapse and acute bronchopneumonia, which was most severe from Days 1 to 3. Pure cultures of B. parapertussis were consistently recovered from nasal swabs of infected lambs throughout the study. Viable bacterial counts of bronchoalveolar lavage fluid showed a rapid elimination of this organism from the lower respiratory tract between Days 1 to 5. Bordetella parapertussis infection in the respiratory tract of lambs has the potential to compromise pulmonary defence mechanisms and allow other pathogenic organisms to become established in the lower respiratory tract.

  11. [Presence of Bordetella holmesii in an outbreak of pertussis in Chile].

    PubMed

    Miranda, Carolina; Wozniak, Aniela; Castillo, Claudia; Geoffroy, Enrique; Zumarán, Cecilia; Porte, Lorena; Román, Juan C; Potin, Marcela; García, Patricia

    2013-06-01

    The incidence of whooping cough in Chile ranges from 4.1 and 7.5 per hundred thousand inhabitants. B. pertussis detection is performed by Real Time PCR (Q-PCR) directed to the insertion sequence IS481. However, this sequence is also found in the genome of B. bronchiseptica and B. holmesii. The latter is also a respiratory pathogen whose clinical features are similar to B. pertussis. However, it is important to differentiate between these species because in immunosuppressed patients B. holmesii is more likely to cause bacteremia and is less susceptible to erythromycin. The goal of this work is to measure prospectively and retrospectively the presence of B. holmesii in samples reported positive for B. pertussis in the period 2010-2011. During this period, 1994 nasopharyngeal specimens entered the laboratory for Bordetella sp. PCR, of which 224 were positive. The analysis by Q-PCR directed to the recA gene of B. holmesii of all 224 positive samples determined a prevalence of B. holmesii of 0.6% (12/1994). Because of its more aggressive behavior in immunosupressed patients and its different resistance pattern, routine screening of B. pertussis and B. holmesii is currently performed for all samples in which Bordetella sp PCR is initially detected.

  12. Cyclic-di-GMP signalling regulates motility and biofilm formation in Bordetella bronchiseptica

    PubMed Central

    Sisti, Federico; Ha, Dae-Gon; O'Toole, George A.; Hozbor, Daniela

    2013-01-01

    The signalling molecule bis-(3′–5′)-cyclic-dimeric guanosine monophosphate (c-di-GMP) is a central regulator of diverse cellular functions, including motility, biofilm formation, cell cycle progression and virulence, in bacteria. Multiple diguanylate cyclase and phosphodiesterase-domain-containing proteins (GGDEF and EAL/HD-GYP, respectively) modulate the levels of the second messenger c-di-GMP to transmit signals and obtain such specific cellular responses. In the genus Bordetella this c-di-GMP network is poorly studied. In this work, we evaluated the expression of two phenotypes in Bordetella bronchiseptica regulated by c-di-GMP, biofilm formation and motility, under the influence of ectopic expression of Pseudomonas aeruginosa proteins with EAL or GGDEF domains that regulates the c-di-GMP level. In agreement with previous reports for other bacteria, we observed that B. bronchiseptica is able to form biofilm and reduce its motility only when GGDEF domain protein is expressed. Moreover we identify a GGDEF domain protein (BB3576) with diguanylate cyclase activity that participates in motility and biofilm regulation in B. bronchiseptica. These results demonstrate for the first time, to our knowledge, the presence of c-di-GMP regulatory signalling in B. bronchiseptica. PMID:23475948

  13. Microarray and functional analysis of growth phase-dependent gene regulation in Bordetella bronchiseptica.

    PubMed

    Nicholson, Tracy L; Buboltz, Anne M; Harvill, Eric T; Brockmeier, Susan L

    2009-10-01

    Growth phase-dependent gene regulation has recently been demonstrated to occur in Bordetella pertussis, with many transcripts, including known virulence factors, significantly decreasing during the transition from logarithmic to stationary-phase growth. Given that B. pertussis is thought to have derived from a Bordetella bronchiseptica-like ancestor, we hypothesized that growth phase-dependent gene regulation would also occur in B. bronchiseptica. Microarray analysis revealed and quantitative real-time PCR (qRT-PCR) confirmed that growth phase-dependent gene regulation occurs in B. bronchiseptica, resulting in prominent temporal shifts in global gene expression. Two virulence phenotypes associated with these gene expression changes were tested. We found that growth-dependent increases in expression of some type III secretion system (TTSS) genes led to a growth phase-dependent increase in a TTSS-dependent function, cytotoxicity. Although the transcription of genes encoding adhesins previously shown to mediate adherence was decreased in late-log and stationary phases, we found that the adherence of B. bronchiseptica did not decrease in these later phases of growth. Microarray analysis revealed and qRT-PCR confirmed that growth phase-dependent gene regulation occurred in both Bvg(+) and Bvg(-) phase-locked mutants, indicating that growth phase-dependent gene regulation in B. bronchiseptica can function independently from the BvgAS regulatory system.

  14. BipA Is Associated with Preventing Autoagglutination and Promoting Biofilm Formation in Bordetella holmesii

    PubMed Central

    Hiramatsu, Yukihiro; Saito, Momoko; Otsuka, Nao; Suzuki, Eri; Watanabe, Mineo; Shibayama, Keigo; Kamachi, Kazunari

    2016-01-01

    Bordetella holmesii causes both invasive and respiratory diseases in humans. Although the number of cases of pertussis-like respiratory illnesses due to B. holmesii infection has increased in the last decade worldwide, little is known about the virulence factors of the organism. Here, we analyzed a B. holmesii isolate that forms large aggregates and precipitates in suspension, and subsequently demonstrated that the autoagglutinating isolate is deficient in Bordetella intermediate protein A (BipA) and that this deletion is caused by a frame-shift mutation in the bipA gene. A BipA-deficient mutant generated by homologous recombination also exhibited the autoagglutination phenotype. Moreover, the BipA mutant adhered poorly to an abiotic surface and failed to form biofilms, as did two other B. holmesii autoagglutinating strains, ATCC 51541 and ATCC 700053, which exhibit transcriptional down-regulation of bipA gene expression, indicating that autoagglutination indirectly inhibits biofilm formation. In a mouse intranasal infection model, the BipA mutant showed significantly lower levels of initial lung colonization than did the parental strain (P < 0.01), suggesting that BipA might be a critical virulence factor in B. holmesii respiratory infection. Together, our findings suggest that BipA production plays an essential role in preventing autoagglutination and indirectly promoting biofilm formation by B. holmesii. PMID:27448237

  15. The vaccine potential of Bordetella pertussis biofilm-derived membrane proteins.

    PubMed

    de Gouw, Daan; Serra, Diego O; de Jonge, Marien I; Hermans, Peter Wm; Wessels, Hans Jct; Zomer, Aldert; Yantorno, Osvaldo M; Diavatopoulos, Dimitri A; Mooi, Frits R

    2014-08-01

    Pertussis is an infectious respiratory disease of humans caused by the gram-negative pathogen Bordetella pertussis. The use of acellular pertussis vaccines (aPs) which induce immunity of relative short duration and the emergence of vaccine-adapted strains are thought to have contributed to the recent resurgence of pertussis in industrialized countries despite high vaccination coverage. Current pertussis vaccines consist of antigens derived from planktonic bacterial cultures. However, recent studies have shown that biofilm formation represents an important aspect of B. pertussis infection, and antigens expressed during this stage may therefore be potential targets for vaccination. Here we provide evidence that vaccination of mice with B. pertussis biofilm-derived membrane proteins protects against infection. Subsequent proteomic analysis of the protein content of biofilm and planktonic cultures yielded 11 proteins which were ≥three-fold more abundant in biofilms, of which Bordetella intermediate protein A (BipA) was the most abundant, surface-exposed protein. As proof of concept, mice were vaccinated with recombinantly produced BipA. Immunization significantly reduced colonization of the lungs and antibodies to BipA were found to efficiently opsonize bacteria. Finally, we confirmed that bipA is expressed during respiratory tract infection of mice, and that anti-BipA antibodies are present in the serum of convalescent whooping cough patients. Together, these data suggest that biofilm proteins and in particular BipA may be of interest for inclusion into future pertussis vaccines.

  16. Bordetella pertussis FbpA binds both unchelated iron and iron siderophore complexes.

    PubMed

    Banerjee, Sambuddha; Weerasinghe, Aruna J; Parker Siburt, Claire J; Kreulen, R Timothy; Armstrong, Sandra K; Brickman, Timothy J; Lambert, Lisa A; Crumbliss, Alvin L

    2014-06-24

    Bordetella pertussis is the causative agent of whooping cough. This pathogenic bacterium can obtain the essential nutrient iron using its native alcaligin siderophore and by utilizing xeno-siderophores such as desferrioxamine B, ferrichrome, and enterobactin. Previous genome-wide expression profiling identified an iron repressible B. pertussis gene encoding a periplasmic protein (FbpABp). A previously reported crystal structure shows significant similarity between FbpABp and previously characterized bacterial iron binding proteins, and established its iron-binding ability. Bordetella growth studies determined that FbpABp was required for utilization of not only unchelated iron, but also utilization of iron bound to both native and xeno-siderophores. In this in vitro solution study, we quantified the binding of unchelated ferric iron to FbpABp in the presence of various anions and importantly, we demonstrated that FbpABp binds all the ferric siderophores tested (native and xeno) with μM affinity. In silico modeling augmented solution data. FbpABp was incapable of iron removal from ferric xeno-siderophores in vitro. However, when FbpABp was reacted with native ferric-alcaligin, it elicited a pronounced change in the iron coordination environment, which may signify an early step in FbpABp-mediated iron removal from the native siderophore. To our knowledge, this is the first time the periplasmic component of an iron uptake system has been shown to bind iron directly as Fe(3+) and indirectly as a ferric siderophore complex.

  17. Bordetella pertussis FbpA Binds Both Unchelated Iron and Iron Siderophore Complexes

    PubMed Central

    2015-01-01

    Bordetella pertussis is the causative agent of whooping cough. This pathogenic bacterium can obtain the essential nutrient iron using its native alcaligin siderophore and by utilizing xeno-siderophores such as desferrioxamine B, ferrichrome, and enterobactin. Previous genome-wide expression profiling identified an iron repressible B. pertussis gene encoding a periplasmic protein (FbpABp). A previously reported crystal structure shows significant similarity between FbpABp and previously characterized bacterial iron binding proteins, and established its iron-binding ability. Bordetella growth studies determined that FbpABp was required for utilization of not only unchelated iron, but also utilization of iron bound to both native and xeno-siderophores. In this in vitro solution study, we quantified the binding of unchelated ferric iron to FbpABp in the presence of various anions and importantly, we demonstrated that FbpABp binds all the ferric siderophores tested (native and xeno) with μM affinity. In silico modeling augmented solution data. FbpABp was incapable of iron removal from ferric xeno-siderophores in vitro. However, when FbpABp was reacted with native ferric-alcaligin, it elicited a pronounced change in the iron coordination environment, which may signify an early step in FbpABp-mediated iron removal from the native siderophore. To our knowledge, this is the first time the periplasmic component of an iron uptake system has been shown to bind iron directly as Fe3+ and indirectly as a ferric siderophore complex. PMID:24873326

  18. Differential expression of Bordetella pertussis iron transport system genes during infection

    PubMed Central

    Brickman, Timothy J.; Hanawa, Tomoko; Anderson, Mark T.; Suhadolc, Ryan J.; Armstrong, Sandra K.

    2008-01-01

    Summary Temporal expression patterns of the Bordetella pertussis alcaligin, enterobactin, and heme iron acquisition systems were examined using alcA-, bfeA-, and bhuR-tnpR recombinase fusion strains in a mouse respiratory infection model. The iron systems were differentially expressed in vivo, showing early induction of the alcaligin and enterobactin siderophore systems, and delayed induction of the heme system in a manner consistent with predicted changes in host iron source availability during infection. Previous mixed infection competition studies established the importance of alcaligin and heme utilization for B. pertussis in vivo growth and survival. In this study, the contribution of the enterobactin system to the fitness of B. pertussis was confirmed using wild-type and enterobactin receptor mutant strains in similar competition infection experiments. As a correlate to the in vivo expression studies of B. pertussis iron systems in mice, sera from uninfected and B. pertussis-infected human donors were screened for antibody reactivity with Bordetella iron-repressible cell envelope proteins. Pertussis patient sera recognized multiple iron-repressible proteins including the known outer membrane receptors for alcaligin, enterobactin, and heme, supporting the hypothesis that B. pertussis is iron-starved and responds to the presence of diverse iron sources during natural infection. PMID:18554331

  19. Clearance of Bordetella parapertussis from the Lower Respiratory Tract Requires Humoral and Cellular Immunity

    PubMed Central

    Wolfe, Daniel N.; Kirimanjeswara, Girish S.; Harvill, Eric T.

    2005-01-01

    Bordetella parapertussis and Bordetella pertussis are closely related species that cause whooping cough, an acute, immunizing disease. Their coexistence in the same host populations at the same time and vaccine studies showing that B. pertussis vaccines have little effect on B. parapertussis infection or disease suggest that the protective immunity induced by each does not efficiently cross protect against the other. Although the mechanisms of protective immunity to B. pertussis have been well studied, those of B. parapertussis have not. The present study explores the mechanism by which B. parapertussis is cleared from the lower respiratory tract by anamnestic immunity. Serum antibodies are necessary and sufficient for elimination of this bacterium, and CD4+ T cells, complement, and neutrophils are required for serum antibody-mediated clearance. Mice lacking immunoglobulin A had no defect in their ability to control or clear infection. Interestingly, serum antibody-mediated clearance of B. parapertussis did not require Fc receptors that are required for antibody-mediated clearance of B. pertussis. Together these data support a model for the mechanism of protective immunity to B. parapertussis that is similar but distinct from that of B. pertussis. PMID:16177324

  20. Effect of simulated stress on susceptibility of bighorn sheep neutrophils to Pasteurella haemolytica leukotoxin.

    PubMed

    Kraabel, B J; Miller, M W

    1997-07-01

    We examined the effects of simulated stress on susceptibility of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) neutrophils to Pasteurella haemolytica leukotoxin in a blocked, crossover experiment. Ten captive-raised bighorn sheep were sampled 10 hr after separate administrations of long-acting adrenocorticotrophic hormone (ACTH) gel and normal saline (control). We then compared in vitro leukotoxin-dependent neutrophil death rates after exposure to culture supernatants from four unique P. haemolytica isolates (one from domestic and three from bighorn sheep). Simulated stress effects were evidenced by elevated (P = 0.002) mean plasma cortisol concentrations, more neutrophils (P = 0.037), and fewer lymphocytes and eosinophils (P < or = 0.043) in ACTH-treated bighorn sheep. Maximum leukotoxin-dependent neutrophil death rates were > or = 61% for three of four P. haemolytica isolates tested. For all three cytotoxic isolates, neutrophil death rates at 150 micrograms/50 microliters supernatant were about 1.13 times higher (P = 0.0001) after bighorns received ACTH; for two of these, overall neutrophil death rates were higher (P < or = 0.001) in ACTH-treated bighorn sheep. Although variable leukotoxin production among P. haemolytica strains appeared principally responsible for differences in leukotoxin-dependent neutrophil death rates, susceptibility of bighorn sheep neutrophils to leukotoxin was increased by prior exposure to elevated plasma cortisol concentrations. It follows that if similar processes occur in neutrophils and alveolar macrophages in vivo, they could contribute to greater susceptibility of stressed bighorn sheep to pneumonic pasteurellosis.

  1. Susceptibility of phagocytes from elk, deer, bighorn sheep, and domestic sheep to Pasteurella haemolytica cytotoxins.

    PubMed

    Silflow, R M; Foreyt, W J

    1994-10-01

    Alveolar macrophages and peripheral blood neutrophils from elk (Cervus elaphus), bighorn sheep (Ovis canadensis canadensis), and domestic sheep were exposed to culture supernatants from Pasteurella haemolytica isolated from bighorn sheep and domestic sheep. In a second experiment, peripheral blood neutrophils from mule deer (Odocoileus hemionus), elk, and bighorn sheep were exposed to culture supernatants from P. haemolytica isolated from elk, bighorn sheep and domestic sheep. Alveolar macrophages from elk, bighorn sheep and domestic sheep were resistant to killing by P. haemolytica supernatants from bighorn sheep and domestic sheep; susceptibility of neutrophils to cell death, as measured by release of lactate dehydrogenase, differed significantly (P < 0.05) between the four species tested. Bighorn sheep and domestic sheep neutrophils were susceptible to cytotoxin damage by the P. haemolytica isolates used; bighorn sheep neutrophils were four- to eight-fold more susceptible to cytotoxin damage than domestic sheep neutrophils. Neutrophils from deer and elk were resistant to killing by P. haemolytica cytotoxins from any species tested.

  2. THE SURVIVAL OF PASTEURELLA PESTIS IN MATERIALS PRESERVED BY SOLID CARBON DIOXIDE (DRY-ICE).

    PubMed

    GOLDENBERG, M I; QUAN, S F; PRINCE, F M

    1964-01-01

    In the investigation of outbreaks of plague it is frequently more satisfactory to send specimens of suspected material collected in the field to a central laboratory, where usually better facilities for their more detailed examination are available. In the present study the authors have investigated the suitability of solid CO(2) (dry-ice) for the preservation of such material during transit. Three types of preparation were tested: broth suspensions of Pasteurella pestis, the livers and spleens of guinea-pigs dying after being infected with two different strains of P. pestis, and whole carcasses of mice and ground-squirrels infected with the organism. An additional test to ascertain the rate at which animal specimens became frozen and thawed was also carried out.These studies showed that (1) organisms in the various tissues frozen in dry-ice were not adversely affected by such treatment; (2) the survival of P. pestis cells did not depend on the number of organisms present in broth cultures or tissue suspensions, small numbers surviving equally well as large; and (3) plague bacilli contained in whole carcasses, even when present in small numbers, were also successfully preserved. It is concluded from these results, and also from the authors' practical use of the method over several years, that for the transport of plague-suspect materials from the field to the laboratory freezing with dry-ice can be confidently recommended.

  3. Pasteurella haemolytica serotype 2 contains the gene for a noncapsular serotype 1-specific antigen.

    PubMed Central

    Gonzalez, C T; Maheswaran, S K; Murtaugh, M P

    1995-01-01

    An ssa1-homologous genomic fragment cloned from Pasteurella haemolytica serotype 2 (ST2) enabled transformation of Escherichia coli DH5 alpha to a serotype 1 (ST1) phenotype through expression of the ST1-specific antigen (Ssa1). The Ssa1 protein expressed by ssa1-transformed E. coli was susceptible to heat and protease treatment and was distinct from P. haemolytica ST1-specific capsular polysaccharide. Electrophoretic analysis of in vitro-translated proteins, as well as the predicted amino acid sequence, demonstrated that Ssa1 proteins encoded from either ST1- or ST2-derived ssa1 genes were essentially identical. A comparison of the nucleotide sequences of ssa1 genes derived from P. haemolytica ST1 and ST2 revealed greater than 99% homology. Amino acid sequence homology of the predicted products of ST1- and ST2-derived ssa1 genes was greater than 98%. Northern (RNA) blot studies revealed that the presence of an increased level of ssa1 transcript in P. haemolytica ST1 grown as surface-adherent cultures on solid medium was correlated with a serologically detectable Ssa1 protein. Expression of the ssa1 transcript in ST1 was similarly upregulated by a high iron concentration in the growth medium. PMID:7890392

  4. Cloning, nucleotide sequence, and expression of the Pasteurella haemolytica A1 glycoprotease gene.

    PubMed Central

    Abdullah, K M; Lo, R Y; Mellors, A

    1991-01-01

    Pasteurella haemolytica serotype A1 secretes a glycoprotease which is specific for O-sialoglycoproteins such as glycophorin A. The gene encoding the glycoprotease enzyme has been cloned in the recombinant plasmid pH1, and its nucleotide sequence has been determined. The gene (designated gcp) codes for a protein of 35.2 kDa, and an active enzyme protein of this molecular mass can be observed in Escherichia coli clones carrying pPH1. In vivo labeling of plasmid-encoded proteins in E. coli maxicells demonstrated the expression of a 35-kDa protein from pPH1. The amino-terminal sequence of the heterologously expressed protein corresponds to that predicted from the nucleotide sequence. The glycoprotease is a neutral metalloprotease, and the predicted amino acid sequence of the glycoprotease contains a putative zinc-binding site. The gene shows no significant homology with the genes for other proteases of procaryotic or eucaryotic origin. However, there is substantial homology between gcp and an E. coli gene, orfX, whose product is believed to function in the regulation of macromolecule biosynthesis. Images PMID:1885539

  5. Sialoglycoprotease of Pasteurella haemolytica A1: detection of antisialoglycoprotease antibodies in sera of calves.

    PubMed Central

    Lee, C W; Shewen, P E; Cladman, W M; Conlon, J A; Mellors, A; Lo, R Y

    1994-01-01

    Log phase culture supernate from Pasteurella haemolytica biotype A, serotype 1 contains a proteolytic enzyme specific for O-sialoglycoproteins. Using two methods, Western immunoblotting and enzyme neutralization assay, it was demonstrated that certain bovine sera from two previous P. haemolytica A1 vaccination and challenge trials contained antibodies (Ab) (isotypes IgG1 and IgG2 on Western immunoblot) to the sialoglycoprotease (Gcp). In these trials, selected calves were vaccinated twice with either the commercial culture supernate vaccine Presponse or given phosphate-buffered saline (PBS). One trial was conducted during spring, P. haem XIX, and the other during the winter, P. haem XXI. Although there was no clear evidence for induction of anti-Gcp in response to vaccination, several calves seroconverted following intrapulmonary challenge with live P. haemolytica A1. This is the first report of anti-Gcp Ab in bovine sera. The results indicated that the Gcp is immunogenic and that the bacterium produces the enzyme in vivo. Further, animals with an anti-Gcp response had less pneumonia at necropsy, suggesting the Gcp may induce protective immunity. Images Fig. 1. Fig. 2. PMID:8004547

  6. Molecular studies of Ssa1, a serotype-specific antigen of Pasteurella haemolytica A1.

    PubMed Central

    Lo, R Y; Strathdee, C A; Shewen, P E; Cooney, B J

    1991-01-01

    A serotype-specific antigen of Pasteurella haemolytica A1 encoded on the recombinant plasmid pSSA1 is characterized. Nucleotide sequence analysis of the insert DNA in pSSA1 identified the gene ssaI, which codes for a protein of approximately 100 kDa. In vivo labeling of pSSA1-encoded protein in Escherichia coli maxicells showed the expression of a 100-kDa protein from the insert DNA on the recombinant plasmid. Northern blot and primer extension analyses were used to identify the mRNA transcript in P. haemolytica A1 and the putative promoter of ssaI. The antigen (designated Ssa1) could be localized to the outer membrane of P. haemolytica A1 and E. coli clones carrying pSSA1. A rabbit serum against Ssa1 was produced by using whole cells of E. coli expressing Ssa1 on the surface as the immunogen, demonstrating that Ssa1 is immunogenic in rabbits. The results from colony immunoblot analysis with calf serum from animals that were resistant to P. haemolytica A1-induced pneumonia suggest indirectly that Ssa1 is also immunogenic in the animals. Images PMID:1840576

  7. Pulmonary lesions induced by Pasteurella haemolytica in neutrophil sufficient and neutrophil deficient calves.

    PubMed Central

    Breider, M A; Walker, R D; Hopkins, F M; Schultz, T W; Bowersock, T L

    1988-01-01

    The role of neutrophils in the development of peracute lung lesions of bovine pneumonic pasteurellosis was investigated. Eight calves were divided into two groups of four calves each. Group I was treated with intravenous phosphate-buffered saline and served as the neutrophil sufficient calves. Group II was treated with intravenous hydroxyurea which produced a state of neutropenia. When peripheral blood neutrophil numbers dropped below 300 cells/microL in group II, all calves were challenged with an intrabronchial bolus of Pasteurella haemolytica in the log phase of growth. An acute inflammatory process occurred in both groups of calves indicated by a rise in body temperature. While pulmonary lesions occurred in both groups by six hours postinoculation, they varied in pathological characteristics. Pulmonary lesions in the neutrophil sufficient calves consisted of fibrinopurulent alveolitis-bronchiolitis with associated alveolar septal necrosis, interlobular edema, and intravascular thrombi. The neutrophil deficient calves had extensive intra-alveolar edema, interlobular edema, intraalveolar hemorrhage, atelectasis, and focal areas of alveolar septal necrosis. These results show that P. haemolytica can induce severe pulmonary tissue damage through both neutrophil dependent and neutrophil independent mechanisms. Images Fig. 1. Fig. 2. PMID:3370555

  8. Application of ozone disinfection to remove Enterococcus seriolicida, Pasteurella piscicida, and Vibrio anguillarum from seawater.

    PubMed Central

    Sugita, H; Asai, T; Hayashi, K; Mitsuya, T; Amanuma, K; Maruyama, C; Deguchi, Y

    1992-01-01

    Survival of bacterial fish pathogens, including Enterococcus seriolicida, Vibrio anguillarum, and Pasteurella piscicida, in ozonated seawater was determined in a batch system. Bacterial counts of all fish pathogens decreased at more than 0.040 to 0.060 mg of total residual oxidants (TROs) per liter, whereas no decrease in viable counts was observed at less than 0.018 to 0.028 mg of TROs per liter. The 99% inactivation point was achieved at concentrations of 0.111 mg/liter for E. seriolicida, 0.063 mg/liter for P. piscicida, and 0.064 mg/liter for V. anguillarum within 1 min. Moreover, the mean 99 and 99.9% killing concentration-contact time (C.t) products were 0.123 and 0.186 mg.min/liter for E. seriolicida, 0.056 and 0.084 mg.min/liter for P. piscicida, and 0.081 and 0.123 mg.min/liter for V. anguillarum, respectively. However, the mean 99 and 99.9% C.t products for the mixed population in coastal seawater were 0.200 and 0.621 mg.min/liter. These results strongly suggest that ozone treatment at more than 1.0 mg of TROs per liter for several minutes is able to disinfect seawater for mariculture efficiently. PMID:1476447

  9. Pneumonia in Calves Produced with Aerosols of Bovine Herpesvirus 1 and Pasteurella haemolytica

    PubMed Central

    Jericho, K. W. F.; Langford, E. V.

    1978-01-01

    In each of 11 experiments, four calves were exposed first to an aerosol of bovine herpesvirus 1 (BHV1, virus of infectious bovine rhinotracheitis) and second to an aerosol of Pasteurella haemolytica. The interval between aerosols was three to five days. In two other experiments, calves were exposed only to a bacterial aerosol. Climate was controlled for all experiments from the day of viral exposure and for eight of the experiments it was also controlled for four to six days before the first aerosol. The concentration of infectious doses of virus in the aerosols and the number of bacteria in the aerosols of each calf were determined. Macroscopically recognizable rhinitis, tonsillitis, laryngitis, tracheitis and pneumonia of lobar distribution in 42 lobes from 11 calves were seen in five experiments in which bacterial aerosol followed the viral aerosol by at least four days. One calf died with marked respiratory disease in each of four experiments within four days of exposure to the bacterial aerosol. Production of pneumonia was dependent on an interval between aerosols of at least four days but not on the condition of controlled climate on the environmental chamber either before or after the viral aerosol nor on the period of habituation allowed calves of some experiments. ImagesFig. 1.Fig. 2.Fig. 3. PMID:210912

  10. The Bordetella avium BAV1965-1962 fimbrial locus is regulated by temperature and produces fimbriae involved in adherence to turkey tracheal tissue.

    PubMed

    Loker, Stewart B; Temple, Louise M; Preston, Andrew

    2011-06-01

    Bordetella pertussis, Bordetella parapertussis, and Bordetella bronchiseptica cause respiratory tract disease in mammals, whereas Bordetella avium causes respiratory tract disease in avian hosts. While there are striking similarities between the diseases caused by the mammalian- and avian-adapted bordetellae, differences at the genetic level may account for their different host tropisms. Bacterial pathogens utilize the chaperone-usher pathway to assemble extracellular multisubunit structures (fimbriae) that play a role in virulence. Fimbriae of the mammalian bordetellae mediate attachment to the host respiratory epithelium. They are assembled by a single chaperone/usher system encoded by the fimbrial biogenesis operon fimA-D. B. avium contains a homologous fimbrial operon (BAV1965-1962), and we report here the functionality of this locus. Reverse transcription (RT)-PCR and quantitative PCR analyses demonstrated that transcription of the locus is regulated by temperature. By immuno-transmission electron microscopy (TEM), BAV1965-containing fimbriae were observed on bacteria grown at 37°C but not those grown at 22°C. A mutant in which BAV1965-1962 was deleted displayed significantly lower levels of adherence to turkey tracheal rings than the wild type. Thus, the BAV1965-1962 fimbrial locus is functional, its expression is regulated in response to temperature, and it produces fimbriae involved in adherence to host respiratory tract tissue.

  11. Unusual Aetiology of Pasteurella canis Biovar 2 Causing Dacryocystitis in HIV Patient: A Case Report and Review of Literature

    PubMed Central

    Negi, Sanjay Singh; Gade, Neeta

    2017-01-01

    Pasteurella species are zoonotic bacterial pathogens implicated very infrequently in various human infections following animal bites or licks usually of dogs and cats. This case report described a rare clinical presentation of dacryocystitis caused by P.canis in a Human Immunodeficiency Virus (HIV) positive young male patient involved in caring of cattle. It advocates the utmost need of recognizing the wide clinical manifestation spectrum of P.canis even without prior penetrating injury. P.canis associated clinical infection is more extensive than had been thought previously especially in immunocompromised patient. Early accurate identification and evidence based anti-microbial therapy may prove crucial in preventing further potential complications. PMID:28384864

  12. Phenotypic, antigenic, and molecular characterization of Pasteurella piscicida strains isolated from fish.

    PubMed Central

    Magariños, B; Romalde, J L; Bandín, I; Fouz, B; Toranzo, A E

    1992-01-01

    We compared Pasteurella piscicida strains isolated from different fish species in several European countries with strains isolated in Japan and the United States. The taxonomic analysis revealed that, regardless of the geographic origin and source of isolation, all the strains exhibited the same biochemical and physiological characteristics. Serological assays with different rabbit antisera demonstrated a high level of antigenic similarity among strains, with cross-agglutination titers of 20,480 to 40,960. This serological homogeneity was supported by the lipopolysaccharide (LPS) and membrane protein profiles. All the P. piscicida strains had the same electrophoretic LPS pattern, showing O side chains with a ladder-like structure, and shared at least four major outer membrane proteins, of 20, 30, 42, and 53 kDa. Western blot (immunoblot) analysis with LPS and protein indicated that all the P. piscicida strains are immunologically related. In addition, the chromosomal DNA fingerprint patterns obtained for the European strains with the enzymes EcoRI and BamHI were practically identical to those of the Japanese and U.S. strains. Although some differences were found in the plasmid profiles of P. piscicida, a large number of strains possessed in common plasmid bands of 20 and 7 MDa. In addition, a plasmid of 50 MDa was present in the majority of the European strains. Restriction endonuclease analysis demonstrated the genetic homology of the plasmid bands shared by most of the European strains. All the P. piscicida strains had the same drug resistance patterns, indicating that a correlation between plasmid carriage and resistance to a specific antimicrobial agent cannot be established. The high levels of phenotypic, serological, and genetic homogeneity found among the P. piscicida strains should facilitate the development of DNA probes with diagnostic purposes as well as the design of effective vaccines. Images PMID:1444366

  13. Hematological changes in calves exposed to a mixture of lipopolysaccharide and crude leukotoxin of Pasteurella haemolytica.

    PubMed Central

    Bowersock, T L; Walker, R D; Maddux, J M; Fenner, D; Moore, R N

    1990-01-01

    The purpose of this investigation was to determine if culture supernatants of Pasteurella haemolytica containing crude leukotoxin and lipopolysaccharide (CLCL) causes disseminated intravascular coagulopathy (DIC) when injected into calves. The effect of intraduodenal (ID) exposure followed by a subsequent subcutaneous (SC) inoculation of either heat-treated or untreated CLCL was evaluated. The relative contribution of the crude leukotoxin and lipopolysaccharide (LPS) to the virulence of P. haemolytica was evaluated. One group of calves received an ID inoculation of CLCL followed two weeks later by a SC inoculation of CLCL; one group received an ID inoculation of tissue culture medium followed two weeks later by a SC inoculation of CLCL; and a third group received an ID inoculation of CLCL followed two weeks later by a SC inoculation of heat-treated CLCL. Hematological parameters used to evaluate DIC included white cell count, platelet count, neutrophil number, fibrinogen, fibrin degradation products, one stage prothrombin time (OSPT), activated partial thromboplastin time, body temperature and clinical signs. Each parameter was measured in calves at 0, 2, 4, 6, 12 and 24 h following the SC inoculation of CLCL. Each group had significant changes over time in all parameters except body temperature. Calves that received a SC inoculation of heat-treated CLCL had smaller changes in all parameters except OSPT compared to the other groups. Results suggest that the LPS and leukotoxin of P. haemolytica exert additive effects on the coagulation cascade and number of peripheral leukocytes, and that the ID inoculation of CLCL does not affect the response of calves to a SC inoculation of toxin. PMID:2249175

  14. Phenotypic, antigenic, and molecular characterization of Pasteurella piscicida strains isolated from fish.

    PubMed

    Magariños, B; Romalde, J L; Bandín, I; Fouz, B; Toranzo, A E

    1992-10-01

    We compared Pasteurella piscicida strains isolated from different fish species in several European countries with strains isolated in Japan and the United States. The taxonomic analysis revealed that, regardless of the geographic origin and source of isolation, all the strains exhibited the same biochemical and physiological characteristics. Serological assays with different rabbit antisera demonstrated a high level of antigenic similarity among strains, with cross-agglutination titers of 20,480 to 40,960. This serological homogeneity was supported by the lipopolysaccharide (LPS) and membrane protein profiles. All the P. piscicida strains had the same electrophoretic LPS pattern, showing O side chains with a ladder-like structure, and shared at least four major outer membrane proteins, of 20, 30, 42, and 53 kDa. Western blot (immunoblot) analysis with LPS and protein indicated that all the P. piscicida strains are immunologically related. In addition, the chromosomal DNA fingerprint patterns obtained for the European strains with the enzymes EcoRI and BamHI were practically identical to those of the Japanese and U.S. strains. Although some differences were found in the plasmid profiles of P. piscicida, a large number of strains possessed in common plasmid bands of 20 and 7 MDa. In addition, a plasmid of 50 MDa was present in the majority of the European strains. Restriction endonuclease analysis demonstrated the genetic homology of the plasmid bands shared by most of the European strains. All the P. piscicida strains had the same drug resistance patterns, indicating that a correlation between plasmid carriage and resistance to a specific antimicrobial agent cannot be established. The high levels of phenotypic, serological, and genetic homogeneity found among the P. piscicida strains should facilitate the development of DNA probes with diagnostic purposes as well as the design of effective vaccines.

  15. Histological changes in the nasal ventral conchae of piglets infected with Bordetella bronchiseptica.

    PubMed Central

    Martineau-Doizé, B; Trépanier, H; Martineau, G P

    1991-01-01

    Piglets infected intranasally with Bordetella bronchiseptica were injected with two fluorochrome markers. Transverse sections of undecalcified nasal conchae (cut between the third incisor and the third premolar teeth) were examined by microradiography and fluorescence microscopy; surface-stained sections were evaluated by light microscopy. The fluorescent surface of the nasal ventral conchae from the infected piglets was increased as compared with the controls. This was due to an increased amount of fluorescent mineralization fronts as well as to the presence of abnormal fluorescent areas within trabeculae. Trabecular mineral content of the microradiographs was irregular and varied from hypo- to hypermineralized. When compared with the corresponding surface-stained sections, no correlation could be made between the mineral content and the type of tissue. These findings suggest that an increased number of osteoblasts which secrete prebone matrix but are modified so that mineralization does not occur normally. Images Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. Fig. 8. PMID:1884283

  16. Pneumonia in mice produced by cell-free extract of cultures of Bordetella parapertussis.

    PubMed

    Chen, W; Alley, M R; Manktelow, B W

    1990-01-01

    A single dose of culture fluid of Bordetella parapertussis freed from cells (CFCF) given intranasally to four-week-old mice free from intercurrent respiratory disease produced a subacute bronchopneumonia, which was similar to that induced by whole cells of ovine isolates of B parapertussis, except that the lesions were less severe and less extensive. From eight hours to 17 days after inoculation, the mice exhibited marked infiltration of neutrophils and macrophages into the alveolar septa, bronchiolar and alveolar spaces, and hyperplasia of peribronchiolar and perivascular lymphoid tissue. Electron microscopy showed damage to ciliated cells, type 1 pneumocytes and alveolar macrophages. These results suggest that extracellular toxic substance(s) produced by ovine isolates of B parapertussis might be involved in the initiation and development of lesions in ovine chronic non-progressive pneumonia.

  17. Morbidity and Mortality Due to Bordetella pertussis: A Significant Pathogen in West Africa?

    PubMed Central

    Kampmann, Beate; Mackenzie, Grant

    2016-01-01

    In the absence of specific surveillance platforms for pertussis and availability of suitable diagnostics at the hospital level, reliable data that describe morbidity and mortality from pertussis are difficult to obtain in any setting, as is the case in West Africa. Here, we summarize the available evidence of the burden of pertussis in the region, given historical data, and describe recent and ongoing epidemiological studies that offer opportunities for additional data collection. The available seroepidemiological data provide evidence of ongoing circulation of Bordetella pertussis in the region. Due to the lack of systematic and targeted surveillance with laboratory confirmation of B. pertussis infection, we cannot definitively conclude that pertussis disease is well controlled in West Africa. However, based on observations by clinicians and ongoing demographic surveillance systems that capture morbidity and mortality data in general terms, currently there is no evidence that pertussis causes a significant burden of disease in young children in West Africa. PMID:27838666

  18. Microbiological and clinical aspects of respiratory infections associated with Bordetella bronchiseptica.

    PubMed

    García-de-la-Fuente, Celia; Guzmán, Laura; Cano, María Eliecer; Agüero, Jesús; Sanjuán, Carmen; Rodríguez, Cristina; Aguirre, Amaia; Martínez-Martínez, Luis

    2015-05-01

    Bordetella bronchiseptica is a well-known veterinary pathogen, but its implication in human disease is probably not fully recognized. The purpose of this study was to determine the clinical significance of 36 B. bronchiseptica isolates from respiratory samples of 22 patients. Therefore, we describe microbiological characteristics, including phenotypic and genotypic identification as well as antimicrobial susceptibilities of the isolates. Clonal relatedness was evaluated using pulsed-field gel electrophoresis (PFGE). Most of the patients had some underlying immunosuppressive condition. Eighteen out of 22 (82%) patients had respiratory symptoms, and the death of 2 patients was associated with respiratory infection.All strains were correctly identified at species level by the simultaneous use of phenotypic methods and were confirmed by specific amplification of the upstream region of the fla gene. Tigecycline, minocycline, doxycycline, colistin, and meropenem were the most active agents tested. PFGE analysis revealed that repeated infections involving each patient had been caused by the same strain.

  19. Prevalence of Bordetella hinzii in mice in experimental facilities in Japan.

    PubMed

    Hayashimoto, Nobuhito; Morita, Hanako; Yasuda, Masahiko; Ishida, Tomoko; Kameda, Shuko; Takakura, Akira; Itoh, Toshio

    2012-10-01

    To reveal the current status of the prevalence of Bordetella hinzii in mice in experimental facilities in Japan, a survey of this agent was performed by culture of tracheal swabs from a total of 12,923 mice from 1699 facilities (12,192 mice from 1572 facilities in universities and research institutes and 731 mice from 127 facilities in pharmaceutical companies) in total. In the results, 195 out of 12,192 mice (1.6%) from 44 out of 1572 facilities (2.8%) in universities and research institutes were positive for B. hinzii. No B. hinzii-positive mice were found in 127 pharmaceutical companies surveyed. Gross lesions in the lungs with isolation of B. hinzii were observed in seven mice from four universities, and the lesions were identified as bronchopneumonia histopathologically. To our knowledge, this is the first report to reveal the prevalence of B. hinzii in laboratory mice.

  20. Bordetella dermonecrotic toxin binds to target cells via the N-terminal 30 amino acids.

    PubMed

    Fukui-Miyazaki, Aya; Ohnishi, Shinya; Kamitani, Shigeki; Abe, Hiroyuki; Horiguchi, Yasuhiko

    2011-03-01

    Bordetella dermonecrotic toxin (DNT) affects the biological function of host cells by activating intracellular Rho GTPases. The toxin binds to unidentified receptor(s) via 54 N-terminal amino acids, undergoes intramolecular cleavage on the C-terminal side of Arg(44) by furin or furin-like protease, and eventually enters the cytoplasm where the Rho GTPases reside. The binding to the receptor(s) and intramolecular cleavage are essential for DNT to intoxicate cells, and the 54 amino-acid binding domain encompasses the cleavage site, however, it is unclear whether these two events are related. In this study, we could narrow down the cell-binding domain to the N-terminal amino acids 2-30. The region does not contain the furin-recognition site, indicating that the cell binding and the intramolecular cleavage are independent events.

  1. Bordetella filamentous hemagglutinin and fimbriae: critical adhesins with unrealized vaccine potential.

    PubMed

    Scheller, Erich V; Cotter, Peggy A

    2015-11-01

    Pertussis, or whooping cough, is a highly contagious respiratory disease that is caused by the Gram-negative bacterium Bordetella pertussis, which is transmitted exclusively from human to human. While vaccination against B. pertussis has been successful, replacement of the whole cell vaccine with an acellular component vaccine has correlated with reemergence of the disease, especially in adolescents and infants. Based on their presumed importance in mediating adherence to host tissues, filamentous hemagglutinin (FHA) and fimbria (FIM) were selected as components of most acellular pertussis vaccines. In this review, we describe the biogenesis of FHA and FIM, recent data that show that these factors do, in fact, play critical roles in adherence to respiratory epithelium, and evidence that they also contribute to persistence in the lower respiratory tract by modulating the host immune response. We also discuss shortcomings of whole cell and acellular pertussis vaccines and the possibility that FHA and FIM could serve as effective protective antigens in next-generation vaccines.

  2. Pertactin deficient Bordetella pertussis present a better fitness in mice immunized with an acellular pertussis vaccine.

    PubMed

    Hegerle, N; Dore, G; Guiso, N

    2014-11-20

    Bordetella pertussis is the etiologic agent of whooping cough and has been the target of vaccination for over fifty years. The latest strategies include the use of acellular pertussis vaccines that induce specific immunity against few virulence factors amongst which pertactin is included in three and five component acellular pertussis vaccines. Recently, it has been reported that B. pertussis clinical isolates loose the production of this adhesin in regions reaching high vaccine coverage with vaccines targeting this virulence factor. We here demonstrate that isolates not producing pertactin are capable of sustaining longer infection as compared to pertactin producing isolates in an in vivo model of acellular pertussis immunization. Loosing pertactin production might thus provide a selective advantage to these isolates in this background, which could account for the upraise in prevalence of these pertactin deficient isolates in the population.

  3. Bordetella holmesii meningitis in an asplenic patient with systemic lupus erythematosus.

    PubMed

    Livovsky, Dan M; Leibowitz, David; Hidalgo-Grass, Carlos; Temper, Violeta; Salameh, Shaden; Korem, Maya

    2012-08-01

    Bordetella holmesii is a slow-growing, Gram-negative, non-oxidizing bacillus with colonies that produce a brown soluble pigment and was originally described by Weyant et al. (1995) as CDC nonoxidizer group 2 (NO-2). It has recently been shown that B. holmesii may be isolated from nasopharyngeal specimens of up to 20% of patients with pertussis-like symptoms. However, invasive B. holmesii has rarely been reported and in the vast majority of cases the patients were immune deficient, mostly as a result of splenectomy or functional asplenia. Clinical presentations have included endocarditis, pneumonia, cellulitis, suppurative arthritis, pyelonephritis and septicaemia but no previous reports have documented meningitis secondary to this organism. Here we report what we believe to be the first clinical description of an adult with B. holmesii meningitis and bacteraemia with a brief review of published cases.

  4. Complete genome sequence of a clinical Bordetella pertussis isolate from Brazil

    PubMed Central

    Andrade, Bruno Gabriel N; Marin, Michel F Abanto; Cambuy, Diego Duque; Fonseca, Erica Lourenço; Souza, Nadjla Ferreira; Vicente, Ana Carolina P

    2014-01-01

    There has been a resurgence in the number of pertussis cases in Brazil and around the world. Here, the genome of a clinical Bordetella pertussis strain (Bz181) that was recently isolated in Brazil is reported. Analysis of the virulence-associated genes defining the pre- and post-vaccination lineages revealed the presence of the prn2-ptxS1A-fim3B-ptxP3 allelic profile in Bz181, which is characteristic of the current pandemic lineage. A putative metallo-β-lactamase gene presenting all of the conserved zinc-binding motifs that characterise the catalytic site was identified, in addition to a multidrug efflux pump of the RND family that could confer resistance to erythromycin, which is the antibiotic of choice for treating pertussis disease. PMID:25411007

  5. Complete genome sequence of a clinical Bordetella pertussis isolate from Brazil.

    PubMed

    Andrade, Bruno Gabriel N; Marin, Michel F Abanto; Cambuy, Diego Duque; Fonseca, Erica Lourenço; Souza, Nadjla Ferreira; Vicente, Ana Carolina P

    2014-11-01

    There has been a resurgence in the number of pertussis cases in Brazil and around the world. Here, the genome of a clinical Bordetella pertussis strain (Bz181) that was recently isolated in Brazil is reported. Analysis of the virulence-associated genes defining the pre- and post-vaccination lineages revealed the presence of the prn2-ptxS1A-fim3B-ptxP3 allelic profile in Bz181, which is characteristic of the current pandemic lineage. A putative metallo-β-lactamase gene presenting all of the conserved zinc-binding motifs that characterise the catalytic site was identified, in addition to a multidrug efflux pump of the RND family that could confer resistance to erythromycin, which is the antibiotic of choice for treating pertussis disease.

  6. Pertactin is required for Bordetella species to resist neutrophil-mediated clearance.

    PubMed

    Inatsuka, Carol S; Xu, Qian; Vujkovic-Cvijin, Ivan; Wong, Sandy; Stibitz, Scott; Miller, Jeff F; Cotter, Peggy A

    2010-07-01

    Pertactin (PRN) is an autotransporter protein produced by all members of the Bordetella bronchiseptica cluster, which includes B. pertussis, B. parapertussis, and B. bronchiseptica. It is a primary component of acellular pertussis vaccines, and anti-PRN antibody titers correlate with protection. In vitro studies have suggested that PRN functions as an adhesin and that an RGD motif located in the center of the passenger domain is important for this function. Two regions of PRN that contain sequence repeats (region 1 [R1] and R2) show polymorphisms among strains and have been implicated in vaccine-driven evolution. We investigated the role of PRN in pathogenesis using B. bronchiseptica and natural-host animal models. A Deltaprn mutant did not differ from wild-type B. bronchiseptica in its ability to adhere to epithelial and macrophage-like cells in vitro or to establish respiratory infection in rats but was cleared much faster than wild-type bacteria in a mouse lung inflammation model. Unlike wild-type B. bronchiseptica, the Deltaprn mutant was unable to cause a lethal infection in SCID-Bg mice, but, like wild-type bacteria, it was lethal for neutropenic mice. These results suggest that PRN plays a critical role in allowing Bordetella to resist neutrophil-mediated clearance. Mutants producing PRN proteins in which the RGD motif was replaced with RGE or in which R1 and R2 were deleted were indistinguishable from wild-type bacteria in all assays, suggesting that these sequences do not contribute to PRN function.

  7. Outer membrane protein OmpQ of Bordetella bronchiseptica is required for mature biofilm formation.

    PubMed

    Cattelan, Natalia; Villalba, María Inés; Parisi, Gustavo; Arnal, Laura; Serra, Diego Omar; Aguilar, Mario; Yantorno, Osvaldo

    2016-02-01

    Bordetella bronchiseptica, an aerobic Gram-negative bacterium, is capable of colonizing the respiratory tract of diverse animals and chronically persists inside the hosts by forming biofilm. Most known virulence factors in Bordetella species are regulated by the BvgAS two-component transduction system. The Bvg-activated proteins play a critical role during host infection. OmpQ is an outer membrane porin protein which is expressed under BvgAS control. Here, we studied the contribution of OmpQ to the biofilm formation process by B. bronchiseptica. We found that the lack of expression of OmpQ did not affect the growth kinetics and final biomass of B. bronchiseptica under planktonic growth conditions. The ΔompQ mutant strain displayed no differences in attachment level and in early steps of biofilm formation. However, deletion of the ompQ gene attenuated the ability of B. bronchiseptica to form a mature biofilm. Analysis of ompQ gene expression during the biofilm formation process by B. bronchiseptica showed a dynamic expression pattern, with an increase of biofilm culture at 48 h. Moreover, we demonstrated that the addition of serum anti-OmpQ had the potential to reduce the biofilm biomass formation in a dose-dependent manner. In conclusion, we showed for the first time, to the best of our knowledge, evidence of the contribution of OmpQ to a process of importance for B. bronchiseptica pathobiology. Our results indicate that OmpQ plays a role during the biofilm development process, particularly at later stages of development, and that this porin could be a potential target for strategies of biofilm formation inhibition.

  8. Comparative genomic analysis of the swine pathogen Bordetella bronchisepticastrain KM22.

    PubMed

    Nicholson, Tracy L; Shore, Sarah M; Register, Karen B; Bayles, Darrell O; Kingsley, Robert A; Brunelle, Brain W

    2016-01-01

    The well-characterized Bordetella bronchiseptica strain KM22, originally isolated from a pig with atrophic rhinitis, has been used to develop a reproducible swine respiratory disease model. The goal of this study was to identify genetic features unique to KM22 by comparing the genome sequence of KM22 to the laboratory reference strain RB50. To gain a broader perspective of the genetic relationship of KM22 among other B. bronchiseptica strains, selected genes of KM22 were then compared to five other B. bronchiseptica strains isolated from different hosts. Overall, the KM22 genome sequence is more similar to the genome sequences of the strains isolated from animals than the strains isolated from humans. The majority of virulence gene expression in Bordetella is positively regulated by the two-component sensory transduction system BvgAS. bopN, bvgA, fimB, and fimC were the most highly conserved BvgAS-regulated genes present in all seven strains analyzed. In contrast, the BvgAS-regulated genes present in all seven strains with the highest sequence divergence werefimN, fim2, fhaL, andfhaS. A total of eight major fimbrial subunit genes were identified in KM22. Quantitative real-time PCR data demonstrated that seven of the eight fimbrial subunit genes identified in KM22 are expressed and regulated by BvgAS. The annotation of the KM22 genome sequence, coupled with the comparative genomic analyses reported in this study, can be used to facilitate the development of vaccines with improved efficacy towards B. bronchiseptica in swine to decrease the prevalence and disease burden caused by this pathogen.

  9. In Vitro Susceptibilities of Bordetella pertussis and Bordetella parapertussis to Two Ketolides (HMR 3004 and HMR 3647), Four Macrolides (Azithromycin, Clarithromycin, Erythromycin A, and Roxithromycin), and Two Ansamycins (Rifampin and Rifapentine)

    PubMed Central

    Hoppe, Jörg E.; Bryskier, André

    1998-01-01

    When tested by agar dilution on Mueller-Hinton agar supplemented with 5% horse blood, the ketolides HMR 3004 and HMR 3647 were slightly more active (MIC at which 90% of the isolates were inhibited [MIC90], 0.03 μg/ml) against Bordetella pertussis than azithromycin, clarithromycin, erythromycin A, and roxithromycin. Azithromycin (MIC90, 0.06 μg/ml) was the most active compound against B. parapertussis. Rifampin and rifapentine were considerably less active. PMID:9559823

  10. Cross-Species Protection Mediated by a Bordetella bronchiseptica Strain Lacking Antigenic Homologs Present in Acellular Pertussis Vaccines▿

    PubMed Central

    Sukumar, Neelima; Sloan, Gina Parise; Conover, Matt S.; Love, Cheraton F.; Mattoo, Seema; Kock, Nancy D.; Deora, Rajendar

    2010-01-01

    The Bordetella species are Gram-negative bacterial pathogens that are characterized by long-term colonization of the mammalian respiratory tract and are causative agents of respiratory diseases in humans and animals. Despite widespread and efficient vaccination, there has been a world-wide resurgence of pertussis, which remains the leading cause of vaccine-preventable death in developed countries. It has been proposed that current acellular vaccines (Pa) composed of only a few bacterial proteins may be less efficacious because of vaccine-induced antigenic shifts and adaptations. To gain insight into the development of a newer generation of vaccines, we constructed a Bordetella bronchiseptica strain (LPaV) that does not express the antigenic homologs included in any of the Pa vaccines currently in use. This strain also lacks adenylate cyclase toxin, an essential virulence factor, and BipA, a surface protein. While LPaV colonized the mouse nose as efficiently as the wild-type strain, it was highly deficient in colonization of the lower respiratory tract and was attenuated in induction of inflammation and injury to the lungs. Strikingly, to our surprise, we found that in an intranasal murine challenge model, LPaV elicited cross-species protection against both B. bronchiseptica and Bordetella pertussis. Our data suggest the presence of immunogenic protective components other than those included in the pertussis vaccine. Combined with the whole-genome sequences of many Bordetella spp. that are available, the results of this study should serve as a platform for strategic development of the next generation of acellular pertussis vaccines. PMID:20176797

  11. Septic arthritis caused by a gram-negative bacterium representing a new species related to the Bordetella-Alcaligenes complex.

    PubMed

    Kronvall, G; Hanson, H S; von Stedingk, L V; Törnqvist, E; Falsen, E

    2000-03-01

    A knee-joint exudate culture yielded on two occasions a gram-negative bacterium. Regular methods for speciation did not provide an identification. The infection was successfully treated with ciprofloxacin. The unknown isolate, CCUG 36768, was subjected to further investigation, including 16S rDNA sequencing, protein profiling, cellular fatty acid analysis, and various biochemical tests, in order to produce a species identification. The 1469 bp-long 16S rDNA sequence did not reveal identity with any known species sequence. CCUG 36768 clustered in a group of species, including Alcaligenes defragrans, Denitrobacter permanens, Taylorella equigenitalis, Alcaligenes faecalis, and four strains of Alcaligenes species without a specific species name. Bordetella species also showed a high degree of similarity with CCUG 36768. Protein profiling, cellular fatty acid analysis and computer-assisted analysis of biochemical profiles indicated similarity with Bordetella-Alcaligenes species, often close to B. holmesii and B. avium. API 20 NE indicated the profile of Moraxella species of poor identity. It is concluded that CCUG 36768 represents a new bacterial species of pathogenic potential in humans. It is related to the Bordetella-Alcaligenes group. Powerful new methods for speciation are available and it is recommended that unknown isolates from normally sterile sites be submitted for further analysis. Several isolates are required for the definition of new species.

  12. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin

    PubMed Central

    Sager, Martin; Benten, W. Peter M.; Engelhardt, Eva; Gougoula, Christina; Benga, Laurentiu

    2015-01-01

    [Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However, mechanisms contributing to their high prevalence such as the ability to form biofilms have not been studied yet. In the present investigation we analyze if these bacterial species can produce biofilms in vitro and investigate whether proteins, extracellular DNA and polysaccharides are involved in the biofilm formation and structure by inhibition and dispersal assays using proteinase K, DNase I and sodium periodate. Finally, the capacity of the biofilms to confer resistance to antibiotics is examined. We demonstrate that both [P.] pneumotropica biotypes but not [A.] muris are able to form robust biofilms in vitro, a phenotype which is widely spread among the field isolates. The biofilm inhibition and dispersal assays by proteinase and DNase lead to a strong inhibition in biofilm formation when added at the initiation of the biofilm formation and dispersed pre-formed [P.] pneumotropica biofilms, revealing thus that proteins and extracellular DNA are essential in biofilm formation and structure. Sodium periodate inhibited the bacterial growth when added at the beginning of the biofilm formation assay, making difficult the assessment of the role of β-1,6-linked polysaccharides in the biofilm formation, and had a biofilm stimulating effect when added on pre-established mature biofilms of [P.] pneumotropica biotype Heyl and a majority of [P.] pneumotropica biotype Jawetz strains, suggesting that the presence of β-1,6-linked polysaccharides on the bacterial surface might attenuate the biofilm production. Conversely, no effect or a decrease in the biofilm quantity was observed by biofilm dispersal using sodium periodate on further biotype Jawetz isolates, suggesting that polysaccharides might be incorporated in the biofilm structure. We additionally show that [P.] pneumotropica cells enclosed in biofilms

  13. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin.

    PubMed

    Sager, Martin; Benten, W Peter M; Engelhardt, Eva; Gougoula, Christina; Benga, Laurentiu

    2015-01-01

    [Pasteurella] pneumotropica biotypes Jawetz and Heyl and [Actinobacillus] muris are the most prevalent Pasteurellaceae species isolated from laboratory mouse. However, mechanisms contributing to their high prevalence such as the ability to form biofilms have not been studied yet. In the present investigation we analyze if these bacterial species can produce biofilms in vitro and investigate whether proteins, extracellular DNA and polysaccharides are involved in the biofilm formation and structure by inhibition and dispersal assays using proteinase K, DNase I and sodium periodate. Finally, the capacity of the biofilms to confer resistance to antibiotics is examined. We demonstrate that both [P.] pneumotropica biotypes but not [A.] muris are able to form robust biofilms in vitro, a phenotype which is widely spread among the field isolates. The biofilm inhibition and dispersal assays by proteinase and DNase lead to a strong inhibition in biofilm formation when added at the initiation of the biofilm formation and dispersed pre-formed [P.] pneumotropica biofilms, revealing thus that proteins and extracellular DNA are essential in biofilm formation and structure. Sodium periodate inhibited the bacterial growth when added at the beginning of the biofilm formation assay, making difficult the assessment of the role of β-1,6-linked polysaccharides in the biofilm formation, and had a biofilm stimulating effect when added on pre-established mature biofilms of [P.] pneumotropica biotype Heyl and a majority of [P.] pneumotropica biotype Jawetz strains, suggesting that the presence of β-1,6-linked polysaccharides on the bacterial surface might attenuate the biofilm production. Conversely, no effect or a decrease in the biofilm quantity was observed by biofilm dispersal using sodium periodate on further biotype Jawetz isolates, suggesting that polysaccharides might be incorporated in the biofilm structure. We additionally show that [P.] pneumotropica cells enclosed in biofilms

  14. The Bordetella type III secretion system effector BteA contains a conserved N-terminal motif that guides bacterial virulence factors to lipid rafts.

    PubMed

    French, Christopher T; Panina, Ekaterina M; Yeh, Sylvia H; Griffith, Natasha; Arambula, Diego G; Miller, Jeff F

    2009-12-01

    The Bordetella type III secretion system (T3SS) effector protein BteA is necessary and sufficient for rapid cytotoxicity in a wide range of mammalian cells. We show that BteA is highly conserved and functionally interchangeable between Bordetella bronchiseptica, Bordetella pertussis and Bordetella parapertussis. The identification of BteA sequences required for cytotoxicity allowed the construction of non-cytotoxic mutants for localization studies. BteA derivatives were targeted to lipid rafts and showed clear colocalization with cortical actin, ezrin and the lipid raft marker GM1. We hypothesized that BteA associates with the cytoplasmic face of lipid rafts to locally modulate host cell responses to Bordetella attachment. B. bronchiseptica adhered to host cells almost exclusively to GM1-enriched lipid raft microdomains and BteA colocalized to these same sites following T3SS-mediated translocation. Disruption of lipid rafts with methyl-beta-cyclodextrin protected cells from T3SS-induced cytotoxicity. Localization to lipid rafts was mediated by a 130-amino-acid lipid raft targeting domain at the N-terminus of BteA, and homologous domains were identified in virulence factors from other bacterial species. Lipid raft targeting sequences from a T3SS effector (Plu4750) and an RTX-type toxin (Plu3217) from Photorhabdus luminescens directed fusion proteins to lipid rafts in a manner identical to the N-terminus of BteA.

  15. Glucosamine found as a substituent of both phosphate groups in Bordetella lipid A backbones: role of a BvgAS-activated ArnT ortholog.

    PubMed

    Marr, Nico; Tirsoaga, Alina; Blanot, Didier; Fernandez, Rachel; Caroff, Martine

    2008-06-01

    Endotoxins are amphipathic lipopolysaccharides (LPSs), major constituents of the outer membrane of gram-negative bacteria. They consist of a lipid region, covalently linked to a core oligosaccharide, to which may be linked a repetitive glycosidic chain carrying antigenic determinants. Most of the biological activities of endotoxins have been associated with the lipid moiety of the molecule: unique to gram-negative bacteria, LPS is a ligand of the mammalian TLR4-MD2-CD14 pathogen recognition receptor complex. Lipid A preparations are often heterogeneous with respect to both the numbers and the lengths of fatty acids and the natures of substituents on the phosphate groups when present. The variants can significantly affect host immune responses. Nine species in the Bordetella genus have been described, and the fine LPS structures of seven of them have been published. In this report, lipids A from Bordetella pertussis Tohama I and B. bronchiseptica strain 4650 were further characterized and revealed to have a glucosamine substituting both lipid A phosphate groups of the diglucosamine backbone. These substitutions have not been previously described for bordetellae. Moreover, a B. pertussis transposon mutation that maps within a gene encoding a Bordetella ArnT (formerly PmrK) glycosyl transferase ortholog does not carry this substitution, thus providing a genetic basis for the modification. Reverse transcriptase PCR of this locus showed that it is Bvg regulated, suggesting that the ability of Bordetella to modify lipid A via this glucosamine modification is a potential virulence trait.

  16. Differentially Expressed Genes in Bordetella pertussis Strains Belonging to a Lineage Which Recently Spread Globally

    PubMed Central

    de Gouw, Daan; Hermans, Peter W. M.; Bootsma, Hester J.; Zomer, Aldert; Heuvelman, Kees; Diavatopoulos, Dimitri A.; Mooi, Frits R.

    2014-01-01

    Pertussis is a highly contagious, acute respiratory disease in humans caused by the Gram-negative pathogen Bordetella pertussis. Pertussis has resurged in the face of intensive vaccination and this has coincided with the emergence of strains carrying a particular allele for the pertussis toxin promoter, ptxP3, which is associated with higher levels of pertussis toxin (Ptx) production. Within 10 to 20 years, ptxP3 strains have nearly completely replaced the previously dominant ptxP1 strains resulting in a worldwide selective sweep. In order to identify B. pertussis genes associated with the selective sweep, we compared the expression of genes in ptxP1 and ptxP3 strains that are under control of the Bordetella master virulence regulatory locus (bvgASR). The BvgAS proteins comprise a two component sensory transduction system which is regulated by temperature, nicotinic acid and sulfate. By increasing the sulfate concentration, it is possible to change the phase of B. pertussis from virulent to avirulent. Until recently, the only distinctive phenotype of ptxP3 strains was a higher Ptx production. Here we identify additional phenotypic differences between ptxP1 and ptxP3 strains which may have contributed to its global spread by comparing global transcriptional responses under sulfate-modulating conditions. We show that ptxP3 strains are less sensitive to sulfate-mediated gene suppression, resulting in an increased production of the vaccine antigens pertactin (Prn) and Ptx and a number of other virulence genes, including a type III secretion toxin, Vag8, a protein involved in complement resistance, and lpxE involved in lipid A modification. Furthermore, enhanced expression of the vaccine antigens Ptx and Prn by ptxP3 strains was confirmed at the protein level. Identification of genes differentially expressed between ptxP1 and ptxP3 strains may elucidate how B. pertussis has adapted to vaccination and allow the improvement of pertussis vaccines by identifying novel

  17. Enhanced recovery of airborne T3 coliphage and Pasteurella pestis bacteriophage by means of a presampling humidification technique.

    PubMed

    Hatch, M T; Warren, J C

    1969-05-01

    This paper reports a series of experiments in which two methods of collecting airborne bacteriophage particles were compared. A standard aerosol sampler, the AGI-30, was evaluated for its competence in measuring the content of bacteriophage aerosols. It was used alone or with a prewetting or humidification device (humidifier bulb) to recover T(3) coliphage and Pasteurella pestis bacteriophage particles from aerosols maintained at 21 C and varied relative humidity. Collection of bacteriophage particles via the humidifier bulb altered both the initial recovery level and the apparent biological decay. Sampling airborne bacteriophage particles by the AGI-30 alone yielded data that apparently underestimated the maximal number of potentially viable particles within the aerosol, sometimes by as much as 3 logs.

  18. Restriction endonuclease analysis and ribotyping differentiate Pasteurella haemolytica serotype A1 isolates from cattle within a feedlot.

    PubMed Central

    Murphy, G L; Robinson, L C; Burrows, G E

    1993-01-01

    Pasteurella haemolytica serotype A1 isolates were collected from cattle within a feedlot during an outbreak of bovine respiratory disease. Genetic heterogeneity among the isolates was examined by restriction endonuclease analysis (REA), ribotyping, and analysis of plasmid content. The susceptibilities of isolates to several antibiotics were also examined. Five different REA patterns and three different ribotypes were observed among the isolates. Fifty percent of the isolates had an identical REA type, ribotype, and plasmid profile. Examination of the plasmid content of the isolates revealed that most (73%) carry a single plasmid which encodes beta-lactamase, 13.5% carry two plasmids, and 13.5% carry no plasmid. The data reveal the presence of genetic differences among isolates of P. haemolytica A1, associated with shipping fever pneumonia within a closed feedlot, and suggest that a combination of REA, ribotyping, plasmid analysis, and antibiotic susceptibility determination will be useful in analyzing the molecular epidemiology of this disease. Images PMID:7691872

  19. Evaluation of Level of Agreement in Bordetella Species Identification in Three U.S. Laboratories during a Period of Increased Pertussis.

    PubMed

    Burgos-Rivera, Brunilís; Lee, Adria D; Bowden, Katherine E; Faulkner, Amanda E; Seaton, Brent L; Lembke, Bryndon D; Cartwright, Charles P; Martin, Stacey W; Tondella, M Lucia

    2015-06-01

    While PCR is the most common method used for detecting Bordetella pertussis in the United States, most laboratories use insertion sequence 481 (IS481), which is not specific for B. pertussis; therefore, the relative contribution of other Bordetella species is not understood. The objectives of this study were to evaluate the proportion of other Bordetella species misidentified as B. pertussis during a period of increased pertussis incidence, determine the level of agreement in Bordetella species detection between U.S. commercial laboratories and the CDC, and assess the relative diagnostic sensitivity of CDC's PCR assay when using a different PCR master mix. Specimens collected between May 2012 and May 2013 were tested at two U.S. commercial laboratories for B. pertussis and B. parapertussis detection. Every fifth specimen positive for IS481 and/or IS1001 with cycle threshold (CT) values of ≤35 was sent to CDC for PCR testing that identifies Bordetella species. Specimens with indeterminate or negative results in the CDC PCR were tested using an alternate PCR master mix. Of 755 specimens, there was agreement in species identification for 83.4% (n = 630). Of the specimens with different identifications (n = 125), 79.2% (n = 99) were identified as indeterminate B. pertussis at CDC. Overall, 0.66% (n = 5) of the specimens were identified as B. holmesii or B. bronchiseptica at CDC. Of 115 specimens with indeterminate or negative results, 46.1% (n = 53) were B. pertussis positive when tested by an alternate master mix, suggesting a possible increase in assay sensitivity. This study demonstrates good agreement between the two U.S. commercial laboratories and CDC and little misidentification of Bordetella species during the 2012 U.S. epidemic.

  20. Characterization of a Membrane-active Peptide from the Bordetella pertussis CyaA Toxin*

    PubMed Central

    Subrini, Orso; Sotomayor-Pérez, Ana-Cristina; Hessel, Audrey; Spiaczka-Karst, Johanna; Selwa, Edithe; Sapay, Nicolas; Veneziano, Rémi; Pansieri, Jonathan; Chopineau, Joel; Ladant, Daniel; Chenal, Alexandre

    2013-01-01

    Bordetella pertussis, the pathogenic bacteria responsible for whooping cough, secretes several virulence factors, among which is the adenylate cyclase toxin (CyaA) that plays a crucial role in the early stages of human respiratory tract colonization. CyaA invades target cells by translocating its catalytic domain directly across the plasma membrane and overproduces cAMP, leading to cell death. The molecular process leading to the translocation of the catalytic domain remains largely unknown. We have previously shown that the catalytic domain per se, AC384, encompassing residues 1–384 of CyaA, did not interact with lipid bilayer, whereas a longer polypeptide, AC489, spanning residues 1–489, binds to membranes and permeabilizes vesicles. Moreover, deletion of residues 375–485 within CyaA abrogated the translocation of the catalytic domain into target cells. Here, we further identified within this region a peptidic segment that exhibits membrane interaction properties. A synthetic peptide, P454, corresponding to this sequence (residues 454–485 of CyaA) was characterized by various biophysical approaches. We found that P454 (i) binds to membranes containing anionic lipids, (ii) adopts an α-helical structure oriented in plane with respect to the lipid bilayer, and (iii) permeabilizes vesicles. We propose that the region encompassing the helix 454–485 of CyaA may insert into target cell membrane and induce a local destabilization of the lipid bilayer, thus favoring the translocation of the catalytic domain across the plasma membrane. PMID:24064217