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Sample records for experimental pasteurella multocida

  1. New sites of localisation of Pasteurella multocida B:2 in buffalo surviving experimental haemorrhagic septicaemia.

    Science.gov (United States)

    Annas, Salleh; Zamri-Saad, Mohammad; Jesse, Faez Firdaus Abdullah; Zunita, Zakaria

    2014-04-11

    Haemorrhagic septicaemia (HS) is an acute septicaemic disease of buffalo and cattle caused by Pasteurella multocida B:2 and E:2. Field outbreaks of HS are known to result in localisation of bacteria in the tonsils of surviving buffalo, confirming that animals can become carriers and the role of respiratory tract in the transmission of the disease. This report describes additional sites of localisation of P. multocida B:2 in surviving buffalo following experimental induction of HS. Following P. multocida B:2 infection, all calves in group 1 and one calf in group 2 that was allowed to commingle with infected calves from group 1 were euthanised within 48 h. Pasteurella multocida B:2 was detected from the nasal and rectal swab samples on days 5 and 6 from the remaining calves in group 2. The first injection of dexamethasone into the carrier animals resulted in reemergence in samples from the nose, rectum and vagina. However, subsequent dexamethasone injections failed to re-activate P. multocida B:2. When surviving carrier calves in group 2 were euthanised at the end of the experiment, P. multocida B:2 was detected in the lungs and various organs of the respiratory, gastrointestinal and urinary tracts. Commingling naive buffalo calves with calves acutely infected with P. multocida B:2 resulted in carriers among surviving buffalo. Pasteurella was found in various organs of the respiratory, gastrointestinal and urinary tracts, suggesting their role in the pathogenesis of HS.

  2. Treatment of pigs experimentally infected with Mycoplasma hyopneumoniae, Pasteurella multocida, and Actinobacillus pleuropneumoniae with various antibiotics.

    OpenAIRE

    Stipkovits, L.; Miller, D; Glavits, R; Fodor, L; Burch, D

    2001-01-01

    The authors have performed a comparative study of the efficacy of various in-feed medications for the treatment of 5- to 6-week-old specific pathogen-free (SPF) piglets experimentally infected on day 1 with Mycoplasma hyopneumoniae, on day 8 with Pasteurella multocida (serotype A), and on day 15 with Actinobacillus pleuropneumoniae (serotype 2). The treatment started on day 9 and continued for 12 consecutive days, then the piglets were euthanized for examination of macroscopic, histologic, an...

  3. Quantification of Pasteurella multocida in experimentally infected pigs using a real-time PCR assay.

    Science.gov (United States)

    Tocqueville, V; Kempf, I; Paboeuf, F; Marois-Créhan, C

    2017-06-01

    The aim of the study was to quantify Pasteurella multocida in experimentally infected pigs using a new qPCR assay based on the sodA gene and validated with 35 P. multocida strains, including strains isolated from pigs with pneumonia, clinically healthy pigs (nasal cavities), and human infections. The specificity of the test was verified with a collection of 60 strains of bacterial species other than P. multocida. The estimated detection threshold was 10 genome equivalents per microliter. The amplification efficiency and value of the correlation coefficients were 95.5% (±3.5%) and 0.995 (±0.005), respectively. Analysis of P. multocida suspensions in Buffered Peptone Water Broth and of samples prepared from lungs experimentally spiked with P. multocida revealed detection thresholds of 1.4CFU/μl and 8.4CFU/μl, respectively. In live pigs, experimentally-infected, approximately 10(5), 10(7) and 10(8)genomeequivalents/ml of P. multocida DNA was detected on Day 8 post-infection in the nasal cavities, tonsils and trachea samples, respectively. In dead pigs, approximatively 10(7)genomeequivalents/ml of P. multocida DNA was detected in the lung tissue with pneumonia. The qPCR assay's diagnostic specificity and sensitivity were 100% and 96%, respectively. This new qPCR assay should be a very useful tool for controlling enzootic pneumonia and studying the dynamics of infections in pig herds. Copyright © 2017 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

    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. Copyright © 2016. Published by Elsevier B.V.

  5. Treatment of pigs experimentally infected with Mycoplasma hyopneumoniae, Pasteurella multocida, and Actinobacillus pleuropneumoniae with various antibiotics.

    Science.gov (United States)

    Stipkovits, L; Miller, D; Glavits, R; Fodor, L; Burch, D

    2001-10-01

    The authors have performed a comparative study of the efficacy of various in-feed medications for the treatment of 5- to 6-week-old specific pathogen-free (SPF) piglets experimentally infected on day 1 with Mycoplasma hyopneumoniae, on day 8 with Pasteurella multocida (serotype A), and on day 15 with Actinobacillus pleuropneumoniae (serotype 2). The treatment started on day 9 and continued for 12 consecutive days, then the piglets were euthanized for examination of macroscopic, histologic, and pathologic lesions and for the presence of mycoplasmas and bacteria in the lungs. Based on the results of clinical observations (respiratory signs, rectal temperature, body weight gain, and feed conversion efficiency), macroscopic and histologic lesions of the lungs, and microbiologic findings, the best results were obtained by treatment of pigs with Econor + chlortetracycline, followed by Tetramutin, Pulmotil, Cyfac, and lincomycin + chlortetracycline.

  6. 9 CFR 113.121 - Pasteurella Multocida Bacterin.

    Science.gov (United States)

    2010-01-01

    ... OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.121 Pasteurella Multocida Bacterin. Pasteurella Multocida...

  7. Hematogenous Pasteurella multocida brain abscess

    Energy Technology Data Exchange (ETDEWEB)

    Wallace, M.; Lipsky, B.A.

    1985-10-01

    A case of hematogenously acquired brain abscess caused by Pasteurella multocida is described. CT scans of the head revealed the lesions in a 67 year old man with mild alcoholic liver disease and severe chronic obstructive pulmonary disease. Ultrasound examinations of the abdomen and chest and an echocardiogram failed to reveal a source for the abscess. On autopsy examination three encapsulated brain abscesses were found. 34 references, 2 figures, 1 table.

  8. Genome characterization of Pasteurella multocida subspecies septica and comparison with Pasteurella multocida subspecies multocida and gallicida.

    Science.gov (United States)

    Peng, Zhong; Liang, Wan; Liu, Wenjing; Chen, Huanchun; Wu, Bin

    2017-05-01

    Pasteurella multocida: subspecies multocida, gallicida, and septica are noted to have a contribution to fowl cholera, a life-threatening disease to both domestic and wild bird species. The genome sequences of avian P. multocida subspecies gallicida and multocida have been well analyzed and compared. However, the genome characterization of avian P. multocida subspecies septica is rarely discussed. In this study, we determined the draft genome sequence of a virulent P. multocida subspecies septica serogroup A strain HB02. The genome of P. multocida HB02 was composed of a single chromosome of 2,213,604 bp with a 40.27% G + C content, which showed a similar genome size and %GC content to the P. multocida subspecies multocida and gallicida genomes. The entire sequence specified 2002 putative coding DNA sequences plus 2 encoded rRNAs and 46 encoded tRNAs. In addition, the subspecies septica had a similar content of genes coding for metabolic traits that found in the subspecies gallicida and multocida. In addition, comparison of virulent versus avirulent avian P. multocida genomes identified 657 unique genes in either of the virulent strains HB02, P1059 and/or X73 compared to the avirulent strain Pm70. These genes should be the potential virulence-associated genes. Our work may add a novel genome sequence for the avian P. multocida genome database and shed a light on the pathogenesis of P. multocida.

  9. Pasteurella multocida and immune cells.

    Science.gov (United States)

    Kubatzky, Katharina F

    2012-01-01

    Pasteurella multocida was first discovered by Perroncito in 1878 and named after Louis Pasteur who first isolated and described this Gram-negative bacterium as the cause of fowl disease in 1880. Subsequently, P. multocida was also found to cause atrophic rhinitis in pigs, haemorrhagic septicaemia in cattle and respiratory diseases in many other animals. Among other factors such as lipopolysaccharide, outer membrane proteins and its capsule, the protein toxin (PMT) of P. multocida is an important virulence factor that determines the immunological response of the host's immune system. However, the exact molecular mechanisms taking place in cells of the innate and adaptive immune system are largely unknown for any of these virulence factors. Due to the obvious function of PMT on cells of the porcine skeletal system where it causes bone destruction, PMT was regarded as an osteolytic protein toxin. However, it remained unclear what the actual benefit for the bacteria would be. Recently, more attention was drawn to the osteoimmunological effects of PMT and the interplay between bone and immune cells. This review summarises the knowledge of effects of P. multocida virulence factors on the host's immune system.

  10. Pathology of experimental infection by Pasteurella multocida serotype A: 1 in buffalo calves.

    Science.gov (United States)

    Praveena, P E; Periasamy, S; Kumar, A A; Singh, N

    2014-11-01

    Pasteurella multocida serotype A:3 has been mostly implicated in pneumonic pasteurellosis in ruminants. In contrast, our previous studies have reported that both serotypes A:1 and A:3 were responsible for respiratory diseases in cattle and buffaloes. However, the pathology and pathogenesis of P. multocida serotype A:1 (Pm A:1) infection have not been studied in ruminants. In the present study, 12- to 15-week-old buffalo calves (Bubalus bubalis) infected by Pm A:1 had fibrinous and suppurative bronchopneumonia with focal areas of coagulation necrosis typical of pneumonic pasteurellosis. For the first time, this study reports the lung pathology and pathogenecity of Pm A:1 infection in calves. © The Author(s) 2014.

  11. Experimental Study of the Pathogenicity of Pasteurella multocida Capsular Type B in Rabbits.

    Science.gov (United States)

    Katoch, S; Verma, L; Sharma, M; Asrani, R K; Kumar, S; Chahota, R; Verma, S

    2015-01-01

    The increased frequency of isolation of Pasteurella multocida capsular type B from rabbitries in north-western India prompted this investigation into the role of this organism in inducing disease in rabbits. Ten rabbits were divided into two groups of five animals. Group I rabbits were infected intranasally (IN) with 1 ml of inoculum containing 2 × 10(5) colony forming units/ml, while rabbits in group II were given 1 ml phosphate buffered saline IN. The rabbits in group I developed respiratory distress, increased rectal temperature and severe dyspnoea, with death occurring 24-48 h post infection. The main pathological findings were severe congestion and haemorrhage in the trachea, fibrinopurulent pneumonia, bacteraemia and septicaemia. The nasal secretions of all group I animals contained P. multocida. These observations indicate that in addition to P. multocida capsular types A and D, P. multocida capsular type B can also be highly pathogenic for rabbits. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Genome wide host gene expression analysis in mice experimentally infected with Pasteurella multocida.

    Science.gov (United States)

    Priya, G Bhuvana; Nagaleekar, Viswas Konasagara; Milton, A Arun Prince; Saminathan, M; Kumar, Amod; Sahoo, Amit Ranjan; Wani, Sajad Ahmad; Kumar, Amit; Gupta, S K; Sahoo, Aditya P; Tiwari, A K; Agarwal, R K; Gandham, Ravi Kumar

    2017-01-01

    Pasteurella multocida causes acute septicemic and respiratory diseases, including haemorrhagic septicaemia, in cattle and buffalo with case fatality of 100%. In the present study, mice were infected with P. multocida (1.6 × 103 cfu, intraperitoneal) to evaluate host gene expression profile at early and late stages of infection using high throughput microarray transcriptome analyses. Several differentially expressed genes (DEGs) at both the time points were identified in P.multocida infected spleen, liver and lungs. Functional annotation of these DEGs showed enrichment of key pathways such as TLR, NF-κB, MAPK, TNF, JAK-STAT and NOD like receptor signaling pathways. Several DEGs overlapped across different KEGG pathways indicating a crosstalk between them. The predicted protein-protein interaction among these DEGs suggested, that the recognition of P. multocida LPS or outer membrane components by TLR4 and CD14, results in intracellular signaling via MyD88, IRAKs and/or TRAF6 leading to activation of NFκB and MAPK pathways and associated cytokines.

  13. Infective Exacerbation of Pasteurella multocida

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    Mayumi Hamada

    2016-01-01

    Full Text Available An 89-year-old lady presented with a one-day history of shortness of breath as well as a cough productive of brown sputum. Her medical history was significant for chronic obstructive pulmonary disease (COPD. She was in severe type one respiratory failure and blood tests revealed markedly raised inflammatory markers; however her chest X-ray was clear. On examination there was bronchial breathing with widespread crepitations and wheeze. She was treated as per an infective exacerbation of COPD. Subsequent blood cultures grew Pasteurella multocida, a common commensal in the oropharynx of domesticated animals. The patient was then asked about any contact with animals, after which she revealed she had a dog and was bitten on her left hand the day before admission. We should not forget to enquire about recent history of injuries or animal bites when patients present acutely unwell. She made a complete recovery after treatment with penicillin.

  14. Mannan-binding lectin (MBL) in two chicken breeds and the correlation with experimental Pasteurella multocida infection

    DEFF Research Database (Denmark)

    Schou, Torben Wilde; Permin, Anders; Christensen, Jens Peter

    2010-01-01

    The present study is the first demonstration of an association of the genetic serum Mannan-binding lectin (MBL) concentration with bacterial infections in chickens. The genetic serum MBL concentration was determined in two chicken breeds, and the association with the specific Pasteurella multocida...... of the chickens, suggesting that MBL plays an important role against P. multocida. A statistically significant negative correlation was found between the specific antibody response and the genetic serum MBL concentration for both breeds. This indicates that MBL in chickens is capable of acting as the first line...... of defence against P. multocida by diminishing the infection before the adaptive immune response takes over. Udgivelsesdato: May...

  15. Granulomatous hepatitis in pasteurella multocida infection

    Energy Technology Data Exchange (ETDEWEB)

    Chateil, J.F.; Brun, M.; Perel, Y.; Sananes, J.C.; Castell, J.F.; Diard, F. [Service de Radiologie A, Hopital Pellegrin, Bordeaux (France)

    1998-05-01

    Numerous diseases can lead to multilocular lesions of the liver. The authors report a rare pediatric case of hepatic granulomas due to Pasteurella multocida: a 7-year-old girl with chronic fever was investigated by sonography and CT scan, demonstrating mesenteric lymph node enlargement and numerous small hepatic lesions. After surgical biopsy, histopathology of the liver specimens showed pyogenic granuloma, with serologic testing positive for Pasteurella multocida. Treatment with a tetracycline and corticosteroids was successful. Pasteurella multocida infection, despite its habitual benign course, should be suspected among differential diagnoses of lymphogranulomatous affections with hepatic involvement. No case of liver and lymph node foci in a child has been previously described. (orig.) With 4 figs., 8 refs.

  16. The Myriad Properties of Pasteurella multocida Lipopolysaccharide

    OpenAIRE

    Harper, Marina; Boyce, John Dallas

    2017-01-01

    Pasteurella multocida is a heterogeneous species that is a primary pathogen of many different vertebrates. This Gram-negative bacterium can cause a range of diseases, including fowl cholera in birds, haemorrhagic septicaemia in ungulates, atrophic rhinitis in swine, and lower respiratory tract infections in cattle and pigs. One of the primary virulence factors of P. multocida is lipopolysaccharide (LPS). Recent work has shown that this crucial surface molecule shows significant structural var...

  17. Phenotypic variability among strains of Pasteurella multocida ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-02

    May 2, 2008 ... septica was demonstrated in all but the leporine species while P.m. gallicida was demonstrated only in the avian and ovine ... Key words: Animal species, Nigeria, Pasteurella multocida, phenotypic, subspeciation. INTRODUCTION .... waterfowl and associated birds (Hirsh et al., 1990), in poultry (Fegan et ...

  18. Lipase activity from strains of Pasteurella multocida.

    Science.gov (United States)

    Pratt, J; Cooley, J D; Purdy, C W; Straus, D C

    2000-05-01

    Thirteen clinical isolates of Pasteurella multocida from a variety of different animals and humans were examined for their ability to produce lipase. Lipase substrates used included Tween 20, Tween 40, Tween 80, and Tween 85. Lipase activity was detected in the filtrates of organisms grown to the exponential phase in Roswell Park Memorial Institute-1640 defined media (RPMI-1640), but activity increased in the filtrates when the cultures were allowed to proceed to the stationary phase. All strains examined (except for serotype 2) showed lipase activity against at least one of the Tweens. Tween 40 was the best substrate to demonstrate lipase activity. Pasteurella multocida serotype 8 produced the most active lipase against Tween 40 (3,561.7 units of activity/microgram of protein). This activity continued to increase after P. multocida entered a stationary growth phase. P. multocida lipase activity was optimal at pH 8.0. Lipase activity of P. multocida serotype 8 was eluted from a Sepharose 2B column at several points, indicating that several lipases may be produced in vitro by this organism. These data demonstrate that clinical isolates of P. multocida produce lipase; therefore, this enzyme should be considered a potential virulence factors for this organism.

  19. Pasteurella multocida pathogenesis: 125 years after Pasteur.

    Science.gov (United States)

    Harper, Marina; Boyce, John D; Adler, Ben

    2006-12-01

    Pasteurella multocida was first shown to be the causative agent of fowl cholera by Louis Pasteur in 1881. Since then, this Gram-negative bacterium has been identified as the causative agent of many other economically important diseases in a wide range of hosts. The mechanisms by which these bacteria can invade the mucosa, evade innate immunity and cause systemic disease are slowly being elucidated. Key virulence factors identified to date include capsule and lipopolysaccharide. The capsule is clearly involved in bacterial avoidance of phagocytosis and resistance to complement, while complete lipopolysaccharide is critical for bacterial survival in the host. A number of other virulence factors have been identified by both directed and random mutagenesis, including Pasteurella multocida toxin (PMT), putative surface adhesins and iron acquisition proteins. However, it is likely that many key virulence factors are yet to be identified, including those required for initial attachment and invasion of host cells and for persistence in a relatively nutrient poor and hostile environment.

  20. Isolation of Pasteurella multocida from chickens, preparation of formalin killed fowl cholera vaccine, and determination of efficacy in experimental chickens

    Directory of Open Access Journals (Sweden)

    Mahmuda Akhtar

    2016-03-01

    Full Text Available Objectives: The objectives of this study were to isolate and identify Pasteurella multocida from fowl cholera (FC suspected chicken, and to prepare and efficacy determination of formalin killed fowl cholera vaccine using the isolated P. multocida strain. Materials and methods: A total of five suspected dead chickens were collected from Brothers Poultry Farm located at Gazipur district, Bangladesh. The samples were processed and the P. multocida was isolated through conventional bacteriological techniques, were finally confirmed by polymerase chain reaction using P. multocida specific primers targeting cap gene. The P. multocida isolate was used to develop a formalin killed fowl cholera vaccine. The efficacy of the newly prepared vaccine was determined in Starcross-579 chickens (n=30 aging 15 weeks either by injecting 1 mL (group-A; n=10 or 0.5 mL (group-B; n=10 vaccine containing approximately 3.2x108 CFU/mL P. multocida organism; 10 birds were kept as unvaccinated control. The sera from the vaccinated and control birds were collected and were subjected for antibody titre determination by enzyme-linked immunosorbent assay (ELISA. Finally the vaccinated birds were challenged using virulent strains of P. multocida to confer the protection against FC. Results: P. multocida could be isolated from both the samples. The formalin killed vaccine prepared from the isolated bacteria was subjected for the determination of antibody titre in chicken, and found that the antibody titres in the birds of group A and group B were 4.513 and 4.07 respectively after primary vaccination, and 4.893 and 4.37 respectively after booster vaccination. Most of the vaccinated birds were found to be survived after challenging with virulent strain of P. multocida. Conclusion: It is concluded that the causal agent of FC (P. multocida was successfully isolated from FC affected dead chickens. The prepared formalin killed fowl cholera vaccine induces protective immune response and

  1. Isolation of Pasteurella multocida from broiler chickens

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    Sri Poernomo

    1996-06-01

    Full Text Available Pasteurella multocida, the etiological agent of fowl cholera, was isolated from five, 32 days oldbroilerchickens in the late of 1992. The chickens were from a farm located in Bogor area, raised in cages and each flock consisted of 1,550 broilers . Therewere 230 birds, aging from 28-31 days old, died with clinical signs of lameness and difficulty in breathing. Serological test of the isolate revealed serotype Aof Carter classification . To prove its virulences, the isolate was then inoculated into 3 mice subcutaneously. The mice died less then 24 hours postinoculation and P. multocida can be reisolated . The sensitivity test to antibiotics and sulfa preparations showed that the isolate was sensitive to ampicillin, doxycyclin, erythromycin, gentamycin, sulfamethoxazol-trimethoprim and baytril, but resistance to tetracyclin, kanamycin and oxytetracyclin. This is the first report of P. multocida isolation in broiler chickens in Indonesia, and it is intended to add information on bacterial diseases in poultry in Indonesia.

  2. Field Trial of a Pasteurella multocida Extract Vaccine in Rabbits

    OpenAIRE

    Suckow, Mark A; Haab, Rudolph W; Miloscio, Lawrence J; Guilloud, Norman B

    2008-01-01

    Pasteurella multocida is a bacterial pathogen that can cause significant disease and subsequent effects on research activities involving rabbits. Although several vaccines have been tested under laboratory conditions, field trials of vaccines for the control of P. multocida in rabbits are few. We used a potassium thiocyanate extract (PTE) produced from P. multocida serotype D:3,12,15 to vaccinate Pasteurella-free rabbits at their introduction into a colony having endemic infection with P. mul...

  3. Pasteurella multocida infection in solid organ transplantation.

    Science.gov (United States)

    Christenson, Eric S; Ahmed, Haitham M; Durand, Christine M

    2015-02-01

    We present a case of fulminant Pasteurella multocida sepsis in a 66-year-old man who had undergone a renal transplant. Our patient lived with two dogs and a cat with which he was very close. We propose that his bacteraemia might have resulted from direct inoculation of P multocida via his cat licking the venous stasis ulcers on his legs. The patient's clinical course was complicated by cardiopulmonary failure and he ultimately succumbed to his infection. P multocida is a rare cause of infections in immunocompromised hosts, epidemiologically linked to exposure to cats, dogs, and other animals. This case of P multocida shows the importance of considering this organism in immunocompromised hosts presenting with severe infections, especially if their history shows exposure to domesticated or wild animals known to be potential carriers of this disease. In this Grand Round, we review the clinical features, epidemiology, treatment, and prognosis of P multocida infections with a focus on these features in patients who are immunosuppressed. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. The Myriad Properties of Pasteurella multocida Lipopolysaccharide.

    Science.gov (United States)

    Harper, Marina; Boyce, John Dallas

    2017-08-21

    Pasteurella multocida is a heterogeneous species that is a primary pathogen of many different vertebrates. This Gram-negative bacterium can cause a range of diseases, including fowl cholera in birds, haemorrhagic septicaemia in ungulates, atrophic rhinitis in swine, and lower respiratory tract infections in cattle and pigs. One of the primary virulence factors of P. multocida is lipopolysaccharide (LPS). Recent work has shown that this crucial surface molecule shows significant structural variability across different P. multocida strains, with many producing LPS structures that are highly similar to the carbohydrate component of host glycoproteins. It is likely that this LPS mimicry of host molecules plays a major role in the survival of P. multocida in certain host niches. P. multocida LPS also plays a significant role in resisting the action of chicken cathelicidins, and is a strong stimulator of host immune responses. The inflammatory response to the endotoxic lipid A component is a major contributor to the pathogenesis of certain infections. Recent work has shown that vaccines containing killed bacteria give protection only against other strains with identical, or nearly identical, surface LPS structures. Conversely, live attenuated vaccines give protection that is broadly protective, and their efficacy is independent of LPS structure.

  5. Pasteurella multocida and bovine respiratory disease.

    Science.gov (United States)

    Dabo, S M; Taylor, J D; Confer, A W

    2007-12-01

    Pasteurella multocida is a pathogenic Gram-negative bacterium that has been classified into three subspecies, five capsular serogroups and 16 serotypes. P. multocida serogroup A isolates are bovine nasopharyngeal commensals, bovine pathogens and common isolates from bovine respiratory disease (BRD), both enzootic calf pneumonia of young dairy calves and shipping fever of weaned, stressed beef cattle. P. multocida A:3 is the most common serotype isolated from BRD, and these isolates have limited heterogeneity based on outer membrane protein (OMP) profiles and ribotyping. Development of P. multocida-induced pneumonia is associated with environmental and stress factors such as shipping, co-mingling, and overcrowding as well as concurrent or predisposing viral or bacterial infections. Lung lesions consist of an acute to subacute bronchopneumonia that may or may not have an associated pleuritis. Numerous virulence or potential virulence factors have been described for bovine respiratory isolates including adherence and colonization factors, iron-regulated and acquisition proteins, extracellular enzymes such as neuraminidase, lipopolysaccharide, polysaccharide capsule and a variety of OMPs. Immunity of cattle against respiratory pasteurellosis is poorly understood; however, high serum antibodies to OMPs appear to be important for enhancing resistance to the bacterium. Currently available P. multocida vaccines for use in cattle are predominately traditional bacterins and a live streptomycin-dependent mutant. The field efficacy of these vaccines is not well documented in the literature.

  6. The Myriad Properties of Pasteurella multocida Lipopolysaccharide

    Science.gov (United States)

    Harper, Marina; Boyce, John Dallas

    2017-01-01

    Pasteurella multocida is a heterogeneous species that is a primary pathogen of many different vertebrates. This Gram-negative bacterium can cause a range of diseases, including fowl cholera in birds, haemorrhagic septicaemia in ungulates, atrophic rhinitis in swine, and lower respiratory tract infections in cattle and pigs. One of the primary virulence factors of P. multocida is lipopolysaccharide (LPS). Recent work has shown that this crucial surface molecule shows significant structural variability across different P. multocida strains, with many producing LPS structures that are highly similar to the carbohydrate component of host glycoproteins. It is likely that this LPS mimicry of host molecules plays a major role in the survival of P. multocida in certain host niches. P. multocida LPS also plays a significant role in resisting the action of chicken cathelicidins, and is a strong stimulator of host immune responses. The inflammatory response to the endotoxic lipid A component is a major contributor to the pathogenesis of certain infections. Recent work has shown that vaccines containing killed bacteria give protection only against other strains with identical, or nearly identical, surface LPS structures. Conversely, live attenuated vaccines give protection that is broadly protective, and their efficacy is independent of LPS structure. PMID:28825691

  7. Pasteurella multocida Bacteremia in an Immunocompromised Patient

    Directory of Open Access Journals (Sweden)

    Shweta Kukrety

    2016-01-01

    Full Text Available 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.

  8. Pasteurella multocida: from Zoonosis to Cellular Microbiology

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    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

  9. Pasteurella multocida: from zoonosis to cellular microbiology.

    Science.gov (United States)

    Wilson, Brenda A; Ho, Mengfei

    2013-07-01

    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.

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Hydrogen peroxide as an effective disinfectant for Pasteurella multocida.

    Science.gov (United States)

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

    2014-07-01

    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.

  12. Meningitis bacteriémica por Pasteurella multocida Pasteurella multocida bacteremic meningitis

    Directory of Open Access Journals (Sweden)

    R. Soloaga

    2008-12-01

    Full Text Available Las infecciones por Pasteurella multocida en seres humanos habitualmente están relacionadas con mordeduras o arañazos de perros y gatos. Muchas de ellas se acompañan de otros microorganismos de la orofaringe de estos animales. Se presenta un caso de meningitis bacteriémica por P. multocida en una mujer de 86 años que convivía con siete gatos. Si bien no se documentó una infección de piel o de partes blandas, es posible que ésta haya pasado inadvertida inicialmente y que fuera la causa de la bacteriemia con impacto en meninges, o bien que la meningitis se haya producido luego de la colonización nasofaríngea (no demostrada. Los aislamientos de hemocultivos y de líquido cefalorraquídeo fueron identificados como P. multocida por medio de API 20NE, API 20E y Vitek 1. La cepa aislada presentó sensibilidad a penicilina, cefotaxima, levofloxacina y tetraciclinas, en coincidencia con lo descrito en la literatura.Human infections by Pasteurella multocida are usually associated with bites or scratches from dogs and cats. Many of them are accompanied by other oropharyngeal microorganisms of these animals. We herein present a case of bacteremic meningitis by P. multocida in an 86-year-old woman who was living with seven cats. Even though no skin or soft tissue infection was recorded, it is possible that a mild infection had gone undetected and a subsequent bacteremia had impacted on the meninges, or that meningitis could have occurred after nasopharyngeal colonization (not demonstrated. The isolates obtained from blood cultures and cerebrospinal fluid were identified as P. multocida by API 20NE, API 20E, and Vitek 1. In agreement with findings in the literature, this strain was susceptible to penicillin, cefotaxime, levofloxacin and tetracyclines.

  13. Pheno- and genotyping of Pasteurella multocida isolated from goat ...

    African Journals Online (AJOL)

    In this study, phenotype, capsular type and some virulence factors (pfha1, tbpA, and toxA) of caprine Pasteurella multocida were described in 10 isolates from goat. Two biochemical biovars were recognized among the isolates based on dulcitol and sorbitol fermentation. The isolates belonged to biovars P. multocida sub.

  14. Draft genome sequences of two virulent serotypes of avian Pasteurella multocida

    Science.gov (United States)

    Here we report the draft genome sequences of two virulent avian strains of Pasteurella multocida. Comparative analyses of these genomes were done with the published genome sequence of avirulent Pasteurella multocida strain Pm70....

  15. Pathology, Tissue Metalloproteinase Transcription and Haptoglobin Responses in Mice after Experimental Challenge with Different Isolates of Pasteurella multocida Obtained from Cases of Porcine Pneumonia

    DEFF Research Database (Denmark)

    Pors, S. E.; Chadfield, M. S.; Sørensen, D. B.

    2011-01-01

    Pasteurella multocida is a major cause of porcine pneumonia, but the pathogenesis of the disease is poorly defined. The aim of this study was to further understand the host response to infection by use of a mouse model of P. multocida pneumonia. Twenty female mice were divided into four groups (n=5......). Three groups were infected with one of three isolates of P. multocida isolated from clinical cases of chronic porcine pneumonia with necrotizing, suppurative and non-suppurative lesions, respectively. The fourth group served as uninfected controls. Mice were killed 24h postinfection and samples were...... differences in the severity and nature of lesions were seen according to the isolate of P. multocida used for infection. Intranasal infection of mice can therefore be used to evaluate the host response and lesions caused by P. multocida obtained from porcine pneumonic infections. The inflammatory response...

  16. Pasteurella multocida bacterial meningitis caused by contact with pigs

    Directory of Open Access Journals (Sweden)

    C. López

    2013-01-01

    Full Text Available 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.

  17. A cryopreservation method for Pasteurella multocida from wetland samples

    Science.gov (United States)

    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.

  18. Pasteurella multocida bacterial meningitis caused by contact with pigs.

    Science.gov (United States)

    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.

  19. Hematology of layers chickens vaccinated with fowl cholera vaccine and experimentally inoculated with virulent Pasteurella multocida serotypes in Zaria, Nigeria

    Directory of Open Access Journals (Sweden)

    Yusuf Madaki Lekko

    2017-09-01

    Conclusion The PCV significantly decrease P≤0.05 in layers vaccinated and inoculated with P. multocida but increase in unvaccinated layers inoculated P. multocida. The mean serum ALP concentration significantly increase P≤0.05 in unvaccinated layers inoculated with P. multocida when compared to layers vaccinated and inoculated with P. multocida. [J Adv Vet Anim Res 2017; 4(3.000: 234-240

  20. A Case of Polyarticular Pasteurella multocida Septic Arthritis

    Directory of Open Access Journals (Sweden)

    Sarah Nitoslawski

    2016-01-01

    Full Text Available 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.

  1. Signaling Cascades of Pasteurella multocida Toxin in Immune Evasion

    Science.gov (United States)

    Kubatzky, Katharina F.; Kloos, Bianca; Hildebrand, Dagmar

    2013-01-01

    Pasteurella multocida toxin (PMT) is a protein toxin found in toxigenic strains of Pasteurella multocida. PMT is the causative agent for atrophic rhinitis in pigs, a disease characterized by loss of nasal turbinate bones due to an inhibition of osteoblast function and an increase in osteoclast activity and numbers. Apart from this, PMT acts as a strong mitogen, protects from apoptosis and has an impact on the differentiation and function of immune cells. Many signaling pathways have been elucidated, however, the effect of these signaling cascades as a means to subvert the host’s immune system are just beginning to unravel. PMID:24064721

  2. MHC haplotype and susceptibility to experimental infections (Salmonella Enteritidis, Pasteurella multocida or Ascaridia galli) in a commercial and an indigenous chicken breed

    DEFF Research Database (Denmark)

    Schou, Torben Wilde; Labouriau, R.; Permin, A.

    2010-01-01

    In three independent experimental infection studies, the susceptibility and course of infection of three pathogens considered of importance in most poultry production systems, Ascaridia galli, Salmonella Enteritidis and Pasteurella multocida was compared in two chicken breeds, the indigenous Viet...... might be associated with resistance. These results demonstrate the presence of local genetic resources in Vietnamese chickens, which could be utilized in breeding programmes aiming at improving disease resistance...

  3. Effect of aerial ammonia on porcine infection of the respiratory tract with toxigenic Pasteurella multocida

    DEFF Research Database (Denmark)

    Andreasen, Morten; Bækbo, P.; Nielsen, J.P.

    1999-01-01

    The objective of the experimental study was to examine whether aerial ammonia alone could predispose the respiratory system of pigs to infection with toxigenic Pasteurella multocida type A. Two groups of 5 pigs each were continuously exposed to 50 ppm ammonia and less than 5 ppm ammonia...

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

    Science.gov (United States)

    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...

  5. Fatal multifocal Pasteurella multocida infection: a case report.

    Science.gov (United States)

    Guilbart, Mathieu; Zogheib, Elie; Hchikat, Abdel Hakim; Kirat, Kahina; Ferraz, Linda; Guerin-Robardey, Anne-Marie; Trojette, Faouzi; Moubarak-Daher, Mona; Dupont, Hervé

    2015-07-02

    In humans, Pasteurella multocida infections are usually limited to the soft tissues surrounding a lesion. However, P. multocida can also cause systemic infections (such as pneumonia, lung abscess, peritonitis, endocarditis, meningitis and sepsis)-especially in patients with other underlying medical conditions. We report on a case of fulminant P. multocida bacteremia at several sites (soft tissues, endocarditis and joints) on a white European man. Despite surgery and intensive medical care, the patient died. The present case emphasizes the importance of appropriate initial treatment of skin wounds. Patients at risk should be aware of the possible consequences of being bitten, scratched or licked by their pet.

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

    Science.gov (United States)

    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.

  7. Immunohistochemical study of porcine lung lesions associated with Pasteurella multocida

    DEFF Research Database (Denmark)

    Pors, Susanne Elisabeth; Hansen, Mette Sif; Bisgaard, Magne

    2013-01-01

    Infectious bronchopneumonia is a widespread disease in modern commercial pig production and Pasteurella multocida is frequently associated with the lesions. To evaluate porcine lung lesions associated with P. multocida, populations of inflammatory cells were examined by immunohistochemistry...... by immunohistochemistry were T-lymphocytes (CD3(+), CD4(+) and CD8(+) subsets), B-lymphocytes, neutrophils, macrophages, and IgA(+), IgM(+) and IgG(+) cells. The results showed that (1) a significant increase in all inflammatory cells was found in lesions associated with P. multocida, (2) necrotic lesions had a larger....... The results show that P. multocida significantly alters the inflammatory response in the lung and that lesions associated with P. multocida display diverse inflammatory responses according to their distinct morphological pattern....

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

    Science.gov (United States)

    2010-01-01

    ... pure, safe, and immunogenic shall be used for vaccine production. All serials of vaccine shall be... Master Seed used for vaccine production shall be tested for immunogenicity. The immunogenicity of a... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Multocida Vaccine, Bovine...

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

    Science.gov (United States)

    2010-01-01

    ... immunogenic shall be used for vaccine production. (a) The Master Seed shall meet the applicable general... vaccine production shall be tested for immunogenicity in each species and for each serotype for which the... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Multocida Vaccine, Avian...

  10. Pasteurella multocida-toxin induced atrophic rhinitis in piglets

    NARCIS (Netherlands)

    Diemen, van P.M.

    1995-01-01


    Progressive atrophic rhinitis (AR) is a complex of disease symptoms caused by infection with toxigenic Pasteurella multocida . Environmental and animal factors contribute to the severity of the disease. Their impact and relationship with severity of disease are

  11. and genotyping of Pasteurella multocida isolated from goat in Iran

    African Journals Online (AJOL)

    STORAGESEVER

    2009-08-18

    Aug 18, 2009 ... In this study, phenotype, capsular type and some virulence factors (pfha1, tbpA, and toxA) of caprine. Pasteurella multocida were described in 10 isolates from goat. Two biochemical biovars were recognized among the isolates based on dulcitol and sorbitol fermentation. The isolates belonged to biovars P.

  12. Isolation and characterization of Pasteurella multocida from caprine ...

    African Journals Online (AJOL)

    By conventional microbiological methods, investigation was carried out in Nsukka and Enugu areasof Enugu State, Nigeria to determne the percentage frequency of occurrence of Pasteurella multocida from caprine pneumonic lungs in Nsukka and Enugu areas of Enugu State, Nigeria. In this study that spanned 12 years, ...

  13. Preliminary Findings on the Carrier Status of Pasteurella multocida ...

    African Journals Online (AJOL)

    One hundred and twenty three indigenous chickens and 24 ducks reared under free range scavenging system were examined for the carrier status of. Pasteurella multocida. Both the oropharynyngeal and cloacal swab samples were examined for the presence of the organisms by means of mouse passage and inoculation ...

  14. Isolation of Pasteurella multocida subspec. Multocida from chronic periapical lesion: First isolation in ex-Yugoslavia

    Directory of Open Access Journals (Sweden)

    Suvajdžić Ljiljana Ð.

    2006-01-01

    Full Text Available This study presents five isolates of Pasteurella multocida subsp. multo-cida isolated from chronic periapical inflammatory lesion. We described the methods of sampling and cultivation as well as diagnostic criteria. Pasteurella multocida was diagnosed on the basis of characteristic cultural and tinctorial properties and the facts that all strains produced indole and induced ornithine decarboxilation, glucose, saccharose and manitole fermentation. Isolates produced neither urease, nor fermented lactose and maltose. Further classification to subspecies multocida was based on the fact that all investigated isolates fermented trechalose, xylose and sorbitol the traits which are diagnostically significant for the species. Patients deny any contact with farm animals or pets, which indicates a possible aerosol transport and animal-human as well as human-human infection. We consider that this organism should be paid more attention by dentist, oral surgeons and microbiologists.

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

    Science.gov (United States)

    Blanchong, Julie A.; Samuel, M.D.; Goldberg, Diana 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.

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

    Science.gov (United States)

    Blanchong, Julie A; Samuel, Michael D; Goldberg, Diana R; Shadduck, Daniel J; Lehr, Margaret 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 1996-99. 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.

  17. Characterization of Pasteurella multocida strains isolated from geese.

    Science.gov (United States)

    Varga, Zsuzsanna; Volokhov, Dmitriy V; Stipkovits, László; Thuma, Akos; Sellyei, Boglárka; Magyar, Tibor

    2013-04-12

    Phenotypic and genotypic diversity of forty-two Pasteurella multocida isolates from geese were characterized by analysis of their capsular type, Heddleston serotype, biotype, fimbrial gene allele type, comparative outer membrane protein (OMP) electrophoresis patterns, and were analyzed using PCR for the presence of virulence-associated genes (toxA, tbpA, pfhA, hgbA, hgbB, nanH, nanB, fimA, hsf-1, and pmHAS). A sequence comparison of the thdF and rpoB housekeeping genes of twenty representative P. multocida strains from three different OMP groups demonstrated that seventeen strains were closely related phylogenetically to previously published strains of P. multocida subsp. multocida and P. multocida subsp. gallicida, and only three strains from geese were grouped with previously published strains of P. multocida subsp. septica. This study is the first report regarding the characterization of phenotypic and genotypic features of different P. multocida field strains isolated from geese with the different clinical representation of pasteurellosis and provide our overview on the usefulness of these in vitro tests for primary characterization of P. multocida strains for the epidemiological studies of fowl cholera. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Pasteurella multocida peritonitis after cat scratch in a patient with cirrhotic ascites

    Directory of Open Access Journals (Sweden)

    Roshan Gunathilake

    2015-06-01

    Full Text Available 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.

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

    Science.gov (United States)

    2010-01-01

    ... SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.116 Pasteurella Multocida Bacterin, Avian...

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

    Science.gov (United States)

    2010-01-01

    ... SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.118 Pasteurella Multocida Bacterin, Avian...

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

    Science.gov (United States)

    2010-01-01

    ... SERVICE, DEPARTMENT OF AGRICULTURE VIRUSES, SERUMS, TOXINS, AND ANALOGOUS PRODUCTS; ORGANISMS AND VECTORS STANDARD REQUIREMENTS Inactivated Bacterial Products § 113.117 Pasteurella Multocida Bacterin, Avian...

  2. Transcriptomic Analysis on Responses of Murine Lungs to Pasteurella multocida Infection

    OpenAIRE

    Chenlu Wu; Xiaobin Qin; Pan Li; Tingting Pan; Wenkai Ren; Nengzhang Li; Yuanyi Peng

    2017-01-01

    Pasteurella multocida infection in cattle causes serious epidemic diseases and leads to great economic losses in livestock industry; however, little is known about the interaction between host and P. multocida in the lungs. To explore a fully insight into the host responses in the lungs during P. multocida infection, a mouse model of Pasteurella pneumonia was established by intraperitoneal infection, and then transcriptomic analysis of infected lungs was performed. P. multocida localized and ...

  3. Immunogenicity of Pasteurella multocida and Mannheimia haemolytica outer membrane vesicles

    Science.gov (United States)

    Roier, Sandro; Fenninger, Judith C.; Leitner, Deborah R.; Rechberger, Gerald N.; Reidl, Joachim; Schild, Stefan

    2013-01-01

    Pasteurella multocida is able to cause disease in humans and in a wide range of animal hosts, including fowl cholera in birds, atrophic rhinitis in pigs, and snuffles in rabbits. Together with Mannheimia haemolytica, P. multocida also represents a major bacterial causative agent of bovine respiratory disease (BRD), which is one of the most important causes for economic losses for the cattle backgrounding and feedlot industry. Commercially available vaccines only partially prevent infections caused by P. multocida and M. haemolytica. Thus, this study characterized the immunogenicity of P. multocida and M. haemolytica outer membrane vesicles (OMVs) upon intranasal immunization of BALB/c mice. Enzyme-linked immunosorbent assays (ELISA) revealed that OMVs derived from P. multocida or M. haemolytica are able to induce robust humoral and mucosal immune responses against the respective donor strain. In addition, also significant cross-immunogenic potential was observed for both OMV types. Colonization studies showed that a potential protective immune response against P. multocida is not only achieved by immunization with P. multocida OMVs, but also by immunization with OMVs derived from M. haemolytica. Immunoblot and immunoprecipitation analyses demonstrated that M. haemolytica OMVs induce a more complex immune response compared to P. multocida OMVs. The outer membrane proteins OmpA, OmpH, and P6 were identified as the three major immunogenic proteins of P. multocida OMVs. Amongst others, the serotype 1-specific antigen, an uncharacterized outer membrane protein, as well as the outer membrane proteins P2 and OmpA were found to be the most important antigens of M. haemolytica OMVs. These findings are useful for the future development of broad-spectrum OMV based vaccines against BRD and other infections caused by P. multocida or M. haemolytica. PMID:23731905

  4. Prolonged incubation period in neonatal Pasteurella multocida meningitis and bacteremia.

    Science.gov (United States)

    Yamaguchi, Hiroshi; Tamura, Takuya; Abe, Michiko; Ogiwara, Shigetoshi; Sai, Shuji; Kosugiyama, Kiyotaka; Sugihara, Akemi; Nagumo, Kiyoshi; Iwata, Seido; Kinugawa, Yoshikazu

    2014-12-01

    Pasteurella multocida, often found as part of the human oral flora and in finger/toenails, also exists in many animals, especially cats, dogs, and pigs. Although rare, pasteurella infection in neonates can cause serious systemic disease, such as meningitis. In this article, a 23-day-old girl presented with decreased appetite and irritability for >2 days. Eighteen days previously her pet cat had jumped onto the left side of her head while she was sleeping. On laboratory data C-reactive protein was high, and on cerebrospinal fluid (CSF) analysis leukocyte count was extremely high, with low glucose and high protein. P. multocida grew out of the blood and CSF cultures, and she was successfully treated with antibiotics for 3 weeks. Although pasteurellosis rarely occurs, it can sometimes lead to life-threatening situations, so parents should exercise caution when having pets around their children. © 2014 Japan Pediatric Society.

  5. Disseminated Pasteurella multocida infection: Cellulitis, osteomyelitis, and myositis.

    Science.gov (United States)

    Marcantonio, Yasmin C; Kulkarni, Prathit A; Sachs, Shira; Ting, Kevin; Lee, Jennifer; Mendoza, Daniel

    2017-01-01

    A 67-year-old man with poorly controlled type II diabetes mellitus was evaluated for right lower extremity erythema and swelling and left-sided lower back pain. He was found to have Pasteurella multocida bacteremia; magnetic resonance imaging showed osteomyelitis of the lumbar spine with myositis in the adjacent left paraspinal muscles. He was initially treated with intravenous antibiotics and was later transitioned to oral amoxicillin. He recovered completely with six weeks of antimicrobial therapy.

  6. Identification of Pasteurella multocida CHAPS-soluble outer membrane proteins.

    Science.gov (United States)

    Tabatabai, Louisa B

    2008-03-01

    Fowl cholera continues to be of concern to the poultry industry, especially for turkey growers. This disease costs the turkey industry millions of dollars annually. In order to develop improved live attenuated vaccines or subunit vaccines, the outer-membrane proteins of Pasteurella multocida were examined with the use of proteomics. Of the 11 proteins total present in an outer-membrane subfraction of P. multocida, four additional proteins were identified, completing the composition of the detergent-soluble cross-protective protein fraction. These additional four proteins include protective bacterial surface antigen, OMA87 (Accession no. 15603857); heme-hemopexin receptor, HemR (Accession no. 15602441); lactate permease, LctP (Accession no. 15603717); and heptosyl transferase F, RfaF (Accession no. 15603709). Both the Oma87 and the HemR proteins would be of interest for subunit and modified live vaccine studies, respectively, because of their purported roles as virulence factors for P. multocida.

  7. Conditions for transformation of Pasteurella multocida by electroporation.

    Science.gov (United States)

    Jablonski, L; Sriranganathan, N; Boyle, S M; Carter, G R

    1992-01-01

    Conditions for electroporation of plasmid DNA into Pasteurella multocida were determined for use in developing a cloning system to study virulence factors of P. multocida. The highest efficiency of transformation (1.25 x 10(7) cfu/micrograms DNA) was obtained when 7.6 x 10(10) cells of P. multocida strain R473 were electroporated at 12.5 kV/cm (10 ms, 5 ng of pVM109). Transformation efficiencies of cells prepared at mid-log-phase were approximately 0.5 log10 lower than early, late, or stationary phases. Neither pBR322 nor pUC-19 were able to transform strain R473 under these conditions, even when DNA concentrations were increased to 1 microgram. When pBR322 was ligated with a Pasteurella plasmid, pLAR-1, the hybrid was able to transform strain R473 at an efficiency between 4.5 x 10(2) and 8 x 10(4) cfu/micrograms DNA. Six strains of P. multocida including serotypes A, B, D, and E were transformed successfully.

  8. Immunohistochemical study of porcine lung lesions associated with Pasteurella multocida.

    Science.gov (United States)

    Pors, Susanne E; Hansen, Mette S; Bisgaard, Magne; Jensen, Henrik E; Iburg, Tine M

    2013-08-01

    Infectious bronchopneumonia is a widespread disease in modern commercial pig production and Pasteurella multocida is frequently associated with the lesions. To evaluate porcine lung lesions associated with P. multocida, populations of inflammatory cells were examined by immunohistochemistry in necrotic lung lesions from nine pigs and exudative lung lesions from eleven pigs. Lungs from five pigs served as controls. All cases were selected from naturally infected pigs using co-infection based criteria to make them as comparable as possible. The inflammatory cells demonstrated by immunohistochemistry were T-lymphocytes (CD3(+), CD4(+) and CD8(+) subsets), B-lymphocytes, neutrophils, macrophages, and IgA(+), IgM(+) and IgG(+) cells. The results showed that (1) a significant increase in all inflammatory cells was found in lesions associated with P. multocida, (2) necrotic lesions had a larger number of CD3(+) T-lymphocytes and IgA(+) cells, and (3) cases with exudative lesions had a more CD8(+) T-lymphocytes, B-lymphocytes, macrophages and neutrophils. No differences in the numbers of CD4(+) T-lymphocytes, IgG(+) and IgM(+) positive cells were found between necrotic and exudative cases. The results show that P. multocida significantly alters the inflammatory response in the lung and that lesions associated with P. multocida display diverse inflammatory responses according to their distinct morphological pattern. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Pasteurella multocida lipopolysaccharide: the long and the short of it.

    Science.gov (United States)

    Harper, Marina; Cox, Andrew D; Adler, Ben; Boyce, John D

    2011-11-21

    Pasteurella multocida is a capsulated, gram-negative cocco-bacillus that can cause serious disease in a wide range of mammals and birds. P. multocida strains are classified into 16 serovars based on lipopolysaccharide (LPS) antigens. LPS is an essential virulence factor of P. multocida; mutants expressing severely truncated LPS are completely attenuated in chickens. LPS is also a major immunogen of P. multocida and protection against infections caused by P. multocida is generally considered to be serovar specific. In this review we summarize current knowledge of the structure and genetics of LPS assembly of P. multocida strains belonging to five different serovars. These include strains belonging to serovars 1 and 3, the most common serovars found in the poultry industry, and strains belonging serovars 2 and 5, the serovars associated with bovine haemorrhagic septicaemia outbreaks. A number of the serovars are genetically related; serovars 1 and 14 share the same LPS outer core biosynthesis locus, but due to a mutation within the phosphocholine biosynthesis gene, pcgA, the serovar 14 strain produces a truncated LPS structure. Similarly serovars 2 and 5 share an identical LPS outer core locus and express near-identical LPS structures. However, due to a single point mutation in the phosphoethanolamine (PEtn) transferase gene, lpt_3, the serovar 2 strain does not elaborate a PEtn residue on heptose II. Knowledge of the genetic basis for the LPS structures expressed by P. multocida will facilitate the development of rapid molecular methods for typing and diagnosis and will be essential for a rational approach to vaccine formulation. Copyright © 2011 Elsevier B.V. All rights reserved.

  10. Clinical Features and Outcomes of Pasteurella multocida Infection.

    Science.gov (United States)

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

    2015-09-01

    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.

  11. Two novel cross-protective antigens for bovine Pasteurella multocida

    Science.gov (United States)

    Du, Huihui; Wu, Chenlu; Li, Chunming; Fang, Rendong; Ma, Jianwei; Ji, Jiale; Li, Zhihong; Li, Nengzhang; Peng, Yuanyi; Zhou, Zeyang

    2017-01-01

    Pasteurella multocida is an important pathogen that leads to a range of diseases that have severe economic consequences on cattle production. In order to develop an effective cross-protective component vaccine, an immunoproteomics approach was used to analyze outer membrane proteins (OMPs) of the P. multocida serotype A, B and F strains. Candidate antigen molecules from the whole genome were screened via linear trap quadrupole mass spectrometry and bioinformatics analysis, and the reactogenicity of the candidate antigen molecules was analyzed via cloning, expression, and ELISA or protein immunoblotting, and the vaccine efficacy of the candidate molecules was determined in infective animal models and cross-protective antigens may be obtained. rPmCQ2_2g0128, rPmCQ2_1g0327 and rPmCQ2_1g0020 proteins were selected. Their protective rates were 40/30/20% (rPmCQ2_2g0128), 50/40/0% (rPmCQ2_1g0327) and 0/40/30% (rPmCQ2_1g0020) against ten-fold median lethal dose (10LD50) of the P. multocida serotypes A, B and F in a mouse model, respectively. The results suggested that the two proteins rPmCQ2_2g0128 and rPmCQ2_1g0327 may be used as vaccine candidates against bovine P. multocida serotypes A, B. To the best of our knowledge, the present study was the first to identify cross-protective antigens, extracted OMPs from bovine P. multocida. PMID:28791368

  12. Development of a typing system for epidemiological studies of porcine toxin-producing Pasteurella multocida ssp. multocida in Denmark

    DEFF Research Database (Denmark)

    Fussing, V.; Nielsen, Jens; Bisgaard, M.

    1999-01-01

    The aim of the present study was to evaluate capsular-typing, plasmid-profiling, phage-typing and ribotyping for epidemiological studies of toxin-producing Pasteurella multocida ssp. multocida in Denmark. The evaluation of methods was based on 68 strains from nasal swabs and 14 strains from...... by HindIII ribotyping, as 85% of isolates from all herds were assigned to one ribotype. In conclusion, HindIII ribotyping seems to represent a useful tool for epidemiological studies of toxigenic P. multocida ssp. multocida....

  13. Proximity-Dependent Inhibition of Growth of Mannheimia haemolytica by Pasteurella multocida

    Science.gov (United States)

    Bavananthasivam, Jegarubee; Dassanayake, Rohana P.; Kugadas, Abirami; Shanthalingam, Sudarvili; Call, Douglas R.; Knowles, Donald P.

    2012-01-01

    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. haemolytica has been isolated by culture less frequently than the other bacteria. We hypothesized that the growth of M. haemolytica is inhibited by other bacteria in the lungs of BHS. The objective of this study was to determine whether P. multocida inhibits the growth of M. haemolytica. Although in monoculture both bacteria exhibited similar growth characteristics, in coculture with P. multocida there was a clear inhibition of growth of M. haemolytica. The inhibition was detected at mid-log phase and continued through the stationary phase. When cultured in the same medium, the growth of M. haemolytica was inhibited when both bacteria were separated by a membrane that allowed contact (pore size, 8.0 μm) but not when they were separated by a membrane that limited contact (pore size, 0.4 μm). Lytic bacteriophages or bactericidal compounds could not be detected in the culture supernatant fluid from monocultures of P. multocida or from P. multocida-M. haemolytica cocultures. These results indicate that P. multocida inhibits the growth of M. haemolytica by a contact- or proximity-dependent mechanism. If the inhibition of growth of M. haemolytica by P. multocida occurs in vivo as well, it could explain the inconsistent isolation of M. haemolytica from the lungs of pneumonic BHS. PMID:22798357

  14. Kwantitatief onderzoek naar de gevoeligheid van Bordetella bronchiseptica en Pasteurella multocida voor sulfonamiden

    NARCIS (Netherlands)

    Klingeren; B.van; Dessens-Kroon; M.; Verheuvel; M.

    1985-01-01

    Teneinde kwantitatieve informatie te verkrijgen over de gevoeligheid en het resistentieniveau van Bordetella bronchiseptica en Pasteurella multocida voor sulfonamiden werd voor resp. 119 en 151 bij biggen/varkens geisoleerde stammen van deze species uit een vijftal veterinair bacteriologische

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

    Science.gov (United States)

    Samuel, M.D.; Shadduck, D.J.; Goldberg, Diana 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.

  16. Pasteurella multocida involved in respiratory disease of wild chimpanzees.

    Directory of Open Access Journals (Sweden)

    Sophie Köndgen

    Full Text Available Pasteurella multocida can cause a variety of diseases in various species of mammals and birds throughout the world but nothing is known about its importance for wild great apes. In this study we isolated P. multocida from wild living, habituated chimpanzees from Taï National Park, Côte d'Ivoire. Isolates originated from two chimpanzees that died during a respiratory disease outbreak in 2004 as well as from one individual that developed chronic air-sacculitis following this outbreak. Four isolates were subjected to a full phenotypic and molecular characterisation. Two different clones were identified using pulsed field gel electrophoresis. Multi Locus Sequence Typing (MLST enabled the identification of previous unknown alleles and two new sequence types, ST68 and ST69, were assigned. Phylogenetic analysis of the superoxide dismutase (sodA gene and concatenated sequences from seven MLST-housekeeping genes showed close clustering within known P. multocida isolated from various hosts and geographic locations. Due to the clinical relevance of the strains described here, these results make an important contribution to our knowledge of pathogens involved in lethal disease outbreaks among endangered great apes.

  17. Pasteurella multocida Involved in Respiratory Disease of Wild Chimpanzees

    Science.gov (United States)

    Köndgen, Sophie; Leider, Michaela; Lankester, Felix; Bethe, Astrid; Lübke-Becker, Antina; Leendertz, Fabian H.; Ewers, Christa

    2011-01-01

    Pasteurella multocida can cause a variety of diseases in various species of mammals and birds throughout the world but nothing is known about its importance for wild great apes. In this study we isolated P. multocida from wild living, habituated chimpanzees from Taï National Park, Côte d'Ivoire. Isolates originated from two chimpanzees that died during a respiratory disease outbreak in 2004 as well as from one individual that developed chronic air-sacculitis following this outbreak. Four isolates were subjected to a full phenotypic and molecular characterisation. Two different clones were identified using pulsed field gel electrophoresis. Multi Locus Sequence Typing (MLST) enabled the identification of previous unknown alleles and two new sequence types, ST68 and ST69, were assigned. Phylogenetic analysis of the superoxide dismutase (sodA) gene and concatenated sequences from seven MLST-housekeeping genes showed close clustering within known P. multocida isolated from various hosts and geographic locations. Due to the clinical relevance of the strains described here, these results make an important contribution to our knowledge of pathogens involved in lethal disease outbreaks among endangered great apes. PMID:21931664

  18. Pharmacokinetic/pharmacodynamic integration and modelling of florfenicol for the pig pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida

    National Research Council Canada - National Science Library

    Lucy Dorey; Ludovic Pelligand; Zhangrui Cheng; Peter Lees

    2017-01-01

    Pharmacokinetic-pharmacodynamic (PK/PD) integration and modelling were used to predict dosage schedules for florfenicol for two pig pneumonia pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida...

  19. Fatal peritonitis caused by Pasteurella multocida capsular type F in calves.

    Science.gov (United States)

    Catry, Boudewijn; Chiers, Koen; Schwarz, Stefan; Kehrenberg, Corinna; Decostere, Annemie; de Kruif, Aart

    2005-03-01

    A fatal case of atypical septicemia of pasteurellosis in veal calves is described. The causative organism was identified as a multiresistant Pasteurella multocida capsular type F isolate. The outbreak was characterized by fibrinous peritonitis and mortality, which are hitherto unreported features of P. multocida capsular type F infections.

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

    Science.gov (United States)

    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 ...

  1. [Pasteurella multocida meningitis in a 93-year-old woman following a cat bite].

    Science.gov (United States)

    Dutheil, F; Wahl, D; Chamoux, A

    2009-01-01

    Pasteurella multocida is commensal of the normal nasopharynx or gastrointestinal flora in many domestic and wild animals. Cat and dog bites usually cause soft tissue infections. P. multocida is an unusual causative agent of meningitis which tends to affect neonates or aged patients. The authors report a case (a 93-year-old woman) of P. multocida meningitis induced by a cat bite with dramatic sequels leading to complete patient disability.

  2. Infection-stage adjusted dose of beta-lactams for parsimonious and efficient antibiotic treatments: A Pasteurella multocida experimental pneumonia in mice.

    Directory of Open Access Journals (Sweden)

    Maleck V Vasseur

    Full Text Available In this study, the impact of infection stage on clinically and microbiologically efficacious doses and on antibiotic consumption was assessed during a naturally evolving infectious disease, using an original mouse model of pulmonary infection produced by air-borne contamination. When Pasteurella multocida was administered as pathogenic agent to immunocompetent mice, 60% of the animals exhibited clinical symptoms of pneumonia 2 to 4 days after bacterial contamination of the lungs. Two beta-lactam antibiotics were evaluated: amoxicillin and cefquinome, a fourth generation cephalosporin developed for food animals. First, a pharmacokinetic study was performed in infected mice to determine the exposure to amoxicillin or cefquinome required to treat clinically affected animals, based on the targeted values of PK/PD indices for beta-lactams. We then confirmed that these doses resulted in a 100% clinical cure rate in animals exhibiting clinical signs of infection and harboring a high pathogenic inoculum. More interestingly, we also showed that the same 100% clinical cure could be obtained in our model with 10-fold lower doses in animals at pre-patent stages of infection i.e. when harboring a low pathogenic inoculum. At the group level, antimicrobial drug consumption was reduced by treating animals at an early stage of the infection course with a pre-patent tailored dose. These results suggest that early treatment with a dose suitably adjusted to the stage of infection might help to reduce both overall antibiotic consumption and resistance selection pressure in the animals and in the environment.

  3. Homology modeling, functional annotation and comparative genomics of outer membrane protein H of Pasteurella multocida.

    Science.gov (United States)

    Ganguly, Bhaskar; Tewari, Kamal; Singh, Rashmi

    2015-12-07

    Pasteurella multocida is an important pathogen of animals and humans. Outer Membrane Protein (Omp) H is a major conserved protein in the envelope of P. multocida and has been commonly targeted as a protective antigen. However, not much is known about its structure and function due to the difficulties that are typically associated with obtaining sufficient amounts of purified prokaryotic transmembrane proteins. The present work is aimed at studying the OmpH using an in silico approach and consolidate the findings in light of existing experimental evidences. Our study describes the first 3D model of the P. multocida OmpH obtained through a combination of several in silico modeling approaches. From our results, OmpH of P. multocida could be classified as a homotrimeric, 16 stranded, β-barrel porin involved in the non-specific transport of small, hydrophilic molecules, serving essential osmoregulatory function. Moreover, very small homologous sequences could be identified in the host proteome, strengthening the probability of a successful OmpH-based vaccine against the pathogen with remote chances of cross-reaction to host proteins. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Pasteurella multocida thymidine kinase 1 efficiently activates pyrimidine nucleoside analogs.

    Science.gov (United States)

    Clausen, A R; Al Meani, S A L; Piskur, J

    2010-06-01

    In the Pasteurella multocida genome only one putative deoxyribonucleoside kinase encoding gene, for thymidine kinase 1 (PmTK1), was identified. The PmTK1 gene was sub-cloned into Escherichia coli KY895 and it sensitized the host towards 2',2'-difluoro-deoxycytidine (gemcitabine, dFdC), 3'-azido-thymidine (AZT) and 5-fluoro-deoxyuridine (5F-dU). PmTK1 was over-expressed and purified with two different tags. Apparently, deoxyuridine (dU), and not thymidine (dT), is the preferred substrate. We suggest that PmTK1s could be employed as a species-specific activator of uracil-based nucleoside antibiotics.

  5. Characterization of Pasteurella multocida mutants of low virulence.

    Science.gov (United States)

    Lee, M D; Glisson, J R; Wooley, R E; Brown, J

    1990-01-01

    Ten temperature-sensitive mutants of the Clemson University (CU) vaccine strain of Pasteurella multocida have been developed and were characterized by phenotypic attributes such as carbohydrate fermentation, antibiotic resistance, and membrane protein profiles. Some mutants were found to have lost the ability to utilize some substrates, notably xylose and gluconate, whereas others were able to ferment additional carbohydrates such as arabinose and rhamnose. CU was found to be resistant to sulfisoxazole, of intermediate resistance to bacitracin, and sensitive to rifampin; the sensitivity to these three antibiotics varied among the mutant strains, but 60% were resistant to rifampin. Membrane protein profiles demonstrated some changes in major bands, and there was variation in 50% of the mutants in proteins in the 31 kilodalton range. All strains were assayed for the presence of several virulence factors, and many were found to produce siderophore and to exhibit some degree of complement resistance.

  6. Biofilm formation of Pasteurella multocida on bentonite clay.

    Science.gov (United States)

    Rajagopal, Ramachandranpillai; Nair, Govindapillai Krishnan; Mini, Mangattumuruppel; Joseph, Leo; Saseendranath, Mapranath Raghavan; John, Koshy

    2013-06-01

    Biofilms are structural communities of bacterial cells enshrined in a self produced polymeric matrix. The studies on biofilm formation of Pasteurella multocida have become imperative since it is a respiratory pathogen and its biofilm mode could possibly be one of its virulence factors for survival inside a host. The present study describes a biofilm assay for P. multocida on inert hydrophilic material called bentonite clay. The potential of the organism to form in vitro biofilm was assessed by growing the organism under nutrient restriction along with the inert substrate bentonite clay, which will provide a surface for attachment. For quantification of biofilm, plate count by the spread plate method was employed. Capsule production of the attached bacteria was demonstrated by light microscopic examination following Maneval staining and capsular polysaccharide estimation was done using standard procedures. The biofilm formation peaked on the third day of incubation (1.54 ×10(6) cfu/g of bentonite clay) while the planktonic cells were found to be at a maximum on day one post inoculation (8.10 ×10(8) cfu/ml of the broth). Maneval staining of late logarithmic phase biofilm cultures revealed large aggregates of bacterial cells, bacteria appearing as chains or as a meshwork. The capsular polysaccharide estimation of biofilm cells revealed a 3.25 times increase over the planktonic bacteria. The biofilm cells cultured on solid media also produced some exclusive colony morphotypes.

  7. Occurrence of Pasteurella multocida and related species in village free ranging chickens and their animal contacts in Tanzania.

    Science.gov (United States)

    Muhairwa, A P; Mtambo, M M; Christensen, J P; Bisgaard, M

    2001-01-26

    Investigation was done to determine the presence of Pasteurella multocida and related species in free ranging chickens and ducks, dogs, cats and pigs in three climatic zones (cool, warm and hot) of rural Morogoro, Tanzania. A total of 153 isolates of P. multocida ssp. multocida and related species were obtained by direct culture on blood agar, selective medium and mouse inoculation. P. multocida ssp. multocida was isolated from 0.7% of chickens and 7% of ducks. In dogs and cats, P. multocida ssp. multocida was isolated from 1 and 68%, respectively. One isolate of Pasteurella gallinarum was isolated from a duck. Other species obtained were; P. multocida ssp. septica, Pasteurella stomatis and taxon 16 from dogs and cats, while Pasteurella dagmatis and Pasteurella canis were found in dogs only. Prevalence of P. multocida ssp. multocida was significantly higher (PPasteurella spp. were also observed in the study, but further characterization is required before the final classification can be made. This paper reports for the first time the isolation of unclassified Pasteurella from dogs and cats in Africa. The results implies that fowl cholera might be occurring in free ranging poultry, and dogs and cats kept in contact might serve as sources of P. multocida to chickens and ducks. Subsequent applications of molecular techniques to analyse the epidemiological relatedness of clones isolated from different host species is indicated.

  8. Transcriptomic Analysis on Responses of Murine Lungs to Pasteurella multocida Infection

    Directory of Open Access Journals (Sweden)

    Chenlu Wu

    2017-06-01

    Full Text Available Pasteurella multocida infection in cattle causes serious epidemic diseases and leads to great economic losses in livestock industry; however, little is known about the interaction between host and P. multocida in the lungs. To explore a fully insight into the host responses in the lungs during P. multocida infection, a mouse model of Pasteurella pneumonia was established by intraperitoneal infection, and then transcriptomic analysis of infected lungs was performed. P. multocida localized and grew in murine lungs, and induced inflammation in the lungs, as well as mice death. With transcriptomic analysis, approximately 107 clean reads were acquired. 4236 differently expressed genes (DEGs were detected during P. multocida infection, of which 1924 DEGs were up-regulated. By gene ontology (GO and Kyoto encyclopedia of genes and genomes (KEGG enrichments, 5,303 GO enrichments and 116 KEGG pathways were significantly enriched in the context of P. multocida infection. Interestingly, genes related to immune responses, such as pattern recognition receptors (PRRs, chemokines and inflammatory cytokines, were significantly up-regulated, suggesting the key roles of these genes in P. multocida infection. Transcriptomic data showed that IFN-γ/IL-17-related genes were increased, which were validated by qRT-PCR, ELISA, and immunoblotting. Our study characterized the transcriptomic profile of the lungs in mice upon Pasteurella infection, and our findings could provide valuable information with respect to better understanding the responses in mice during P. multocida infection.

  9. Transcriptomic Analysis on Responses of Murine Lungs to Pasteurella multocida Infection.

    Science.gov (United States)

    Wu, Chenlu; Qin, Xiaobin; Li, Pan; Pan, Tingting; Ren, Wenkai; Li, Nengzhang; Peng, Yuanyi

    2017-01-01

    Pasteurella multocida infection in cattle causes serious epidemic diseases and leads to great economic losses in livestock industry; however, little is known about the interaction between host and P. multocida in the lungs. To explore a fully insight into the host responses in the lungs during P. multocida infection, a mouse model of Pasteurella pneumonia was established by intraperitoneal infection, and then transcriptomic analysis of infected lungs was performed. P. multocida localized and grew in murine lungs, and induced inflammation in the lungs, as well as mice death. With transcriptomic analysis, approximately 107 clean reads were acquired. 4236 differently expressed genes (DEGs) were detected during P. multocida infection, of which 1924 DEGs were up-regulated. By gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichments, 5,303 GO enrichments and 116 KEGG pathways were significantly enriched in the context of P. multocida infection. Interestingly, genes related to immune responses, such as pattern recognition receptors (PRRs), chemokines and inflammatory cytokines, were significantly up-regulated, suggesting the key roles of these genes in P. multocida infection. Transcriptomic data showed that IFN-γ/IL-17-related genes were increased, which were validated by qRT-PCR, ELISA, and immunoblotting. Our study characterized the transcriptomic profile of the lungs in mice upon Pasteurella infection, and our findings could provide valuable information with respect to better understanding the responses in mice during P. multocida infection.

  10. Pasteurella multocida Septicemia in a Patient with Cirrhosis: An Autopsy Report

    Directory of Open Access Journals (Sweden)

    Takuma Yamamoto

    2015-01-01

    Full Text Available More people are keeping pets in their homes but may not be sufficiently aware of the potential danger from infections. We report an autopsy case of a 57-year-old man affected by cirrhosis. Septic shock with Pasteurella multocida pneumonia was the cause of his death. P. multocida was the source of infection via the respiratory tract and caused pneumonia. Cirrhosis is one of the risk factors for P. multocida infection. A detailed patient history about animal exposure should be obtained and a differential diagnosis of P. multocida infection must be kept in mind.

  11. Fowl Cholera and Its Control Prospect With Locally isolated Pasteurella multocida Bivalent Vaccines

    Directory of Open Access Journals (Sweden)

    Tati Ariyanti

    2008-03-01

    Full Text Available Pasteurellosis or fowl cholera disease which associated with Pasteurella multocida group A and D infections occurred sporadically in many parts of the world, including in Indonesia. The pathogenic activity of P. multocida in chickens were based on lipopolysacharide (LPS antigens associated with group A and D capsules, and the resistance factor of complement mediated bacteriolysis in animals. In order to reduce common bacterial infections, antibiotics were routinely used as feed additive or by drinking water, but fowl cholera cases still occur. Fowl cholera control by vaccinations have been used more than a hundred years ago by means of inactive vaccine, but imported inactive vaccine was reported not effective due to lack of cross protection against heterologous serotype. At present, many local P. multocida isolates from chicken and ducks from many areas in Indonesia were characterised for their antigenicity, immunogenicity and prepared as monovalent or bivalent vaccine. Only the monovalent vaccine prepared from BCC 2331 or DY2 demonstrated the presence of immunoprotection against homologous and heterologous challenged with live bacteria. The prototype bivalent vaccine consisting of BCC 2331 + DY2 demonstrated high degree of cross protection against challenged individual with or mixed of BCC 2331 + DY2 at average of 60 – 75% and 75 – 100%, respectively. Monovalent and bivalent vaccine prepared from other isolates including imported reference strains of P. multocida demonstrated no protection in experimentally vaccinated ducks and chicken against challenged with live bacteria of neither BCC 2331 nor with DY2. From these retrospective studies, it was concluded that the local isolates P. multocida designated as BCC 2331 and DY2 could be used as candidates of prototype vaccine or master seed vaccine but their effectiveness still need to be evaluated under field conditions.

  12. Pasteurella multocida toxin- induced osteoclastogenesis requires mTOR activation.

    Science.gov (United States)

    Kloos, Bianca; Chakraborty, Sushmita; Lindner, Sonja G; Noack, Katrin; Harre, Ulrike; Schett, Georg; Krämer, Oliver H; Kubatzky, Katharina F

    2015-09-14

    Pasteurella multocida toxin (PMT) is a potent inducer of osteoclast formation. Pigs suffering from an infection with toxigenic Pasteurella multocida strains develop atrophic rhinitis characterised by a loss of turbinate bones and conchae. However, on the molecular level the process of bone loss remains largely uncharacterised. Recently it was found that PMT activates the serine/threonine kinase mammalian target of rapamycin (mTOR) in fibroblasts. Using RAW264.7 macrophages, we investigated the role of the mTOR complex 1 (mTORC1) in PMT-mediated osteoclast formation. PMT induces the differentiation of RAW264.7 macrophages into multinucleated, tartrate resistant acid phosphatase (TRAP) positive osteoclasts that are capable to resorb bone. In the presence of the mTORC1 inhibitor rapamycin, PMT was significantly less able to induce the formation of TRAP-positive osteoclasts. Accordingly, the resulting resorption of bone was strongly reduced. A major target of mTOR is the 70 kDa ribosomal protein S6 kinase 1 (p70 S6K1). Activated p70 S6K1 decreases the expression of programmed cell death protein 4 (PDCD4), a negative transcriptional regulator of osteoclastogenesis, at the protein and gene level. Ultimately this results in the activation of c-Jun, a component of the activator protein 1 (AP-1) complex, which is a major transcription factor for the induction of osteoclast-specific genes. We now demonstrate that c-Jun and its downstream target, the osteoclast-specific bone degrading protease cathepsin K, are upregulated upon PMT treatment in an mTOR-dependent manner. Activation of mTOR signalling plays a central role in the formation of osteoclasts through the bacterial toxin PMT. On the molecular level, PMT-induced activation of mTOR leads to down regulation of PDCD4, a known repressor of AP-1 complex, culminating in the activation of c-Jun, an essential transcription factor for triggering osteoclastogenesis.

  13. Lack of effect of aerial ammonia on atrophic rhinitis and pneumonia induced by Mycoplasma hyopneumoniae and toxigenic Pasteurella multocida

    DEFF Research Database (Denmark)

    Andreasen, Morten; Bækbo, P.; Nielsen, Jens

    2000-01-01

    The objective of this experimental study was to determine the effects of aerial ammonia on disease development and bacterial colonization in weaned pigs inoculated with toxigenic Pasteurella multocida and Mycoplasma hyopneumoniae. Two groups of 10 pigs each were continuously exposed to 50 and 100 p.......p.m ammonia, respectively, and compared to a non-exposed control, group of 20 pigs. Following aerosol inoculation with M. hyopneumoniae at day 9, ail pigs were aerosol-inoculated with toxigenic P. multocida type A at days 28, 42 and 56. Ac day 63 they were euthanized. Clinical signs including coughing...... and respiratory distress were present in all groups following inoculation. No significant differences could be established in the extent or frequency of pneumonia between ammonia-exposed pigs and controls, or in the: extent of conchal atrophy, the frequency of isolation of toxigenic P. multocida from conchae...

  14. Phenotypic and genotypic diversity of organisms previously classified as maltose positive Pasteurella multocida.

    Science.gov (United States)

    Petersen, K D; Christensen, J P; Bisgaard, M

    1998-07-01

    Fifteen isolates tentatively classified as maltose positive Pasteurella multocida have been characterized in 79 biochemical tests and by ribotyping using HindIII and HpaII for digestion of DNA. Phenotypic and genotypic results were analysed using the computer programmes NTSYS and GelCompar, respectively. Two strains were classified as maltose positive P. multocida ssp. multocida while six strains were classified as maltose positive P. multocida ssp. septica. The remaining strains clustered with P. volantium and P. gallinarum, but remained unclassified. With the exception of a single isolate correlation was observed between phenotypic and genotypic results. The unclassified isolates which represented three different sources were heterogenous according to both phenotypic and genotypic results. The findings obtained support the 16S rRNA sequencing results indicating that the genus Pasteurella sensu stricto might represent two or more genera.

  15. Outer membrane vesicles of Pasteurella multocida contain virulence factors

    Science.gov (United States)

    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

  16. Outer membrane vesicles of Pasteurella multocida contain virulence factors.

    Science.gov (United States)

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

    2014-10-01

    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 Amp(r) (serogroup A). Pm OMVs may be involved in different aspects of disease pathogenesis. © 2014 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  17. Genome-Wide Analyses Reveal Genes Subject to Positive Selection in Pasteurella multocida

    OpenAIRE

    Peili Cao; Dongchun Guo; Jiasen Liu; Qian Jiang; Zhuofei Xu; Liandong Qu

    2017-01-01

    Pasteurella multocida, a Gram-negative opportunistic pathogen, has led to a broad range of diseases in mammals and birds, including fowl cholera in poultry, pneumonia and atrophic rhinitis in swine and rabbit, hemorrhagic septicemia in cattle, and bite infections in humans. In order to better interpret the genetic diversity and adaptation evolution of this pathogen, seven genomes of P. multocida strains isolated from fowls, rabbit and pigs were determined by using high-throughput sequencing a...

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

    Science.gov (United States)

    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%.

  19. Isolation, Antimicrobial Resistance, and Virulence Genes of Pasteurella multocida Strains from Swine in China▿

    OpenAIRE

    Tang, Xibiao; Zhao, Zhanqin; Hu, Junyong; Wu, Bin; Cai, Xuwang; He, Qigai; Chen, Huanchun

    2009-01-01

    A total of 233 isolates of Pasteurella multocida were obtained from 2,912 cases of clinical respiratory disease in pigs in China, giving an isolation rate of 8.0%. Serogroup A P. multocida isolates were isolated from 92 cases (39.5%), and serogroup D isolates were isolated from 128 cases (54.9%); 12 isolates (5.2%) were untypeable. P. multocida was the fourth most frequent pathogenic bacterium recovered from the respiratory tract, after Streptococcus suis, Haemophilus parasuis, and Escherichi...

  20. Utilization of L-aspartate, L-malate and fumarate by Pasteurella multocida

    Energy Technology Data Exchange (ETDEWEB)

    Hoefer, M.; Flossmann, K.D. (Akademie der Landwirtschaftswissenschaften der DDR, Jena. Inst. fuer Bakterielle Tierseuchenforschung)

    1981-01-01

    Strains of Pasteurella multocida use L-aspartate, L-malate and furmarate, respectively, as substrates for production of succinic acid which accumulates in the medium. As was established by studies with /sup 14/C- and /sup 3/H-labelled substrates, the degradation of these substances proceeds analogously via the citric acid cycle.

  1. Results of the in vivo application of /sup 125/I-labelled Pasteurella multocida antigens

    Energy Technology Data Exchange (ETDEWEB)

    Flossmann, K.D.; Heilmann, P.; Mueller, G. (Akademie der Landwirtschaftswissenschaften der DDR, Jena. Inst. fuer Bakterielle Tierseuchenforschung)

    1981-09-01

    In in vivo experiments iodine was separated rapidly from /sup 125/I-labelled Pasteurella multocida germs. By means of direct radioactivity measurement as well as by autoradiography could be demonstrated that the iodine was enriched in the thyroid following application of labelled antigen to mice.

  2. In vitro synthesis of heparosan using recombinant Pasteurella multocida heparosan synthase PmHS2

    NARCIS (Netherlands)

    Chavaroche, A.A.E.; Springer, J.; Kooy, F.K.; Boeriu, C.G.; Eggink, G.

    2010-01-01

    In vertebrates and bacteria, heparosan the precursor of heparin is synthesized by glycosyltransferases via the stepwise addition of UDP-N-acetylglucosamine and UDP-glucuronic acid. As heparin-like molecules represent a great interest in the pharmaceutical area, the cryptic Pasteurella multocida

  3. Cat induced Pasteurella multocida peritonitis in peritoneal dialysis: a case report and review of the literature.

    NARCIS (Netherlands)

    Sol, P.M.; Kar, N.C.A.J. van de; Schreuder, M.F.

    2013-01-01

    A 7-year-old girl treated with peritoneal dialysis developed a peritonitis due to Pasteurella multocida after physical contact of the domestic cat with the dialysis machine. Only 25 of such cases have been reported, mostly concerning adults. We report the third case involving a child, together with

  4. Sepsis by Pasteurella multocida in an Elderly Immunocompetent Patient after a Cat Bite

    Directory of Open Access Journals (Sweden)

    Lara Caserza

    2017-01-01

    Full Text Available Pasteurella multocida colonizes animal scratches and bites. This bacterium was described to cause sepsis or endocarditis mainly in immunocompromised patients. We report the case of a 92-year-old woman presenting at the Emergency Department with coma and fever a week after the bite of her cat. The cat bite was misdiagnosed at admission partly due to an underestimation of this event by the patient’s relatives. An inflamed area localized at perimalleolar skin of the right leg was detected. Laboratory biomarkers of inflammation were elevated. The cerebral computed tomography (CT scan with angiographic sequences showed a complete occlusion of right intracranial vertebral artery. Total body CT scan and abdominal echocardiography were negative for foci of infection. Three consecutive blood cultures were positive for Pasteurella multocida. A diagnosis of sepsis by Pasteurella multocida was made, and the patient recovered after a specific antimicrobial treatment. In order to confirm the animal transmission, the cat saliva was cultured and found positive for Pasteurella multocida with a similar antibiotic sensitivity to that isolated from the patient. In conclusion, the case of a patient with coma and fever after a cat bite was presented. The transmission of pathogens from pets has to be carefully considered as an important route of infection in immunocompetent patients.

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

    Science.gov (United States)

    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...

  6. Pasteurella multocida infection, a rare cause of mycotic abdominal aortic aneurysm

    NARCIS (Netherlands)

    Koelemay, Mark J. W.

    2009-01-01

    A 64-year-old man was admitted with abdominal pain 6 weeks after treatment with intravenous flucloxacillin for cellulitis of his right leg. Urgent operation was necessary for a mycotic aneurysm of the abdominal aorta due to infection with Pasteurella multocida, a microorganism residing in the oral

  7. Synthesis of heparosan oligosaccharides by Pasteurella multocida PmHS2 single-action transferases

    NARCIS (Netherlands)

    Chavaroche, A.A.E.; Broek, van den L.A.M.; Boeriu, C.G.; Eggink, G.

    2012-01-01

    Pasteurella multocida heparosan synthase PmHS2 is a dual action glycosyltransferase that catalyzes the polymerization of heparosan polymers in a non-processive manner. The two PmHS2 single-action transferases, obtained previously by site-directed mutagenesis, have been immobilized on

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

    Science.gov (United States)

    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.

  9. Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran

    National Research Council Canada - National Science Library

    Khamesipour, Faham; Momtaz, Hassan; Azhdary Mamoreh, Morteza

    2014-01-01

    A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence-associated genes by polymerase chain reaction (PCR...

  10. Development of a novel multiplex PCR assay for rapid detection of virulence associated genes of Pasteurella multocida from pigs

    National Research Council Canada - National Science Library

    Rajkhowa, S

    2015-01-01

    Significance and Impact of the Study: The study reports the development and evaluation of a novel multiplex PCR assay for the rapid detection of 11 important VAGs of Pasteurella multocida isolates from pigs...

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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.

  14. In vitro and in vivo pathogenicity studies of Pasteurella multocida strains harbouring different ompA.

    Science.gov (United States)

    Katoch, Shailja; Sharma, Mandeep; Patil, R D; Kumar, Sandeep; Verma, Subhash

    2014-09-01

    Pasteurella multocida is a pathogenic, Gram-negative bacterium that is commonly found as normal flora in nasopharynx of variety of wild and domestic animals. Numerous virulence factors have been described for P. multocida isolates which include adherence and colonization factors, iron-regulated and acquisition proteins, extracellular enzymes such as neuraminidase, lipopolysaccharide (LPS), capsule and a variety of outer membrane proteins (Omp). OmpA has a significant role in stabilizing the cell envelope structure by providing physical linkage between the outer membrane & peptidoglycan. It has been shown to mediate P. multocida -host cells interaction via heparin and/or fibronectin binding and therefore act as an important invasive molecule which could determine the final outcome of initial infection. Comparative nucleotide sequence analysis of ompA gene of P. multocida has revealed that despite extensive genetic diversity in ompA of P. multocida, most sequences could be classified into two major allele classes namely ompA allele (I) and allele (II). The P. multocida recovered from nasal cavity of bovine and belonging to two ompA classes were tested for their differential virulence. In vitro pathogenicity studies on Madin Darby Bovine Kidney (MDBK) cell line employing adhesion and invasion assays indicated that P. multocida strain with ompA (I) is more invasive than P. multocida strain with ompA (II). In vivo studies in mice further reiterated that the isolates harbouring ompA(I) were comparatively more virulent to isolates harbouring ompA (II).

  15. Crystallization and preliminary crystallographic studies of the Pasteurella multocida toxin catalytic domain

    Energy Technology Data Exchange (ETDEWEB)

    Miyazawa, Masayuki [Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita-shi, Osaka 565-0871 (Japan); Kitadokoro, Kengo [Research Center for Low Temperature and Materials Sciences, Kyoto University, Sakyo-ku, Kyoto 606-8502 (Japan); Kamitani, Shigeki; Shime, Hiroaki; Horiguchi, Yasuhiko, E-mail: horiguti@biken.osaka-u.ac.jp [Research Institute for Microbial Diseases, Osaka University, 3-1 Yamada-oka, Suita-shi, Osaka 565-0871 (Japan)

    2006-09-01

    The C-terminal catalytic domain of P. multocida toxin, which is the virulence factor of the organism in P. multocida, has been expressed, purified and subsequently crystallized using the sitting-drop vapour-diffusion technique. The C-terminal catalytic domain of Pasteurella multocida toxin, which is the virulence factor of the organism in P. multocida, has been expressed, purified and subsequently crystallized using the sitting-drop vapour-diffusion technique. Native diffraction data to 1.9 Å resolution were obtained at the BL44XU beamline of SPring-8 from a flash-frozen crystal at 100 K. The crystals belong to space group C2, with unit-cell parameters a = 111.0, b = 150.4, c = 77.1 Å, β = 105.5°, and are likely to contain one C-PMT (726 residues) per asymmetric unit.

  16. Prevalence, characterization and antibiotic resistance of Pasteurella multocida isolated from bovine respiratory infection.

    Science.gov (United States)

    Jamali, Hossein; Rezagholipour, Mojtaba; Fallah, Sepideh; Dadrasnia, Arezoo; Chelliah, Shamini; Velappan, Rita Devi; Wei, Kelvin Swee Chuan; Ismail, Salmah

    2014-11-01

    The objectives of this study were to determine the prevalence, characterization and antibiotic resistance of Pasteurella multocida isolated from calves with respiratory infection in Iran. P. multocida was detected in 141/169 bovine respiratory infection cases on Iranian dairy and beef farms. P. multocida were grouped into serogroups A (126/141), D (12/141), and B (3/141). Of the P.  multocida isolates, all harboured the psl, ompH, oma87, fimA, ptfA, nanB, and nanH genes, 139/141 had hsf-2, and 115/141 pfhA, and tadD. The isolates were most frequently resistant to penicillin G (43/141 resistant isolates; 30.5%) and streptomycin (31/141; 22%). Copyright © 2014. Published by Elsevier Ltd.

  17. The ability of lipopolysaccharide (LPS) of Pasteurella multocida B:2 to induce clinical and pathological lesions in the nervous system of buffalo calves following experimental inoculation.

    Science.gov (United States)

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

    2017-03-01

    Lipopolysaccharide (LPS) of P. multocida B:2, a causative agent of haemorrhagic septicaemia (HS) in cattle and buffaloes, is considered as the main virulence factor and contribute in the pathogenesis of the disease. Recent studies provided evidences about the involvement of the nervous system in pathogenesis of HS. However, the role of P. multocida B:2 immunogens, especially the LPS is still uncovered. Therefore, this study was designed to investigate the role of P. multocida B:2 LPS to induce pathological changes in the nervous system. Nine eight-month-old, clinically healthy buffalo calves were used and distributed into three groups. Calves of Group 1 and 2 were inoculated orally and intravenously with 10 ml of LPS broth extract represent 1 × 10(12) cfu/ml of P. multocida B:2, respectively, while calves of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. Significant differences were found in the mean scores for clinical signs, post mortem and histopathological changes especially in Group 2, which mainly affect different anatomic regions of the nervous system, mainly the brain. On the other hand, lower scores have been recorded for clinical signs, gross and histopathological changes in Group 1. These results provide for the first time strong evidence about the ability of P. multocida B:2 LPS to cross the blood brain barrier and induce pathological changes in the nervous system of the affected buffalo calves. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Evidence of circulation of an epidemic strain of Pasteurella multocida in Jiangsu, China by multi-locus sequence typing (MLST).

    Science.gov (United States)

    Wang, Yanhong; Zhu, Jie; Lu, Chengping; Wu, Bai; Liu, Dandan; Hang, Wei; Liu, Huimou; Liu, Xiufan

    2013-12-01

    Pasteurella multocida, the causative agent of fowl cholera, is a serious threat to poultry farming. In this study, we isolated and identified 40 P. multocida strains in fowl cholera outbreaks in Jiangsu province, China. The identified P. multocida was further characterized using multi-locus sequence typing (MLST). All of the 40 P. multocida strains studied are genetically identical and belong to the ST129 sequence type based on seven MLST loci. Our study provides evidence of a circulating epidemic strain of P. multocida in Jiangsu, China. Copyright © 2013. Published by Elsevier B.V.

  19. Non-zoonotic Pasteurella multocida infection as a cause of septic shock in a patient with liver cirrhosis: A case report and review of the literature

    Directory of Open Access Journals (Sweden)

    Jose Orsini

    2013-01-01

    Full Text Available Pasteurella multocida is a Gram-negative organism characterized morphologically as coccobacillus. It is the commonest organism infecting pet bites. However, severe infections may occur in the absence of animal bites or scratches. Pasteurella multocida serves as an opportunistic pathogen in humans, especially in patients with depressed immune system. Few cases in the literature identify Pasteurella multocida as the causative agent of septic shock, especially in cirrhotic patients. We report a patient who presented with septic shock as a result of Pasteurella multocida bacteremia, without prior history of animal exposure.

  20. Interaction between Pasteurella multocida B:2 and its derivatives with bovine aortic endothelial cell (BAEC).

    Science.gov (United States)

    Kamal, Nuriqmaliza M; Zamri-Saad, M; Masarudin, Mas Jaffri; Othman, Sarah

    2017-06-19

    Pasteurella multocida B:2 causes bovine haemorrhagic septicaemia (HS), leading to rapid fatalities in cattle and buffaloes. An attenuated derivative of P. multocida B:2 GDH7, was previously constructed through mutation of the gdhA gene and proved to be an effective live attenuated vaccine for HS. Currently, only two potential live attenuated vaccine candidates for HS are being reported; P. multocida B:2 GDH7 and P. multocida B:2 JRMT12. This study primarily aims to investigate the potential of P. multocida B:2 GDH7 strain as a delivery vehicle for DNA vaccine for future multivalent applications. An investigation on the adherence, invasion and intracellular survival of bacterial strains within the bovine aortic endothelial cell line (BAEC) were carried out. The potential vaccine strain, P. multocida B:2 GDH7, was significantly better (p ≤ 0.05) at adhering to and invading BAEC compared to its parent strain and to P. multocida B:2 JRMT12 and survived intracellularly 7 h post treatment, with a steady decline over time. A dual reporter plasmid, pSRGM, which enabled tracking of bacterial movement from the extracellular environment into the intracellular compartment of the mammalian cells, was subsequently transformed into P. multocida B:2 GDH7. Intracellular trafficking of the vaccine strain, P. multocida B:2 GDH7 was subsequently visualized by tracking the reporter proteins via confocal laser scanning microscopy (CLSM). The ability of P. multocida B:2 GDH7 to model bactofection represents a possibility for this vaccine strain to be used as a delivery vehicle for DNA vaccine for future multivalent protection in cattle and buffaloes.

  1. Molecular characterization of the capsular antigens of Pasteurella multocida isolates using multiplex PCR

    Directory of Open Access Journals (Sweden)

    Khalid S. Al-Maary

    2017-02-01

    Full Text Available The use of molecular techniques for detection and characterization of the Pasteurella multocida is very important for rapid and specific detection and characterization of the organism. During the period from 15th February, 2014 to 15th April, 2015, 425 nasopharyngeal swabs and 175 lung and spleen samples were collected and examined by conventional methods, 80 strains (18.82% of P. multocida were isolated from the calves, sheep and goat with respiratory manifestation. Meanwhile, 77 strains (44% were isolated from emergency slaughtered animals. All the recovered strains were positive for specific PCR for detection of P. multocida strains previously identified as P. multocida by standard microbiological techniques. Multiplex PCR for molecular typing of the capsular antigens of the recovered P. multocida revealed positive amplification of 1044 bp fragments specific to the capsular antigen type A with 105 strains (66.88%, and amplification 511 bp fragments of the capsular antigen type E with 52 strain (33.12% and absence of B, D and F antigens. Multiplex PCR for molecular typing of the capsular antigens of P. multocida can be used as a simple, sensitive, rapid, reliable technique instead of the serological techniques for identification of the capsular antigens of P. multocida

  2. Septicaemic pasteurellosis in free-range pigs associated with an unusual biovar 13 of Pasteurella multocida.

    Science.gov (United States)

    Cardoso-Toset, Fernando; Gómez-Laguna, Jaime; Callejo, María; Vela, Ana Isabel; Carrasco, Librado; Fernández-Garayzábal, Jose Francisco; Maldonado, Alfonso; Luque, Inmaculada

    2013-12-27

    Biochemical profiles, PFGE typing and MLST analysis were used to investigate an outbreak of septicaemic pasteurellosis in a free-range pig farm in Spain. Signs of coughing, dyspnoea and a visible inflammation of the ventral area of the neck (jowl), which acquired a cyanotic and necrotic appearance, were the characteristic findings in affected animals, associated with a high morbidity (70%) and case mortality (95%). Diffuse, haemorrhagic and fibrinous pleuroneumonia and acute, focally extensive and haemorrhagic myositis and panniculitis were observed in the histopathological analysis from three analyzed animals. Pasteurella multocida subsp. multocida, capsular type B, biovar 13 was isolated in pure culture from lung, submandibular tissue (jowl), liver, spleen and kidney tissue from diseased pigs. After PFGE typing, all P. multocida isolates displayed undistinguishable macrorestriction patterns with Bsp120I restriction enzyme demonstrating that the infection was caused by a single strain. With the multihost P. multocida MLST database, all P. multocida isolates were assigned to the new sequence type ST47 which was highly related with other bovine isolates of P. multocida type B associated with haemorrhagic septicaemia. This is the first description of an outbreak of septicaemic pasteurellosis in free-range pigs associated with P. multocida type B of the unusual biovar 13. The communication and complete diagnosis of cases of swine septicaemia and the possible role of pigs as reservoirs of this new pathogen must be evaluated to determine the importance of this disease for pigs. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Genomic characterization of Pasteurella multocida HB01, a serotype A bovine isolate from China.

    Science.gov (United States)

    Peng, Zhong; Liang, Wan; Liu, Wenjing; Wu, Bin; Tang, Biao; Tan, Chen; Zhou, Rui; Chen, Huanchun

    2016-04-25

    Pasteurella multocida infects various domestic and feral animals, generally causing clinical disease. To investigate P. multocida disease in cattle, we sequenced the complete genome of P. multocida HB01 (GenBank accession CP006976), a serotype A organism isolated from a cow in China. The genome is composed of a single circular chromosome of 2,416,068 base pairs containing 2212 protein-coding sequences, 6 ribosomal rRNA operons, and 56 tRNA genes. The present study confirms that P. multocida HB01 possesses a more complete metabolic pathway with an intact trichloroacetic acid cycle for anabolism compared with A. pleuropneumoniae and Haemophilus parasuis. This is the first time that this metabolic mechanism of P. multocida has been described. We also identified a full spectrum of genes related to known virulence factors of P. multocida. The differences in virulence factors between strains of different serotypes and origins were also compared. This comprehensive comparative genome analysis will help in further studies of the metabolic pathways, genetic basis of serotype, and virulence of P. multocida. Copyright © 2016 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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

  5. Pasteurella multocida Toxin Interaction with Host Cells: Entry and Cellular Effects

    Science.gov (United States)

    Ho, Mengfei

    2015-01-01

    The mitogenic dermonecrotic toxin from Pasteurella multocida (PMT) is a 1285-residue multipartite protein that belongs to the A-B family of bacterial protein toxins. Through its G-protein-deamidating activity on the α subunits of heterotrimeric Gq-, Gi- and G12/13-proteins, PMT potently stimulates downstream mitogenic, calcium, and cytoskeletal signaling pathways. These activities lead to pleiotropic effects in different cell types, which ultimately result in cellular proliferation, while inhibiting cellular differentiation, and account for the myriad of physiological outcomes observed during infection with toxinogenic strains of P. multocida. PMID:22552700

  6. Crystallization and preliminary crystallographic studies of the Pasteurella multocida toxin catalytic domain.

    Science.gov (United States)

    Miyazawa, Masayuki; Kitadokoro, Kengo; Kamitani, Shigeki; Shime, Hiroaki; Horiguchi, Yasuhiko

    2006-09-01

    The C-terminal catalytic domain of Pasteurella multocida toxin, which is the virulence factor of the organism in P. multocida, has been expressed, purified and subsequently crystallized using the sitting-drop vapour-diffusion technique. Native diffraction data to 1.9 A resolution were obtained at the BL44XU beamline of SPring-8 from a flash-frozen crystal at 100 K. The crystals belong to space group C2, with unit-cell parameters a = 111.0, b = 150.4, c = 77.1 A, beta = 105.5 degrees, and are likely to contain one C-PMT (726 residues) per asymmetric unit.

  7. Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine

    OpenAIRE

    Furian, Thales Quedi; Borges,Karen Apellanis; Laviniki,Vanessa; da Silveira Rocha, Silvio Luis; Almeida,Camila Neves de; do Nascimento, Vladimir Pinheiro; Salle, Carlos Tadeu Pippi; de Souza Moraes, Hamilton Luiz

    2016-01-01

    Abstract Pasteurella multocida causes atrophic rhinitis in swine and fowl cholera in birds, and is a secondary agent in respiratory syndromes. Pathogenesis and virulence factors involved are still poorly understood. The aim of this study was to detect 22 virulence-associated genes by PCR, including capsular serogroups A, B and D genes and to evaluate the antimicrobial susceptibility of P. multocida strains from poultry and swine. ompH, oma87, plpB, psl, exbD-tonB, fur, hgbA, nanB, sodA, sodC,...

  8. Invasive Pasteurella multocida Infections - Report of Five Cases at a Minnesota Hospital, 2014.

    Science.gov (United States)

    Talley, P; Snippes-Vagnone, P; Smith, K

    2016-09-01

    During October 2014, the Minnesota Department of Health was notified of five Hospital A patients with Pasteurella multocida bacteraemia; three had died. Human soft tissue infection with P. multocida typically results from cat or dog bites or scratches. Invasive infection, defined as a P. multocida isolate from a usually sterile site, is rare. We evaluated P. multocida isolations at Hospital A, compared with other Minnesota hospitals to understand invasive infection trends. A case was defined as clinically confirmed P. multocida in a Minnesota resident during 2012-2014. All hospital laboratories were queried; Fisher's exact test was used for comparison. Medical charts were reviewed for 2014 Hospital A patients with P. multocida infections. The Minnesota clinical laboratories survey response rate was 79% (63/80). At Hospital A, proportion of P. multocida isolates from usually sterile sites increased from 0% (0/2) during 2012 to 11% (1/9) during 2013, and to 86% (5/6) during 2014. The proportion of patients with P. multocida isolated from sterile sites was 35% (6/17) at Hospital A compared with 10% (58/583) statewide during 2012-2014 combined (P pet exposure. Collaborative educational efforts of chronically ill pet owners by physicians and veterinarians can acknowledge the health benefits of pet ownership, while minimizing risk for serious invasive zoonotic infections, including those caused by P. multocida. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  9. Characterization of sucrose-negative Pasteurella multocida variants, including isolates from large-cat bite wounds

    DEFF Research Database (Denmark)

    Christensen, Henrik Grimmig; Bisgaard, Magne; Angen, Øystein

    2005-01-01

    To validate the identification of Pasteurella multocida-like bacteria negative for acid formation from sucrose, including isolates from bite wounds caused by large cats, 17 strains were phenotypically and genotypically characterized. Phylogenetic analysis of partially sequenced rpoB and infB genes...... and the reference strains of P. multocida. Two strains isolated from leopard bite wounds were related to the type strain of P. dagmatis; however, they represented a new taxon (taxon 46 of Bisgaard), in accordance with their distinct phenotypic and genotypic identifications. The present study documents that sucrose...... care should be taken in the identification of Pasteurella-like bacteria isolated from bite wounds caused by large cats. The evidence of phenotypic and genotypic divergence calls for the further development of PCR tests and DNA sequencing to document doubtful isolates....

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

    OpenAIRE

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

    2014-01-01

    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 tren...

  11. Multiplex PCR To Identify Macrolide Resistance Determinants in Mannheimia haemolytica and Pasteurella multocida

    OpenAIRE

    Rose, Simon; Desmolaize, Benoit; Jaju, Puneet; Wilhelm, Cornelia; Warrass, Ralf; Douthwaite, Stephen

    2012-01-01

    The bacterial pathogens Mannheimia haemolytica and Pasteurella multocida are major etiological agents in respiratory tract infections of cattle. Although these infections can generally be successfully treated with veterinary macrolide antibiotics, a few recent isolates have shown resistance to these drugs. Macrolide resistance in members of the family Pasteurellaceae is conferred by combinations of at least three genes: erm(42), which encodes a monomethyltransferase and confers a type I MLSB ...

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

    Science.gov (United States)

    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). Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Hyperimmune serum from rabbits immunized with potassium thiocyanate extract of Pasteurella multocida protects against homologous challenge.

    OpenAIRE

    Lu, Y S; Pakes, S P; Massey, L

    1987-01-01

    Hyperimmune rabbit sera directed to the KSCN extract of 3:A Pasteurella multocida were characterized by enzyme-linked immunosorbent assay (ELISA), presolubilized cell radioimmunoprecipitation, and immunoblotting analysis. The results showed that the hyperimmune serum had a very high titer of immunoglobulin G ELISA antibody and a negligible immunoglobulin A ELISA antibody, precipitated 10 different outer membrane protein antigens by radioimmunoprecipitation, and reacted to 10 different membran...

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

    Directory of Open Access Journals (Sweden)

    Changfu eCao

    2015-07-01

    Full Text Available 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 Pasteurella multocida(type B, serotype 2, then intramuscularly received a range of marbofloxacin doses 24h after inoculation. The ratio of 24h area under the concentration-time curve divided by the minimum inhibitory concentration or the mutant prevention concentration (AUC24h/MIC or AUC24h/MPC was the pharmacokinetic-pharmacodynamic (PK/PD index that best described the effectiveness of marbofloxacin against Pasteurella multocida (R2=0.8514 by nonlinear regression analysis. Marbofloxacin exhibited a good antimicrobial activity in vivo. The levels of AUC24h/MIC and AUC24h/MPC that produced 50% (1.5log10CFU/mL reduction and 90% (3log10CFU/mL reduction of maximum response were 18.60h and 50.65h, 4.67h and 12.89h 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.

  15. Effects of Subminimum Inhibitory Concentrations of Antibiotics on the Pasteurella multocida Proteome: A Systems Approach

    Directory of Open Access Journals (Sweden)

    Bindu Nanduri

    2008-01-01

    Full Text Available To identify key regulators of subminimum inhibitory concentration (sub-MIC antibiotic response in the Pasteurella multocida proteome, we applied systems approaches. Using 2D-LC-ESI-MS2, we achieved 53% proteome coverage. To study the differential protein expression in response to sub-MIC antibiotics in the context of protein interaction networks, we inferred P. multocida Pm70 protein interaction network from orthologous proteins. We then overlaid the differential protein expression data onto the P. multocida protein interaction network to study the bacterial response. We identified proteins that could enhance antimicrobial activity. Overall compensatory response to antibiotics was characterized by altered expression of proteins involved in purine metabolism, stress response, and cell envelope permeability.

  16. Use of Molecular Pathogenicity Indices to Identify Pathogenic Strains of Pasteurella multocida.

    Science.gov (United States)

    Furian, Thales Quedi; Borges, Karen Apellanis; Pilatti, Roberta Marmitt; de Almeida, Camila Neves; Streck, André Felipe; de Emery, Brunna Dias; Nascimento, Vladimir Pinheiro do; Salle, Carlos Tadeu Pippi; de Souza Moraes, Hamilton Luiz

    2016-12-01

    In addition to being the causative agent of fowl cholera (FC), Pasteurella multocida is also one of the most prevalent opportunistic pathogens associated with respiratory diseases in various hosts. However, understanding of the traits that distinguish the virulent isolates that cause FC is still limited. The objective of this study was to characterize P. multocida isolates of Brazil by PCR-restriction fragment length polymorphism (PCR-RFLP) analysis in order to determine if strain-type correlates with virulence or with 22 previously studied virulence genes. The PCR-RFLP was used to classify the isolates into seven strain types, and the isolates in Profile II had a higher pathogenicity index (P multocida .

  17. Cell culture assay for toxigenic Pasteurella multocida from atrophic rhinitis of pigs.

    Science.gov (United States)

    Rutter, J M; Luther, P D

    1984-04-21

    A toxin produced by strains of Pasteurella multocida isolated from pigs with atrophic rhinitis caused a cytopathic effect in cell cultures derived from embryonic bovine lung. The toxin was produced during the late logarithmic phase of bacterial growth and inactivated by heating for 30 minutes at 56 degrees C. The cell culture assay was reproducible and 10(3) to 10(4) times more sensitive than a lethal assay in BALB/c mice. There was complete agreement between results in the two tests with 76 isolates of P multocida. Neutralising activity was demonstrated in both assays with sera from infected gnotobiotic piglets. It was concluded that embryonic bovine lung cell cultures provided a sensitive in vitro test for the differentiation of toxigenic from non toxigenic isolates of P multocida. The assay could be used in diagnostic laboratories and for characterisation of the toxin.

  18. Identificación, biotipificación y caracterización de cepas de Pasteurella multocida aisladas en la Argentina Identification, biotypification and characterization of Pasteurella multocida strains isolated in Argentina

    Directory of Open Access Journals (Sweden)

    G. A. Leotta

    2006-09-01

    Full Text Available Treinta cepas de Pasteurella multocida aisladas en la Argentina a partir de muestras de origen humano y animal fueron identificadas, biotipificadas y caracterizadas. Veintidós de ellas (73% correspondieron a P. multocida subsp. multocida; cinco (17% a P. multocida subsp. gallicida y tres (10% a P. multocida subsp. septica. Todas las cepas fueron agrupadas en 8 biotipos; el 70% presentó el tipo capsular A. Los serotipos somáticos más frecuentes fueron el 1 (n:11 y el 3 (n:9. Las cepas de origen porcino fueron resistentes a tiamulina, estreptomicina y tetraciclina. La caracterización de las cepas de P. multocida aisladas en la Argentina es el primer paso para concretar futuros estudios destinados a la prevención y al tratamiento de la pasteurelosis en medicina humana y veterinaria.Thirty Pasteurella multocida strains isolated in Argentina from human and animal samples were identified, biotypified and characterized. Twenty-two (73% strains were identified as P. multocida subsp. multocida, 5 (17% as P. multocida subsp. gallicida, and 3 (10% as P. multocida subsp. septica. All strains were grouped in 8 biotypes, and 70% of the strains presented capsular type A. The most frequent somatic serotypes were 1 (n:11 and 3 (n:9. P. multocida strains from swine source were resistant to tiamulin, streptomycin and tetracycline. Characterization of P. multocida strains isolated in Argentina is the first step to conduct future studies intended for the prevention and treatment of pasteurellosis in human and veterinary medicine.

  19. Molecular study on Pasteurella multocida and Mannheimia granulomatis from Kenyan Camels (Camelus dromedarius).

    Science.gov (United States)

    Gluecks, Ilona V; Bethe, Astrid; Younan, Mario; Ewers, Christa

    2017-08-22

    Outbreaks of a Haemorrhagic Septicaemia (HS) like disease causing large mortalities in camels (Camelus dromedarius) in Asia and in Africa have been reported since 1890. Yet the aetiology of this condition remains elusive. This study is the first to apply state of the art molecular methods to shed light on the nasopharyngeal carrier state of Pasteurellaceae in camels. The study focused on HS causing Pasteurella multocida capsular types B and E. Other Pasteurellaceae, implicated in common respiratory infections of animals, were also investigated. In 2007 and 2008, 388 nasopharyngeal swabs were collected at 12 locations in North Kenya from 246 clinically healthy camels in 81 herds that had been affected by HS-like disease. Swabs were used to cultivate bacteria on blood agar and to extract DNA for subsequent PCR analysis targeting P. multocida and Mannheimia-specific gene sequences. Forty-five samples were positive for P. multocida genes kmt and psl and for the P. multocida Haemorrhagic Septicaemia (HS) specific sequences KTSP61/KTT72 but lacked HS-associated capsular type B and E genes capB and capE. This indicates circulation of HS strains in camels that lack established capsular types. Sequence analysis of the partial 16S rRNA gene identified 17 nasal swab isolates as 99% identical with Mannheimia granulomatis, demonstrating a hitherto unrecognised active carrier state for M. granulomatis or a closely related Mannheimia sp. in camels. The findings of this study provide evidence for the presence of acapsular P. multocida or of hitherto unknown capsular types of P. multocida in camels, closely related to P. multocida strains causing HS in bovines. Further isolations and molecular studies of camelid P. multocida from healthy carriers and from HS-like disease in camels are necessary to provide conclusive answers. This paper is the first report on the isolation of M. granulomatis or a closely related new Mannheimia species from camelids.

  20. The transcriptional response of Pasteurella multocida to three classes of antibiotics.

    Science.gov (United States)

    Nanduri, Bindu; Shack, Leslie A; Burgess, Shane C; Lawrence, Mark L

    2009-07-14

    Pasteurella multocida is a gram-negative bacterial pathogen that has a broad host range. One of the diseases it causes is fowl cholera in poultry. The availability of the genome sequence of avian P. multocida isolate Pm70 enables the application of functional genomics for observing global gene expression in response to a given stimulus. We studied the effects of three classes of antibiotics on the P. multocida transcriptome using custom oligonucleotide microarrays from NimbleGen Systems. Hybridizations were conducted with RNA isolated from three independent cultures of Pm70 grown in the presence or absence of sub-minimum inhibitory concentration (sub-MIC) of antibiotics. Differentially expressed (DE) genes were identified by ANOVA and Dunnett's test. Biological modeling of the differentially expressed genes (DE) was conducted based on Clusters of Orthologous (COG) groups and network analysis in Pathway Studio. The three antibiotics used in this study, amoxicillin, chlortetracycline, and enrofloxacin, collectively influenced transcription of 25% of the P. multocida Pm70 genome. Some DE genes identified were common to more than one antibiotic. The overall transcription signatures of the three antibiotics differed at the COG level of the analysis. Network analysis identified differences in the SOS response of P. multocida in response to the antibiotics. This is the first report of the transcriptional response of an avian strain of P. multocida to sub-lethal concentrations of three different classes of antibiotics. We identified common adaptive responses of P. multocida to antibiotic stress. The observed changes in gene expression of known and putative P. multocida virulence factors establish the molecular basis for the therapeutic efficacy of sub-MICs. Our network analysis demonstrates the feasibility and limitations of applying systems modeling to high throughput datasets in 'non-model' bacteria.

  1. Multilocus sequence analysis of Pasteurella multocida demonstrates a type species under development.

    Science.gov (United States)

    Bisgaard, Magne; Petersen, Andreas; Christensen, Henrik

    2013-03-01

    The aim of the present study was to use multilocus sequence typing (MLST) of a diverse collection of Pasteurella multocida with regard to animal source, place and date of collection, including all available serovars of Carter, Heddleston, Little & Lyon, Namioka, Cornelius and Roberts, to further investigate the evolution of this species with a focus on two lineages, A (P. multocida subsp. multocida and P. multocida subsp. gallicida) and B (P. multocida subsp. septica), previously reported. Isolates of P. multocida (n = 116) including reference strains of major serotyping systems were investigated by MLST based on partial sequences of the genes adk, est, gdh, mdh, pgi, pmi and zwf, and 67 sequence types (STs) were observed. Phylogenetic analysis of these concatenated sequences confirmed the separation of groups A (41 STs, 71 isolates) and B (22 STs, 38 isolates) out of the 67 STs. All Carter serovars, 12 Heddleston serovars, all three Little-Lyon types, six out of seven Namioka serovars, all five Roberts types and all four Cornelius serovars were allocated to the A group, while group B included the remaining four Heddleston serovars, 6, 7, 8 and 13, in addition to Namioka type 8 : A. The overrepresentation of reference strains of serotyping systems in the A group contrasts with the high number of isolates obtained from diseased birds in the B group, the effect of which should be addressed in future vaccine development. Isolates from birds (25) dominated the B group, which also included four isolates from Felidae, whereas group A included isolates from all types of hosts. The evolutionary implications of the lack of capsular type D, pig and bovine isolates in group B, as well as its association with Aves and Felidae that also applied to the whole Rural Industries Research and Development Corporation (RIRDC) MLST database, need further investigation. The combination of rpoB and 16S rRNA gene sequence comparison as well as the developed PCR test assigned

  2. Characterization of an outer membrane protein of Pasteurella multocida belonging to the OmpA family.

    Science.gov (United States)

    Marandi, M; Mittal, K R

    1996-12-01

    The outer membrane vesicle and N-lauroylsarcosine-insoluble protein preparations of Pasteurella multocida 656 were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. A major outer membrane protein (OMP) was found to be heat-modifiable, having a molecular mass of 28 kDa when the OMP preparation was solubilized at 60 degrees C and a molecular mass of 37 kDa when it was solubilized at 100 degrees C. A monoclonal antibody, designated mAb MT4.1, was generated against heat-modifiable OMP of P. multocida. This mAb reacted with the heat-modifiable OMP irrespective of the temperature at which it was solubilized, as demonstrated by immunoblot results. The heat-modifiable OMP of P. multocida showed a significant N-terminal amino acid sequence homology with OmpA family. Immunoelectron microscopic study revealed that the mAb Mt4.1 epitope was not surface exposed on the intact bacterium. The mAb MT4.1 reacted with all the reference strains of 5 capsular and 16 somatic serotypes, as well as with 75 field strains of P. multocida in immunoblot assay. This mAb MT4.1 also reacted with strains of various other Pasteurella species such as P. stomatis, P. aerogenes P. gallinarum, P. betti, P. sp, B, P. SP-g and P. canis, but not with strains of 12 other Gram-negative bacteria. These results indicated that this protein carried a genus-specific epitope and mAb MT4.1 may be useful for identification of Pasteurella species. This is the first report in which a major heat-modifiable OMP has been identified and characterized using a mAb, and has been shown belonging to the OmpA family.

  3. Plasmid DNA Analysis of Pasteurella multocida Serotype B isolated from Haemorrhagic Septicaemia outbreaks in Malaysia

    Directory of Open Access Journals (Sweden)

    Jamal, H.

    2005-01-01

    Full Text Available A total of 150 purified isolates of Pasteurella multocida serotype B were used (Salmah, 2004 for plasmid DNA curing experiment to determine hyaluronidase activity, antibiotic resistance pattern (ARP and mice lethality test (LD50 for their role of pathogenicity. A plasmid curing experiment was carried out by using the intercalating agent; ethidium bromide and rifampicin, where it was found all the plasmids had been cured (plasmidless from Pasteurella multocida. All of these plasmidless isolates maintained their phenotypic characteristics. They showed the same antibiotic resistancepattern as before curing, produced hyaluronidase and possessed lethality activity in mice when injected intraperitoneally(i.p. Based on this observation, the antibiotic resistance, hyaluronidase activity and mice virulence could probably be chromosomal-mediated. Plasmids were detected 100% in all P. multocida isolates with identical profile of 2 plasmids size 3.0 and 5.5 kb. No large plasmids could be detected in all isolates. Since all the isolates appeared to have identicalplasmid profiles, they were subjected to restriction enzyme(RE analysis. From RE analysis results obtained, it can be concluded that the plasmid DNA in serotype B isolates are identical. Only 4 of 32 REs were found to cleave these plasmids with identical restriction fingerprints; BglII, HaeIII, RsaI and SspI. From RE analysis results, it can be concluded that the plasmid DNA isolates are identical. This plasmid might not played any role in pathogenicity of Pasteurella multocida serotype B, however this information is important for the construction of shuttle vectors in genetic studies of the pathogenicity of haemorrhagic septicaemia(HS.

  4. Ultrastructural Changes and Bacterial Localization in Buffalo Calves Following Oral Exposure to Pasteurella multocida B:2

    Directory of Open Access Journals (Sweden)

    M.S. Abubakar, M. Zamri-Saad* and S. Jasni

    2013-01-01

    Full Text Available This study describes the ultrastructural changes and distribution of P. multocida B:2 in the respiratory and gastrointestinal tracts of buffalo calves following oral exposure and compared with intratracheal exposure. Nine 8-month old buffalo calves were divided into 3 groups. At the start of the experiment, dexamethasone (1mg/kg was injected intramuscularly into buffaloes of all groups for three consecutive days. Then, calves of Group 1 were exposed orally to 50ml of the inoculum containing 109 colony-forming unit (CFU/mL of live P. multocida B:2. Calves of Group 2 were exposed intra-tracheal to the same inoculum while calves of Group 3 were exposed orally to PBS. Transient and mild clinical signs were observed in calves of Groups 1 and 2. Pasteurella multocida B:2 was isolated from the nasal mucosa for up to 5 days post-infection. Only calf that was killed at 48 h post-infection had P. multocida B:2 in both respiratory and gastro-intestinal tracts showing ultrastructural changes typical of acute cellular injury, with degeneration of endothelium and vascular walls. There were deciliation in the respiratory and microvilli degeneration in the gastrointestinal tracts. The study concludes that oral exposure may not play major role in the development of hemorrhagic septicemia. Nevertheless, the buffalo calf that succumbed to hemorrhagic septicemia carried P. multocida B:2 in the gastro-intestinal organs.

  5. Regeneration of toxigenic Pasteurella multocida induced severe turbinate atrophy in pigs detected by computed tomography.

    Science.gov (United States)

    Magyar, Tibor; Donkó, Tamás; Repa, Imre; Kovács, Melinda

    2013-10-30

    Atrophic rhinitis is a widely prevalent infectious disease of swine caused by Bordetella bronchiseptica and Pasteurella multocida. The course of the disease is considered to be different depending on the principal aetiological agents distinguishing B. bronchiseptica induced non-progressive and toxigenic P. multocida produced progressive forms. In order to compare the pathological events of the two forms of the disease, the development of nasal lesions has longitudinally been studied in pigs infected by either B. bronchiseptica alone or B. bronchiseptica and toxigenic P. multocida together using computed tomography to visualise the nasal structures. B. bronchiseptica infection alone caused moderately severe nasal turbinate atrophy and these lesions completely regenerated by the time of slaughter. Unexpectedly, complete regeneration of the bony structures of the nasal cavity was also observed in pigs infected by B. bronchiseptica and toxigenic P. multocida together in spite of seeing severe turbinate atrophy in most of the infected animals around the age of six weeks. B. bronchiseptica mono-infection has been confirmed to cause only mild to moderate and transient lesions, at least in high health status pigs. Even severe turbinate atrophy induced by B. bronchiseptica and toxigenic P. multocida combined infection is able to be reorganised to their normal anatomical structure. Computed tomography has further been verified to be a useful tool to examine the pathological events of atrophic rhinitis in a longitudinal manner.

  6. Phenotypic characterization and random amplified polymorphic DNA (RAPD) analysis of Pasteurella multocida isolated from Korean pigs.

    Science.gov (United States)

    Lee, Ki-Eun; Jeoung, Hye-Young; Lee, Ji-Youn; Lee, Myoung-Heon; Choi, Hwan-Won; Chang, Kyung-Soo; Oh, Young-Hee; An, Dong-Jun

    2012-05-01

    Pasteurella multocida causes various respiratory disease symptoms in pigs, including atrophic rhinitis and pneumonia. In the present study, 69 strains of P. multocida were isolated from 443 pigs with respiratory clinical symptoms at 182 farms located throughout South Korea from 2009 to 2010. A multiplex capsular PCR typing assay revealed that 69 strains of P. multocida isolated in this study had the biosynthetic locus of the capsules of either serogroup A (47 strains, 68.1%) or serogroup D (22 strains, 31.9%). The 22 strains positive for serogroup D-specific primers were divided into four clusters and the 47 strains positive for serogroup A-specific primers were divided into 12 clusters according to the results of Random Amplified Polymorphic DNA (RAPD) analysis. P. multocida strains in the present study were susceptible to most of the antimicrobial agents used. An analysis of antimicrobial resistance and virulence gene pattern combined with RAPD indicated that a certain P. multocida strain appeared to be genetically identical, implying the persistence of the strain within a single farm.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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...

  9. Genome-Wide Analyses Reveal Genes Subject to Positive Selection in Pasteurella multocida

    Science.gov (United States)

    Cao, Peili; Guo, Dongchun; Liu, Jiasen; Jiang, Qian; Xu, Zhuofei; Qu, Liandong

    2017-01-01

    Pasteurella multocida, a Gram-negative opportunistic pathogen, has led to a broad range of diseases in mammals and birds, including fowl cholera in poultry, pneumonia and atrophic rhinitis in swine and rabbit, hemorrhagic septicemia in cattle, and bite infections in humans. In order to better interpret the genetic diversity and adaptation evolution of this pathogen, seven genomes of P. multocida strains isolated from fowls, rabbit and pigs were determined by using high-throughput sequencing approach. Together with publicly available P. multocida genomes, evolutionary features were systematically analyzed in this study. Clustering of 70,565 protein-coding genes showed that the pangenome of 33 P. multocida strains was composed of 1,602 core genes, 1,364 dispensable genes, and 1,070 strain-specific genes. Of these, we identified a full spectrum of genes related to virulence factors and revealed genetic diversity of these potential virulence markers across P. multocida strains, e.g., bcbAB, fcbC, lipA, bexDCA, ctrCD, lgtA, lgtC, lic2A involved in biogenesis of surface polysaccharides, hsf encoding autotransporter adhesin, and fhaB encoding filamentous haemagglutinin. Furthermore, based on genome-wide positive selection scanning, a total of 35 genes were subject to strong selection pressure. Extensive analyses of protein subcellular location indicated that membrane-associated genes were highly abundant among all positively selected genes. The detected amino acid sites undergoing adaptive selection were preferably located in extracellular space, perhaps associated with bacterial evasion of host immune responses. Our findings shed more light on conservation and distribution of virulence-associated genes across P. multocida strains. Meanwhile, this study provides a genetic context for future researches on the mechanism of adaptive evolution in P. multocida. PMID:28611758

  10. Isolation, Antimicrobial Resistance, and Virulence Genes of Pasteurella multocida Strains from Swine in China▿

    Science.gov (United States)

    Tang, Xibiao; Zhao, Zhanqin; Hu, Junyong; Wu, Bin; Cai, Xuwang; He, Qigai; Chen, Huanchun

    2009-01-01

    A total of 233 isolates of Pasteurella multocida were obtained from 2,912 cases of clinical respiratory disease in pigs in China, giving an isolation rate of 8.0%. Serogroup A P. multocida isolates were isolated from 92 cases (39.5%), and serogroup D isolates were isolated from 128 cases (54.9%); 12 isolates (5.2%) were untypeable. P. multocida was the fourth most frequent pathogenic bacterium recovered from the respiratory tract, after Streptococcus suis, Haemophilus parasuis, and Escherichia coli. All isolates were characterized for their susceptibilities to 20 antibiotics and the presence of 19 genes for virulence factors (VFs). The frequency of antimicrobial resistance among P. multocida isolates from swine in China was higher than that reported among P. multocida isolates from swine in from other countries, and 93.1% of the isolates showed multiple-drug resistance. There was a progressive increase in the rate of multiresistance to more than seven antibiotics, from 16.2% in 2003 to 62.8% in 2007. The resistance profiles suggested that cephalosporins, florfenicol, and fluoroquinolones were the drugs most likely to be active against P. multocida. Use of PCR showed that colonization factors (ptfA, fimA, and hsf-2), iron acquisition factors, sialidases (nanH), and outer membrane proteins occurred in most porcine strains. The VFs pfhA, tadD, toxA, and pmHAS were each present in multocida isolates and suggest that the potential threat of such multiresistant bacteria in food-producing animals should not be neglected. PMID:19158260

  11. Genome-Wide Analyses Reveal Genes Subject to Positive Selection in Pasteurella multocida

    Directory of Open Access Journals (Sweden)

    Peili Cao

    2017-05-01

    Full Text Available Pasteurella multocida, a Gram-negative opportunistic pathogen, has led to a broad range of diseases in mammals and birds, including fowl cholera in poultry, pneumonia and atrophic rhinitis in swine and rabbit, hemorrhagic septicemia in cattle, and bite infections in humans. In order to better interpret the genetic diversity and adaptation evolution of this pathogen, seven genomes of P. multocida strains isolated from fowls, rabbit and pigs were determined by using high-throughput sequencing approach. Together with publicly available P. multocida genomes, evolutionary features were systematically analyzed in this study. Clustering of 70,565 protein-coding genes showed that the pangenome of 33 P. multocida strains was composed of 1,602 core genes, 1,364 dispensable genes, and 1,070 strain-specific genes. Of these, we identified a full spectrum of genes related to virulence factors and revealed genetic diversity of these potential virulence markers across P. multocida strains, e.g., bcbAB, fcbC, lipA, bexDCA, ctrCD, lgtA, lgtC, lic2A involved in biogenesis of surface polysaccharides, hsf encoding autotransporter adhesin, and fhaB encoding filamentous haemagglutinin. Furthermore, based on genome-wide positive selection scanning, a total of 35 genes were subject to strong selection pressure. Extensive analyses of protein subcellular location indicated that membrane-associated genes were highly abundant among all positively selected genes. The detected amino acid sites undergoing adaptive selection were preferably located in extracellular space, perhaps associated with bacterial evasion of host immune responses. Our findings shed more light on conservation and distribution of virulence-associated genes across P. multocida strains. Meanwhile, this study provides a genetic context for future researches on the mechanism of adaptive evolution in P. multocida.

  12. Isolation, antimicrobial resistance, and virulence genes of Pasteurella multocida strains from swine in China.

    Science.gov (United States)

    Tang, Xibiao; Zhao, Zhanqin; Hu, Junyong; Wu, Bin; Cai, Xuwang; He, Qigai; Chen, Huanchun

    2009-04-01

    A total of 233 isolates of Pasteurella multocida were obtained from 2,912 cases of clinical respiratory disease in pigs in China, giving an isolation rate of 8.0%. Serogroup A P. multocida isolates were isolated from 92 cases (39.5%), and serogroup D isolates were isolated from 128 cases (54.9%); 12 isolates (5.2%) were untypeable. P. multocida was the fourth most frequent pathogenic bacterium recovered from the respiratory tract, after Streptococcus suis, Haemophilus parasuis, and Escherichia coli. All isolates were characterized for their susceptibilities to 20 antibiotics and the presence of 19 genes for virulence factors (VFs). The frequency of antimicrobial resistance among P. multocida isolates from swine in China was higher than that reported among P. multocida isolates from swine in from other countries, and 93.1% of the isolates showed multiple-drug resistance. There was a progressive increase in the rate of multiresistance to more than seven antibiotics, from 16.2% in 2003 to 62.8% in 2007. The resistance profiles suggested that cephalosporins, florfenicol, and fluoroquinolones were the drugs most likely to be active against P. multocida. Use of PCR showed that colonization factors (ptfA, fimA, and hsf-2), iron acquisition factors, sialidases (nanH), and outer membrane proteins occurred in most porcine strains. The VFs pfhA, tadD, toxA, and pmHAS were each present in multocida isolates and suggest that the potential threat of such multiresistant bacteria in food-producing animals should not be neglected.

  13. Genome-Wide Analyses Reveal Genes Subject to Positive Selection in Pasteurella multocida.

    Science.gov (United States)

    Cao, Peili; Guo, Dongchun; Liu, Jiasen; Jiang, Qian; Xu, Zhuofei; Qu, Liandong

    2017-01-01

    Pasteurella multocida, a Gram-negative opportunistic pathogen, has led to a broad range of diseases in mammals and birds, including fowl cholera in poultry, pneumonia and atrophic rhinitis in swine and rabbit, hemorrhagic septicemia in cattle, and bite infections in humans. In order to better interpret the genetic diversity and adaptation evolution of this pathogen, seven genomes of P. multocida strains isolated from fowls, rabbit and pigs were determined by using high-throughput sequencing approach. Together with publicly available P. multocida genomes, evolutionary features were systematically analyzed in this study. Clustering of 70,565 protein-coding genes showed that the pangenome of 33 P. multocida strains was composed of 1,602 core genes, 1,364 dispensable genes, and 1,070 strain-specific genes. Of these, we identified a full spectrum of genes related to virulence factors and revealed genetic diversity of these potential virulence markers across P. multocida strains, e.g., bcbAB, fcbC, lipA, bexDCA, ctrCD, lgtA, lgtC, lic2A involved in biogenesis of surface polysaccharides, hsf encoding autotransporter adhesin, and fhaB encoding filamentous haemagglutinin. Furthermore, based on genome-wide positive selection scanning, a total of 35 genes were subject to strong selection pressure. Extensive analyses of protein subcellular location indicated that membrane-associated genes were highly abundant among all positively selected genes. The detected amino acid sites undergoing adaptive selection were preferably located in extracellular space, perhaps associated with bacterial evasion of host immune responses. Our findings shed more light on conservation and distribution of virulence-associated genes across P. multocida strains. Meanwhile, this study provides a genetic context for future researches on the mechanism of adaptive evolution in P. multocida.

  14. Isolation, characterization, antibiogram and pathology of Pasteurella multocida isolated from pigs

    Directory of Open Access Journals (Sweden)

    Mamta Tigga

    2014-05-01

    Full Text Available Aim: Isolation, characterization and antibiogram of Pasteurella multocida from diseased pigs of district Durg of Chhattisgarh, and to study pathological changes caused by swine pasteurellosis. Materials and Methods: An outbreak of swine pasteurellosis was suspected in pigs of Ruwabandha (Bhilai, Anjora, Somni, Tedesara, Tirgajhola villages of Durg district in Chhattisgarh, India during August and September of 2011. Nasal Swabs and blood samples from ailing pigs and heart blood and impression smears from morbid pigs were processed for detection and isolation of P. multocida by bacteriological methods. Detailed necropsy was conducted and gross and histopathological lesions were recorded. The test Isolates were subjected to antimicrobial sensitivity profile by disc-diffusion method. Results: The blood smears from heart blood and tissue impression smears revealed teaming of bipolar organisms indicating the presence of Pasteurella spp. The isolates obtained were subjected to Gram's staining for checking the purity and bipolar morphology and characterized biochemically. Gross lesions included severe acute pneumonia and haemorrhages in lungs, petechial haemorrhages on serous membranes and other visceral organs. On histopathological examination, lungs showed typical fibrinous bronchopneumonia, multifocal suppuration. All the isolates of P. multocida were 100% sensitive to Amoxicillin, Gentamicin, Enrofloxacin and showed100% resistance to Ceftizoxim and Cloxacillin. Conclusion: Gross and microscopic lesions in dead animals are of great diagnostic value and are of characteristic of P. multocida infection. Cultural, morphological and biochemical characters are useful to rule out the causative agent as P. multocida. Antibiotic sensitivity pattern of the isolates should routinely be carried out for knowing the antibiotic resistance trends in an endemic area.

  15. The pathogenesis of Pasteurella multocida local isolates in mice and chicken

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    Supar

    2000-03-01

    Full Text Available Avian cholera or fowl cholera is a bacterial disease caused by Pasteurella multocida strain of serogroup A, has been recognized as important disease in domestic poultry such as ducks and chicken. P. multocida strains derived from overseas and local isolates are stored as freeze dried and kept at the Research hlstitute for Veterinary Science (BALITVET culture collection (BCC. Some of those bacteria are still alive and can be used as vaccine candidates. Each strain or isolate was activated in brain heart infusion broth containing foetal calf serum and incubated at 37°C then it was identitied by biochemical reactions. Field surveys Were conducted in Central Java and South Kalimantan to observe fowl cholera problems and sample collections for isolation of pathogens. Of the 14 of Pasteurella multocida strains or isolates from BCC, II strains (9 imported 2 local isolates were still alive. In addition to this 2 isolates trom chicken and duck were viable. Seven out of 9 imported strains killed mice within 3 x 24 hours, similarly for the local isolates (BCC 299, 2331, DYI, DY2, 12TG, 15TG. However, the only BCC 2331 and DY2 isolates were able to kill two week old chicken witIlin 6 x 24 hours post inoculation. From this experiment indicated that the P. multocida local isolates (BCC 2,331 and DY2 are more pathogenic than that of imported strains. Two strains of imported P. multocida BCC 2331, 1362 and 6 local isolates (BCC 299, 2331, DYI, DY2, 12TG and 15TG would be selected for mono- and polyvalent vaccine candidates in the following experiments and the highly patogenic BCC 2331 and DY2 isolates would be used to challenge the vaccinated animals.

  16. Involvement of nervous system in cattle and buffaloes due to Pasteurella multocida B:2 infection: A review of clinicopathological and pathophysiological changes

    Directory of Open Access Journals (Sweden)

    Ali Dhiaa Marza

    2015-09-01

    Full Text Available Hemorrhagic septicemia (HS is an acute septicemic disease principally affecting cattle and buffaloes caused by specific serotypes B:2 and E:2 of Pasteurella multocida in Asia and Africa, respectively. Despite continuing researches on pathogenesis of P. multocida for several decades, the mechanisms by which these bacteria develop the diseases are poorly understood. Although the involvement of the nervous system in the disease progress of HS is rare under natural conditions, few reports indicated the involvement of the nervous system in outbreaks of HS in cattle and buffaloes. Additionally, recent pathogenesis studies in both mouse and buffalo experimental models reported the involvement of nervous system due to P. multocida B:2, with bacteriological and histopathological evidences. In this review, we summarized and discussed the updates on the involvement of the nervous system in pathogenesis of HS focusing on clinical signs, pathological and pathophysiological changes. [J Adv Vet Anim Res 2015; 2(3.000: 252-262

  17. Molecular analysis of Pasteurella multocida strains isolated from fowl cholera infection in backyard chickens.

    Science.gov (United States)

    Mohamed, Mohamed-Wael Abdelazeem; Mageed, Moemen Abdel Azeem Mohamed Abdel

    2014-01-01

    To characterize Pasteurella isolated from backyard chickens using whole cell protein lysate profiles and random amplified polymorphic DNA (RAPD) techniques to show their genetic relationship because Pasteurella multocida (P. multocida) is an important cause of fatal infections in backyard chickens. Twenty one P. multocida isolates were recovered previously from clinical cases of fowl cholera belonging to individual owners and phenotypically analyzed using biochemical tests and serotyping were used for the genetic characterization. Phylogenetic study based on both methods revealed that the recovered population of P. multocida isolated from backyard chickens differs markedly, constituting a well-separated cluster and appearance of 3 distinguishing lineages with greater discrimination shown by RAPD-PCR that resulted in two suclusters in cluster A and three subclusters in cluster B and were related greatly with capsular serogroups for the examined strains. The whole cell protein revealed the presence of dominant protein bands at approximately 41 and 61 kDa in all of the examined isolates that may be a virulent proteins share in the increasing of its pathogenicity. Clear distinctive bands ranged from 123 to 1554 bp. Based on the previous findings, there are three spreading clusters that may indicate the association of a small number of P. multocida variants with the majority of cases suggesting that certain clones of P. multocida are able to colonize the examined backyard chickens. Also, the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PAGE system for strain differentiation and epidemiological studies of avian P. multocida. Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P. multocida genotypes, to identify affiliations

  18. Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

    DEFF Research Database (Denmark)

    Christensen, Henrik; Angen, Øystein; Olsen, John E.

    2004-01-01

    , xylose and mannitol, 18 strains of Pasteurella canis biovar 2 and variants of this taxon, five strains of P multocida subsp. septica showing variations in indole and ornithine decarboxylase, nine strains of P. multocida subsp. multocida showing variation in ornithine decarboxylase and mannitol, and type...... and that the existence of the biovar 2 variants of P. avium and P. canis is highly questionable. It is concluded that the redefined P. multocida is genotypically homogeneous, although phenotypically diverse lineages exist with respect to ornithine decarboxylase, indole and mannitol, characters that have been regarded...

  19. Host response in rabbits to infection with Pasteurella multocida serogroup F strains originating from fowl cholera

    Science.gov (United States)

    Jaglic, Zoran; Jeklova, Edita; Christensen, Henrik; Leva, Lenka; Register, Karen; Kummer, Vladimir; Kucerova, Zdenka; Faldyna, Martin; Maskova, Jarmila; Nedbalcova, Katerina

    2011-01-01

    Although Pasteurella multocida serogroup F has been described as an avian-adapted serogroup, it was recently found in rabbit nests in the Czech Republic. Therefore, the ability of 2 avian P. multocida serogroup F strains to induce disease in rabbits was investigated. Two groups of 18 Pasteurella-free rabbits were intranasally challenged with strains isolated from chickens and turkeys. Half of the animals in each challenge group were immunosuppressed using dexamethasone. All of the challenged rabbits exhibited clinical signs of peracute septicemic disease, ending with shock, and died or were euthanized in the terminal stages of the disease 1 to 2 d post-infection. Gross pathological changes included systemic vascular collapse and vascular leak syndrome. Hyperemia, hemorrhage, edema, inflammatory cell infiltrates, focal necrosis, and degenerative changes were observed histologically in parenchymatous organs. This is the first study directly demonstrating that avian P. multocida serogroup F strains are highly virulent in rabbits and that avian hosts cannot be excluded as a possible source of rabbit infection with serogroup F. PMID:22210996

  20. Host response in rabbits to infection with Pasteurella multocida serogroup F strains originating from fowl cholera.

    Science.gov (United States)

    Jaglic, Zoran; Jeklova, Edita; Christensen, Henrik; Leva, Lenka; Register, Karen; Kummer, Vladimir; Kucerova, Zdenka; Faldyna, Martin; Maskova, Jarmila; Nedbalcova, Katerina

    2011-07-01

    Although Pasteurella multocida serogroup F has been described as an avian-adapted serogroup, it was recently found in rabbit nests in the Czech Republic. Therefore, the ability of 2 avian P. multocida serogroup F strains to induce disease in rabbits was investigated. Two groups of 18 Pasteurella-free rabbits were intranasally challenged with strains isolated from chickens and turkeys. Half of the animals in each challenge group were immunosuppressed using dexamethasone. All of the challenged rabbits exhibited clinical signs of peracute septicemic disease, ending with shock, and died or were euthanized in the terminal stages of the disease 1 to 2 d post-infection. Gross pathological changes included systemic vascular collapse and vascular leak syndrome. Hyperemia, hemorrhage, edema, inflammatory cell infiltrates, focal necrosis, and degenerative changes were observed histologically in parenchymatous organs. This is the first study directly demonstrating that avian P. multocida serogroup F strains are highly virulent in rabbits and that avian hosts cannot be excluded as a possible source of rabbit infection with serogroup F.

  1. Characterization of Pasteurella multocida involved in rabbit infections

    DEFF Research Database (Denmark)

    Massacci, Francesca Romana; Magistrali, Chiara Francesca; Cucco, Lucilla

    2018-01-01

    In rabbit, P. multocida is considered a predominant pathogenic agent; despite this, few data on the molecular epidemiology are available so far. The aim of this work was to characterize P. multocida isolates from rabbit affected by various diseases in Italy. Comparison was made to reference strains...... belonged to the LPS genotypes 3 (22/39) or 6 (17/39). The clonal relationships of the Italian strains from rabbit had similarity to previously reported rabbit isolates that belonged to ST9, ST74, ST204 and ST206, however, they differed from other rabbit references strains that belonged to six other STs....... In particular, ST9 with capsular type F has been previously reported from diseased rabbit in Czech Republic and ST74 has been observed for older rabbit isolates. ST50 has probably been reported from Spain. ST9 and ST50 have previously also been reported from birds and pig, respectively, whereas ST74 has...

  2. Signature-Tagged Mutagenesis of Pasteurella multocida Identifies Mutants Displaying Differential Virulence Characteristics in Mice and Chickens

    OpenAIRE

    Harper, Marina; Boyce, John D.; Wilkie, Ian W.; Adler, Ben

    2003-01-01

    Pasteurella multocida is the causative agent of fowl cholera in birds. Signature-tagged mutagenesis (STM) was used to identify potential virulence factors in a mouse septicemia disease model and a chicken fowl cholera model. A library of P. multocida mutants was constructed with a modified Tn916 and screened for attenuation in both animal models. Mutants identified by the STM screening were confirmed as attenuated by competitive growth assays in both chickens and mice. Of the 15 mutants ident...

  3. Endophytic fungi from tropical ethnoveterinary plants and their antibacterial efficacy against Pasteurella multocida Capsular Type A strain

    OpenAIRE

    Nath, Archana; Joshi, Santaram

    2016-01-01

    Pasteurella multocida is an important veterinary pathogen causing infections in animals and birds. Nowadays, different reports have described the severity of infections, increasing resistance of micro-organisms to antibiotics, and the contribution of ethnoveterinary practices towards the treatment of various ailments of animals. The aim of the present study was to investigate the antibacterial efficacy of the ethanolic extracts of endophytic fungi against P. multocida Capsular Type A strains....

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

    Science.gov (United States)

    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. © The Author(s) 2016.

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

    Science.gov (United States)

    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. Poultry Science Association Inc.

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

    Science.gov (United States)

    Rocke, T.E.; Smith, S.R.; Miyamoto, A.; Shadduck, D.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.

  7. Capsular Polysaccharide Interferes with Biofilm Formation by Pasteurella multocida Serogroup A.

    Science.gov (United States)

    Petruzzi, Briana; Briggs, Robert E; Swords, W Edward; De Castro, Cristina; Molinaro, Antonio; Inzana, Thomas J

    2017-11-21

    Pasteurella multocida is an important multihost animal and zoonotic pathogen that is capable of causing respiratory and multisystemic diseases, bacteremia, and bite wound infections. The glycosaminoglycan capsule of P. multocida is an essential virulence factor that protects the bacterium from host defenses. However, chronic infections (such as swine atrophic rhinitis and the carrier state in birds and other animals) may be associated with biofilm formation, which has not been characterized in P. multocida Biofilm formation by clinical isolates was inversely related to capsule production and was confirmed with capsule-deficient mutants of highly encapsulated strains. Capsule-deficient mutants formed biofilms with a larger biomass that was thicker and smoother than the biofilm of encapsulated strains. Passage of a highly encapsulated, poor-biofilm-forming strain under conditions that favored biofilm formation resulted in the production of less capsular polysaccharide and a more robust biofilm, as did addition of hyaluronidase to the growth medium of all of the strains tested. The matrix material of the biofilm was composed predominately of a glycogen exopolysaccharide (EPS), as determined by gas chromatography-mass spectrometry, nuclear magnetic resonance, and enzymatic digestion. However, a putative glycogen synthesis locus was not differentially regulated when the bacteria were grown as a biofilm or planktonically, as determined by quantitative reverse transcriptase PCR. Therefore, the negatively charged capsule may interfere with biofilm formation by blocking adherence to a surface or by preventing the EPS matrix from encasing large numbers of bacterial cells. This is the first detailed description of biofilm formation and a glycogen EPS by P. multocidaIMPORTANCEPasteurella multocida is an important pathogen responsible for severe infections in food animals, domestic and wild birds, pet animals, and humans. P. multocida was first isolated by Louis Pasteur in

  8. Demonstration of bacteriocin activity in bovine and bison strains of Pasteurella multocida. [UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Chengappa, M.M.; Carter, G.R.

    1977-08-01

    Of 33 strains of Pasteurella multocida examined, 14 showed bacteriocin activity and 17 were susceptible to bacteriocin. The activity was increased by about twofold if the cultures were induced with ultraviolet radiation; however, no increase in bacteriocin activity was observed if the potential producer strains were induced with mitomycin C. The bacteriocin activity of potential producer strains was increased if CaCl/sub 2/ was incorporated in the medium. The patterns of bacteriocin susceptibility indicate that these substances may ultimately contribute to a typing scheme for the species. An extra-chromosomal genetic element was not detected when a potential producer strain was not detected when a potential producer strain was tested by the dye-buoyant density gradient method. This fact suggests that the genetic material responsible for bacteriocin activity in P multocida is located on the host chromosome proper.

  9. Meningencefalite por Pasteurella multocida: estudo clínico-laboratorial de um caso em lactente

    Directory of Open Access Journals (Sweden)

    C. E. Levy

    1989-12-01

    Full Text Available Os autores apresentam descrição clínico-laboratorial e evolutiva do caso de lactente com o diagnóstico de meningencefalite por Pasteurella multocida que apresentou na evolução atraso neuromotor, manifestações epilépticas, surdez neurossensorial e paresia crural à esquerda. Fazem também breve revisão do papel deste agente etiológico na patologia humana. Ressaltam a importância da P. multocida em casos de meningites bacterianas, fazendo-se o diagnóstico laboratorial diferencial com o Haemophilus influenzae e Neisseria meningitidis em processos infecciosos conseqüentes a arranhadura ou mordida de animais e nas bacteremias ou septicemias em pacientes com hepatopatias crônicas ou em estados de imunodepressão.

  10. Draft Genome Sequence of a Virulent Strain of Pasteurella Multocida Isolated From Alpaca.

    Science.gov (United States)

    Hurtado, Raquel Enma; Aburjaile, Flavia; Mariano, Diego; Canário, Marcus Vinicius; Benevides, Leandro; Fernandez, Daniel Antonio; Allasi, Nataly Olivia; Rimac, Rocio; Juscamayta, Julio Eduardo; Maximiliano, Jorge Enrique; Rosadio, Raul Hector; Azevedo, Vasco; Maturrano, Lenin

    2017-01-01

    Pasteurella multocida is one of the most frequently isolated bacteria in acute pneumonia cases, being responsible for high mortality rates in Peruvian young alpacas, with consequent social and economic costs. Here we report the genome sequence of P. multocida strain UNMSM, isolated from the lung of an alpaca diagnosed with pneumonia, in Peru. The genome consists of 2,439,814 base pairs assembled into 82 contigs and 2,252 protein encoding genes, revealing the presence of known virulence-associated genes (ompH, ompA, tonB, tbpA, nanA, nanB, nanH, sodA, sodC, plpB and toxA). Further analysis could provide insights about bacterial pathogenesis and control strategies of this disease in Peruvian alpacas.

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

    Science.gov (United States)

    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.

  12. Pasteurella multocida virulence factors: selection of fowl cholera-inducing and non-inducing strains.

    Science.gov (United States)

    Rhoades, K R; Rimler, R B

    1993-01-01

    Relatively little information is available on Pasteurella multocida virulence factors involved in producing fowl cholera. Because of the complex nature of bacterial pathogenesis, the recommended approach for ascertaining these factors is to compare biological attributes of high- and low-virulence strains. To permit use of this approach for fowl cholera, P. multocida strains of high and low virulence were identified. Turkey poults were exposed intrapharyngeally and intravenously (IV) to two antigenically and biochemically similar strains. Based on mortality, strain P-1059 was highly virulent and strain P-1062 was avirulent. Microbiological examination indicated that only the virulent strain infected the pharyngeal mucosa of intrapharyngeally exposed poults and survived and multiplied in IV-exposed poults. These findings indicate strain differences in those virulence factors concerned with the colonization and multiplication stages of disease development.

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

    Directory of Open Access Journals (Sweden)

    Thais Sebastiana Porfida Ferreira

    2012-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    Samuel, M.D.; Shadduck, D.J.; Goldberg, Diana 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%).

  15. Detection and genetic characterization of Pasteurella multocida from alpaca (Vicugna pacos) pneumonia cases.

    Science.gov (United States)

    Rímac, Rocío; Luna, Luis; Hurtado, Raquel; Rosadio, Raúl; Maturrano, Lenin

    2017-08-01

    Pasteurella multocida is a common constituent of upper respiratory tract microbiota but is frequently isolated of alpaca lung tissues from pulmonary infections. Despite its importance, very little is known about this bacteria at molecular level. In order to characterize P. multocida isolates, 24 isolates recovered from 46 mortal acute cases in young alpacas with suspected pneumonia were analyzed, using biochemical and molecular tests for capsule and LPS typing, virulence factors detection, and ERIC-PCR genetic diversity analysis. All the P. multocida isolates belonged to the capsular type A, LPS genotype L6 (related to serotypes 10, 11, 12, and 15), and possessed virulence factors gene toxA and tbpA. ERIC-PCR analysis revealed two electrophoretic profiles, and the majority of isolates (23/24) shared the same fingerprint, indicating strong evidence that there was a common source of infection for all the affect animals. This study revealed the detection of P. multocida type A, LPS genotype L6, and toxA+ and tbpA+ from dead young alpacas with pneumonia in Peru.

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

    Science.gov (United States)

    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.

  17. Activity of florfenicol for Actinobacillus pleuropneumoniae and Pasteurella multocida using standardised versus non-standardised methodology.

    Science.gov (United States)

    Dorey, L; Hobson, S; Lees, P

    2016-12-01

    Four indices of antimicrobial potency were determined for florfenicol and the pig pneumonia pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), mutant prevention concentration (MPC) and time-kill curves were determined in two matrices, broth and pig serum. Five overlapping sets of two-fold dilutions were used to increase accuracy of the measurements. MIC and MBC serum:broth ratios for A. pleuropneumoniae were 0.96:1 and 1.07:1, respectively, and corresponding values for P. multocida were 0.72:1 and 0.50:1. The percentage binding of florfenicol to serum protein was 65.4%, and fraction unbound (fu) serum MICs were significantly lower, by 2.71-fold and 3.82-fold, respectively, than predicted for free serum concentrations for A. pleuropneumoniae and P. multocida. Similar culture medium differences were obtained for MBC and MPC. MICs in serum and broth were increased significantly and progressively for high, medium and low initial inoculum counts. Serum MPC:MIC ratios for A. pleuropneumoniae and P. multocida were 12.5:1 and 13.6:1, respectively; ratios for broth were similar. The killing action of florfenicol had the characteristics of concentration dependency for both species in both growth media. These data indicate the value of using a biological medium, when determining microbiological potency indices, to predict dosage for clinical use. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Pheno- and genotypic characterization of Pasteurella multocida isolated from cats, dogs and rabbits from Brazil.

    Science.gov (United States)

    Ferreira, Thais Sebastiana Porfida; Moreno, Luisa Zanolli; Felizardo, Maria Roberta; de Gobbi, Débora Dirani Sena; Filsner, Pedro Henrique de Lima Nogueira; de Moura Gomes, Vasco Tulio; Moreno, Marina; Moreno, Andrea Micke

    2016-04-01

    Pasteurella multocida is the causative agent of many diseases of economic importance in veterinary medicine and is characterized by high zoonotic potential. Pet animals can be infected and play a major role as carriers. This study aimed to characterize the genetic diversity of P. multocida isolated from dogs, cats and rabbits, and to evaluate their antimicrobial susceptibility profiles. A total of 620 animals were studied; 51 were positive for P. multocida and 92 strains were isolated. 60.9% of the strains belonged to the capsular type A, while the remaining were classified as non-typeable. The hgbA, ptfA, sodC, tadD and hsf2 genes were more frequent among the rabbit strains. Sulfonamides and cotrimoxazole presented the highest resistance rate, followed by erythromycin. PFGE clustered strains according to host species. Our results indicate that P. multocida from companion animals carry several virulence factors and are resistant to antimicrobials commonly used in human and veterinary medicine. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Potency of marbofloxacin for pig pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida: Comparison of growth media.

    Science.gov (United States)

    Dorey, L; Hobson, S; Lees, P

    2017-04-01

    Pharmacodynamic properties of marbofloxacin were established for six isolates each of the pig respiratory tract pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida. Three in vitro indices of potency were determined; Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and Mutant Prevention Concentration (MPC). For MIC determination Clinical Laboratory Standards Institute guidelines were modified in three respects: (1) comparison was made between two growth media, an artificial broth and pig serum; (2) a high inoculum count was used to simulate heavy clinical bacteriological loads; and (3) five overlapping sets of two-fold dilutions were used to improve accuracy of determinations. Similar methods were used for MBC and MPC estimations. MIC and MPC serum:broth ratios for A. pleuropneumoniae were 0.79:1 and 0.99:1, respectively, and corresponding values for P. multocida were 1.12:1 and 1.32:1. Serum protein binding of marbofloxacin was 49%, so that fraction unbound (fu) serum MIC values were significantly lower than those predicted by correction for protein binding; fu serum:broth MIC ratios were 0.40:1 (A. pleuropneumoniae) and 0.50:1 (P. multocida). For broth, MPC:MIC ratios were 13.7:1 (A. pleuropneumoniae) and 14.2:1 (P. multocida). Corresponding ratios for serum were similar, 17.2:1 and 18.8:1, respectively. It is suggested that, for dose prediction purposes, serum data might be preferable to potency indices measured in broths. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Identification of Pasteurella multocida virulence genes in a septicemic mouse model using signature-tagged mutagenesis.

    Science.gov (United States)

    Fuller, T E; Kennedy, M J; Lowery, D E

    2000-07-01

    P. multocida is the causative agent of several economically significant veterinary diseases occurring in numerous species worldwide. Signature-tagged mutagenesis (STM) is a powerful genetic technique used to simultaneously screen multiple transposon mutants of a pathogen for their inability to survive in vivo. We have designed an STM system based on a mini-Tn10 transposon, chemiluminescent detection and semi-quantitative analysis and have identified transposon insertions into genes of Pasteurella multocida that attenuate virulence in a septicemic mouse model. A bank of 96 transposons containing strongly-hybridizing tags was used to create 19 pools of P. multocida transposon mutants containing approximately 70-90 mutants/pool. A total of 62 mutants were attenuated when checked individually, and 25 unique single transposon insertion mutations were identified from this group. The sequence of the disrupted ORF for each attenuated mutant was determined by either cloning or PCR-amplifying and sequencing the flanking regions. The attenuated mutants contained transposon insertions in genes encoding biosynthetic enzymes, virulence factors, regulatory components and unknown functions. This study should contribute to an understanding of the pathogenic mechanisms by which P. multocida and other pathogens in the Pasteurellaceae family cause disease and identify novel live vaccine candidates and new potential antibiotic targets. Copyright 2000 Academic Press.

  1. A novel Erm monomethyltransferase in antibiotic-resistant isolates of Mannheimia haemolytica and Pasteurella multocida

    DEFF Research Database (Denmark)

    Desmolaize, Benoit; Rose, Simon; Warrass, Ralf

    2011-01-01

    Mannheimia haemolytica and Pasteurella multocida are aetiological agents commonly associated with respiratory tract infections in cattle. Recent isolates of these pathogens have been shown to be resistant to macrolides and other ribosome-targeting antibiotics. Direct analysis of the 23S r......RNAs by mass spectrometry revealed that nucleotide A2058 is monomethylated, consistent with a Type I erm phenotype conferring macrolide-lincosamide resistance. The erm resistance determinant was identified by full genome sequencing of isolates. The sequence of this resistance determinant, now termed erm(42...

  2. Alternative treatment of serious and mild Pasteurella multocida infection in New Zealand White rabbits.

    Science.gov (United States)

    Palócz, Orsolya; Gál, János; Clayton, Paul; Dinya, Zoltán; Somogyi, Zoltán; Juhász, Csaba; Csikó, György

    2014-11-25

    Pasteurella multocida causes numerous economically relevant diseases in livestock including rabbits. Immunisation is only variably effective. Prophylactic antibiotics are used in some species but are contra-indicated in rabbits, due to their adverse effects on the rabbit microbiota. There is therefore a substantial need for alternative forms of infection control in rabbits; we investigated the effect of oral β-glucan on P. multocida infection in this species. Thirthy-five New Zealand White rabbits were randomly divided into five groups of seven animals. Three groups were inoculated with Pasteurella multocida intranasally (in.), a physiologically appropriate challenge which reproduces naturally acquired infection, and received either (1-3), (1-6) β-glucans or placebo. Four other groups were inoculated both in. and intramuscularly (im.), representing a supra-physiological challenge, and received either (1-3), (1-6) β-glucans, antibiotic or placebo. β-glucans given prophylactically were highly effective in protecting against physiological (in.) bacterial challenge. They were less effective in protecting against supra-physiological bacterial challenge (in. and im.), although they extended survival times. This latter finding has practical relevance to breeders as it extends the window in which heavily infected and symptomatic animals can be salvaged with antibiotics. In our study, (1-3), (1-6) β-glucans were highly effective in protecting against a model of naturally acquired P. multocida infection and extended survival times in the supra-physiological model. Enrofloxacin was effective in protecting against supra-physiological infection. We are currently reviewing the use of combined prophylaxis.

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

    Science.gov (United States)

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

    2016-01-01

    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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. Prosthetic joint infection caused by Pasteurella multocida: a case series and review of literature.

    Science.gov (United States)

    Honnorat, Estelle; Seng, Piseth; Savini, Hélène; Pinelli, Pierre-Olivier; Simon, Fabrice; Stein, Andreas

    2016-08-20

    Pasteurella multocida is a well-recognized zoonotic agent following dog or cat bites or scratches. Nevertheless, prosthetic joint infection caused by P. multocida are rarely reported. We report here a series of six cases of prosthetic joint infection caused by P. multocida managed at a referral centre for the treatment of bone and joint infection in southern France. We also reviewed the 26 cases reported in literature. The mean age of our cases was 74 years [±8.2, range 63-85]. In majority of our cases (5 cases) were associated with knee prostheses and one case with a hip prosthesis. Most of cases occurred after cat or dog scratches or licks or contact. Diagnoses of prosthetic joint infection caused by P. multocida were made by positive cultures of surgical biopsies or needle aspiration. Mean time delay between prosthetic joint implantation and infection onset was 7.6 years (±5.12 years, range 2-17). Local inflammation, which occurred in all six cases, was the most frequent clinical symptom, followed by pain in five cases, fever and swollen joints in four cases, and a fistula with purulent discharge inside the wound in two cases. The mean time of antibiotic therapy was 8 months. Surgical treatment with prosthesis removal was performed in three cases. Six of our cases were in remission without apparent relapse at 3 years after end of treatment. Prosthetic joint infections caused by P. multocida usually occur after animal scratches or bites, but can occasionally occur after a short animal lick. These infections are usually resulting from a contiguous infection and localized in the knee. An early antibiotic therapy after surgical debridement could avoid prosthetic withdrawal, notably in elderly patients. Patients with prosthetic joints should be warned that animals are potential sources of serious infection and urgent medical advice should be sought if they are bitten or scratched.

  5. A compendium of antibiotic-induced transcription profiles reveals broad regulation of Pasteurella multocida virulence genes.

    Science.gov (United States)

    Melnikow, E; Schoenfeld, C; Spehr, V; Warrass, R; Gunkel, N; Duszenko, M; Selzer, P M; Ullrich, H J

    2008-10-15

    The transcriptional responses of Pasteurella multocida to eight antibiotics with known mode of actions (MoAs) and one novel antibiotic compound with an unknown MoA were collected to create a compendium of transcriptional profiles for MoA studies. At minimal inhibitory concentration the three bactericidal compounds enrofloxacin, cefquinome and the novel compound had a minor impact on gene regulation with approximately 1% of the P. multocida genome affected, whilst the bacteriostatic compounds florfenicol, tilmicosin, rifampin, trimethoprim and brodimoprim regulated 20% of the genome. Novobiocin was special in that it regulated 40% of all P. multocida genes. Regulation of target genes was observed for novobiocin, rifampin, florfenicol and tilmicosin and signature genes were identified for most antibiotics. The transcriptional profile induced by the novel compound was unrelated to the compendium profiles suggesting a new MoA. The transcription of many P. multocida virulence factors, particularly genes involved in capsule synthesis and export, LPS synthesis, competence, adherence and iron transport were altered in the presence of antibiotics. Virulence gene transcription was mainly negatively affected, however the opposite effect was also observed in the case of rifampin where the up-regulation of the tad locus involved in tight adherence was seen. Novobiocin and trimethoprim caused a marked reduction in the transcription of capsule genes, which correlated with a concomitant reduction of the capsular layer on the surface of P. multocida. The broad negative impact on virulence gene transcription supports the notion that the therapeutic effect of some antibiotics could be a combination of growth and virulence inhibition.

  6. Genital form of pasteurellosis in breeding turkeys infected during artificial insemination and isolation of an unusual strain of Pasteurella multocida.

    Science.gov (United States)

    Cariou, Nadine; Christensen, Henrik; Salandre, Olivier; Albaric, Olivier; Bisgaard, Magne; Malher, Xavier

    2013-09-01

    A genital and potentially fatal form of Pasteurella multocida infection was reported on two turkey-breeding farms on which birds were vaccinated against Pasteurella multocida. Both outbreaks were linked to the use of semen from young vaccinated toms with a history of respiratory pasteurellosis followed by treatment during rearing. Typing by agar gel immunodiffusion and rapid slide agglutination of P. multocida isolated from cloacal swabs was completed by multilocus sequence typing. Restriction enzyme analysis showed that that the isolates were clonal. They belonged to sequence type (ST) 30, described in chickens, cats, and ducks. This strain differed in sequence type from the ones used in the vaccine (ST8, ST60, ST53, and ST235), which might have limited its effectiveness. No contamination of the semen (n = 30) was found, suggesting fecal contamination during semen collection.

  7. Identification and characterization of a protective antigen, PlpB of bovine Pasteurella multocida strain LZ-PM.

    Science.gov (United States)

    Wei, Xiaoqin; Wang, Yugang; Luo, Runbo; Qian, Wei; Sizhu, Suolang; Zhou, Hongbo

    2017-06-01

    The Pasteurella multocida lipoprotein B (PlpB) was cloned from Pasteurella multocida (P. multocida) strain LZ-PM (serotype A) and expressed in Escherichia coli (E. coli). Sequence analysis showed that PlpB from different strains of P. multocida exhibited 80.8-99.4% sequence identity to each other, suggesting that PlpB might serve as a cross-protective antigen. The purified PCR product of PlpB gene consisting of 831 base pairs was inserted into the pET-32a (+) plasmid, and then transferred into E. coli. The protective immunity conferred by recombinant PlpB (rPlpB) on mice was evaluated. The results showed that mice immunized with 200 μg of purified rPlpB were protected (60% survival rate) against challenge infection with 1 MLD of P. multocida strain LZ-PM. In conclusion, our data indicated that the PlpB protein may be a potential target as a candidate subunit vaccine for P. multocida infection. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Occurrence of virulence-associated genes in Pasteurella multocida isolates obtained from different hosts.

    Science.gov (United States)

    Shirzad Aski, Hesamaddin; Tabatabaei, Mohammad

    2016-07-01

    Pasteurella multocida infects a wide range of animals and the infection may spread to human through animal bites and scratches. Pasteurella multocida isolates, obtained from several clinically healthy and diseased animals (bovine, sheep, goat, poultry, dog and cat), were investigated for capsule biosynthesis (capA, B, D, E and F) and expression of 22 virulence-associated genes using Polymerase Chain Reaction (PCR). Multiplex PCR results revealed expression of capA, capD and capB genes in 81 (61.83%), 30 (22.90%) and 10 isolates (7.29%), respectively. However, neither of the isolates harbored capE or capF genes and ten isolates (7.29%) were negative for all cap genes. The expression of the capB gene was observed in small ruminant isolates. The occurrence of the ompA, ompH, oma87, sodA and sodC genes was noticed in all of the samples. More than 90% of the isolates harbored hgbA (96.18%), ptfA (95.41%), exbBD-tonB (93.12%), nanB (93.12%) and plbB genes (90.83%). The transferrin binding protein encoding gene tbpA was exclusively detected in the ruminant isolates. The limited number of isolates (25.95%) harbored dermonecrotoxin gene (toxA) and the highest occurrence was noted in the small ruminants, and the capsular type D isolates. This study highlights that the toxA, tbpA, and pfhA genes can be considered as important epidemiological markers for the characterization of P. multocida isolates. Copyright © 2016 Elsevier Ltd. All rights reserved.

  9. Pharmacokinetics/pharmacodynamics of marbofloxacin in a Pasteurella multocida serious murine lung infection model.

    Science.gov (United States)

    Qu, Ying; Qiu, Zhenzhen; Cao, Changfu; Lu, Yan; Sun, Meizhen; Liang, Chaoping; Zeng, Zhenling

    2015-12-02

    Marbofloxacin is a third-generation fluoroquinolone developed solely for veterinary medicine with a broad spectrum of antibacterial activity against some Gram-positive and most Gram-negative bacteria, including the bovine respiratory tract pathogen, Pasteurella multocida. The objective of our study was to elucidate the pharmacokinetics and pharmacodynamics of marbofloxacin in a Pasteurella multocida infected murine lung model, and to estimate the magnitudes of pharmacokinetics-pharmacodynamics parameters associated with various effects. The pharmacokinetic studies revealed marbofloxacin kinetics in infected mice were linear over a dose ranging from 1.25 to 10 mg/kg of body weight. The protein binding in the plasma of neutropenic infected mice was 29.77 %. The magnitudes of the ratio of the free-drug area under the concentration-time curve over 24 h to MIC (fAUC 0-24h/MIC) associated with static effect, a 2 log10 reduction and a 3 log10 reduction of bacterial counts were 40.84, 139.34, and 278.08 h, respectively. Based on the dose range study, marbofloxacin exhibited concentration-dependent killing and the fAUC/MIC was the PK/PD index that correlated best with efficacy (R(2) = 83 %). On the basis of a bactericidal effect goal of fAUC 0-24h/MIC of 278.08 h, if marbofloxacin is used for the treatment of P. multocida serious lung infection with an MIC90 of 0.12 μg/ml, the current dose (2 mg/kg) would fail to achieve a bactericidal effect. It would benefit from higher doses (4 ~ 6 mg/kg) than those commonly used in clinical practice.

  10. Pasteurella multocida type A as the primary agent of pneumonia and septicaemia in pigs

    Directory of Open Access Journals (Sweden)

    João X. de Oliveira Filho

    2015-08-01

    Full Text Available Abstract: In order to understand better the pathological aspects and spread of Pasteurella multocida type A as the primary cause of pneumonia in pigs, was made an experiment with intranasal inoculation of different concentrations of inocula [Group (G1: 108 Colony Forming Units (CFU/ml; G2: 107 CFU/ml; G3: 106 CFU/ml and G4: 105 CFU/ml], using two pigs per group. The pigs were obtained from a high health status herd. Pigs were monitored clinically for 4 days and subsequently necropsied. All pigs had clinical signs and lesions associated with respiratory disease. Dyspnoea and hyperthermia were the main clinical signs observed. Suppurative cranioventral bronchopneumonia, in some cases associated with necrosuppurative pleuropneumonia, fibrinous pericarditis and pleuritic, were the most frequent types of lesion found. The disease evolved with septicaemia, characterized by septic infarctions in the liver and spleen, with the detection of P. multocida type A. In this study, P. multocida type A strain #11246 was the primary agent of fibrinous pleuritis and suppurative cranioventral bronchopneumonia, pericarditis and septicaemia in the pigs. All concentrations of inoculum used (105-108 CFU/ml were able to produce clinical and pathological changes of pneumonia, pleuritis, pericarditis and septicemia in challenged animals.

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

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    Magda Patricia Carrillo

    2015-01-01

    Full Text Available 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.

  12. Immune response to dna vaccine expressing transferrin binding protein a gene of Pasteurella multocida

    Directory of Open Access Journals (Sweden)

    Satparkash Singh

    2011-06-01

    Full Text Available Haemorrhagic Septicaemia (HS, an acute and fatal disease of cattle and buffalo is primarily caused by serotype B:2 or E:2 of Pasteurella multocida. The transferrin binding protein A (TbpA has been found to act as immunogen and potent vaccine candidate in various Gram negative bacteria including P. multocida. The present study was carried out to evaluate the potential of this antigen as a DNA vaccine against HS in mice model. The tbpA gene of P. multocida serotype B:2 was cloned in a mammalian expression vector alone and along with murine IL2 gene as immunological adjuvant to produce monocistronic and bicistronic DNA vaccine constructs, respectively. The immune response to DNA vaccines was evaluated based on serum antibody titres and lymphocyte proliferation assay. A significant increase in humoral and cell mediated immune responses was observed in mice vaccinated with DNA vaccines as compared to non immunized group. Additionally, the bicistronic DNA vaccine provided superior immune response and protection level following challenge as compared to monocistronic construct. The study revealed that DNA vaccine presents a promising approach for the prevention of HS.

  13. Multiplex PCR to identify macrolide resistance determinants in Mannheimia haemolytica and Pasteurella multocida.

    Science.gov (United States)

    Rose, Simon; Desmolaize, Benoit; Jaju, Puneet; Wilhelm, Cornelia; Warrass, Ralf; Douthwaite, Stephen

    2012-07-01

    The bacterial pathogens Mannheimia haemolytica and Pasteurella multocida are major etiological agents in respiratory tract infections of cattle. Although these infections can generally be successfully treated with veterinary macrolide antibiotics, a few recent isolates have shown resistance to these drugs. Macrolide resistance in members of the family Pasteurellaceae is conferred by combinations of at least three genes: erm(42), which encodes a monomethyltransferase and confers a type I MLS(B) (macrolide, lincosamide, and streptogramin B) phenotype; msr(E), which encodes a macrolide efflux pump; and mph(E), which encodes a macrolide-inactivating phosphotransferase. Here, we describe a multiplex PCR assay that detects the presence of erm(42), msr(E), and mph(E) and differentiates between these genes. In addition, the assay distinguishes P. multocida from M. haemolytica by amplifying distinctive fragments of the 23S rRNA (rrl) genes. One rrl fragment acts as a general indicator of gammaproteobacterial species and confirms whether the PCR assay has functioned as intended on strains that are negative for erm(42), msr(E), and mph(E). The multiplex system has been tested on more than 40 selected isolates of P. multocida and M. haemolytica and correlated with MICs for the veterinary macrolides tulathromycin and tilmicosin, and the newer compounds gamithromycin and tildipirosin. The multiplex PCR system gives a rapid and robustly accurate determination of macrolide resistance genotypes and bacterial genus, matching results from microbiological methods and whole-genome sequencing.

  14. Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine

    Directory of Open Access Journals (Sweden)

    Thales Quedi Furian

    2016-03-01

    Full Text Available Abstract Pasteurella multocida causes atrophic rhinitis in swine and fowl cholera in birds, and is a secondary agent in respiratory syndromes. Pathogenesis and virulence factors involved are still poorly understood. The aim of this study was to detect 22 virulence-associated genes by PCR, including capsular serogroups A, B and D genes and to evaluate the antimicrobial susceptibility of P. multocida strains from poultry and swine. ompH, oma87, plpB, psl, exbD-tonB, fur, hgbA, nanB, sodA, sodC, ptfA were detected in more than 90% of the strains of both hosts. 91% and 92% of avian and swine strains, respectively, were classified in serogroup A. toxA and hsf-1 showed a significant association to serogroup D; pmHAS and pfhA to serogroup A. Gentamicin and amoxicillin were the most effective drugs with susceptibility higher than 97%; however, 76.79% of poultry strains and 85% of swine strains were resistant to sulphonamides. Furthermore, 19.64% and 36.58% of avian and swine strains, respectively, were multi-resistant. Virulence genes studied were not specific to a host and may be the result of horizontal transmission throughout evolution. High multidrug resistance demonstrates the need for responsible use of antimicrobials in animals intended for human consumption, in addition to antimicrobial susceptibility testing to P. multocida.

  15. Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine

    Science.gov (United States)

    Furian, Thales Quedi; Borges, Karen Apellanis; Laviniki, Vanessa; da Silveira Rocha, Silvio Luis; de Almeida, Camila Neves; do Nascimento, Vladimir Pinheiro; Salle, Carlos Tadeu Pippi; de Souza Moraes, Hamilton Luiz

    2016-01-01

    Pasteurella multocida causes atrophic rhinitis in swine and fowl cholera in birds, and is a secondary agent in respiratory syndromes. Pathogenesis and virulence factors involved are still poorly understood. The aim of this study was to detect 22 virulence-associated genes by PCR, including capsular serogroups A, B and D genes and to evaluate the antimicrobial susceptibility of P. multocida strains from poultry and swine. ompH, oma87, plpB, psl, exbD-tonB, fur, hgbA, nanB, sodA, sodC, ptfA were detected in more than 90% of the strains of both hosts. 91% and 92% of avian and swine strains, respectively, were classified in serogroup A. toxA and hsf-1 showed a significant association to serogroup D; pmHAS and pfhA to serogroup A. Gentamicin and amoxicillin were the most effective drugs with susceptibility higher than 97%; however, 76.79% of poultry strains and 85% of swine strains were resistant to sulphonamides. Furthermore, 19.64% and 36.58% of avian and swine strains, respectively, were multi-resistant. Virulence genes studied were not specific to a host and may be the result of horizontal transmission throughout evolution. High multidrug resistance demonstrates the need for responsible use of antimicrobials in animals intended for human consumption, in addition to antimicrobial susceptibility testing to P. multocida. PMID:26887247

  16. Variability of Pasteurella multocida isolated from Icelandic sheep and detection of the toxA gene.

    Science.gov (United States)

    Einarsdottir, Thorbjorg; Gunnarsson, Eggert; Sigurdardottir, Olof G; Jorundsson, Einar; Fridriksdottir, Vala; Thorarinsdottir, Gudridur E; Hjartardottir, Sigridur

    2016-09-01

    Pasteurella multocida can be part of the upper respiratory flora of animals, but under conditions of stress or immunocompromisation, the bacteria can cause severe respiratory symptoms. In this study, we compared 10 P. multocida isolates from Icelandic sheep with respiratory symptoms and 19 isolates from apparently healthy abattoir sheep. We examined capsule type, genetic variability and the presence of the toxA gene in the two groups. Surprisingly, we found that all ovine P. multocida isolates examined in this study carried the toxA gene, which markedly differs from what has been published from other studies. Interestingly, all isolates from abattoir animals were capsule type D, whilst bacteria isolated from animals with clinical respiratory symptoms had capsule type A, D or F. Examination of seven housekeeping genes indicated that the clinical respiratory isolates were significantly more heterogeneous than the abattoir isolates (Pmultocida in sheep - a genetically homogeneous group that resides in the respiratory tract and a genetically heterogeneous group that is the predominant cause of disease.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Virulence genes and antimicrobial resistance of Pasteurella multocida isolated from poultry and swine.

    Science.gov (United States)

    Furian, Thales Quedi; Borges, Karen Apellanis; Laviniki, Vanessa; Rocha, Silvio Luis da Silveira; de Almeida, Camila Neves; do Nascimento, Vladimir Pinheiro; Salle, Carlos Tadeu Pippi; Moraes, Hamilton Luiz de Souza

    2016-01-01

    Pasteurella multocida causes atrophic rhinitis in swine and fowl cholera in birds, and is a secondary agent in respiratory syndromes. Pathogenesis and virulence factors involved are still poorly understood. The aim of this study was to detect 22 virulence-associated genes by PCR, including capsular serogroups A, B and D genes and to evaluate the antimicrobial susceptibility of P. multocida strains from poultry and swine. ompH, oma87, plpB, psl, exbD-tonB, fur, hgbA, nanB, sodA, sodC, ptfA were detected in more than 90% of the strains of both hosts. 91% and 92% of avian and swine strains, respectively, were classified in serogroup A. toxA and hsf-1 showed a significant association to serogroup D; pmHAS and pfhA to serogroup A. Gentamicin and amoxicillin were the most effective drugs with susceptibility higher than 97%; however, 76.79% of poultry strains and 85% of swine strains were resistant to sulphonamides. Furthermore, 19.64% and 36.58% of avian and swine strains, respectively, were multi-resistant. Virulence genes studied were not specific to a host and may be the result of horizontal transmission throughout evolution. High multidrug resistance demonstrates the need for responsible use of antimicrobials in animals intended for human consumption, in addition to antimicrobial susceptibility testing to P. multocida. Copyright © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

  20. Multiplex PCR To Identify Macrolide Resistance Determinants in Mannheimia haemolytica and Pasteurella multocida

    Science.gov (United States)

    Rose, Simon; Desmolaize, Benoit; Jaju, Puneet; Wilhelm, Cornelia; Warrass, Ralf

    2012-01-01

    The bacterial pathogens Mannheimia haemolytica and Pasteurella multocida are major etiological agents in respiratory tract infections of cattle. Although these infections can generally be successfully treated with veterinary macrolide antibiotics, a few recent isolates have shown resistance to these drugs. Macrolide resistance in members of the family Pasteurellaceae is conferred by combinations of at least three genes: erm(42), which encodes a monomethyltransferase and confers a type I MLSB (macrolide, lincosamide, and streptogramin B) phenotype; msr(E), which encodes a macrolide efflux pump; and mph(E), which encodes a macrolide-inactivating phosphotransferase. Here, we describe a multiplex PCR assay that detects the presence of erm(42), msr(E), and mph(E) and differentiates between these genes. In addition, the assay distinguishes P. multocida from M. haemolytica by amplifying distinctive fragments of the 23S rRNA (rrl) genes. One rrl fragment acts as a general indicator of gammaproteobacterial species and confirms whether the PCR assay has functioned as intended on strains that are negative for erm(42), msr(E), and mph(E). The multiplex system has been tested on more than 40 selected isolates of P. multocida and M. haemolytica and correlated with MICs for the veterinary macrolides tulathromycin and tilmicosin, and the newer compounds gamithromycin and tildipirosin. The multiplex PCR system gives a rapid and robustly accurate determination of macrolide resistance genotypes and bacterial genus, matching results from microbiological methods and whole-genome sequencing. PMID:22564832

  1. Establishment of a Pathogenicity Index for One-day-old Broilers to Pasteurella multocida Strains Isolated from Clinical Cases in Poultry and Swine

    Directory of Open Access Journals (Sweden)

    RM Pilatti

    Full Text Available ABSTRACT Although Pasteurella multocida is a member of the respiratory microbiota, under some circumstances, it is a primary agent of diseases , such as fowl cholera (FC, that cause significant economic losses. Experimental inoculations can be employed to evaluate the pathogenicity of strains, but the results are usually subjective and knowledge on the pathogenesis of this agent is still limited. The objective of this study was to establish a new methodology for classifying the pathogenicity of P. multocida by formulating a standard index. Strains isolated from FC cases and from swine with respiratory problems were selected. One hundred mL of a bacterial culture of each strain, containing 106 CFU, was inoculated in 10 one-day-old broilers. Mortality after inoculation, time of death (TD, and the presence of six macroscopic lesions were evaluated over a period of seven days post-inoculation (dpi. A Pathogenicity Index Per Bird (IPI, ranging 0 to 10, was calculated. Liver and heart fragments were collected to reisolate the bacteria. Blood was collected from the surviving birds, and an ELISA test was carried out to detect specific antibodies. The median of the pathogenicity indices, the number of lesions and the rate of bacteria reisolation were significantly different (p<0.05 among the origins of the isolates (p<0.05. The pathogenicity index developed in this study allows the classification of Pasteurella multocida pathogenicity and may be an alternative to the pathogenicity models currently used for screening.

  2. The use of filter paper as a transport device for serology of Pasteurella multocida infection : Analysis and comparison ofprotein composition of filter paper extract and serum

    Directory of Open Access Journals (Sweden)

    Lily Natalia

    1998-10-01

    Full Text Available Two methods for collecting blood specimens for measuring antibody to Pasteurella multocida were compared. Blood was collected on filter-paper strips, air-dried and stored at 4°C along with paired samples collected by venepumeture . Analysis using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE showed that the protein composition of filter paper extract and serum was similar. Both samples had common proteins of 67, 52-58 and 27 kDa. However, there are two proteins bands of 14 and 30 kDa that were only found in, filter-paper extract. Westernblot analysis also showed that samples from both sampling techniques reacted to P. multocida proteins of 43 kDa. Samples from experimental and field animals were also collected by the two techniques and assayed by enzyme-linked immunosorbent assay (ELISA for P. multocida antibodies . The agreement between samples from experimental animals and the field using ELISA was analyzed . Samples from experimental animals, showed a very high correlation (r = 0.931 in ELISA results among samples collected by the two techniques. However, the correlation was lower (r = 0.799 in samples collected from the field. Cost analysis showed that filter-paper collection technique was 100 times more economical compared to venepuncture technique. It was concluded that eluates of whole blood dried on filter paper can be used as an alternative to sera in ELISA for measuring antibodies to P. multocida.

  3. Profiling of virulence associated genes of Pasteurella multocida isolated from cattle.

    Science.gov (United States)

    Verma, Subhash; Sharma, Mandeep; Katoch, Shailja; Verma, Lovit; Kumar, Sandeep; Dogra, Vishal; Chahota, Rajesh; Dhar, Prasenjit; Singh, Geetanjali

    2013-03-01

    Pasteurella multocida is a causative agent of many major diseases of which haemorrhagic septiciemia (HS) in cattle & a buffalo is responsible for significant losses to livestock sector in India and south Asia. The disease outcome is affected by various host- and pathogen-specific determinants. Several bacterial species-specific putative virulence factors including the capsular and virulence associated genes have been proposed to play a key role in this interaction. A total of 23 isolates of P. multocida were obtained from 335 cases of various clinically healthy and diseased cattle. These isolates were examined for capsule synthesis genes (capA, B, D, E and F) and eleven virulence associated genes (tbpA, pfhA, toxA, hgbB, hgbA, nanH, nanB, sodA, sodC, oma87 and ptfA) by PCR. A total of 19 P. multocida isolates belonging to capsular type B and 4 of capsular type A were isolated. All isolates of capsular type B harboured the virulence associated genes: tbpA, pfhA, hgbA, sodC and nanH, coding for transferrin binding protein, filamentous hemagglutinin, haemoglobin binding protein, superoxide dismutase and neuraminidases, respectively; while isolates belonging to capsular type A also carried tbpA, pfhA, hgbA and nanH genes. Only 50 % of capsular type A isolates contained sodC gene while 100 % of capsular type B isolates had sodC gene. The gene nanB and toxA were absent in all the 23 isolates. In capsular type A isolates, either sodA or sodC gene was present & these genes did not occur concurrently. The presence of virulence associated gene ptfA revealed a positive association with the disease outcome in cattle and could therefore be an important epidemiological marker gene for characterizing P. multocida isolates.

  4. Antimicrobial resistance genes in Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida isolated from Australian pigs.

    Science.gov (United States)

    Dayao, Dae; Gibson, J S; Blackall, P J; Turni, C

    2016-07-01

    To identify genes associated with the observed antimicrobial resistance in Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida isolated from Australian pigs. Isolates with known phenotypic resistance to β-lactams, macrolides and tetracycline were screened for the presence of antimicrobial resistance genes. A total of 68 A. pleuropneumoniae, 62 H. parasuis and 20 P. multocida isolates exhibiting phenotypic antimicrobial resistance (A. pleuropneumoniae and P. multocida) or elevated minimal inhibitory concentrations (MICs) (H. parasuis) to any of the following antimicrobial agents - ampicillin, erythromycin, penicillin, tetracycline, tilmicosin and tulathromycin - were screened for a total of 19 associated antimicrobial resistance genes (ARGs) by PCR. The gene bla ROB-1 was found in all ampicillin- and penicillin-resistant isolates, but none harboured the bla TEM-1 gene. The tetB gene was found in 76% (74/97) of tetracycline-resistant isolates, 49/53 A. pleuropneumoniae, 17/30 H. parasuis and 8/14 P. multocida. One A. pleuropneumoniae isolate harboured the tetH gene, but none of the 97 isolates had tetA, tetC, tetD, tetE, tetL, tetM or tetO. A total of 92 isolates were screened for the presence of macrolide resistance genes. None was found to have ermA, ermB, ermC, erm42, mphE, mefA, msrA or msrE. The current study has provided a genetic explanation for the resistance or elevated MIC of the majority of isolates of Australian porcine respiratory pathogens to ampicillin, penicillin and tetracycline. However, the macrolide resistance observed by phenotypic testing remains genetically unexplained and further studies are required. © 2016 Australian Veterinary Association.

  5. Carboxyl terminus heterogeneity of type IV fimbrial subunit protein of Pasteurella multocida isolates.

    Science.gov (United States)

    Shivachandra, Sathish Bhadravati; Kumar, Abhinendra; Yogisharadhya, Revanaiah; Ramakrishnan, M A; Viswas, K N

    2013-12-01

    Pasteurella multocida, a Gram-negative bacterial pathogen, known to affect a wide range of domestic as well as wild animal and avian species throughout the world by causing either systemic or localized infections termed as 'pasteurellosis'. P. multocida isolates are known to possess type IV fimbriae (pili) as one of the major virulence factors based on their role in adhesion to host surfaces and subsequent pathogenesis. In the present study, ptfA gene of Indian P. multocida isolates (n = 8) originated from different animal (buffalo, sheep, goat, pig) and avian host species (chicken, turkey, duck, quail) were amplified, cloned, sequenced and compared with available ptfA/fimbrial protein sequences in GenBank/publications (n = 22) to understand its variability with respect to geography/host/serogroup/disease specific patterns. Multiple sequence alignment revealed highly conserved N-terminus α-1 helix region and heterogeneous C-terminus (68-137 aa) comprised of β-strand regions (β1, β2, β3, β4) with conserved two pairs of cysteine residues. Interestingly, an existence of absolute homogeneity among the P. multocida isolates that caused haemorrhagic septicaemia in bovines and septicaemic pasteurellosis in sheep and goats was noticed. Pig isolates had 99.3% homogeneity. On contrary, more diversity (35.8%) was observed among isolates that caused fowl cholera in avians irrespective of identical capsular/somatic serogroup and similar host species. Phylogenetic analysis based on nucleotide sequences of ptfA gene revealed formation of mixed clusters with isolates representing different disease conditions as well as serogroups irrespective of country of origin which indicated the possible role of cross-species transmission among different animal/avian species. The study indicated highly conserved and host specific fimbriae among animal species than relatively divergent fimbriae among avian species.

  6. 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

    DEFF Research Database (Denmark)

    Christensen, J.P.; Dietz, Hans-Henrik; Bisgaard, M.

    1998-01-01

    Two outbreaks of fowl cholera in the avifauna in Denmark, affecting primarily elders 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 Hha...

  7. An ST11 clone of Pasteurella multocida, widely spread among farmed rabbits in the Iberian Peninsula, demonstrates respiratory niche association.

    Science.gov (United States)

    García-Alvarez, Andrés; Chaves, Fernando; Fernández, Ana; Sanz, Celia; Borobia, Marta; Cid, Dolores

    2015-08-01

    Pasteurella multocida is a veterinary pathogen causing diseases with considerable economic repercussions in a wide range of animal hosts. In rabbits, P. multocida infections cause a variety of clinical manifestations including rhinitis, pneumonia, septicemia, abscesses, mastitis, and pyometra. In this study, 100 P. multocida isolates from different commercial rabbit farms located throughout the Iberian Peninsula were molecularly characterized by capsular typing, detection of four virulence-associated genes (tbpA, toxA, hgbB, and pfhA), and multilocus sequence typing (MLST). Rabbit P. multocida isolates belonged to three different capsular types: A (47.0%), D (28.0%), and F (25.0%). One group of P. multocida isolates of capsular type D and positive for the hgbB gene was significantly associated with the clinical presentation of respiratory disease (OR 5.91; 95%CI, 1.63-21.38). These isolates belonged to same sequence type, ST11, in the P. multocida Multi-host MLST database. The ST11 clone also includes isolates from porcine and avian pneumonia. This clonal group of epidemiologically unrelated P. multocida isolates could be a virulent clone with some degree of specificity for respiratory disease. These findings could be relevant in the development of vaccines for pasteurellosis prevention, especially respiratory disease. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Alteration of DNA adenine methylase (Dam) activity in Pasteurella multocida causes increased spontaneous mutation frequency and attenuation in mice.

    Science.gov (United States)

    Chen, Liang; Paulsen, Daniel B; Scruggs, Daniel W; Banes, Michelle M; Reeks, Brenda Y; Lawrence, Mark L

    2003-08-01

    Pasteurella multocida is one of the primary bacterial pathogens associated with bovine respiratory disease (BRD) complex. Relatively few virulence factors of P. multocida have been characterized, and there is a need for improved vaccines for prevention of BRD. In other Gram-negative species, DNA adenine methylase (Dam) regulates the expression of virulence genes, and appropriate expression of Dam is required for virulence. In this study, the authors cloned and sequenced the P. multocida A1 dam gene and demonstrated that it is able to restore Dam function in an Escherichia coli dam mutant. When P. multocida dam was placed under the control of a constitutively expressed promoter on a plasmid, it caused an increased spontaneous mutation rate in P. multocida. In addition, the plasmid-mediated alteration of Dam production in P. multocida caused it to be highly attenuated in mice. These findings indicate that appropriate expression of Dam is required for virulence of P. multocida, which is believed to be the first report that Dam is required for virulence of a species in the Pasteurellaceae. Therefore, Dam may function as a virulence gene regulator in the Pasteurellaceae, similar to previously reported findings from other Gram-negative species.

  9. Dietary L-glutamine supplementation increases Pasteurella multocida burden and the expression of its major virulence factors in mice.

    Science.gov (United States)

    Ren, Wenkai; Liu, Shuping; Chen, Shuai; Zhang, Fengmei; Li, Nengzhang; Yin, Jie; Peng, Yuanyi; Wu, Li; Liu, Gang; Yin, Yulong; Wu, Guoyao

    2013-10-01

    This study was conducted to determine the effects of graded doses of L-glutamine supplementation on the replication and distribution of Pasteurella multocida, and the expression of its major virulence factors in mouse model. Mice were randomly assigned to the basal diet supplemented with 0, 0.5, 1.0 or 2.0 % glutamine. Pasteurella multocida burden was detected in the heart, liver, spleen, lung and kidney after 12 h of P. multocida infection. The expression of major virulence factors, toll-like receptors (TLRs), proinflammatory cytokines (interleukin-1 beta, interleukin-6, and tumor necrosis factor alpha) and anti-oxidative factors (GPX1 and CuZnSOD) was analyzed in the lung and spleen. Dietary 0.5 % glutamine supplementation has little significant effect on these parameters, compared to those with basal diet. However, results showed that a high dose of glutamine supplementation increased the P. multocida burden (P multocida burden and the expression of its major virulence factors, while affecting the functions of the lung and spleen.

  10. WHOLE CELL PROTEIN PROFILING OF PASTEURELLA MULTOCIDA FIELD ISOLATES IN PAKISTAN

    Directory of Open Access Journals (Sweden)

    I. NAWAZ, R. MUNIR, U. FAROOQ, R. KAUSAR AND A. KHANUM1

    2006-10-01

    Full Text Available Eight haemorrhagic septicaemia (HS related Pasteurella multocida isolates, collected from different localities of Pakistan, were characterized by sodium-dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE technique. After sonification, the bacterial proteins were separated by centrifugation. Proteins from sonicated supernatant were salted out by ammonium sulfate. Sonicated supernatant, as well as ammonium sulfate precipitated proteins, were analyzed. Molecular weights of proteins were determined from graph between Rf value and log of molecular weight. The ammonium sulfate treated samples showed fewer bands with low molecular weights of 54, 45, 42, 40 and 20.5 kDa, while total 31 visible bands were observed in sonicated supernatants ranging from 126 to 11 kDa. The common protein bands in both preparations were of 54 and 23 kDa molecular weight.

  11. Nasal isolation of Mannheimia haemolytica and Pasteurella multocida as predictors of respiratory disease in shipped calves.

    Science.gov (United States)

    Taylor, J D; Holland, B P; Step, D L; Payton, M E; Confer, A W

    2015-04-01

    Three hundred ninety five calves were purchased from sale barns and delivered to the Willard Sparks Beef Research Center. Nasal swabs were collected to determine if presence of Mannheimia haemolytica and Pasteurella multocida in the upper respiratory tract (URT) can facilitate diagnosis of bovine respiratory disease (BRD). Samples were collected at arrival and at treatment for BRD. Clinically healthy control calves were sampled at time of treatment of sick calves. M. haemolytica was more commonly isolated from calves at treatment than at time of arrival or from control calves. M. haemolytica was more common in calves requiring treatment than in those never treated. Need for treatment and number of treatments were negatively associated with average daily gain, supporting the accuracy of diagnosis. These results suggest that URT sampling, when combined with clinical diagnosis, may assist in providing greater diagnostic accuracy, improving ability to evaluate risk factors, interventions, and treatments. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Biofilm inhibitors targeting the outer membrane protein A of Pasteurella multocida in swine.

    Science.gov (United States)

    Kubera, Anchanee; Thamchaipenet, Arinthip; Shoham, Menachem

    2017-01-01

    Pasteurella multocida (Pm) is the causative agent of atrophic rhinitis in swine. This study aimed to discover biofilm inhibitors against swine Pm to counteract antibiotic resistance and decrease virulence. The virulence factor outer membrane protein A (OmpA) was targeted. A library of drugs approved by the Food and Drug Administration (FDA) was used to perform virtual screening against PmOmpA. The top-scoring compounds had no effect on the growth of Pm serotype A or D. Mycophenolate mofetil showed the highest efficacy in inhibiting biofilm formation by Pm serotype A, with an IC50 of 7.3 nM. For Pm serotype D, indocyanine green showed the highest effect at an IC50 of 11.7 nM. Nevertheless, these compounds had no effect on an established biofilm of Pm. This study offers an alternative way to prevent biofilm formation by Pm that could also be applied to other pathogens.

  13. Recent insights into Pasteurella multocida toxin and other G-protein-modulating bacterial toxins.

    Science.gov (United States)

    Wilson, Brenda A; Ho, Mengfei

    2010-08-01

    Over the past few decades, our understanding of the bacterial protein toxins that modulate G proteins has advanced tremendously through extensive biochemical and structural analyses. This article provides an updated survey of the various toxins that target G proteins, ending with a focus on recent mechanistic insights in our understanding of the deamidating toxin family. The dermonecrotic toxin from Pasteurella multocida (PMT) was recently added to the list of toxins that disrupt G-protein signal transduction through selective deamidation of their targets. The C3 deamidase domain of PMT has no sequence similarity to the deamidase domains of the dermonecrotic toxins from Escherichia coli (cytotoxic necrotizing factor [CNF]1-3), Yersinia (CNFY) and Bordetella (dermonecrotic toxin). The structure of PMT-C3 belongs to a family of transglutaminase-like proteins, with active site Cys-His-Asp catalytic triads distinct from E. coli CNF1.

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

    Directory of Open Access Journals (Sweden)

    Everlon Cid Rigobelo

    2013-01-01

    Full Text Available 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%.

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

    Science.gov (United States)

    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

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

    Science.gov (United States)

    Samuel, M.D.; Shadduck, D.J.; Goldberg, Diana 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.

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

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    Paula Esquinas

    2013-01-01

    Full Text Available 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.

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

    Science.gov (United States)

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. Factors influencing the potency of marbofloxacin for pig pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida.

    Science.gov (United States)

    Dorey, L; Hobson, S; Lees, P

    2017-04-01

    For the pig respiratory tract pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida, Minimum Inhibitory Concentration (MIC) of marbofloxacin was determined in recommended broths and pig serum at three inoculum strengths. MICs in both growth matrices increased progressively from low, through medium to high starting inoculum counts, 10(4), 10(6) and 10(8)CFU/mL, respectively. P. multocida MIC ratios for high:low inocula were 14:4:1 for broth and 28.2:1 for serum. Corresponding MIC ratios for A. pleuropneumoniae were lower, 4.1:1 (broth) and 9.2:1 (serum). MIC high:low ratios were therefore both growth matrix and bacterial species dependent. The effect of alterations to the chemical composition of broths and serum on MIC were also investigated. Neither adjusting broth or serum pH in six increments over the range 7.0 to 8.0 nor increasing calcium and magnesium concentrations of broth in seven incremental steps significantly affected MICs for either organism. In time-kill studies, the killing action of marbofloxacin had the characteristics of concentration dependency against both organisms in both growth matrices. It is concluded that MIC and time-kill data for marbofloxacin, generated in serum, might be preferable to broth data, for predicting dosages of marbofloxacin for clinical use. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. The origin of Pasteurella multocida impacts pathology and inflammation when assessed in a mouse model.

    Science.gov (United States)

    Pors, Susanne E; Chadfield, Mark S; Sørensen, Dorte B; Offenberg, Hanne; Bisgaard, Magne; Jensen, Henrik E

    2016-04-01

    Host-pathogen interactions of Pasteurella multocida isolates of different origin were studied in a mouse model, focusing on pathology, bacterial load and expression of the metalloproteinase MMP9 and its inhibitor TIMP1. Intranasal inoculation with one of three doses (10(6), 10(4), 10(2)CFU) of an isolate from porcine pneumonia or fowl cholera showed marked differences between the two isolates. The avian isolate was highly pathogenic with severe signs of necrotizing pneumonia, liver necrosis and high bacterial load in lung and liver. Clinical signs and pathology related to the porcine isolate were dose dependent and consisted of exudative bronchopneumonia, abscess formation in liver and a lower bacterial load in lung and liver. Both isolates caused increased expression of MMP9 and TIMP1. In conclusion, evaluation and comparison of pathogenicity and host-pathogen interaction of P. multocida isolates from different hosts is possible in the intranasal murine model. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Identification and molecular cloning of a heparosan synthase from Pasteurella multocida type D.

    Science.gov (United States)

    DeAngelis, Paul L; White, Carissa L

    2002-03-01

    Pasteurella multocida Type D, a causative agent of atrophic rhinitis in swine and pasteurellosis in other domestic animals, produces an extracellular polysaccharide capsule that is a putative virulence factor. It was reported previously that the capsule was removed by treating microbes with heparin lyase III. We molecularly cloned a 617-residue enzyme, pmHS, which is a heparosan (nonsulfated, unepimerized heparin) synthase. Recombinant Escherichia coli-derived pmHS catalyzes the polymerization of the monosaccharides from UDP-GlcNAc and UDP-GlcUA. Other structurally related sugar nucleotides did not substitute. Synthase activity was stimulated about 7-25-fold by the addition of an exogenous polymer acceptor. Molecules composed of approximately 500-3,000 sugar residues were produced in vitro. The polysaccharide was sensitive to the action of heparin lyase III but resistant to hyaluronan lyase. The sequence of the pmHS enzyme is not very similar to the vertebrate heparin/heparan sulfate glycosyltransferases, EXT1 and 2, or to other Pasteurella glycosaminoglycan synthases that produce hyaluronan or chondroitin. The pmHS enzyme is the first microbial dual-action glycosyltransferase to be described that forms a polysaccharide composed of beta4GlcUA-alpha4GlcNAc disaccharide repeats. In contrast, heparosan biosynthesis in E. coli K5 requires at least two separate polypeptides, KfiA and KfiC, to catalyze the same polymerization reaction.

  3. Cellular defense of the avian respiratory system: influx and nonopsonic phagocytosis by respiratory phagocytes activated by Pasteurella multocida.

    OpenAIRE

    Toth, T. E.; Pyle, R H; Caceci, T; Siegel, P B; Ochs, D.

    1988-01-01

    Poultry have a very limited number of resident macrophages in the normal steady-state respiratory tract. Thus, poultry must rely heavily on active migration of phagocytic cells to the lungs and air sacs in defending against respiratory pathogens. Intratracheal administration of a live, apathogenic Pasteurella multocida vaccine (Choloral; Clemson University strain) increased the number of avian respiratory phagocytes (ARP; obtained by lavage of lungs and air sacs) within 24 h by 3 orders of ma...

  4. [Clinicobacteriological study of Pasteurella multocida as a zoonosis (1). Condition of dog and cat carriers of Pasteurella, and the influence for human carrier rate by kiss with the pets].

    Science.gov (United States)

    Arashima, Y; Kumasaka, K; Okuyama, K; Kawabata, M; Tsuchiya, T; Kawano, K; Asano, R; Hokari, S

    1992-02-01

    Pasteurella multocida is a gram-negative short rod-shaped bacteria, which is a part of the indigenous flora of the oral cavity of many animals other than man. The number of reports on cases of infections with this bacterium due to animal bites and/or scratches, bacterial infections of the respiratory tract, sepsis due to this organism and death caused by the bacteria have been increasing in recent years. We investigated P. multocida in the hair and oral cavity of 3 dogs and 29 cats according to the classification of Mutters et al.. We also studied the relationship between the carrier rate for Pasteurella in the oral cavity and kissing of pets in 24 pet owners (3 dogs and 11 cats). No P. multocida was isolated from the hair of neither dogs nor cats. One strain of P. multocida subsp. multocida and two strains of P. stomatis, were isolated from the oral cavity of dogs, and 35 strains of Pasteurella were isolated from the oral cavity of cats. Two strains of P. multocida subsp. multocida, whose biochemical properties were different, were detected in the oral cavity of one cat. In three cats, Pasteurella other than P. multocida subsp. multocida was isolated from the same oral cavity. No Pasteurella was detected in the oral cavity of 19 pet owners who had not kissed their cats, whereas P. stomatis was isolated from the oral cavity of one of 2 pet owners who had kissed their cats and in 2 of 3 pet owners who had kissed their dogs (the same bacteria was isolated from a dog that was being kept by some of these positive pet owners).(ABSTRACT TRUNCATED AT 250 WORDS)

  5. Polymerase chain reaction optimization for the detection of Pasteurella multocida B:2, the causative agent of Haemorrhagic septicaemia

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    Lily Natalia

    2001-12-01

    Full Text Available Specific detection of Pasteurella multocida type B:2 by polymerase chain reaction (PCR, using a set of DNA primers wasoptimised. Effects of the addition of ethylene diamine tetra acetic acid (EDTA to the sample preparation, Escherichia coli contamination and the number of P. multocida on the PCR product was assessed. The PCR test was compared to the standard bacteriological method for the detection of P. multocida B:2 in tonsillar swab samples collected from slaughter houses of various regions in Indonesia. Addition of 100 mM EDTA-saline to P. multocida B:2 spiked tonsillar swab samples inhibits the production 350 base pairs (bp PCR product. The inhibitory effect of the EDT A can be eliminated by three times washing with deionised water. The PCR can detect P. multocida as low as I organism and contanimation of 100 CFU of E. coli does not effect the PCR result. The results show that the DNA primers for P. multocida B:2 is sensitive and specific. The inhibitory effect of EDTA in PCR samples can be eliminated by washings.

  6. Genotyping of Pasteurella multocida ovine and bovine isolates from Iran based on PCR-RFLP of ompH gene

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    A. Ghanizadeh

    2015-10-01

    Full Text Available Pasteurella multocida (P. multocida, A Gram-negative facultative anaerobic bacterium, is a causative animal pathogen in porcine atrophic rhinitis and avian fowl cholera. The outer membrane of Gram-negative bacteria contains of many different protein in very high copy numbers. One of the major outer membrane, the H proteins have functional as high immunogenicity and antigenicity. In this study to increase information about epidemiology of ovine and bovine P. multocida, the 24 isolates from sheep and nine isolates from cattle were investigated by PCR-RFLP analysis of the ompH gene. In all 33 isolates, digestion of the amplified fragment of ompH gene by using EcoRI, cfoI and HindIII produced 3, 5 and 3 different restriction patterns respectively. Sixteen RFLP patterns were found among 33 investigated P.multocida isolates. This study showed that, the PCR RFLP based on ompH gene is potentially a useful method for typing of P. multocida isolates from sheep and cattle. The RFLP patterns of this gene exhibited extensive restriction site heterogeneity, which may be particularly suitable for fingerprinting of P. multocida isolates.Considering ompH protein as a protective immunogenic moiety of P.ultocida, the results of this study showed a heterogenic bacteria and this means the possibility to produce a multivalent vaccine to be protective against diseases caused by this organism in sheep and cattle in Iran.

  7. Isolation, characterization, virulence and immunogenicity testing of field isolates of Pasteurella multocida, Staphylococcus aureus, and Streptococcus agalactiae in laboratory settings.

    Science.gov (United States)

    Qudratullah; Muhammad, G; Saqib, M; Bilal, M Qamar

    2017-08-01

    The present study was designed to investigate isolation, characterization, virulence and immunogenicity testing of field isolates of Pasteurella multocida, Staphylococcus aureus, and Streptococcus agalactiae in rabbits and mice. Isolates of P. multocida, S. aureus and Str. agalactiae recovered from field cases of Hemorragic septicemia and mastitis were scrutinized for virulence/pathogenicity and immunogenicity. Mouse LD50 of P. multocida showed that P. multocida isolate No.1 was more virulent than isolates No. 2 and 3. Virulence of isolate No.1S. aureus and Str. agalactiae revealed that 100, 80% rabbits died within 18h of inoculation. Seven-digit numerical profiles of these 4 isolates with API® Staph test strips isolates, No.1 (6736153) showed good identification (S. aureus id=90.3%). Indirect ELISA-based serum antibody titers to P. multocida isolate No.1, S. aureus No.1, Str. agalactiae, isolate No.1 elicited high antibody titers 1.9, 1.23, 1.12 respectively. All the pathogens of Isolate No. 1 (P. multocida, S. aureus Str. agalactiae), were high antibody than others isolates. Copyright © 2017 Elsevier B.V. All rights reserved.

  8. Detection of Pasteurella multocida isolates from local pigs of India by polymerase chain reaction and their antibiogram.

    Science.gov (United States)

    Rajkhowa, Swaraj; Shakuntala, Ingudam; Pegu, Seema Rani; Das, Rajib Kumar; Das, Anubrata

    2012-10-01

    Pasteurella multocida has been recognized as an important veterinary pathogen for over a century. Conventional methods for diagnosis of pasteurellosis rely on the detection of the organism by microscopy and its isolation and identification. However, as far as pasteurellosis is concerned, it is not just sufficient to know the identity of the organism. To constitute effective control measures, it is important to know the serotype of the organism. A study was undertaken to characterize the Pasteurella isolates from local pigs in India with clinical respiratory disease by determination of their capsule types and presence or absence of toxin gene. Pasteurella could be isolated from 66.70% of pigs with clinical respiratory disease. All the isolates were confirmed through biochemical characterization and P. multocida-specific polymerase chain reaction. It has also been observed that all the isolates belonged to capsular type D. All the isolates were sensitive to chloramphenicol, chlortetracycline, doxycycline, and enrofloxacin, while the rest of the antibiotics were less effective. It has also been observed that all isolates were resistant to cephalexin, penicillin G, and sulphadiazine. The study revealed the detection of P. multocida serotype D from clinical respiratory diseases of local pigs of India, which could be one of the important respiratory tract pathogens responsible for mortality of local pigs in India.

  9. Cloning and sequence analysis of hsf, an outer membrane protein gene of Pasteurella multocida serotype B:2

    Directory of Open Access Journals (Sweden)

    A. Priyadarshini

    2014-12-01

    Full Text Available Aim: The present study was undertaken to clone, sequence and analyze the hsf, an outer membrane protein gene of Pasteurella multocida serotype B:2 Materials and Methods: hsf gene was amplified from genomic DNA of P. multocida. Polymerase chain reaction (PCR product was cloned in pET-32a vector and was characterized. hsf gene was sequenced, analyzed and phylogenetic tree was constructed taking sequences of other strains. Results: Amplicon size was found to be 785 bp. Recombinant got characterized through colony PCR and restriction enzyme analysis. Conclusion: hsf gene of P. multocida serotype B is similar to serotype A, but different from serotype D. Further work is needed to evaluate role of Hsf protein in protection studies and to study the antigenic properties of this recombinant protein as a candidate for vaccine.

  10. Homogeneity of VacJ outer membrane lipoproteins among Pasteurella multocida strains and heterogeneity among members of Pasteurellaceae.

    Science.gov (United States)

    Shivachandra, Sathish Bhadravati; Kumar, Abhinendra; Mohanty, Nihar Nalini; Yogisharadhya, Revanaiah; Chacko, Nirmal; Viswas, K N; Ramakrishnan, Muthannan Andavar

    2014-06-01

    Outer membrane lipoproteins are widely distributed in Gram-negative bacteria which are involved in diverse mechanisms of physiology/pathogenesis. Various pathogenic bacterial strains belonging to the family-Pasteurellaceae have several surface exposed virulence factors including VacJ/VacJ-like lipoproteins. In the present study, vacJ gene encoding for VacJ outer membrane lipoprotein of different Pasteurella multocida strains (n = 10) were amplified, sequenced and compared with available VacJ/VacJ-like sequences (n = 45) of Pasteurellaceae members. Comparative multiple sequence analysis at amino acid level indicated absolute homogeneity of VacJ lipoprotein among different P. multocida strains. However, heterogeneity (18.0-89.9%) of VacJ lipoprotein was noticed among members of Pasteurellaceae. A predicted lipobox motif (L-3-[A/S/T/V]-2-[G/A]-1-C) was found to be conserved between 12-32aa residues at N-terminus among all VacJ sequences. Bioinformatic analysis indicated that VacJ is a chromosomal gene product exposed on the bacterial surface, possibly essential for either physiological or pathogenicity process of Pasteurellae and distributed widely among P. multocida serogroups. The study indicated potential possibilities of using absolutely conserved VacJ lipoprotein either as 'signature gene/protein' in developing diagnostic assay or as a recombinant subunit vaccine for P. multocida infections in livestock. Copyright © 2014 Elsevier Ltd. All rights reserved.

  11. MLST typing of Pasteurella multocida associated with haemorrhagic septicaemia and development of a real-time PCR specific for haemorrhagic septicaemia associated isolates

    DEFF Research Database (Denmark)

    Petersen, Andreas; Bisgaard, Magne; Townsend, Kirsty

    2014-01-01

    Two serovars of Pasteurella multocida, B:2 and E:2, have been reportedly associated with haemorrhagic septicaemia (HS), a peracute and devastating disease mainly affecting cattle and water buffaloes. We multilocus sequence typed (MLST) 64 isolates of P. multocida including 55 associated with HS...... method. The new HS-est-RT-PCR represents a fast and specific detection of the specific types of P. multocida involved in HS. The HS-est-RT-PCR developed in the current study seems to more accurately identify isolates of P. multocida associated with HS compared to PCR detection methods previously...

  12. Development of a novel multiplex PCR assay for rapid detection of virulence associated genes of Pasteurella multocida from pigs.

    Science.gov (United States)

    Rajkhowa, S

    2015-09-01

    As the pathogenicity of Pasteurella multocida is associated with various virulence factors (VFs), the aim of the study was to develop a novel multiplex PCR (m-PCR) assay for the rapid detection of important virulence associated genes (VAGs) of P. multocida isolates from pigs. The target recognized VFs used in the study were diverse adhesins (ptfA and pfhA), toxins (toxA), siderophores (tonB and hgbA), sialidases (nanB, nanH) and outer membrane proteins (ompA, ompH, oma87 and plpB). The primers for the genes encoding these VFs were designed by primer3 software (http://bioinfo.ut.ee/primer3-0.4.0/) using gene sequences available in Genbank. The detection limit of the developed assay was 10(2)  CFU ml(-1) . The m-PCR did not produce any nonspecific amplification products when tested against Bordetella bronchiseptica which also commonly infects pigs. We applied m-PCR to the field samples, and the results obtained were the same as the single PCR results. The developed assay would be very useful for veterinary diagnostic laboratories and for others interested in the rapid virulence profiling of porcine P. multocida isolates circulating in the piggeries. The study reports the development and evaluation of a novel multiplex PCR assay for the rapid detection of 11 important VAGs of Pasteurella multocida isolates from pigs. Rapid and simultaneous detection of recognized VFs of the organism are essential to know the virulo-types of P. multocida isolates circulating in the piggeries. The developed novel assay will be very useful for the rapid detection of VAGs of P. multocida isolates from pigs. © 2015 The Society for Applied Microbiology.

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

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    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 multocida B:2 than buffaloes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Genetic diversity of porcine Pasteurella multocida strains from the respiratory tract of healthy and diseased swine.

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    Bethe, Astrid; Wieler, Lothar H; Selbitz, Hans-J; Ewers, Christa

    2009-10-20

    A total of 382 porcine Pasteurella multocida strains, isolated from cases of pneumonia and progressive atrophic rhinitis (PAR) as well as from clinically healthy pigs of more than 150 German husbandries were characterized by detection of virulence-associated genes (VAGs) and ribotyping to understand the relationships between "commensal" and "pathogenic" strains, enabling a rational choice of vaccine strains. The diversity of the strains according to VAGs was low and mainly limited to capsular type genes (capA: 53.4%; capD: 45.8%; capF: 0.3%; cap-negative: 0.5%; hssB: 95.3%), dermonecrotoxin gene toxA (3.4%), as well as adhesion-related genes pfhaB (20.9%) and hgbB (84.3%). Ribotyping identified 13 patterns, but the vast majority of strains (95.8%) clustered in only three of these, namely IA-1 (45.5%), IA-7 (30.1%), and IIA-1 (20.2%). Pattern IA-1 was associated with capD(+) strains (93.6%) and harboured the majority of toxA(+) strains (84.6%). Pattern IA-7 mostly contained pfhaB(-), toxA(-)capA(+) strains (93.9%), while pattern IIA-1 was predominantly composed of pfhaB(+), toxA(-)capA(+) strains (87.0%). Clinical strains associated with pneumonia or PAR shared the above mentioned major ribotypes in comparable proportions with strains derived from healthy pigs, suggesting P. multocida to act more as an opportunistic than as an obligate pathogen in pigs. The limited number of subpopulations may either reflect a recent evolution of P. multocida in pigs or a selection by means of horizontal transfer of capsular genes, toxA or pfhaB. These data enforce further phylogenetic and epidemiological studies, examining the properties of different subpopulations of porcine P. multocida strains as well as factors of the porcine hosts themselves, which might be involved in disease susceptibility.

  15. Influence of systemic fluoroquinolone administration on the presence of Pasteurella multocida in the upper respiratory tract of clinically healthy calves

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    Kehrenberg Corinna

    2008-09-01

    Full Text Available Abstract The influence of enrofloxacin administration (5 mg/kg for five consecutive days on the occurrence of Pasteurella multocida in the upper respiratory tract of two healthy calves was monitored over a 10-day period. From nasal swabs of two additional healthy control calves, which received a placebo saline administration, P. multocida was isolated throughout the study period. In the enrofloxacin treated calves, P. multocida was not demonstrated in the nasopharynx from 48 h after the first injection until two days after the last administration, when P. multocida reappeared and proved to be clonal in nature to the original isolates. During the experiment, no change in minimal inhibitory concentration for enrofloxacin of the P. multocida isolates was detected (MIC ≤ 0.015 μg/mL. Enrofloxacin concentrations were determined in the plasma by a high-performance liquid chromatography method with fluorescence detection. The PK/PD indices AUC/MIC and Cmax/MIC ratio were calculated and found to be 1157.7 and 129.8, respectively. Remarkably, the respiratory pathogen Arcanobacterium pyogenes became the predominant recovered organism in the nasopharynx of one animal following enrofloxacin therapy throughout the remaining of the experiment.

  16. New interpretive criteria for danofloxacin antibacterial susceptibility testing against Mannheimia haemolytica and Pasteurella multocida associated with bovine respiratory disease.

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    Sweeney, Michael T; Papich, Mark G; Watts, Jeffrey L

    2017-03-01

    Danofloxacin is a fluoroquinolone antibacterial agent approved for use in veterinary medicine to treat and control bovine respiratory disease caused by Mannheimia haemolytica or Pasteurella multocida. Susceptible minimal inhibitory concentration (MIC) breakpoint (≤0.25 µg/mL) and disk diffusion interpretive criteria (≥22 mm) values for danofloxacin against M. haemolytica and P. multocida were first approved by the Clinical and Laboratory Standards Institute (CLSI) in 2003. However, intermediate and resistant breakpoint values were not established because only susceptible wild-type populations were evident at the time of breakpoint approvals. Since then, nonsusceptible isolates of M. haemolytica and P. multocida have been identified. We report danofloxacin intermediate MIC breakpoint (0.5 µg/mL) and disk diffusion interpretive criteria (18-21 mm), as well as danofloxacin-resistant MIC breakpoint (≥1 µg/mL) and disk diffusion interpretive criteria (≤17 mm), based on scattergram plots of MIC values versus disk zone diameters and calculated error-bound rates using M. haemolytica and P. multocida isolates recovered from bovine respiratory disease in North America in 2004-2014. These newly established intermediate and resistant clinical breakpoint values have been endorsed by CLSI and can be used for interpreting results from antibacterial susceptibility testing of danofloxacin against M. haemolytica and P. multocida isolated from bovine respiratory disease.

  17. Serum resistance of Pasteurella multocida in avian and porcine sera, and comparative virulence investigations of selected serum-sensitive and resistant strains in chickens.

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    Muhairwa, Amandus P; Christensen, Jens P; Bisgaard, Magne

    2002-04-01

    Growth in serum of Pasteurella multocida and related species in chicken, turkey, duck and pig sera were compared, and selected serum-resistant and serum-sensitive strains were inoculated into 18-week-old layers. Eighty-seven field strains of Pasteurella spp. and nine reference strains representing different clones defined by restriction endonuclease analysis (REA) profiles were used in the study. Serum activity was measured by changes in the optical density (OD) of the serum after inoculation and incubation at 41 degrees C for chicken, turkey and duck serum and 39 degrees C for pig serum. Serum activity was measured by comparison with previously determined serum-resistant (P-1059) and serum-sensitive (CU vaccine) strains, and classified into highly serum-resistant, moderately serum-resistant and serum-sensitive. Strains of the same REA type were found to have identical growth curves and the same maximum OD values when tested in serum from the same host species. Turkey serum was shown to be less inhibitory to a wide range of P. multocida strains than chicken, duck and pig sera. Serum-resistant strains were demonstrated among avian as well as mammalian strains. Among the avian strains, the proportion of serum-resistant strains was higher in outbreak strains than in strains from apparently healthy carriers. Removal of the capsule from selected strains by hyaluronidase treatment failed to change the serum activity. The most severe lesions in experimentally infected chickens were produced by a serum-resistant strain; however, lesions were also found in chickens infected by serum-sensitive strains, indicating the involvement of multiple factors in the virulence of P. multocida. Further investigations on serum resistance are indicated in order to relate other host and bacterial factors responsible for the development of fowl cholera.

  18. Multilocus sequence typing of a global collection of Pasteurella multocida isolates from cattle and other host species demonstrates niche association

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    Lainson F Alex

    2011-05-01

    Full Text Available Abstract Background Pasteurella multocida causes disease in many host species throughout the world. In bovids, it contributes to bovine respiratory disease (BRD and causes haemorrhagic septicaemia (HS. Previous studies have suggested that BRD-associated P. multocida isolates are of limited diversity. A multilocus sequence typing (MLST scheme for P. multocida was used to determine whether the low levels of diversity reported are due to the limited discriminatory power of the typing method used, restricted sample selection or true niche association. Bovine respiratory isolates of P. multocida (n = 133 from the UK, the USA and France, collected between 1984 and 2008 from both healthy and clinically affected animals, were typed using MLST. Isolates of P. multocida from cases of HS, isolates from other host species and data from the MLST database were used as comparison. Results Bovine respiratory isolates were found to be clonal (ISA 0.45 with 105/128 belonging to clonal complex 13 (CC13. HS isolates were not related to bovine respiratory isolates. Of the host species studied, the majority had their own unique sequence types (STs, with few STs being shared across host species, although there was some cross over between porcine and bovine respiratory isolates. Avian, ovine and porcine isolates showed greater levels of diversity compared to cattle respiratory isolates, despite more limited geographic origins. Conclusions The homogeneity of STs of bovine respiratory P. multocida observed, and the differences between these and P. multocida subpopulations from bovine non-respiratory isolates and non-bovine hosts may indicate niche association.

  19. Ducks as a potential reservoir for Pasteurella multocida infection detected using a new rOmpH-based ELISA.

    Science.gov (United States)

    Liu, Rongchang; Chen, Cuiteng; Cheng, Longfei; Lu, Ronghui; Fu, Guanghua; Shi, Shaohua; Chen, Hongmei; Wan, Chunhe; Lin, Jiansheng; Fu, Qiuling; Huang, Yu

    2017-07-28

    Pasteurella multocida is an important pathogen of numerous domestic poultry and wild animals and is associated with a variety of diseases including fowl cholera. The aim of this study was to develop an indirect enzyme-linked immunosorbent assay (ELISA) based on recombinant outer-membrane protein H (rOmpH) for detection of anti-P. multocida antibodies in serum to determine their prevalence in Chinese ducks. The P. multocida ompH gene was cloned into pET32a, and rOmpH was expressed in Escherichia coli BL21 (DE3). Western blotting revealed that purified rOmpH was recognized by duck antisera against P. multocida, and an indirect ELISA was established. During analysis of serum samples (n=115) from ducks, the rOmpH ELISA showed 95.0% specificity, 100% sensitivity and a 92.0% κ coefficient (95% confidence interval 0.844-0.997) as compared with a microtiter agglutination test. Among 165 randomly selected serum samples, which were collected in 2015 and originated from six duck farms across Fujian Province, China, anti-P. multocida antibodies were detected in 22.42% of apparently healthy ducks, including 25 of 90 sheldrakes (27.8%), eight of 50 Peking ducks (16.0%) and four of 25 Muscovy ducks (16%). Overall, the data suggest that rOmpH is a suitable candidate antigen for the development of an indirect ELISA for detection of P. multocida in ducks; moreover, our results showed that ducks could serve as a potential reservoir for P. multocida infection.

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

    Science.gov (United States)

    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. Copyright © 2016 Elsevier Ltd. All rights reserved.

  1. Virulence genotype of Pasteurella multocida strains isolated from different hosts with various disease status.

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    Ewers, Christa; Lübke-Becker, Antina; Bethe, Astrid; Kiebling, Sabine; Filter, Matthias; Wieler, Lothar H

    2006-05-31

    To learn more about the molecular biology of Pasteurella multocida 289 strains isolated from various clinically healthy and diseased hosts were examined for capsule biosynthesis genes (capA, B, D, E, and F) and 14 virulence associated genes by PCR and DNA-DNA-hybridization. As expected, capsule type A strains were highly adapted to bovines (92.3%) and poultry (85.7%) while we mainly found capA (34.9%)- and capD (58.1%)-positive strains in swine. A noticeable amount of capD-positive strains also originated from small ruminants (34.9%) and capF was detected in wild type strains from diseased cattle (2.2%) and cats (7.4%). None of the isolates harboured capE, while capB was exclusively found in all strains from buffaloes. Nearly all isolates showed a combination of genes encoding outer membrane proteins, colonization factors, iron aquisition factors and superoxid-dismutases without any clue for host specificity. In contrast, the transferrin binding protein encoding gene tbpA (31.5%) was limited to ruminant strains and only 37.0% of all P. multocida strains harboured pfhA, coding for a filamentous hemagglutinin, supposed to be a putative adhesion- und serum resistance factor. PfhA revealed a strong positive association to the outcome of disease in bovine hosts and in combination with toxA to that in swine. The dermonecrotoxin encoding toxA, present in 12.5% of all strains, was detected in isolates from swine, small ruminants, cattle, and poultry. A significant association to the disease status, however, was only existent in swine, although with 66.7% we found a notably high prevalence of the toxin gene among strains from small ruminants. The genes toxA, tbpA and pfhA as well as capsule biosynthesis genes are supposed to be important epidemiological marker genes for characterizing P. multocida field strains.

  2. Development of loop-mediated isothermal amplification-based diagnostic assays for detection of Pasteurella multocida and hemorrhagic septicemia-associated P multocida serotype B:2.

    Science.gov (United States)

    Moustafa, Ahmed M; Bennett, Mark D

    2017-02-01

    OBJECTIVE To develop 2 rapid loop-mediated isothermal amplification (LAMP) assays for detection of Pasteurella multocida DNA (Pm-LAMP assay) and P multocida DNA from strains associated with hemorrhagic septicemia (HS) in cattle and buffalo (HS-LAMP assay). SAMPLE Solutions containing 16 P multocida strains and 9 other bacterial species at various concentrations. PROCEDURES Optimal conditions were determined for running the Pm-LAMP and HS-LAMP assays. The assays were then used to detect DNA of the test organisms. Results of LAMP assays were validated against conventional PCR assays designed for specific detection of P multocida and the B:2 serotype of HS-associated strains. RESULTS Following incubation of sample reaction mixtures for 27 minutes, specificity and sensitivity of the HS-LAMP assay at template DNA amounts as low as 5 pg were 93% and 97%, respectively. When duplicates of each sample were incubated for 28 minutes (a positive result defined as positive results for both reactions of a given sample), specificity and sensitivity of the HS-LAMP assay in the same conditions increased to 100%. The best specificity and sensitivity of Pm-LAMP single (93% and 91%) and duplicate (97% and 98%) reactions at template DNA amounts as low as 10 pg were achieved at 33 and 34 minutes, respectively. CONCLUSIONS AND CLINICAL RELEVANCE These preliminary findings suggested the developed HS-LAMP assay had high sensitivity and specificity for detection of HS-associated P multocida. Additional research is needed to determine the accuracy of the assay for use on clinical specimens obtained in HS-endemic countries such as Pakistan and Thailand.

  3. Cellular fatty acid composition of Haemophilus species, Pasteurella multocida, Actinobacillus Actinomycetemcomitans and Haemophilus vaginalis (Corynebacterium vaginale).

    Science.gov (United States)

    Jantzen, E; Berdal, B P; Omland, T

    1980-04-01

    The fatty acid composition of 35 Haemophilus influenzae strains was found to be grossly similar and characterized by relatively large amounts of 14:0, 3-OH-14:0, 16:1 and 16:0. The three C18 fatty acids 18:2, 18:1 and 18:0 were also present, but in much lower concentrations. This general pattern was also found for most of the other species of Haemophilus examined (H. aegyptius, H. aphrophilus, H. canis, H. gallinarum, H. haemolyticus, and H. parainfluenzae). Small but distinct quantitative discrepancies were detected for H. ducreyi and the haemin-independent species H. paraphrohaemolyticus, H. paraphrophilus and H. suis. Actinobacillus actinomycetemcomitans was found to be indistinguishable from H. influenzae. Pasteurella multocida also exhibited a fatty acid pattern closely related to that of Haemophilus, but could be distinguished by its higher concentration levels of the C18 fatty acids. The fatty acid pattern of H. vaginalis was considerably different from those of the other species examined. This species lacked 3-OH-14:0 and 18:2 and contained small amounts of 14:0 and 16:0, whereas 18:1 and 18:0 were the major constituents.

  4. Virulence genes and antimicrobial susceptibility in Pasteurella multocida isolates from calves.

    Science.gov (United States)

    Katsuda, K; Hoshinoo, K; Ueno, Y; Kohmoto, M; Mikami, O

    2013-12-27

    A total of 378 isolates of Pasteurella multocida from clinically healthy and diseased calves were characterised for their susceptibility to 9 antimicrobial agents and screened by PCR for the presence of antimicrobial resistance genes and 22 genes virulence-associated, including capsule biosynthesis genes. Of the 378 isolates, 102 (27.0%) were resistant to at least one of the 9 tested antimicrobial agents. Resistance to oxytetracycline (21.7%) was the most frequently observed phenotype among the isolates. The tet(H) gene were the primary determinant detected. The resistance rates for thiamphenicol, ampicillin, kanamycin and florfenicol were 13.2%, 5.8%, 9.0% and 0.5%, respectively. Cefazolin, ceftiofur, cefquinome and enrofloxacin were effective antimicrobial agents, with no resistant isolates emerging over the course of the investigation. Most isolates were identified as capsular type A, only 6.3% belonged to capsular type D and no other capsular type was identified. Four of the virulence-associated genes (pfhA, tadD, tbpA and HAS) exhibited associations to the capsular type, and three (pfhA, tbpA and hgbB) were associated with the disease status of the animals. These virulence genes have been considered as epidemiological markers and are hypothesised to have a strong positive association with the outcome of disease in cattle. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Molecular Epidemiology of Pasteurella multocida Circulating in India by Multilocus Sequence Typing.

    Science.gov (United States)

    Sarangi, L N; Thomas, P; Gupta, S K; Kumar, S; Viswas, K N; Singh, V P

    2016-04-01

    Multilocus sequence typing (MLST), a sequence-based typing method for bacterial pathogens, is currently the best method for long-term epidemiological study and to understand the population structure of the bacteria. This investigation was carried out to study the diversity of Pasteurella multocida isolates circulating in India. Ten different sequence types (ST) identified in this study are ST 122 from cattle, goat, mithun and pig; ST 50 from pig; ST 9 from cattle and sheep; ST 229 from cattle and goat; ST 71 and ST 277 from cattle; and ST 129, ST 280, ST 281 and ST 282 from avian species. Of these, ST 277, ST 280, ST 281 and ST 282 were identified for the first time. The analysis of results provides novel epidemiological information on the circulation of multiple STs across India. The majority of STs or their variants identified in this study have already been reported from different parts of the globe. This suggests that probably transboundary spread of strains across countries and continents has occurred across evolutionary time and is still happening. The isolation of ST 122 from small ruminants and pigs suggests that these species may be included in the preventive vaccination policy for effective control of haemorrhagic septicaemia in India. © 2014 Blackwell Verlag GmbH.

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

    Science.gov (United States)

    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.

  7. Factors affecting endotoxin release from the cell surface of avian strains of Pasteurella multocida.

    Science.gov (United States)

    Lee, M D; Glisson, J R; Wooley, R E

    1992-06-15

    Two avian strains of Pasteurella multocida, a vaccine strain and a virulent field isolate, were investigated to determine their propensity to release endotoxin from the cell surface. Both organisms released comparable amounts of endotoxin when plasma complement proteins were present, however the virulent strain did so without the loss of viability that occurred in the vaccine strain. Blocking complement activity decreased the ability of plasma to elicit endotoxin release from the bacteria. When the cells were treated with divalent metal chelators such as trans-1, 2-diaminocyclohexane-N,N,N1,N1-tetraacetic acid (CDTA), more endotoxin was released from the vaccine strain than from the virulent isolate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of purified lipopolysaccharide (LPS) from both strains revealed virtually identical patterns. Both had patterns considered typical of rough LPS. Challenge studies in 8 weeks old turkeys showed that the field strain induced endotoxemia of longer duration than the vaccine strain and produced greater mortality.

  8. ESTUDIOS DE AISLAMIENTO Y FRACCIONAMIENTO DE UN COMPLEJO GLICOPROTEICO DE PASTEURELLA MULTOCIDA

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    Yolanda de Navarro

    2010-07-01

    Full Text Available Mediante la extracción con solución de NaCl 2.5% (p/v a 47' C, se obtuvo un complejo glicoproteíco de Pasteurella multocida que fue parcialmente purificado mediante filtración por gel usando Sephacryl S-200. Las fracciones 1, 2 y 3 presentaron líneas de precipitinas en imnunodifusión contra sueros hiperimnunes de conejos inoculados con extracto salino 2.5% (P/V a 47" C y 66' C. Por electroforesis SDS-PAGE. se determinaron bandas de proteínas con pesos moleculares entre 98.800 y 17.700 daltons. Lafracción lyel extracto salino crudo a 47" C se utilizaron como antígenos adsorbidos en gel de Al(OHj y con ellos se efectuaron ensayos de protección en ratones, teniendo como referencia la vacuna comercial contra Septicemia Hemorrágica producida por VECOL S.A. Mediante el método estadístico de Reed Muench se estableció el índice de protección y se encontró que todos los antígenos fueron considerados prolectores, siendo la fracción 1 la de mayor índice de protección con una dosis inferior.

  9. Increased fucosyl glycoconjugate by Mycoplasma hyopneumoniae enhances adherences of Pasteurella multocida type A in the ciliated epithelial cells of the respiratory tract.

    Science.gov (United States)

    Park, Changhoon; Jeong, Jiwoon; Kang, Ikjae; Choi, Kyuhyung; Park, Su-Jin; Chae, Chanhee

    2016-02-03

    The objective of this study was to elucidate the pathogenic mechanisms of how Mycoplasma hyopneumoniae enhances secondary Pasteurella multocida type A infection which leads to porcine enzootic pneumonia in infected pigs. Sixteen pigs were experimentally infected with M. hyopneumoniae and then euthanized at 7, 14, 21 and 28 days post inoculation. In situ hybridization for M. hyopneumoniae DNA and Ulex europaeus agglutinin-I (UEA-I) lectin histochemistry for fucosyl glycoconjugate, was performed in serial lung sections to determine alteration of fucosyl glycoconjugate in M. hyopneumoniae-infected bronchial and bronchiolar epithelium. Bacterial overlay assay was performed to determine the affinity of P. multocida type A with L-fucose. The luminal surface of bronchial and bronchiolar epithelial cells that were stained with UEA-I always showed hybridization signals for M. hyopneumoniae but it was negative in the unaffected parts of the lung from M. hyopneumoniae-infected pigs and in lung from negative control pigs. Colocalization of M. hyopneumoniae and UEA-I was especially prominent in the luminal surface of bronchial and bronchiolar epithelial cells in serial section of lung. The mean number of M. hyopneumoniae-positive cells correlated with the mean number of UEA-I-positive cells in lungs from infected pigs throughout the experiment. All eight P. multocida type A isolates from naturally occurring enzootic pneumonia, bound strongly at levels of 2 μg and 5 μg of L-fucose. The results of the present study demonstrate that M. hyopneumoniae increases the L-fucose composition to enhance adherence of P. multocida type A to the bronchial and bronchiolar epithelial cells.

  10. Genomic DNA restriction endonuclease from Pasteurella multocida isolated from Indonesia, katha strain and reference strains and analysed by PFGE

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    Supar

    2003-10-01

    Full Text Available Pasteurella multocida strains are the causative disease agents of wide range of domestic and wild animals in Indonesia. The most important serotypes are associated with Hemorrhagic septicaemic (HS diseases in cattle and buffaloes, cholera in ducks and chickens. The HS disease associated with P. multocia in large ruminants in Indonesia is controled by killed whole cell vaccines produced by the use of P. multocida Katha strains. There is no discriminatory data of the molecular biology technique has been applied to investigate P. multocida isolates from different geographic locations in Indonesia. The purpose of this studies were to observe the genetic diversity among P. multocida isolated from various geograpic locations and compared with Katha vaccine strain and other reference strains. A total samples of 38 isolates and strains of P. multocida were analysed by means of pulsed-field gel electrophoresis (PFGE. Each sample was grown in nutrient broth, cells were separeted by centrifugation. Whole cell pellet was mixed with agarose and then prepared agarose plugs. The genomic DNA of each sample was digested in situ (plug with either restriction endonuclease of ApaI and/or BamHI. The digested genomic DNA of each sample was analysed by PFGE, the genomic DNA restricted profile of each sample was compared with others. The use of ApaI restriction endonuclease digestion and analysed by PFGE, demonstrated that 34 out of 38 P. multocia samples could be differentiated into 16 ApaI types, whereas based on the BamHI digestion of these samples were differentiated into 20 BamHI types. Genomic DNA restriction pattern of Indonesian P. multocida isolates originated from cattle and buffaloes associated with haemorrhagic septicaemic diseases demonstrated different pattern to those of vaccine Katha strain, poultry strains as well as the reference strains currenly kept at Balitvet Culture Collection (BCC unit. Two P. multocida isolates derived from ducks with cholera

  11. Signature-tagged mutagenesis of Pasteurella multocida identifies mutants displaying differential virulence characteristics in mice and chickens.

    Science.gov (United States)

    Harper, Marina; Boyce, John D; Wilkie, Ian W; Adler, Ben

    2003-09-01

    Pasteurella multocida is the causative agent of fowl cholera in birds. Signature-tagged mutagenesis (STM) was used to identify potential virulence factors in a mouse septicemia disease model and a chicken fowl cholera model. A library of P. multocida mutants was constructed with a modified Tn916 and screened for attenuation in both animal models. Mutants identified by the STM screening were confirmed as attenuated by competitive growth assays in both chickens and mice. Of the 15 mutants identified in the chicken model, only 5 were also attenuated in mice, showing for the first time the presence of host-specific virulence factors and indicating the importance of screening for attenuation in the natural host.

  12. Cloning and characterisation of the Pasteurella multocida ahpA gene responsible for a haemolytic phenotype in Escherichia coli.

    Science.gov (United States)

    Cox, A J; Hunt, M L; Ruffolo, C G; Adler, B

    2000-03-01

    Haemolysins are membrane-damaging agents which have been described as bacterial virulence factors due to their ability to lyse erythrocytes and other host cells, and therefore inducing a greater inflammatory response (Elliott et al., 1998). Pasteurella multocida was found to be haemolytic under anaerobic conditions. In this study, we cloned and characterised a P. multocida gene, designated ahpA, which conferred a haemolytic phenotype on Escherichia coli when incubated under anaerobic conditions. A deletion was introduced into the ahpA open reading frame which abolished the haemolytic phenotype. The clone containing ahpA showed erythrocyte specificity, causing haemolysis of bovine and equine erythrocytes, and demonstrated weak haemolysis on ovine erythrocytes. Upon further investigation, AhpA was found to affect the expression of the E. coli K-12 latent haemolysin, SheA, under anaerobic conditions.

  13. Macrolide resistance conferred by rRNA mutations in field isolates of Mannheimia haemolytica and Pasteurella multocida.

    Science.gov (United States)

    Olsen, Anders S; Warrass, Ralf; Douthwaite, Stephen

    2015-02-01

    To determine how resistance to macrolides is conferred in field isolates of Pasteurella multocida and Mannheimia haemolytica that lack previously identified resistance determinants for rRNA methylation, efflux and macrolide-modifying enzymes. Isolates of P. multocida and M. haemolytica identified as being highly resistant (MICs >64 mg/L) to the macrolides erythromycin, gamithromycin, tilmicosin, tildipirosin and tulathromycin were screened by multiplex PCR for the previously identified resistance genes erm(42), msr(E) and mph(E). Strains lacking these determinants were analysed by genome sequencing and primer extension on the rRNAs. Macrolide resistance in one M. haemolytica isolate was conferred by the 23S rRNA mutation A2058G; resistance in three P. multocida isolates were caused by mutations at the neighbouring nucleotide A2059G. In each strain, all six copies of the rrn operons encoded the respective mutations. There were no mutations in the ribosomal protein genes rplD or rplV, and no other macrolide resistance mechanism was evident. High-level macrolide resistance can arise from 23S rRNA mutations in P. multocida and M. haemolytica despite their multiple copies of rrn. Selective pressures from exposure to different macrolide or lincosamide drugs presumably resulted in consolidation of either the A2058G or the A2059G mutation. © The Author 2014. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

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

    Science.gov (United States)

    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.

  15. A novel quantitative real-time polymerase chain reaction method for detecting toxigenic Pasteurella multocida in nasal swabs from swine.

    Science.gov (United States)

    Scherrer, Simone; Frei, Daniel; Wittenbrink, Max Michael

    2016-12-01

    Progressive atrophic rhinitis (PAR) in pigs is caused by toxigenic Pasteurella multocida. In Switzerland, PAR is monitored by selective culture of nasal swabs and subsequent polymerase chain reaction (PCR) screening of bacterial colonies for the P. multocida toxA gene. A panel of 203 nasal swabs from a recent PAR outbreak were used to evaluate a novel quantitative real-time PCR for toxigenic P. multocida in porcine nasal swabs. In comparison to the conventional PCR with a limit of detection of 100 genome equivalents per PCR reaction, the real-time PCR had a limit of detection of 10 genome equivalents. The real-time PCR detected toxA-positive P. multocida in 101 samples (49.8%), whereas the conventional PCR was less sensitive with 90 toxA-positive samples (44.3%). In comparison to the real-time PCR, 5.4% of the toxA-positive samples revealed unevaluable results by conventional PCR. The approach of culture-coupled toxA PCR for the monitoring of PAR in pigs is substantially improved by a novel quantitative real-time PCR.

  16. Structural characterization of Fis - A transcriptional regulator from pathogenic Pasteurella multocida essential for expression of virulence factors.

    Science.gov (United States)

    Bagchi, Angshuman

    2015-01-10

    Pasteurella multocida is responsible behind a variety of diseases in animals. The disease causing substance of this bacterium is a capsular polysaccharide. The expression of the gene that codes for the bacterial capsule is regulated by the protein Fis. Fis also regulates the expression of various different genes in P. multocida. So far there have been no previous reports that depict the characterization of Fis from P. multocida from a structural point of view. In the present work, an attempt has been made to characterize Fis by in silico methods. The structure of Fis was built by comparative modeling technique. The model of Fis was then docked onto the corresponding promoter regions of the gene encoding the capsular polysaccharide. The docked complexes of promoter DNA with Fis protein were subjected to molecular dynamics simulations to identify the mode of DNA-protein interactions. The DNA binding amino acid residues from the Fis protein were identified. And a mechanistic detail of the DNA binding interactions was predicted. So far, this is the first report that depicts the mechanistic details of Fis-DNA interactions involved in the regulation of gene expression by Fis protein. This work may therefore be useful to illuminate the still obscure molecular mechanism behind the disease propagation by P. multocida. Copyright © 2014 Elsevier B.V. All rights reserved.

  17. Cloning and characterization of sialidases with 2-6' and 2-3' sialyl lactose specificity from Pasteurella multocida.

    Science.gov (United States)

    Mizan, S; Henk, A; Stallings, A; Maier, M; Lee, M D

    2000-12-01

    Pasteurella multocida is a mucosal pathogen that colonizes the respiratory system of susceptible hosts. Most isolates of P. multocida produce sialidase activity, which may contribute to colonization of the respiratory tract or the production of lesions in an active infection. We have cloned and sequenced a sialidase gene, nanH, from a fowl cholera isolate of P. multocida. Sequence analysis of NanH revealed that it exhibited significant amino acid sequence homology with many microbial sialidases. Insertional inactivation of nanH resulted in a mutant strain that was not deficient in sialidase production. However, this mutant exhibited reduced enzyme activity and growth rate on 2-3' sialyl lactose compared to the wild type. Subsequently, we demonstrated the presence of two sialidases by cloning another sialidase gene that differed from nanH in DNA sequence and substrate specificity. NanB demonstrated activity on both 2-3' and 2-6' sialyl lactose, while NanH demonstrated activity only on 2-3' sialyl lactose. Neither enzyme liberated sialic acid from colominic acid (2-8' sialyl lactose). Recombinant E. coli containing the sialidase genes were able to utilize several sialoconjugants when they were provided as sole carbon sources in minimal medium. These data suggest that sialidases have a nutritional function and may contribute to the ability of P. multocida to colonize and persist on vertebrate mucosal surfaces.

  18. Molecular diversity of Pasteurella multocida isolated from cattle and buffaloes in East Azerbaijan province based on restriction endonuclease analysis

    Directory of Open Access Journals (Sweden)

    jalal shayegh

    2014-05-01

    Full Text Available In order to increase information about the molecular diversity of Pasteurella multocida isolated from cattle and buffalo, 2 buffalo and 8 cattle isolates were investigated by Restriction Endonuclease Analysis (REA. REA was performed with Hha-I Endonuclease which established 2 distinct profiles: I and II.  Cattle and buffalo isolates fell into both REA profiles. Contrary to previous studies, the genetic diversity of the isolates was negligible. Considering the similarity of cattle and buffalo isolates is the present study, further studies witch larger samples should be carried out to investigate the possibility of inter-species transmission.

  19. The Pasteurella multocida nrfE Gene Is Upregulated during Infection and Is Essential for Nitrite Reduction but Not for Virulence

    OpenAIRE

    Boucher, David J.; Adler, Ben; Boyce, John D.

    2005-01-01

    Pasteurella multocida is the causative agent of a range of diseases with economic importance in production animals. Many systems have been employed to identify virulence factors of P. multocida, including in vivo expression technology (IVET), signature-tagged mutagenesis, and whole-genome expression profiling. In a previous study in which IVET was used with P. multocida, nrfE was identified as a gene that is preferentially expressed in vivo. In Escherichia coli, nrfE is part of the formate-de...

  20. Prevalence of different OmpH-types among Pasteurella multocida isolated from lungs of calves with respiratory problems.

    Science.gov (United States)

    Nefedchenko, Alexey V; Glotova, Tatyana I; Glotov, Alexander G; Ternovoy, Vladimir A; Sementsova, Alexandra O

    2017-03-01

    Pasteurella multocida is among the most important respiratory pathogens of cattle. Outer-membrane protein (OmpH) constitutes an essential bacterial antigen and is well studied in avian bacterial strains. Studies on isolates from cattle with signs of respiratory disease caused by Pasteurella multocida serotypes A and D have not yet been covered in the literature. The objective of this study was a comparative analysis of the ompH gene sequences from 83 isolates and four Russian reference strains of P. multocida to assign them to the allelic variants of the gene (OmpH-types). In addition, the above P. multocida strains have been characterized on the basis of capsular serotypes and virulence-associated genes. The isolates were classified into the OmpH -types based on allele specific PCR and gene fragment sequencing. The isolates of capsular serotype A have been subdivided into 6 OmpH -types, of which the most common types identified were A1 and A2. All capsular serotype D isolates belong to the same OmpH-type (D1). On 16 of a total of 23 farms all isolates belong to only one OmpH-type, on 4 farms - to 2, and on 3 farms - to three OmpH-types. The tbpA and pfhA genes were found more often in the isolates of capsular group А as compared to capsular group D (p ≤ 0.05). OmpH-types of serogroup А differ significantly (p ≤ 0.05) among themselves by the prevalence of the pfhA and hgbB genes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Analysis of the polymerization initiation and activity of Pasteurella multocida heparosan synthase PmHS2, an enzyme with glycosyltransferase and UDP-sugar hydrolase activity

    NARCIS (Netherlands)

    Chavaroche, A.A.E.; Broek, van den L.A.M.; Springer, J.; Boeriu, C.; Eggink, G.

    2011-01-01

    Heparosan synthase catalyzes the polymerization of heparosan [-4GlcUAß1-4GlcNAca1-]n by transferring alternatively the monosaccharide units from UDP-GlcUA and UDP-GlcNAc to an acceptor molecule. Details on the heparosan chain initiation by Pasteurella multocida heparosan synthase PmHS2 and its

  2. Antimicrobial susceptibility, serotypes and genotypes of Pasteurella multocida isolates associated with swine pneumonia in Taiwan.

    Science.gov (United States)

    Yeh, Jih-Ching; Lo, Dan-Yuan; Chang, Shao-Kuang; Chou, Chi-Chung; Kuo, Hung-Chih

    2017-09-21

    Pasteurella multocida (PM) can cause progressive atrophic rhinitis and suppurative bronchopneumonia in pigs. The present study performed antimicrobial susceptibility testing and serotype and genotype identification on the 62 PM strains isolated from the lungs of diseased pigs with respiratory symptoms. Antimicrobial susceptibility testing examined 13 antimicrobial agents (amoxicillin, cefazolin, doxycycline, flumequine, enrofloxacin, florfenicol, kanamycin, lincomycin, Linco-Spectin (lincomycin and spectinomycin), erythromycin, tylosin, tilmicosin and tiamulin). Antimicrobial resistance ratios were over 40% in all of the antimicrobial agents except for cefazolin. The highest levels of resistance (100%) were found for kanamycin, erythromycin and tylosin. The majority of isolated strains was serotype D:L6 (n=35) followed by A:L3 (n=17). Comparison of the antimicrobial resistance levels between the two serotypes showed that the antimicrobial resistance rates were higher in D:L6 than in A:L3 for all the tested antimicrobials except for tylosin and tilmicosin. For PM with erm(B), erm(T) or erm(42), the results showed no significant difference compared with non-resistance gene strains in phenotype. Pulsed-field gel electrophoresis genotyping using ApaI restriction digestion of the genomic DNA demonstrated that there were 17 distinct clusters with a similarity of 85% or more, and the genotyping result was similar to that of serotyping. The results of the present study demonstrated that the PM isolated from diseased pigs in Taiwan was resistant to multiple antimicrobials, and the distribution of antimicrobial resistance was associated with pulsotype and serotype. © British Veterinary Association (unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

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

    Science.gov (United States)

    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

  4. Modulating the regioselectivity of a Pasteurella multocida sialyltransferase for biocatalytic production of 3'- and 6'-sialyllactose.

    Science.gov (United States)

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

    2015-10-01

    Several bacterial sialyltransferases have been reported to be multifunctional also catalysing sialidase and trans-sialidase reactions. In this study, we examined the trans-sialylation efficacy and regioselectivity of mutants of the multifunctional Pasteurella multocida sialyltransferase (PmST) for catalysing the synthesis of 3'- and 6'-sialyllactose using casein glycomacropeptide as sialyl-donor and lactose as acceptor. The mutation P34H led to a 980-fold increase in α-2,6-sialyltransferase activity (with cytidine-5'-monophospho-N-acetylneuraminic acid as donor), while its α-2,3-sialyltransferase activity was abolished. Histidine in this position is conserved in α-2,6-sialyltransferases and has been suggested, and recently confirmed, to be the determinant for strict regiospecificity in the sialyltransferase reaction. Our data verified this theorem. In trans-sialidase reactions, the P34H mutant displayed a distinct preference for 6'-sialyllactose synthesis but low levels of 3'-sialyllactose were also produced. The sialyllactose yield was however lower than when using PmSTWT under optimal conditions for 6'-sialyllactose formation. The discrepancy in regiospecificity between the two reactions could indicate subtle differences in the substrate binding site in the two reactions. In contrast, the two mutations E271F and R313Y led to preferential synthesis of 3'-sialyllactose over 6'-sialyllactose and the double mutant (PmSTE271F/R313Y) exhibited the highest α-2,3-regioselectivity via reduced sialidase and α-2,6-trans-sialidase activity. The double mutant PmSTE271F/R313Y thus showed the highest α-2,3-regioselectivity and constitutes an interesting enzyme for regioselective synthesis of α-2,3-sialylated glycans. This study has expanded the understanding of the structure-function relationship of multifunctional, bacterial sialyltransferases and provided new enzymes for regioselective glycan sialylation. Copyright © 2015 Elsevier Inc. All rights reserved.

  5. Florfenicol as a modulator enhancing antimicrobial activity: example using combination with thiamphenicol against Pasteurella multocida

    Directory of Open Access Journals (Sweden)

    Chia-Fong eWei

    2016-03-01

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Eric Lim Teik Chung

    2015-06-01

    Full Text Available 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, gastrointestinal 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

  7. Effects of Ammonia Inhalation and Acetic Acid Pretreatment on Colonization Kinetics of Toxigenic Pasteurella multocida within Upper Respiratory Tracts of Swine

    Science.gov (United States)

    Hamilton, T. D. C.; Roe, J. M.; Hayes, C. M.; Webster, A. J. F.

    1998-01-01

    Pigs reared in intensive production systems are continuously exposed to ammonia released by the microbial degradation of their excrement. Exposure to this gas has been shown to increase the severity of the disease progressive atrophic rhinitis by facilitating colonization of the pig’s upper respiratory tract by Pasteurella multocida. The etiological mechanism responsible for this synergy was investigated by studying the colonization kinetics of P. multocida enhanced by ammonia and comparing them with those evoked by an established disease model. Three-week-old Large White piglets were weaned and allocated to five experimental groups (groups A to E). Pigs in groups A and B were exposed continuously to ammonia at 20 ppm for the first 2 weeks of the study. Pigs in group C were pretreated with 0.5 ml of 1% acetic acid per nostril on days −2 and −1 of the study. On day 0 all the pigs in groups A, C, and D were inoculated with 1.4 × 108 toxigenic P. multocida organisms given by the intranasal route. The kinetics of P. multocida colonization were established by testing samples obtained at weekly intervals throughout the study. The study was terminated on day 37, and the extent of turbinate atrophy was determined by using a morphometric index. The results of the study showed that exposure to aerial ammonia for a limited period had a marked effect on the colonization of toxigenic P. multocida in the nasal cavities of pigs, which resulted in the almost total exclusion of commensal flora. In contrast, ammonia had only a limited effect on P. multocida colonization at the tonsil. The exacerbation of P. multocida colonization by ammonia was restricted to the period of ammonia exposure, and the number of P. multocida organisms colonizing the upper respiratory tract declined rapidly upon the cessation of exposure to ammonia. During the exposure period, the ammonia levels in mucus recovered from the nasal cavity and tonsil were found to be 7- and 3.5-fold higher

  8. 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.

    Science.gov (United States)

    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.

  9. The relationship between capsular type and OmpA of Pasteurella multocida is associated with the outcome of disease.

    Science.gov (United States)

    Verma, Subhash; Salwan, Richa; Katoch, Shailja; Verma, Lovit; Chahota, Rajesh; Dhar, Prasenjit; Sharma, Mandeep

    2016-12-01

    The genes encoding OmpA of Pasteurella multocida recovered from diseased and apparently healthy animals have been characterized. The nucleotide sequence revealed ORFs of 1047-1077 bp encoding proteins of 349-360 amino acids. Domain analysis of OmpA showed signal peptide, N-terminal ompA domain and C-terminal ligand binding domain. The transmembrane topology of OmpA showed short turns at the periplasmic end and longer irregular loops at the extracellular end. The phylogenetic analysis based on OmpA showed affiliation of isolates to 7 groups representing different alleles. The identical segments in OmpA also suggested assortative recombination within classes IV, V and VI of distinct lineages. Principal component analysis separated isolates into groups based on capsular type and PmompA alleles. The alleles belonging to class VI exclusively associated with capsular type A, whereas class I-IV were associated with capsular type B. PmompA alleles in class V were recorded in both serogroups. PmompA6.1, 6.4 were distributed among strains with capsular type A, and PmompA6.2 and 6.3 among capsular type B. Despite internal OmpA variabilty, restrictive and well defined distribution was seen amongst P. multocida. A definitive association of "OmpA-capsular type" was observed with clinical status of animals. A cohort of pasteurellae comprising of OmpA(I-IV)-capB was recovered from diseased animals and OmpA(VI)-capA from healthy subjects. This study concludes that P. multocida with serogroup A and B from healthy and diseased animals represent distinct clusters also differentiated based on their OmpA-types and OmpA-capsular type relationship possibly determine the virulence and disease outcome. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Use of an rRNA probe and restriction endonuclease analysis to fingerprint Pasteurella multocida isolated from turkeys and wildlife.

    Science.gov (United States)

    Snipes, K P; Hirsh, D C; Kasten, R W; Hansen, L M; Hird, D W; Carpenter, T E; McCapes, R H

    1989-01-01

    Twenty-five isolates of the bacterium Pasteurella multocida were characterized (fingerprinted) phenotypically and genotypically in order to compare the abilities of various techniques to differentiate strains for epidemiologic studies of fowl cholera. Isolates were obtained over a 16-month period from turkeys dying from fowl cholera (six outbreak flocks) and from wildlife captured on premises with a history of the disease. The characteristics compared included (i) serotype, (ii) subspecies, (iii) antibiogram, (iv) presence of plasmid DNA, (v) restriction endonuclease analysis patterns of whole-cell DNA, and (vi) ribotype. Ribotyping, a method of highlighting DNA restriction site heterogeneity by using an rRNA probe, worked well for differentiating the strains of P. multocida when the majority of the other techniques could not. Ribotyping results correlated directly with and confirmed results obtained from restriction endonuclease analysis. Ribotyping demonstrated the presence of up to three strains of P. multocida in one outbreak flock, recurrence of a single strain in five of the flocks over an 11-month period, and the presence of common strains in turkeys and wildlife on the premises. Images PMID:2768471

  11. Influence of Pasteurella multocida and high and low environmental temperatures on adrenals and bursa of Fabricius in turkeys

    Energy Technology Data Exchange (ETDEWEB)

    Simensen, E.; Olson, L.D.; Hahn, G.L.

    1980-01-01

    The morphologic changes in the adrenals and bursa of Fabricius were evaluated from turkeys inoculated with Pasteurella multocida either in the palatine air spaces or via drinking water and maintained at high (33.4-37.4 C), low (2.6-5.3 C), and moderate (19.8-22.4 C) temperatures in temperature-controlled chambers. There was a slight hyperplasia of the adrenal cortical cells and a hypertrophy of the nuclei in the uninoculated turkeys maintained at both high and low temperatures, but these changes were more marked in turkeys maintained at low temperatures. Regardless of the temperature to which the turkeys were exposed, there was an increase in adrenal weight, hyperplasia of the cortical cells, hypertrophy of the nuclei of the cortical cells, and depletion of lipid in the cortical cells in the turkeys that became depressed after inoculation with P. multocida. In the uninoculated turkeys exposed to high temperatures there was a reduction in the weight of the bursa of Fabricius, atrophy of the follicles, and a reduction in the number of lymphocytes within the follicle, which did not occur in the bursae from uninoculated turkeys maintained at low temperatures. In the turkeys inoculated with P. multocida, there was a marked reduction in bursal weight, atrophy of the follicles, and reduction in the number of lymphocytes within the follicles.

  12. Virulence gene profiling and antibiotic resistance pattern of Indian isolates of Pasteurella multocida of small ruminant origin.

    Science.gov (United States)

    Sarangi, Laxmi N; Thomas, P; Gupta, S K; Priyadarshini, A; Kumar, S; Nagaleekar, Viswas Konasagara; Kumar, A; Singh, Vijendra P

    2015-02-01

    Pasteurellosis in small ruminants affects the livelihood of small and marginal farmers of India. The present study was undertaken to understand the trends in gene carriage and antibiotic resistance pattern of Pasteurella multocida isolates recovered from small ruminants over a period of 10 years in India. A total of 88 P. multocida isolates of small ruminant origin were subjected to virulence gene profiling for 19 genes by PCR and antibiogram study employing 17 different antibiotics. Virulence genes like exbB, exbD, tonB, oma87, sodA, sodC, nanB and plpB (100% prevalence) and ptfA and hsf-2 (>90% prevalence) were found to be uniformly distributed among isolates. Unexpectedly, a very high prevalence (95.45%) of pfhA gene was observed in the present study. Dermonecrotoxin gene (toxA) was observed in 48.9% of isolates with highest occurrence among serotype A isolates and interestingly, one of each isolate of serotype B and F were found to carry this gene. Antimicrobial susceptibility testing revealed 17.04% isolates to be multidrug resistant. Amongst all the antibiotics tested, most of the P. multocida isolates were found to be susceptible to enrofloxacin and chloramphenicol. This study highlights novel epidemiological information on frequency and occurrence of virulence genes among Indian isolates from small ruminants. Copyright © 2014 Elsevier Ltd. All rights reserved.

  13. Characterization of the ptfA gene of avian Pasteurella multocida strains by allele-specific polymerase chain reaction.

    Science.gov (United States)

    Sellyei, Boglárka; Bányai, Krisztián; Magyar, Tibor

    2010-07-01

    Pasteurella multocida is the causative agent of fowl cholera in domesticated and wild birds. The disease outcome is affected by various host- and pathogen-specific determinants. Several putative virulence factors have been proposed to play a key role in this interaction, including the ptfA gene, the products of which assemble to form type 4 fimbriae on the bacterial surface. One way to understand more precisely how ptfA contributes to pathogenesis is to gather molecular features of this gene in circulating avian P. multocida strains. Therefore, molecular characterization of the ptfA gene of P. multocida strains isolated from domestic poultry was performed using the combination of nucleotide sequence analysis and a newly developed allele-specific polymerase chain reaction assay. Two major ptfA alleles were identified among 31 strains, representing various serogroups and somatic serotypes. It was noteworthy that allele specificity and case severity of a subset of strains correlated with the available gross pathology data. Therefore, the acquisition of comprehensive clinical and epidemiological data together with molecular characteristics of individual strains will help to design and implement adequate preventive and intervention strategies.

  14. 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

    Directory of Open Access Journals (Sweden)

    Lam Philip W

    2015-01-01

    Full Text Available 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.

  15. 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

    Science.gov (United States)

    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

  16. Bronchoalveolar lavage is an ideal tool in evaluation of local immune response of pigs vaccinated with Pasteurella multocida bacterin vaccine

    Directory of Open Access Journals (Sweden)

    Shiney George

    2015-04-01

    Full Text Available Aim: The aim was to study the bronchoalveolar lavage (BAL technique in evaluating the local immune response of pig immunized with Pasteurella multocida bacterin vaccine. Materials and Methods: Weaned piglets were immunized with formalin-inactivated P52 strain of P. multocida bacterin and evaluated for pulmonary immune response in BAL fluid. BAL was performed before vaccination and at different post vaccination days. The BAL fluid was assayed using enzyme-linked immunosorbent assay to study the development of P. multocida specific antibody isotypes and also evaluated for different cell populations using standard protocol. Results: The average recovery percentage of BAL fluid varies from 58.33 to 61.33 in vaccinated and control group of piglets. The BAL fluid of vaccinated pigs showed increase in antibody titer up to 60th days post vaccination (8.98±0.33, IgG being the predominant isotype reached maximum titer of 6.12±0.20 on 45th days post vaccination, followed by IgM and a meager concentration of IgA could be detected. An increased concentration of the lymphocyte population and induction of plasma cells was detected in the BAL fluid of vaccinated pigs. Conclusion: Though intranasal vaccination with P. multocida plain bacterin vaccine could not provoke a strong immune response, but is promising as lymphocyte population was increased and plasma cells were detected. BAL can be performed repeatedly up to 3/4 months of age in pigs to study pulmonary immune response without affecting their health.

  17. Characterization of Two Novel Lipopolysaccharide Phosphoethanolamine Transferases in Pasteurella multocida and Their Role in Resistance to Cathelicidin-2.

    Science.gov (United States)

    Harper, Marina; Wright, Amy; St Michael, Frank; Li, Jianjun; Deveson Lucas, Deanna; Ford, Mark; Adler, Ben; Cox, Andrew D; Boyce, John D

    2017-11-01

    The lipopolysaccharide (LPS) produced by the Gram-negative bacterial pathogen Pasteurella multocida has phosphoethanolamine (PEtn) residues attached to lipid A, 3-deoxy-d-manno-octulosonic acid (Kdo), heptose, and galactose. In this report, we show that PEtn is transferred to lipid A by the P. multocida EptA homologue, PetL, and is transferred to galactose by a novel PEtn transferase that is unique to P. multocida called PetG. Transcriptomic analyses indicated that petL expression was positively regulated by the global regulator Fis and negatively regulated by an Hfq-dependent small RNA. Importantly, we have identified a novel PEtn transferase called PetK that is responsible for PEtn addition to the single Kdo molecule (Kdo1), directly linked to lipid A in the P. multocida glycoform A LPS. In vitro assays showed that the presence of a functional petL and petK, and therefore the presence of PEtn on lipid A and Kdo1, was essential for resistance to the cationic, antimicrobial peptide cathelicidin-2. The importance of PEtn on Kdo1 and the identification of the transferase responsible for this addition have not previously been shown. Phylogenetic analysis revealed that PetK is the first representative of a new family of predicted PEtn transferases. The PetK family consists of uncharacterized proteins from a range of Gram-negative bacteria that produce LPS glycoforms with only one Kdo molecule, including pathogenic species within the genera Vibrio, Bordetella, and Haemophilus We predict that many of these bacteria will require the addition of PEtn to Kdo for maximum protection against host antimicrobial peptides. Copyright © 2017 American Society for Microbiology.

  18. Comparative genome analysis of five Pasteurella multocida strains to decipher the diversification in pathogenicity and host specialization.

    Science.gov (United States)

    Okay, Sezer; Kurt Kızıldoğan, Aslıhan

    2015-08-01

    Pasteurella multocida is a Gram-negative bacterial pathogen causing economically important diseases in distinct animal species. Complete genome sequences of five P. multocida strains (Pm70, HB03, HN06, 3480, and 36950) isolated from poultry, swine or bovine, were retrieved from the GenBank database and compared with each other, for the first time. The missense mutations generating a dissimilar amino acid in the peptide chain, nonsense mutations, and insertion/deletions in the nucleotide sequence were identified due to the potential change in the protein function. A total of 500 putative mutant proteins were identified, and categorized into 10 groups including cellular compartments such as outer membrane, capsule and fimbria, and processes such as carbohydrate, energy, nucleic acid and amino acid metabolisms, transport, and drug resistance. The majority of the mutant proteins were associated with the outer compartments of the bacterial cell. Various mutations were also detected in the genes related with biosynthetic pathways. The highest and the lowest numbers of mutant proteins belonged to 36950 vs. HN06 and Pm70 vs. HB03 comparisons, respectively. The major impact on the diversification of P. multocida strains was observed to be conferred by the mutations related with pathogenicity. To exhibit the outcomes of the mutations in the peptide chains, three sample amino acid sequences belonging to AfuA, MetB, and d,d-heptose 1,7-bisphosphate phosphatase were aligned, and their phylogenetic relationships were shown. These comprehensive analyses improve the understanding of molecular pathogenicity and host specialization of P. multocida, and would have a contribution to the recombinant vaccine development against this pathogen. Copyright © 2015 Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

    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). © 2015 John Wiley & Sons Ltd.

  20. Occurrence Of Virulence Factors And Antimicrobial Resistance In Pasteurella Multocida Strains Isolated From Slaughter Cattle In Iran

    Directory of Open Access Journals (Sweden)

    Faham eKhamesipour

    2014-10-01

    Full Text Available A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence associated genes by polymerase chain reaction. The disc diffusion technique was used to determine antimicrobial resistance profiles among the isolates. Of the isolates, 23 belonged to capsular type A, 5 to capsular type D and two isolates were untypeable. The distribution of the capsular types in pneumonic lungs and in apparently health lungs was statistically similar. All virulence genes tested were detected among the isolates derived from pneumonic lungs; whereas isolates derived from apparently health lungs carried 16 of the 23 genes. The frequently detected genes among isolates from pneumonic lungs were exbD, hgbA, hgbB, ompA, ompH, oma87 and sodC; whereas tadD, toxA and pmHAS genes occurred less frequently. Most of the adhesins and superoxide dismutases; and all of the iron acquisition and protectin proteins occurred at significantly (p≤0.05 higher frequencies in isolates from pneumonic lungs. Isolates from apparently healthy lungs didn’t carry the following genes; hsf-1, hsf-2, tadD, toxA, nanB, nanH and pmHAS. One adhesion (hsf-1 and two iron acquisition (exbD and tonB genes occurred at significantly (p≤0.05 higher frequencies among capA isolates. All the P. multocida isolates were susceptible to ciprofloxacin, co-trimoxazole, doxycycline, enrofloxacin, nitrofurantoin and tetracyclines. Different proportions of the isolates were however resistant to ampicillin, amoxicillin, erythromycin, lincomycin, penicillin, rifampin, streptomycin and florfenicol. Our results reveal presence of virulence factors in P. multocida strains isolated from symptomatic and asymptomatic bovids. A higher frequency of the factors among isolates from symptomatic study animals may suggest their role in pathogenesis of P. multocida-associated bovine respiratory disease. The results further

  1. Cytidine 5'-monophosphate (CMP)-induced structural changes in a multifunctional sialyltransferase from Pasteurella multocida.

    Science.gov (United States)

    Ni, Lisheng; Sun, Mingchi; Yu, Hai; Chokhawala, Harshal; Chen, Xi; Fisher, Andrew J

    2006-02-21

    Sialyltransferases catalyze reactions that transfer a sialic acid from CMP-sialic acid to an acceptor (a structure terminated with galactose, N-acetylgalactosamine, or sialic acid). They are key enzymes that catalyze the synthesis of sialic acid-containing oligosaccharides, polysaccharides, and glycoconjugates that play pivotal roles in many critical physiological and pathological processes. The structures of a truncated multifunctional Pasteurella multocida sialyltransferase (Delta24PmST1), in the absence and presence of CMP, have been determined by X-ray crystallography at 1.65 and 2.0 A resolutions, respectively. The Delta24PmST1 exists as a monomer in solution and in crystals. Different from the reported crystal structure of a bifunctional sialyltransferase CstII that has only one Rossmann domain, the overall structure of the Delta24PmST1 consists of two separate Rossmann nucleotide-binding domains. The Delta24PmST1 structure, thus, represents the first sialyltransferase structure that belongs to the glycosyltransferase-B (GT-B) structural group. Unlike all other known GT-B structures, however, there is no C-terminal extension that interacts with the N-terminal domain in the Delta24PmST1 structure. The CMP binding site is located in the deep cleft between the two Rossmann domains. Nevertheless, the CMP only forms interactions with residues in the C-terminal domain. The binding of CMP to the protein causes a large closure movement of the N-terminal Rossmann domain toward the C-terminal nucleotide-binding domain. Ser 143 of the N-terminal domain moves up to hydrogen-bond to Tyr 388 of the C-terminal domain. Both Ser 143 and Tyr 388 form hydrogen bonds to a water molecule, which in turn hydrogen-bonds to the terminal phosphate oxygen of CMP. These interactions may trigger the closure between the two domains. Additionally, a short helix near the active site seen in the apo structure becomes disordered upon binding to CMP. This helix may swing down upon binding to

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

    DEFF Research Database (Denmark)

    Tjørnehøj, Kirsten; Eriksen, Lizzie; Aalbaek, B

    1992-01-01

    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...

  3. Identification of the crp gene in avian Pasteurella multocida and evaluation of the effects of crp deletion on its phenotype, virulence and immunogenicity.

    Science.gov (United States)

    Zhao, Xinxin; Liu, Qing; Xiao, Kangpeng; Hu, Yunlong; Liu, Xueyan; Li, Yanyan; Kong, Qingke

    2016-06-24

    Pasteurella multocida (P. multocida) is an important veterinary pathogen that can cause severe diseases in a wide range of mammals and birds. The global regulator crp gene has been found to regulate the virulence of some bacteria, and crp mutants have been demonstrated to be effective attenuated vaccines against Salmonella enterica and Yersinia enterocolitica. Here, we first characterized the crp gene in P. multocida, and we report the effects of a crp deletion. The P. multocida crp mutant exhibited a similar lipopolysaccharide and outer membrane protein profile but displayed defective growth and serum complement resistance in vitro compared with the parent strain. Furthermore, crp deletion decreased virulence but did not result in full attenuation. The 50 % lethal dose (LD50) of the Δcrp mutant was 85-fold higher than that of the parent strain for intranasal infection. Transcriptome sequencing analysis showed that 92 genes were up-regulated and 94 genes were down-regulated in the absence of the crp gene. Finally, we found that intranasal immunization with the Δcrp mutant triggered both systematic and mucosal antibody responses and conferred 60 % protection against virulent P. multocida challenge in ducks. The deletion of the crp gene has an inhibitory effect on bacterial growth and bacterial resistance to serum complement in vitro. The P. multocida crp mutant was attenuated and conferred moderate protection in ducks. This work affords a platform for analyzing the function of crp and aiding the formulation of a novel vaccine against P. multocida.

  4. Aislamiento e Identificación Bioquímica de Cepas de Pasteurella multocida y Gallibacterium anatis en Aves de Producción con Signos Respiratorios

    OpenAIRE

    Castillo, Gonzalo; Carrera Profesional de Medicina Veterinaria, Facultad de Ciencias Agropecuarias, Universidad Alas Peruanas, Lima, Perú; Koga, Ysabel; Bioservice SRL, Lima, Perú; Alvarado, Arnaldo; Laboratorio de Patología Clínica y Biología Molecular, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima , Perú; Tinoco, Robert; Bioservice SRL, Lima; Fernández, Daniel; Bioservice SRL, Lima, Perú

    2014-01-01

    El objetivo del estudio fue determinar las biovariedades de Pasteurella multocida y Gallibacterium anatis en aves de producción con signos respiratorios. Estas bacterias fueron aisladas de muestras de secreciones y órganos de pollos de carne, gallinas de postura y patos criollos afectados, provenientes de granjas avícolas de la costa y selva del Perú. De 25 aislamientos se identificaron 13 cepas de P. multocida y 12 de G. anatis mediante características de cultivo, morfología y pruebas bioquí...

  5. Serological evidence of avian encephalomyelitis virus and Pasteurella multocida infections in free-range indigenous chickens in Southern Mozambique.

    Science.gov (United States)

    Taunde, Paula; Timbe, Palmira; Lucas, Ana Felicidade; Tchamo, Cesaltina; Chilundo, Abel; Dos Anjos, Filomena; Costa, Rosa; Bila, Custodio Gabriel

    2017-06-01

    A total of 398 serum samples from free-range indigenous chickens originating from four villages in Southern Mozambique were tested for the presence of avian encephalomyelitis virus (AEV) and Pasteurella multocida (PM) antibodies through commercial enzyme-linked immunosorbent assay (ELISA) kits. AEV and PM antibodies were detected in all villages surveyed. The proportion of positive samples was very high: 59.5% (95% confidence interval (CI) 51.7-67.7%) for AEV and 71.5% (95% CI 67.7-77.3%) for PM. Our findings revealed that these pathogens are widespread among free-range indigenous chickens in the studied villages and may represent a threat in the transmission of AEV and PM to wild, broiler or layer chickens in the region. Further research is warranted on epidemiology of circulating strains and impact of infection on the poultry industry.

  6. Modulating the regioselectivity of a Pasteurella multocida sialyltransferase for biocatalytic production of 3'- and 6'-sialyllactose

    DEFF Research Database (Denmark)

    Guo, Yao; Jers, Carsten; Meyer, Anne S.

    2015-01-01

    ST) for catalysing the synthesis of 3'- and 6'-sialyllactose using casein glycomacropeptide as sialyl-donor and lactose as acceptor. The mutation P34H led to a 980-fold increase in α-2,6-sialyltransferase activity (with cytidine-5'-monophospho-N-acetylneuraminic acid as donor), while its α-2,3-sialyltransferase......Several bacterial sialyltransferases have been reported to be multifunctional also catalysing sialidase and trans-sialidase reactions. In this study, we examined the trans-sialylation efficacy and regioselectivity of mutants of the multifunctional Pasteurella multocida sialyltransferase (Pm...... and α-2,6-trans-sialidase activity. The double mutant PmSTE271F/R313Y thus showed the highest α-2,3-regioselectivity and constitutes an interesting enzyme for regioselective synthesis of α-2,3-sialylated glycans. This study has expanded the understanding of the structure-function relationship...

  7. An abscess due to Pasteurella multocida after a cat scratch: Case report and evaluation of antibiotic prophylaxis

    Directory of Open Access Journals (Sweden)

    Yeşim Alpay

    2014-12-01

    Full Text Available Pasteurella multocida has been isolated from 50% to 70% of healthy cats and most commonly associated with acute skin and soft tissue infections following an animal bite or scratch. As the zone and depth of injury can lead to more serious infections such as deep tissue infections, septic arthritis, osteomyelitis. However, no predictive factor showing which wound would be infected. In our case, patient whom applied with abscess after a cat scratch and P. multocida was found as a causative agent. This situation has caused to review us, once more, that which cases should be taken antibiotic pro­phylaxis in addition to immunoprophylaxis (for rabies post-exposure prophylaxis, and anti-tetanus prophylaxis in the first admission. Antibiotic prophylaxis should be used for 3-5 days in selected cases if they include; moderate to severe crushing injuries especially edematous form, less than 8 hours old, bone or joint penetration, hand wounds, especially emphasizes the importance of hand injuries and deep penetrations. J Microbiol Infect Dis 2014; 4(4: 159-161

  8. Molecular characterization and antimicrobial susceptibility of Pasteurella multocida strains isolated from hosts affected by various diseases in Italy.

    Science.gov (United States)

    Cucco, Lucilla; Massacci, Francesca Romana; Sebastiani, Carla; Mangili, Piermario; Bano, Luca; Cocchi, Monia; Luppi, Andrea; Ortenzi, Roberta; Pezzotti, Giovanni; Magistrali, Chiara Francesca

    2017-03-31

    Pasteurella multocida is a widespread pathogen associated with major animal diseases of economic significance. Despite this, little is known about the capsular types, virulence gene pattern, and antimicrobial susceptibility of isolates from hosts affected by different diseases, and no data are available in Italy. One hundred eighty six isolates of P. multocida, were taken from different species in different states of health in several Italian regions, and were tested for genes encoding for capsular types (cap) and major virulence factors (tbpA, toxA, hgbB and pfhA). Antimicrobial susceptibility was investigated with the agar diffusion test. The majority of isolates was capA+. However, the distribution differed according to species and disease of origin, with a greater heterogeneity in isolates from rabbits; capE was never found, while capB was detected once. Only capA+ and capF+ strains tested positive for pfhA. Conversely, almost all capD+ isolates were hgbB+. In bovine respiratory disease, pfhA+/tbpA+/capA+ isolates predominated, while tbpA+/toxA+/capD+ isolates predominated in sheep. Overall, low levels of resistance were found, with full susceptibility to ceftiofur and florfenicol. Lower susceptibility to older antimicrobials was recorded, since only approximately 1/3 of the isolates showed susceptibility to tylosin and erythromycin, and resistance to tetracycline (7.5%), and trimethoprim - sulphametoxazole (4.8%) was also observed.

  9. Comparison of whole genome sequencing to restriction endonuclease analysis and gel diffusion precipitin-based serotyping of Pasteurella multocida.

    Science.gov (United States)

    LeCount, Karen J; Schlater, Linda K; Stuber, Tod; Robbe Austerman, Suelee; Frana, Timothy S; Griffith, Ronald W; Erdman, Matthew M

    2018-01-01

    The gel diffusion precipitin test (GDPT) and restriction endonuclease analysis (REA) have commonly been used in the serotyping and genotyping of Pasteurella multocida. Whole genome sequencing (WGS) and single nucleotide polymorphism (SNP) analysis has become the gold standard for other organisms, offering higher resolution than previously available methods. We compared WGS to REA and GDPT on 163 isolates of P. multocida to determine if WGS produced more precise results. The isolates used represented the 16 reference serovars, isolates with REA profiles matching an attenuated fowl cholera vaccine strain, and isolates from 10 different animal species. Isolates originated from across the United States and from Chile. Identical REA profiles clustered together in the phylogenetic tree. REA profiles that differed by only a few bands had fewer SNP differences than REA profiles with more differences, as expected. The GDPT results were diverse but it was common to see a single serovar show up repeatedly within clusters. Several errors were found when examining the REA profiles. WGS was able to confirm these errors and compensate for the subjectivity in analysis of REA. Also, results of WGS and SNP analysis correlated more closely with the epidemiologic data than GDPT. In silico results were also compared to a lipopolysaccharide rapid multiplex PCR test. From the data produced in our study, WGS and SNP analysis was superior to REA and GDPT and highlighted some of the issues with the older tests.

  10. Identification of the capsular polysaccharides of Type D and F Pasteurella multocida as unmodified heparin and chondroitin, respectively.

    Science.gov (United States)

    DeAngelis, Paul L; Gunay, Nur Sibel; Toida, Toshihiko; Mao, Wen-jun; Linhardt, Robert J

    2002-09-27

    Pasteurella multocida is a pathogenic Gram-negative bacterial species that infects a wide variety of animals and humans. A notable morphological feature of many isolates is the extracellular capsule. The ability to remove the capsule by treatment with certain glycosidases has been utilized to discern various capsular types called A, D and F. Based on this preliminary evidence, these microbes have capsules made of glycosaminoglycans, linear polysaccharides composed of repeating disaccharide units containing an amino sugar. Glycosaminoglycans are also abundant components of the vertebrate extracellular matrix. It has been shown previously that the major Type A capsular material was hyaluronan (hyaluronic acid). We report that the Type D polymer is an unmodified heparin (N-acetylheparosan) with a -->4)-beta-D-Glcp-UA-(1-->4)-alpha-D-Glcp-NAc-(1--> repeating unit and the Type F polymer is an unmodified chondroitin with a -->4)-beta-D-Glcp-UA-(1-->3)-beta-D-Galp-NAc-(1--> repeating unit. The monosaccharide compositions, disaccharide profiles, and 1H NMR analyses are consistent with these identifications. The molecular size of the Pasteurella polymers is approximately 100-300 kDa as determined by gel electrophoresis and multi-angle laser light scattering; this size is much greater than the 10-30 kDa size of the analogous polymers isolated from animal tissues. The glycosaminoglycan capsular polymers are relatively non-immunogenic virulence factors that enhance microbial pathogenicity.

  11. Predicting the outer membrane proteome of Pasteurella multocida based on consensus prediction enhanced by results integration and manual confirmation

    Directory of Open Access Journals (Sweden)

    E-komon Teerasak

    2012-04-01

    Full Text Available Abstract Background Outer membrane proteins (OMPs of Pasteurella multocida have various functions related to virulence and pathogenesis and represent important targets for vaccine development. Various bioinformatic algorithms can predict outer membrane localization and discriminate OMPs by structure or function. The designation of a confident prediction framework by integrating different predictors followed by consensus prediction, results integration and manual confirmation will improve the prediction of the outer membrane proteome. Results In the present study, we used 10 different predictors classified into three groups (subcellular localization, transmembrane β-barrel protein and lipoprotein predictors to identify putative OMPs from two available P. multocida genomes: those of avian strain Pm70 and porcine non-toxigenic strain 3480. Predicted proteins in each group were filtered by optimized criteria for consensus prediction: at least two positive predictions for the subcellular localization predictors, three for the transmembrane β-barrel protein predictors and one for the lipoprotein predictors. The consensus predicted proteins were integrated from each group into a single list of proteins. We further incorporated a manual confirmation step including a public database search against PubMed and sequence analyses, e.g. sequence and structural homology, conserved motifs/domains, functional prediction, and protein-protein interactions to enhance the confidence of prediction. As a result, we were able to confidently predict 98 putative OMPs from the avian strain genome and 107 OMPs from the porcine strain genome with 83% overlap between the two genomes. Conclusions The bioinformatic framework developed in this study has increased the number of putative OMPs identified in P. multocida and allowed these OMPs to be identified with a higher degree of confidence. Our approach can be applied to investigate the outer membrane proteomes of other Gram

  12. Plasma corticosterone concentrations in turkeys inoculated with Pasteurella multocida and maintained at high and low environmental temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Simensen, E.; Olson, L.D.; Ryan, M.P.; Vanjonack, W.J.; Johnson, H.D.

    1980-01-01

    Radioimmunoassay was used to determine plasma corticosterone concentration (PCC) in turkeys inoculated with Pasteurella multocida via either the palatine air spaces or the drinking water and maintained at high (33.4-37.4 C), low (2.6-5.3 C) and moderate )19.8-22.4 C) temperatures in temperature-controlled chambers. In uninoculated turkeys maintained at high temperatures, the PCC was generally lower than in turkeys maintained at moderate temperatures, whereas the opposite occurred in turkeys maintained at low temperatures. After inoculation with P. multocida, all groups of inoculated turkeys showed an increase in the average PCC, which attained a level in some turkeys of over 40 ng/ml, in relation to the average in the uninoculated turkeys, which ranged from 1.8 to 27.3 ng/ml. This increase was proportional to the severity of the infection that developed. The PCC was found to be a sensitive indicator of an incubating infection of P. multocida, since it was markedly increased in turkeys that were bled one day before the onset of depression. In turkeys that were inoculated via the palatine air spaces and maintained at 20 C, the PCC on the day of inoculation was significantly (P less than 0.05) lower in the turkeys that later died than in those that survived. Generally, the PCC was higher in the turkeys that either died between 5 and 10 days after inoculation or were depressed aa the end of the experiment on day 10, relative to the turkeys that were alert at the end of the experiment.

  13. Genome sequencing of a virulent avian Pasteurella multocida strain GX-Pm reveals the candidate genes involved in the pathogenesis.

    Science.gov (United States)

    Yu, Chengjie; Sizhu, Suolang; Luo, Qingping; Xu, Xuewen; Fu, Lei; Zhang, Anding

    2016-04-01

    Pasteurella multocida (P. multocida) was first shown to be the causative agent of fowl cholera by Louis Pasteur in 1881. First genomic study was performed on an avirulent avian strain Pm70, and until 2013, two genomes of virulent avian strains X73 and P1059 were sequenced. Comparative genome study supplied important information for further study on the pathogenesis of fowl cholera. In the previous study, a capsular serotype A strain GX-Pm was isolated from the liver of a chicken, which died during an outbreak of fowl cholera in 2011. The strain showed multiple drug resistance and was highly virulent to chickens. Therefore, the present study performed the genome sequencing and a comparative genomic analysis to reveal the candidate genes involved in virulence of P. multocida. Sequenced draft genome sequence of GX-Pm was 2,292,886 bp, contained 2941 protein-coding genes, 5 genomic islands, 4 IS elements and 2 prophage regions. Notability, all the predicted drug-resistance genes were included in predicted genomic islands. A comparative genome study on virulent avian strains P1059, X73 and GX-Pm with the avirulent avian strain Pm 70 indicated that 475 unique genes were only identified in either of virulent strains but absent in the avirulent strain. Among these genes, 20 genes were contained within genomes of all three virulent strains, including a few of putative virulence genes. Further characterization of the pathogenic functions of these genes would benefit the understanding of pathogenesis of fowl cholera. Copyright © 2016 Elsevier Ltd. All rights reserved.

  14. Molecular analysis of Pasteurella multocida strains isolated from fowl cholera infection in backyard chickens

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    Mohamed-Wael Abdelazeem Mohamed

    2014-01-01

    Conclusion: Based on the previous findings, there are three spreading clusters that may indicate the association of a small number of P. multocida variants with the majority of cases suggesting that certain clones of P. multocida are able to colonize the examined backyard chickens. Also, the ease and rapidity of RAPD-PCR support the use of this technique as alternative to the more labour-intensive SDS-PAGE system for strain differentiation and epidemiological studies of avian P. multocida. Further application of RAPD technology to the examination of avian cholera outbreaks in commercially available flocks may facilitate more effective management of this disease by providing the potential to investigate correlations of P. multocida genotypes, to identify affiliations between bird types and bacterial genotypes, and to elucidate the role of specific bird species in disease transmission.

  15. Antimikrobna osjetljivost bakterija Escherichia coli, Salmonella spp., Pasteurella multocida, Streptococcus suis i Actinobacillus pleuropneumoniae izdvojenih iz dijagnostičkih uzoraka iz velikih svinjogojskih farmi u Hrvatskoj

    OpenAIRE

    Habrun, Boris; Kompes, Gordan; Cvetnić, Željko; Špičić, Silvio; Benić, Miroslav; Mitak, Mario

    2010-01-01

    Disk difuzijskom metodom istražena je osjetljivost 256 izolata Escherichia (E.) coli, 42 izolata Salmonella spp. i 62 izolata Streptococcus (S.) suis na 13 antimikrobnih lijekova, te 108 izolata Pasteurella (P.) multocida, i 44 izolata Actinobacillus (A.) pleuropneumoniae na 14 antimikrobnih lijekova. Svi su izolati bili izdvojeni iz dijagnostičkoga materijala s 8 velikih svinjogojskih farmi u Hrvatskoj. U E. coli najveći stupanj rezistencije utvrđen je na oksitetraciklin (98% izolata), strep...

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

    Science.gov (United States)

    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 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.

  17. Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran

    Science.gov (United States)

    Khamesipour, Faham; Momtaz, Hassan; Azhdary Mamoreh, Morteza

    2014-01-01

    A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence-associated genes by polymerase chain reaction (PCR). The disc diffusion technique was used to determine antimicrobial resistance profiles among the isolates. Of the isolates, 23 belonged to capsular type A, 5 to capsular type D and two isolates were untypeable. The distribution of the capsular types in pneumonic lungs and in apparently health lungs was statistically similar. All virulence genes tested were detected among the isolates derived from pneumonic lungs; whereas isolates derived from apparently health lungs carried 16 of the 23 genes. The frequently detected genes among isolates from pneumonic lungs were exbD, hgbA, hgbB, ompA, ompH, oma87, and sodC; whereas tadD, toxA, and pmHAS genes occurred less frequently. Most of the adhesins and superoxide dismutases; and all of the iron acquisition and protectin proteins occurred at significantly (p ≤ 0.05) higher frequencies in isolates from pneumonic lungs. Isolates from apparently healthy lungs didn't carry the following genes; hsf-1, hsf-2, tadD, toxA, nanB, nanH, and pmHAS. One adhesion (hsf-1) and two iron acquisition (exbD and tonB) genes occurred at significantly (p ≤ 0.05) higher frequencies among capA isolates. All the P. multocida isolates were susceptible to ciprofloxacin, co-trimoxazole, doxycycline, enrofloxacin, nitrofurantoin, and tetracyclines. Different proportions of the isolates were however resistant to ampicillin, amoxicillin, erythromycin, lincomycin, penicillin, rifampin, streptomycin, and florfenicol. Our results reveal presence of virulence factors (VFs) in P. multocida strains isolated from symptomatic and asymptomatic bovids. A higher frequency of the factors among isolates from symptomatic study animals may suggest their role in pathogenesis of P. multocida-associated bovine respiratory disease (BRD). The results

  18. Occurrence of virulence factors and antimicrobial resistance in Pasteurella multocida strains isolated from slaughter cattle in Iran.

    Science.gov (United States)

    Khamesipour, Faham; Momtaz, Hassan; Azhdary Mamoreh, Morteza

    2014-01-01

    A total of 30 Pasteurella multocida strains isolated from 333 pneumonic and apparently health slaughter cattle were examined for capsule biosynthesis genes and 23 virulence-associated genes by polymerase chain reaction (PCR). The disc diffusion technique was used to determine antimicrobial resistance profiles among the isolates. Of the isolates, 23 belonged to capsular type A, 5 to capsular type D and two isolates were untypeable. The distribution of the capsular types in pneumonic lungs and in apparently health lungs was statistically similar. All virulence genes tested were detected among the isolates derived from pneumonic lungs; whereas isolates derived from apparently health lungs carried 16 of the 23 genes. The frequently detected genes among isolates from pneumonic lungs were exbD, hgbA, hgbB, ompA, ompH, oma87, and sodC; whereas tadD, toxA, and pmHAS genes occurred less frequently. Most of the adhesins and superoxide dismutases; and all of the iron acquisition and protectin proteins occurred at significantly (p ≤ 0.05) higher frequencies in isolates from pneumonic lungs. Isolates from apparently healthy lungs didn't carry the following genes; hsf-1, hsf-2, tadD, toxA, nanB, nanH, and pmHAS. One adhesion (hsf-1) and two iron acquisition (exbD and tonB) genes occurred at significantly (p ≤ 0.05) higher frequencies among capA isolates. All the P. multocida isolates were susceptible to ciprofloxacin, co-trimoxazole, doxycycline, enrofloxacin, nitrofurantoin, and tetracyclines. Different proportions of the isolates were however resistant to ampicillin, amoxicillin, erythromycin, lincomycin, penicillin, rifampin, streptomycin, and florfenicol. Our results reveal presence of virulence factors (VFs) in P. multocida strains isolated from symptomatic and asymptomatic bovids. A higher frequency of the factors among isolates from symptomatic study animals may suggest their role in pathogenesis of P. multocida-associated bovine respiratory disease (BRD). The results

  19. Pharmacokinetic/pharmacodynamic integration and modelling of florfenicol for the pig pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida.

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    Lucy Dorey

    Full Text Available Pharmacokinetic-pharmacodynamic (PK/PD integration and modelling were used to predict dosage schedules for florfenicol for two pig pneumonia pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida. Pharmacokinetic data were pooled for two bioequivalent products, pioneer and generic formulations, administered intramuscularly to pigs at a dose rate of 15 mg/kg. Antibacterial potency was determined in vitro as minimum inhibitory concentration (MIC and Mutant Prevention Concentration in broth and pig serum, for six isolates of each organism. For both organisms and for both serum and broth MICs, average concentration:MIC ratios over 48 h were similar and exceeded 2.5:1 and times greater than MIC exceeded 35 h. From in vitro time-kill curves, PK/PD modelling established serum breakpoint values for the index AUC24h/MIC for three levels of inhibition of growth, bacteriostasis and 3 and 4log10 reductions in bacterial count; means were 25.7, 40.2 and 47.0 h, respectively, for P. multocida and 24.6, 43.8 and 58.6 h for A. pleuropneumoniae. Using these PK and PD data, together with literature MIC distributions, doses for each pathogen were predicted for: (1 bacteriostatic and bactericidal levels of kill; (2 for 50 and 90% target attainment rates (TAR; and (3 for single dosing and daily dosing at steady state. Monte Carlo simulations for 90% TAR predicted single doses to achieve bacteriostatic and bactericidal actions over 48 h of 14.4 and 22.2 mg/kg (P. multocida and 44.7 and 86.6 mg/kg (A. pleuropneumoniae. For daily doses at steady state, and 90% TAR bacteriostatic and bactericidal actions, dosages of 6.2 and 9.6 mg/kg (P. multocida and 18.2 and 35.2 mg/kg (A. pleuropneumoniae were required. PK/PD integration and modelling approaches to dose determination indicate the possibility of tailoring dose to a range of end-points.

  20. Identification of the Avian Pasteurella multocida phoP Gene and Evaluation of the Effects of phoP Deletion on Virulence and Immunogenicity

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    Kangpeng Xiao

    2015-12-01

    Full Text Available Pasteurella multocida (P. multocida is an animal pathogen of worldwide economic significance that causes fowl cholera in poultry and wild birds. Global gene regulators, including PhoP are important in regulating bacterial virulence and are good targets for developing attenuated vaccines against many pathogenic bacteria. However, the biological significance of phoP gene has not been identified in P. multocida. Here, we identified the phoP gene in P. multocida, and we evaluated the roles of phoP in P. multocida by deleting the phoP gene. The P. multocida phoP mutant exhibited similar growth curves and lipopolysaccharide and outer membrane protein profiles but displayed defective polymyxin resistance in vitro compared with the parent strain. Additionally, the phoP deletion resulted in decreased virulence. The LD50 of the ΔphoP mutant was 32- and 154-fold higher than the parent strain via the oral and intranasal routes, respectively. Transcriptome sequencing analysis showed that 161 genes were up-regulated and 173 genes were down-regulated in the absence of the phoP gene. Finally, the immunogenicity and protective efficacy of the ΔphoP mutant were evaluated. Immunized ducks produced significantly higher levels of serum IgY and bile IgA compared to the control ducks, and immunization with the ΔphoP mutant conferred 54.5% protection efficiency against challenge with the virulent P. multocida. This work provides a platform to dissect the function of phoP and develop a new vaccine against P. multocida.

  1. Identification of the Avian Pasteurella multocida phoP Gene and Evaluation of the Effects of phoP Deletion on Virulence and Immunogenicity.

    Science.gov (United States)

    Xiao, Kangpeng; Liu, Qing; Liu, Xueyan; Hu, Yunlong; Zhao, Xinxin; Kong, Qingke

    2015-12-23

    Pasteurella multocida (P. multocida) is an animal pathogen of worldwide economic significance that causes fowl cholera in poultry and wild birds. Global gene regulators, including PhoP are important in regulating bacterial virulence and are good targets for developing attenuated vaccines against many pathogenic bacteria. However, the biological significance of phoP gene has not been identified in P. multocida. Here, we identified the phoP gene in P. multocida, and we evaluated the roles of phoP in P. multocida by deleting the phoP gene. The P. multocida phoP mutant exhibited similar growth curves and lipopolysaccharide and outer membrane protein profiles but displayed defective polymyxin resistance in vitro compared with the parent strain. Additionally, the phoP deletion resulted in decreased virulence. The LD50 of the ΔphoP mutant was 32- and 154-fold higher than the parent strain via the oral and intranasal routes, respectively. Transcriptome sequencing analysis showed that 161 genes were up-regulated and 173 genes were down-regulated in the absence of the phoP gene. Finally, the immunogenicity and protective efficacy of the ΔphoP mutant were evaluated. Immunized ducks produced significantly higher levels of serum IgY and bile IgA compared to the control ducks, and immunization with the ΔphoP mutant conferred 54.5% protection efficiency against challenge with the virulent P. multocida. This work provides a platform to dissect the function of phoP and develop a new vaccine against P. multocida.

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    Carolina Gallego

    2017-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    Gallego, Carolina; Romero, Stefany; Esquinas, Paula; Patiño, Pilar; Martínez, Nhora; Iregui, Carlos

    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.

  5. Development of fowl cholera vaccine: I. Protection of Pasteurella multocida local isolate vaccine against challenge of homologous and heterologous strains.

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    Supar

    2001-03-01

    Full Text Available Pasteurella multocida locally isolated from chicken and ducks (BCC 299, BCC 2331, DY1, DY2, 12TG, 15TG andimported strains (BCC 1359, 1362; HEDDLESTON group 1 and 6 respectively had been tested for its pathogenicity in theprevious study. The aims of this experiment were to study the preparation of local isolate pasteurellosis vaccines and to determine the protective effect of that vaccines in chicken against the highly pathogenic local isolates of P. multocida. Killed monovalent, bivalent and polyvalent pasteurellosis vaccines were prepared and each was adjunvanted with aluminum hydroxide gel at a final concentration of 1.5% and the cell concentration was equal to the No 10 of MacFarland tube standard. Each of the vaccine prepared was used to vaccinated on a group of six week old of layer chicken (8 per group. Each chicken was subcutaneously injected with 0.2 ml of vaccine, four weeks later each was boostered with similar vaccine with the same dose. Two weeks after giving the boostered vaccine each group of chicken were challenged, half with life bacterium of P. Multocida BCC 2331 and other with DY2. Any chick which survive after challenge was designated as protected by vaccination. Before vaccination 1 ml of blood was drawn from each of chicken and then two weeks apart up to challenge. Serum from each sample was separated and kept in deep freeze until tested by enzyme-linked immunosorbent assay (ELISA. Chicken vaccinated with killed whole cell P. multocida vaccines of monovalent (BCC 2331 or DY2 and bivalent (BCC 2331 + DY2 were protected against challenge with live bacterium of either BCC 2331 or DY2 at rate 67-100%. There was no protection in chicken vaccinated with either BCC 299, DY1, 12TG, 15TG, BCC 1359, or 1362 killed vaccine. Similarly no protection of chicken vaccinated with either DY1 + BCC299, 12TG + 15TG or BCC 1359 + BCC 1362 bivalent vaccines. The protection rate of the polyvalent local isolate vaccine was at average 50-75%. All

  6. 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.

    Science.gov (United States)

    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

  7. Multiplex PCR To Identify Macrolide Resistance Determinants in Mannheimia haemolytica and Pasteurella multocida

    DEFF Research Database (Denmark)

    Rose, Simon; Desmolaize, Benoit; Jaju, Puneet

    2012-01-01

    isolates of P. multocida and M. haemolytica and correlated with MICs for the veterinary macrolides tulathromycin and tilmicosin, and the newer compounds gamithromycin and tildipirosin. The multiplex PCR system gives a rapid and robustly accurate determination of macrolide resistance genotypes and bacterial...

  8. Studies on Pasteurella gallinarum.

    Science.gov (United States)

    Mushin, R; Bock, R; Abrams, M

    1977-01-01

    In two outbreaks of disease in chickens associated with Pasteurella gallinarum infection swollen and inflamed wattles were symptoms. A bacteriological study was carried out of 42 P. gallinarum strains isolated from the two outbreaks and from sporadic cases in fowls. Serotyping by the gel diffusion precipitin test indicated that all five strains from one outbreak and 82% of isolants from 22 sporadic cases belonged to one serotype, antigenically unrelated to Pasteurella multocida. All four isolants from another outbreak were of a different serotype and were antigenically related to P. multocida serotypes 6 and 9. Significant titres of agglutinating antibodies against hyaluronidase-treated P. gallinarum antigens and precipitating antibodies were detected in sera from chickens infected under field conditions and experimentally.

  9. Isolation and molecular detection of Pasteurella multocida Type A from naturally infected chickens, and their histopathological evaluation in artificially infected chickens in Bangladesh

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    Sayedun Nahar Panna

    2015-09-01

    Full Text Available Pasteurella multocida type A is the etiologic agent of fowl cholera, a highly contagious and fatal disease of chickens. The present research work was performed for the isolation, identification and molecular detection of P. multocida Type A from chickens. Liver, heart and spleen of suspected dead chicken (n=35 were collected from Gazipur and Pabna districts in Bangladesh. The targeted bacteria from the samples were isolated, identified and characterized based on their morphology, staining, cultural, biochemical characters, pathogenicity test, histopathological study and Polymerase Chain Reaction (PCR. The P. multocida organism was isolated from 11.42% (n=4/35 samples. The organisms were gram negative, non-spore forming rod, non-motile, occurring singly or pairs in Gram staining, whereas in Leishman's stain, bipolar shaped organisms were observed. All the isolates were found positive for oxidase and catalase tests, produced indole, and fermented glucose, mannitol and sucrose. Necrotic foci in liver and congestion with hemorrhages in heart were found on necropsy. After pathogenicity test, the pathological changes were reconfirmed by histopathology depicting congestion, hemorrhage and lymphocyte infiltration in heart, liver and spleen tissues. In type specific PCR reaction, the organisms were confirmed as P. multocida Type A. In conclusion, P. multocida type A is prevalent among poultry in the studied regions; thus, care must be taken to control of the disease. [J Adv Vet Anim Res 2015; 2(3.000: 338-345

  10. High and low-virulent bovine Pasteurella multocida capsular type A isolates exhibit different virulence gene expression patterns in vitro and in vivo.

    Science.gov (United States)

    Li, Nengzhang; Long, Qingshan; Du, Huihui; Zhang, Jixin; Pan, Tingting; Wu, Chenlu; Lei, Guihua; Peng, Yuanyi; Hardwidge, Philip R

    2016-11-30

    Pasteurella multocida capsular type A causes respiratory disease in cattle. P. multocida virulence gene expression patterns, especially among different virulent isolates, during in vitro and in vivo growth are poorly understood. Here we show that the highly virulent bovine P. multocida capsular type A isolate PmCQ2 exhibits a significantly higher growth rate in mice, as compared with a strain of lower virulence, P. multocida capsular type A isolate PmCQ6. Among the six known and potential virulence genes (ompA, ompH, pfhB2, hasR, pm0979, and pm0442) investigated, most genes were expressed more highly in both isolates when grown in vivo as compared with in vitro, with ompH and pm0442 having the highest magnitude of expression. Virulence gene expression was higher in PmCQ6 than in PmCQ2 during in vitro growth. However, in mice, most virulence genes were expressed more highly in PmCQ2 as compared with PmCQ6. Virulence gene expression was highest in the liver and lowest in the lung, but was uncorrelated to bacterial loads. This study indicates that individual pathogenic capacity of P. multocida isolates is associated with the virulence gene expression patterns in vivo growth but not in vitro, and the investigation of virulence gene expression in pathogen should be performed in vivo. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Development of a rapid multiplex PCR assay to genotype Pasteurella multocida strains by use of the lipopolysaccharide outer core biosynthesis locus.

    Science.gov (United States)

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

    2015-02-01

    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. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. The Pasteurella multocida nrfE gene is upregulated during infection and is essential for nitrite reduction but not for virulence.

    Science.gov (United States)

    Boucher, David J; Adler, Ben; Boyce, John D

    2005-04-01

    Pasteurella multocida is the causative agent of a range of diseases with economic importance in production animals. Many systems have been employed to identify virulence factors of P. multocida, including in vivo expression technology (IVET), signature-tagged mutagenesis, and whole-genome expression profiling. In a previous study in which IVET was used with P. multocida, nrfE was identified as a gene that is preferentially expressed in vivo. In Escherichia coli, nrfE is part of the formate-dependent nitrite reductase system involved in utilizing available nitrite as an electron accepter during growth under anaerobic conditions. In this study, we constructed an isogenic P. multocida strain that was unable to reduce nitrite under either aerobic or anaerobic conditions, thereby demonstrating that P. multocida nrfE is essential for nitrite reduction. However, the nrfE mutant was still virulent in mice. Real-time reverse transcription-PCR analysis indicated that nrfE was regulated independently of nrfABCD by an independent promoter that is likely to be upregulated in vivo.

  13. Activation of Galpha (i) and subsequent uncoupling of receptor-Galpha(i) signaling by Pasteurella multocida toxin.

    Science.gov (United States)

    Orth, Joachim H C; Fester, Ines; Preuss, Inga; Agnoletto, Laura; Wilson, Brenda A; Aktories, Klaus

    2008-08-22

    Bacterial protein toxins are powerful tools for elucidating signaling mechanisms in eukaryotic cells. A number of bacterial protein toxins, e.g. cholera toxin, pertussis toxin (PTx), or Pasteurella multocida toxin (PMT), target heterotrimeric G proteins and have been used to stimulate or block specific signaling pathways or to demonstrate the contribution of their target proteins in cellular effects. PMT is a major virulence factor of P. multocida causing pasteurellosis in man and animals and is responsible for atrophic rhinitis in pigs. PMT modulates various signaling pathways, including phospholipase Cbeta and RhoA, by acting on the heterotrimeric G proteins Galpha(q) and Galpha(12/13), respectively. Here we report that PMT is a powerful activator of G(i) protein. We show that PMT decreases basal isoproterenol and forskolin-stimulated cAMP accumulation in intact Swiss 3T3 cells, inhibits adenylyl cyclase activity in cell membrane preparations, and enhances the inhibition of cAMP accumulation caused by lysophosphatidic acid via endothelial differentiation gene receptors. PMT-mediated inhibition of cAMP production is independent of toxin activation of Galpha(q) and/or Galpha(12/13). Although the effects of PMT are not inhibited by PTx, PMT blocks PTx-catalyzed ADP-ribosylation of G(i). PMT also inhibits steady-state GTPase activity and GTP binding of G(i) in Swiss 3T3 cell membranes stimulated by lysophosphatidic acid. The data indicate that PMT is a novel activator of G(i), modulating its GTPase activity and converting it into a PTx-insensitive state.

  14. Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

    Directory of Open Access Journals (Sweden)

    Thales Q. Furian

    2013-02-01

    Full Text Available The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25, the sodA and nanH genes were present in 96% (24/25, ptfA was present in 92% (23/25, and pfhA was present in 60% (15/25. Gene toxA was not identified in any of the samples studied (0/25. Five different genetic profiles were obtained, of which P1 (negative to toxA was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.

  15. Comparison of two indirect ELISA coating antigens for the detection of dairy cow antibodies against Pasteurella multocida.

    Science.gov (United States)

    Tankaew, Pallop; Srisawat, Wanwisa; Singhla, Tawatchai; Tragoolpua, Khajornsak; Kataoka, Yasushi; Sawada, Takuo; Sthitmatee, Nattawooti

    2018-02-01

    The ELISA is recognized as an efficient diagnostic tool for antibody detection, but there is no standard ELISA assay for detection of antibodies against hemorrhagic septicemia (HS) in cattle. The present study reports on an indirect ELISA assay for antibody detection of HS in dairy cows, and evaluates the sensitivity (Se) and specificity (Sp) of the method using a Bayesian approach. An indirect ELISA was developed with two types of heat extract antigens, Pasteurella multocida strains P-1256 and M-1404, as coating antigens. A checkerboard titration was employed using dairy cow sera immunized with P. multocida bacterin and colostrum-deprived calf sera. The concentrations of heat extract antigen (160μg/mL), sample serum (1:100) and goat anti-bovine immunoglobulin G labeled with horseradish peroxidase (1:2000) were optimal for the assay. The cut-off values were 0.147 and 0.128 for P-1256 and M-1404 coating antigens, and there were no differences in the results of tests with positive and negative sera (p<0.05). The characteristics of three diagnostic tests were evaluated using a one-population Bayesian model, assuming conditional dependence between two types of coating antigen-based ELISAs and indirect hemagglutination assay (IHA). A total of 415 sera samples from dairy cows without HS vaccination and no history of disease were tested. The Se and Sp of the P-1256 and M-1404 ELISAs were higher than those of the IHA. The Se and Sp of the P-1256 ELISA were 90.3% and 90.1%, while the Se and Sp of the M-1404 ELISA were 92.1% and 71.9%. The median values of Se and Sp from the IHA were 36.0% and 58.2%. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Meningitis caused by Pasteurella multocida in a dog owner without a dog bite: clonal lineage identification by MALDI-TOF mass spectrometry.

    Science.gov (United States)

    Bardou, Matthieu; Honnorat, Estelle; Dubourg, Gregory; Couderc, Carine; Fournier, Pierre Edouard; Seng, Piseth; Stein, Andreas

    2015-10-31

    Pasteurella multocida meningitis in an immunocompetent patient is rare and commonly occurs after animal bite. To our knowledge, only 48 cases have been reported in the literature since 1989. P. multocida meningitis is commonly linked to animal contagion. Here we report on a new case of P. multocida meningitis in an immunocompetent patient who is a dog owner without a dog bite. We used the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry to investigate the clonal lineage between animal and human isolates. In our case, a 25-year-old immunocompetent French Caucasian woman with nothing notable in her medical history was admitted for meningitis caused by P. multocida. Clonal lineage of P. multocida strains from cerebrospinal fluid and blood culture and her dog's oral cavity has been recognized by MALDI-TOF mass spectrometry dendrograms and clustering of the 21 P. multocida isolates in our centres. She was treated by a combination of intravenous ceftriaxone (2 g/day) and oral levofloxacin (1 g/day). She was discharged on the 6th day of admission. The antimicrobial therapy was conducted for 15 days. The dog was treated by clavulanic-acid amoxicillin for 3 weeks by the veterinarian. The evolution of the patient at the 5th month after the end of the antimicrobial therapy was normal without any neurological after-effects. The meningitis caused by P. multocida could be considered a cause of human meningitis in dog lovers without an animal bite. MALDI-TOF mass spectrometry should be considered as it is an accurate tool to identify clonal lineage between animal and human isolates.

  17. Molecular heterogeneity of plpE gene in Indian isolates of Pasteurella multocida and expression of recombinant PlpE in vaccine strain of P. multocida serotype B: 2

    Science.gov (United States)

    Singh, Ajay Pratap; Singh, Satparkash; Gupta, Santosh Kumar; Singh, Vijendra Pal; Sharma, Bhaskar

    2010-01-01

    Outer membrane proteins of Pasteurella (P.) multocida have been known to be protective immunogens. Pasteurella lipoprotein E (PlpE) has been reported to be an important cross reactive outer membrane protein in P. multocida. The gene encoding the PlpE of P. multocida serotypes A: 3, B: 2 and D: 1 was amplified from the genomic DNA. The amplified products were cloned and the nucleotide sequence was determined. Sequence analysis of the recombinant clones revealed a single open reading frame of 1,011 bp, 1,008 bp and 1,017 bp encoding a protein with a calculated molecular mass of 37.829 kDa, 37.389 kDa and 37.965 kDa for serotypes A: 3, B: 2 and D: 1 respectively. The comparison of the plpE sequence in different capsular types revealed a high degree (>90%) of homology. Furthermore, the plpE gene of Haemorhhagic septicaemia causing serotype (B: 2) was expressed in E. coli and recombinant PlpE was strongly immunostained by antiserum against whole cell antigen, indicating that the protein is expressed in vivo. PMID:20706030

  18. Use of a dual reporter plasmid to demonstrate Bactofection with an attenuated AroA(- derivative of Pasteurella multocida B:2.

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    Sarah Othman

    Full Text Available A reporter plasmid pSRG has been developed which expresses red fluorescent protein (RFP from a constitutive prokaryotic promoter within Pasteurella multocida B:2 and green fluorescent protein (GFP from a constitutive eukaryotic promoter within mammalian cells. This construct has been used to determine the location and viability of the bacteria when moving from the extracellular environment into the intracellular compartment of mammalian cells. Invasion assays with embryonic bovine lung (EBL cells and an attenuated AroA(- derivative of Pasteurella multocida B:2 (strain JRMT12, harbouring the plasmid pSRG, showed that RFP-expressing bacteria could be detected intracellularly at 3 h post-invasion. At this stage, some EBL cells harbouring RFP-expressing bacteria were observed to express GFP simultaneously, indicating release of the plasmid into the intracellular environment. At 5 h post-invasion, more EBL cells were expressing GFP, while still harbouring RFP-expressing bacteria. Concurrently, some EBL cells were shown to express only GFP, indicating loss of viable bacteria within these cells. These experiments proved the functionality of the pSRG dual reporter system and the potential of P. multocida B:2 JRMT12 for bactofection and delivery of a DNA vaccine.

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

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    Stefan Carle

    2017-01-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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.

  2. Crystal structures reveal a thiol protease-like catalytic triad in the C-terminal region of Pasteurella multocida toxin.

    Science.gov (United States)

    Kitadokoro, Kengo; Kamitani, Shigeki; Miyazawa, Masayuki; Hanajima-Ozawa, Miyuki; Fukui, Aya; Miyake, Masami; Horiguchi, Yasuhiko

    2007-03-20

    Pasteurella multocida toxin (PMT), one of the virulence factors produced by the bacteria, exerts its toxicity by up-regulating various signaling cascades downstream of the heterotrimeric GTPases Gq and G12/13 in an unknown fashion. Here, we present the crystal structure of the C-terminal region (residues 575-1,285) of PMT, which carries an intracellularly active moiety. The overall structure of C-terminal region of PMT displays a Trojan horse-like shape, composed of three domains with a "feet"-,"body"-, and "head"-type arrangement, which were designated C1, C2, and C3 from the N to the C terminus, respectively. The C1 domain, showing marked similarity in steric structure to the N-terminal domain of Clostridium difficile toxin B, was found to lead the toxin molecule to the plasma membrane. The C3 domain possesses the Cys-His-Asp catalytic triad that is organized only when the Cys is released from a disulfide bond. The steric alignment of the triad corresponded well to that of papain or other enzymes carrying Cys-His-Asp. PMT toxicities on target cells were completely abrogated when one of the amino acids constituting the triad was mutated. Our results indicate that PMT is an enzyme toxin carrying the cysteine protease-like catalytic triad dependent on the redox state and functions on the cytoplasmic face of the plasma membrane of target cells.

  3. 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

    OpenAIRE

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

    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) molec...

  4. Evaluation of the biofilm formation capacity of Pasteurella multocida strains isolated from cases of fowl cholera and swine lungs and its relationship with pathogenicity

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    Brunna D. de Emery

    Full Text Available ABSTRACT: Pasteurella multocida is a Gram-negative bacillus that causes economic losses due to the development of respiratory diseases in several animal species. Among the mechanisms of virulence, the formation of biofilms is an important factor for bacterial survival in hostile environments. Studies of biofilm formation by P. multocida are needed because P. multocida is an important pathogen involved in respiratory infections. However, in contrast to other microorganisms, few studies of biofilm formation have examined P. multocida. Studies comparing the pathogenicity of microbial strains as a function of their biofilm production capacity are also rare. Consequently, the aim of this study was to evaluate the biofilm formation capacity of 94 P. multocida strains isolated from cases of fowl cholera and from swine lungs on polystyrene plates. The associations of the biofilm formation capacity with the pathogenicity index (PI in vivo and with the presence of four genes (screened by PCR of the tad locus (tadB, tadD, tadE and tadG, described as adhesion markers, were also determined. Strains from both animal origins were able to form biofilms. However, most of the specimens (52.13% were classified as weak producers, and more than 40% of the strains of P. multocida (40.42% did not produce biofilms. There was no significant difference (p>0.05 in the degree of biofilm production between the two sources of isolation. Of the analyzed strains, 56.52% contained all four genes (tadB, tadD, tadE and tadG. The PI arithmetic mean of the strains classified as non-biofilm producers was significantly different (p0.05 with the production of biofilms and with the origin of a given strain. Finally, low virulence strains may suggest a higher biofilm formation capacity on polystyrene plates.

  5. Urinary tract infection with Pasteurella multocida in a patient with cat exposure and abnormal urinary tract physiology: Case report and literature review

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    Joel T. Costanzo, II

    2017-01-01

    Full Text Available Pasteurella multocida is a gram-negative organism that commonly colonizes the mouth of cats and dogs, and is known to cause infection in humans associated with animal bites or scratches. Sites of infection other than skin and soft tissue are rare, but have been reported in patients with specific risk factors including anatomical abnormalities or immunosuppression. Herein, we report a case of a symptomatic urinary tract infection caused by P. multocida in a 59 year old female who presented to the hospital with complaints of systemic symptoms including malaise, rigors, and chills, as well as thick, malodorous urine. The patient self-catheterized multiple times daily due to urostomy with Kock pouch. Treatment with piperacillin/tazobactam followed by amoxicillin resulted in resolution of the infection.

  6. Susceptibilidade in vitro a antimicrobianos da Mannheimia haemolytica e da Pasteurella multocida isoladas de ovinos sadios e com doenças respiratórias

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    Leomar Viana

    2007-12-01

    Full Text Available Pasteurella multocida e Mannheimia haemolytica (P. haemolytica estão associadas a enfermidades no sistema respiratório de ovinos. Com o objetivo de avaliar a susceptibilidade in vitro destes microrganismos frente aos antimicrobianos, foram colhidas amostras de nasofaringe (n=180 e orofaringe (n=82 de ovinos com e sem enfermidade respiratória. Dentre os antimicrobianos testados, a sensibilidade foi maior para enrofloxacina (100% e florfenicol (100%, considerando-se ambas as espécies bacterianas. Observou-se resistência de M. haemolytica e P. multocida à tetraciclina (15,64% e 17,65%, respectivamente e penicilina (1.82% e 4.2%, respectivamente.

  7. Effects of dietary L-glutamine supplementation on specific and general defense responses in mice immunized with inactivated Pasteurella multocida vaccine.

    Science.gov (United States)

    Chen, Shuai; Liu, Shuping; Zhang, Fengmei; Ren, Wenkai; Li, Nengzhang; Yin, Jie; Duan, Jielin; Peng, Yuanyi; Liu, Gang; Yin, Yulong; Wu, Guoyao

    2014-10-01

    Little is known about effects of dietary glutamine supplementation on specific and general defense responses in a vaccine-immunized animal model. Thus, this study determined roles for dietary glutamine supplementation in specific and general defense responses in mice immunized with inactivated Pasteurella multocida vaccine. The measured variables included: (1) the production of pathogen-specific antibodies; (2) mRNA levels for pro-inflammatory cytokines, toll-like receptors and anti-oxidative factors; and (3) the distribution of P. multocida in tissues and the expression of its major virulence factors in vivo. Dietary supplementation with 0.5 % glutamine had a better protective role than 1 or 2 % glutamine against P. multocida infection in vaccine-immunized mice, at least partly resulting from its effects in modulation of general defense responses. Dietary glutamine supplementation had little effects on the production of P. multocida-specific antibodies. Compared to the non-supplemented group, dietary supplementation with 0.5 % glutamine had no effect on bacterial burden in vivo but decreased the expression of major virulence factors in the spleen. Collectively, supplementing 0.5 % glutamine to a conventional diet provides benefits in vaccine-immunized mice by enhancing general defense responses and decreasing expression of specific virulence factors.

  8. Integrated pharmacokinetic-Pharmacodynamic (PK/PD) model to evaluate the in vivo antimicrobial activity of Marbofloxacin against Pasteurella multocida in piglets.

    Science.gov (United States)

    Zeng, Qing Lin; Mei, Xian; Su, Jia; Li, Xiao Hong; Xiong, Wen Guang; Lu, Yan; Zeng, Zhen Ling

    2017-06-15

    Marbofloxacin is a veterinary fluoroquinolone with high activity against Pasteurella multocida. We evaluated it's in vivo activity against P. multocida based on in vivo time-kill data in swine using a tissue-cage model. A series of dosages ranging from 0.15 to 2.5 mg/kg were administered intramuscularly after challenge with P. multocida type B, serotype 2. The ratio of the 24 h area under the concentration-time curve divided by the minimum inhibitory concentration (AUC24TCF/MIC) was the best PK/PD index correlated with the in vivo antibacterial effectiveness of marbofloxacin (R2 = 0.9279). The AUC24TCF/MIC necessary to achieve a 1-log10 CFU/ml reduction and a 3-log10 CFU/ml (90% of the maximum response) reduction as calculated by an inhibitory sigmoid Emax model were 13.48 h and 57.70 h, respectively. Marbofloxacin is adequate for the treatment of swine infected with P. multocida. The tissue-cage model played a significant role in achieving these PK/PD results.

  9. MLST typing of Pasteurella multocida associated with haemorrhagic septicaemia and development of a real-time PCR specific for haemorrhagic septicaemia associated isolates.

    Science.gov (United States)

    Petersen, Andreas; Bisgaard, Magne; Townsend, Kirsty; Christensen, Henrik

    2014-06-04

    Two serovars of Pasteurella multocida, B:2 and E:2, have been reportedly associated with haemorrhagic septicaemia (HS), a peracute and devastating disease mainly affecting cattle and water buffaloes. We multilocus sequence typed (MLST) 64 isolates of P. multocida including 55 associated with HS and found that they mainly included sequence type (ST) 122 (n=50) and rarely ST63 (n=1), ST147 (n=2) and ST162 (n=2) compared to other members of the species isolated from other lesion types and hosts. Single-nucleotide polymorphisms suitable for specific detection of STs associated with HS were detected in the est gene. A new HS-est-RT-PCR (est indicating the target gene) specifically detected ST122, ST63, ST147 and ST162 associated with HS. The new HS-est-RT-PCR did not detect strains of ST151 with capsular type D isolated from pigs that were found positive with a previously published HS PCR detection method. The new HS-est-RT-PCR represents a fast and specific detection of the specific types of P. multocida involved in HS. The HS-est-RT-PCR developed in the current study seems to more accurately identify isolates of P. multocida associated with HS compared to PCR detection methods previously published. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Time-course investigation of infection with a low virulent Pasteurella multocida strain in normal and immune-suppressed 12-week-old free-range chickens.

    Science.gov (United States)

    Mbuthia, P G; Njagi, L W; Nyaga, P N; Bebora, L C; Minga, U; Christensen, J P; Olsen, J E

    2011-12-01

    Twelve-week-old indigenous chickens, either immune-suppressed using dexamethasone (IS) or non-immune-suppressed (NIS), were challenged with a low virulent strain, Pasteurella multocida strain NCTC 10322(T), and developed clinical signs and pathological lesions typical of chronic fowl cholera. NIS birds demonstrated much more severe signs of fowl cholera than IS birds. With few exceptions, signs recorded in IS and NIS birds were of the same types, but significantly milder in the IS birds, indicating that immune suppression does not change the course of infection but rather the severity of signs in fowl cholera. P. multocida signals by fluorescent in situ hybridization (FISH) were observed between 1 h and 14 days in the lungs, trachea, air sacs, liver, spleen, bursa of Fabricius and caecal tonsils, while signals from other organs mostly were observed after 24 h. More organs had FISH signals in NIS birds than in IS birds and at higher frequency per organ. Many organs were positive by FISH even 14 days post infection, and it is suggested that these organs may be likely places for long-term carriage of P. multocida following infection. The present study has demonstrated the spread of P. multocida in different tissues in chickens and distribution of lesions associated with chronic fowl cholera, and pointed to a decrease of pathology in IS birds. Since dexamethasone mostly affects heterophils, the study suggests that these cells play a role in the development of lesions associated with chronic fowl cholera in chickens.

  11. Evaluación de dos técnicas de subtipificación molecular para el estudio de Pasteurella multocida Evaluation of two techniques of molecular subtyping to study Pasteurella multocida

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    G. A. Leotta

    2006-12-01

    Full Text Available Se determinó la tipibilidad, la reproducibilidad y el poder discriminatorio de ERIC-PCR y ApaI-PFGE para establecer la relación genética de cepas de Pasteurella multocida. Se estudiaron 49 cepas de diferente origen, subespecie, biotipo, grupo capsular, serotipo somático y perfil de resistencia antimicrobiana. Por ERIC-PCR se establecieron 31 patrones, los que presentaron entre 10 y 14 bandas en un rango comprendido entre 0,2 y 1,2 kb. Por ApaI-PFGE se detectaron 37 patrones de restricción, los cuales presentaron entre 7 y 15 bandas bien definidas de 34 a 450 kb. La tipibilidad de ERIC-PCR fue del 100% (T=1 y la de ApaI-PFGE del 94% (T=0,94. La reproducibilidad de ambas técnicas fue del 100% (R=1; sin embargo, el poder discriminatorio de ERIC-PCR fue 93% (D=0,93 y el de ApaI-PFGE 98% (D=0,98. Mediante ambas técnicas fue posible agrupar las cepas con relación epidemiológica y diferenciar claramente las cepas no relacionadas. Se demostró el valor de ERIC-PCR y ApaI-PFGE para complementar estudios epidemiológicos, principalmente si las cepas en estudio son analizadas por ambas técnicas.Typeability, reproducibility, and discriminatory power of ERIC-PCR and ApaI-PFGE to establish the genetic relation of P. multocida strains were determined. Forty-nine strains of different source, biotype, capsular group, somatic serotype, and resistance to antimicrobials were studied. By ERIC-PCR, 31 patterns were defined with 10 to 14 bands in a rank of 0.2 and 1.2 kb. By ApaI-PFGE, 37 restriction patterns were established with 7 to 15 bands of 34 to 450 kb. Typeability was 100% (T=1 for ERIC-PCR, and 94% (T=0.94 for ApaI-PFGE. Reproducibility of both techniques was 100% (R=1. Discriminatory power was 93% (D=0.93 for ERIC-PCR, and 98% (D=0.98 for ApaI-PFGE. By using both techniques, epidemiologically related strains were grouped, and unrelated strains were clearly differentiated. The value of ERIC-PCR and ApaI-PFGE as complements to epidemiologic studies

  12. Pasteurella multocida from outbreaks of avian cholera in wild and captive birds in Denmark

    DEFF Research Database (Denmark)

    Pedersen, Karl; Dietz, Hans-Henrik; Jørgensen, J.C.

    2003-01-01

    An outbreak of avian cholera was observed among wild birds in a few localities in Denmark in 2001. The highest mortalities were among breeding ciders (Somateria mollissima) and gulls (Larus spp.). Pulsed-field gel electrophoresis (PFGE) was conducted using ApaI and SmaI as restriction enzymes...... the outbreak strain. Among 68 isolates from wild birds, only one PFGE and one REA pattern were demonstrated, whereas among 23 isolates from domestic poultry, 14 different SmaI, 12 different ApaI, and 10 different HpaII patterns were found. The results suggest that a P. multocida strain has survived during...

  13. The origin of Pasteurella multocida impacts pathology and inflammation when assessed in a mouse model

    DEFF Research Database (Denmark)

    Pors, Susanne E.; Chadfield, Mark S.; Sorensen, Dorte B.

    2016-01-01

    ) of an isolate from porcine pneumonia or fowl cholera showed marked differences between the two isolates. The avian isolate was highly pathogenic with severe signs of necrotizing pneumonia, liver necrosis and high bacterial load in lung and liver. Clinical signs and pathology related to the porcine isolate were...... dose dependent and consisted of exudative bronchopneumonia, abscess formation in liver and a lower bacterial load in lung and liver. Both isolates caused increased expression of MMP9 and TIMP1. In conclusion, evaluation and comparison of pathogenicity and host-pathogen interaction of P. multocida...

  14. Specific detection of Pasteurella multocida in chickens with fowl cholera and in pig lung tissues using fluorescent rRNA in situ hybridization

    DEFF Research Database (Denmark)

    Mbuthia, P.G.; Christensen, H.; Boye, Mette

    2001-01-01

    in formalin-fixed paraffin-embedded lung tissues from experimental fowl cholera in chickens and infections in pigs. In chicken lung tissues P. multocida cells were detected singly, in pairs, as microcolonies, and as massive colonies within air capillaries (septa and lumen), parabronchial septa, and blood...... and fast method for specific detection of P. multocida in histological formalin-fixed tissues. The test was replicable and reproducible and is recommended as a supplementary test for diagnosis and as a tool in pathogenesis studies of fowl cholera and respiratory tract infections in pigs due to P. multocida....

  15. Combinations of macrolide resistance determinants in field isolates of Mannheimia haemolytica and Pasteurella multocida

    DEFF Research Database (Denmark)

    Desmolaize, Benoit; Rose, Simon; Wilhelm, Cornelia

    2011-01-01

    )] that are arranged in tandem and presumably expressed from the same promoter. The third class exhibits the most marked drug phenotype, with high resistance to all of the macrolides tested, and possesses all three resistance determinants. The combinations of erm(42), msr(E), and mph(E) are chromosomally encoded...... of the erm(42) gene. The second class shows no lincosamide resistance and lacks erm(42) and concomitant 23S rRNA methylation. Sequencing of the genome of a representative strain from this class, P. multocida 3361, revealed macrolide efflux and phosphotransferase genes [respectively termed msr(E) and mph(E...... and intermingled with other exogenous genes, many of which appear to have been transferred from other members of the Pasteurellaceae. The presence of some of the exogenous genes explains recent reports of resistance to additional drug classes. We have expressed recombinant versions of the erm(42), msr(E), and mph(E...

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

    Directory of Open Access Journals (Sweden)

    Rebecca C Babb

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

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    Tracy P. M. Chong

    2011-03-01

    Full Text Available 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.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Distribution of the ompA-types among ruminant and swine pneumonic strains of Pasteurella multocida exhibiting various cap-locus and toxA patterns.

    Science.gov (United States)

    Vougidou, C; Sandalakis, V; Psaroulaki, A; Siarkou, V; Petridou, E; Ekateriniadou, L

    2015-05-01

    Pasteurella multocida is an important pathogen in food-producing animals and numerous virulence genes have been identified in an attempt to elucidate the pathogenesis of pasteurellosis. Currently, some of these genes including the capsule biosynthesis genes, the toxA and the OMPs-encoding genes have been suggested as epidemiological markers. However, the number of studies concerning ruminant isolates is limited, while, no attempt has ever been made to investigate the existence of ompA sequence diversity among P. multocida isolates. The aim of the present study was the comparative analysis of 144 P. multocida pneumonic isolates obtained from sheep, goats, cattle and pigs by determining the distribution of the ompA-types in conjunction with the cap-locus and toxA patterns. The ompA genotypes of the isolates were determined using both a PCR-RFLP method and DNA sequence analysis. The most prevalent capsule biosynthesis gene among the isolates was capA (86.1%); a noticeable, however, rate of capD-positive isolates (38.6%) was found among the ovine isolates that had been associated primarily with the capsule type A in the past. Moreover, an unexpectedly high percentage of toxA-positive pneumonic isolates was noticed among small ruminants (93.2% and 85.7% in sheep and goats, respectively), indicating an important epidemiological role of toxigenic P. multocida for these species. Despite their great heterogeneity, certain ompA-genotypes were associated with specific host species, showing evidence of a host preference. The OmpA-based PCR-RFLP method developed proved to be a valuable tool in typing P. multocida strains. Copyright © 2015 Elsevier GmbH. All rights reserved.

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

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    Moemen A. Mohamed

    2012-03-01

    Full Text Available 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.

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

    Science.gov (United States)

    2013-01-01

    Background Pasteurella multocida is the etiologic agent of fowl cholera, a highly contagious and severe disease of poultry causing significant mortality and morbidity throughout the world. All types of poultry are susceptible to fowl cholera. Turkeys are most susceptible to the peracute/acute forms of the disease while chickens are most susceptible to the acute and chronic forms of the disease. The whole genome of the Pm70 strain of P. multocida was sequenced and annotated in 2001. The Pm70 strain is not virulent to chickens and turkeys. In contrast, strains X73 and P1059 are highly virulent to turkeys, chickens, and other poultry species. In this study, we sequenced the genomes of P. multocida strains X73 and P1059 and undertook a detailed comparative genome analysis with the avirulent Pm70 strain. The goal of this study was to identify candidate genes in the virulent strains that may be involved in pathogenicity of fowl cholera disease. Results Comparison of virulent versus avirulent avian P. multocida genomes revealed 336 unique genes among the P1059 and/or X73 genomes compared to strain Pm70. Genes of interest within this subset included those encoding an L-fucose transport and utilization system, several novel sugar transport systems, and several novel hemagglutinins including one designated PfhB4. Additionally, substantial amino acid variation was observed in many core outer membrane proteins and single nucleotide polymorphism analysis confirmed a higher dN/dS ratio within proteins localized to the outer membrane. Conclusions Comparative analyses of highly virulent versus avirulent avian P. multocida identified a number of genomic differences that may shed light on the ability of highly virulent strains to cause disease in the avian host, including those that could be associated with enhanced virulence or fitness. PMID:23672515

  3. Use of radioactively labelled bacteria in animal experiments. 4. Distribution of radioactivity in the mouse after intratracheal administration of labelled Pasteurella multocida germs

    Energy Technology Data Exchange (ETDEWEB)

    Flossmann, K.D.; Mueller, G.; Heilmann, P.; Finsterbusch, L.; Hubald, J. (Akademie der Landwirtschaftswissenschaften der DDR, Jena. Inst. fuer Bakterielle Tierseuchenforschung)

    1981-03-01

    Intratracheal administration of /sup 3/H-, /sup 14/C-, /sup 59/Fe- or /sup 125/I-labelled Pasteurella multocida germs to mice resulted in a more or less differentiated, nuclidedependent distribution of radioactivity in blood, spleen, liver, lung, kidney, and gastrointestinal tract, and was comparable to that following subcutaneous application. The elimination of the antigen from the lungs and other organs could be characterised by an e function, after having reached a certain level of distribution. Some of the antigen was persistent in the lung not less than 14 days. Extremely high activity concentration and persistence was recordable, following the use of /sup 59/Fe complete antigen. Phagocytosis of Pasteurella multocida germs through alveolar macrophages of the lung was secured by autoradiography. Most of the antigen seemed to be discharged from the lungs through the digestive tract. Antigen distribution recorded from immunized and non-immunized mice seemed to suggest that the fate of antigen applied was affected by the kind of immunization. No difference in antigen distribution between non-immunized and subcutaneously immunized animals were provable, following intratracheal antigen application, but is was clearly provable, following intratracheal immunization. Elimination of antigen from the lungs of intratracheally immunized animals was found to occur faster than it did from non-immunized animals.

  4. Use of radioactively labelled bacteria in animal experiments. 5. Fate of radioactively labelled Pasteurella multocida germs, following oral administration to mice

    Energy Technology Data Exchange (ETDEWEB)

    Flossmann, K.D.; Mueller, G.; Heilmann, P. (Akademie der Landwirtschaftswissenschaften der DDR, Jena. Inst. fuer Bakterielle Tierseuchenforschung)

    1981-09-01

    5-/sup 3/H- or 6-/sup 14/C-orotic acid labelled Pasteurella multocida were orally administered, as live or inactivated vaccine, to mice. They were soon discharged through the digestive tract but also led to growing activity in the blood, liver, spleen, and lungs. Using germs with /sup 3/H-1-D mannose-labelled cellular surface have shown that parts of the bacterial cells were easily detached and distributed through the organism, after oral, subcutaneous as well as intratracheal application. Hence, the conclusion seems to be justified that oral administration of Pasteurella multocida cells leads to resorption of bacterial cell fragments which then will readily penetrate the circulatory system, while many of these cells will soon be eliminated from the organism, largely through the digestive tract. Such elimination was found to follow an e function, and it was observed also after intratracheal and subcutaneous application, resp. Some bacteria remained in the digestive tract over a prolonged period (14 days verified). The results are discussed with regard to their possible importance to the immunological development.

  5. Clinico-pathology and hemato-biochemistry responses in buffaloes infected with Pasteurella multocida type B:2 immunogen outer membrane protein.

    Science.gov (United States)

    Chung, Eric Lim Teik; Abdullah, Faez Firdaus Jesse; Marza, Ali Dhiaa; Saleh, Wessam Monther Mohammed; Ibrahim, Hayder Hamzah; Abba, Yusuf; Zamri-Saad, Mohd; Haron, Abd Wahid; Saharee, Abdul Aziz; Lila, Mohd Azmi Mohd; Norsidin, Mohd Jefri

    2017-01-01

    The aim of this study was to investigate the clinico-pathology and haemato-biochemistry alterations in buffaloes inoculated with Pasteurella multocida type B:2 immunogen outer membrane protein via subcutaneous and oral routes. Nine 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 outer membrane protein broth subcutaneously and orally respectively. Group 2 buffaloes showed typical haemorrhagic septicaemia clinical signs and were only able to survive for 72 h of the experiment. However, Group 3 buffaloes were able to survive throughout the stipulated time of 21 days of experiment. There were significant differences (p  0.05) in edema between groups except for the lung. This study was a proof that oral route infection of Pasteurella multocida type B:2 immunogen outer membrane protein can be used to stimulate host cell. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Pasteurella multocida toxin prevents osteoblast differentiation by transactivation of the MAP-kinase cascade via the Gα(q/11--p63RhoGEF--RhoA axis.

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    Peter Siegert

    Full Text Available The 146-kDa Pasteurella multocida toxin (PMT is the main virulence factor to induce P. multocida-associated progressive atrophic rhinitis in various animals. PMT leads to a destruction of nasal turbinate bones implicating an effect of the toxin on osteoblasts and/or osteoclasts. The toxin induces constitutive activation of Gα proteins of the G(q/11-, G₁₂/₁₃- and G(i-family by deamidating an essential glutamine residue. To study the PMT effect on bone cells, we used primary osteoblasts derived from rat calvariae and stromal ST-2 cells as differentiation model. As marker of functional osteoblasts the expression and activity of alkaline phosphatase, formation of mineralization nodules or expression of specific transcription factors as osterix was determined. Here, we show that the toxin inhibits differentiation and/or function of osteoblasts by activation of Gα(q/11. Subsequently, Gα(q/11 activates RhoA via p63RhoGEF, which specifically interacts with Gα(q/11 but not with other G proteins like Gα₁₂/₁₃ and Gα(i. Activated RhoA transactivates the mitogen-activated protein (MAP kinase cascade via Rho kinase, involving Ras, MEK and ERK, resulting in inhibition of osteoblast differentiation. PMT-induced inhibition of differentiation was selective for the osteoblast lineage as adipocyte-like differentiation of ST-2 cells was not hampered. The present work provides novel insights, how the bacterial toxin PMT can control osteoblastic development by activating heterotrimeric G proteins of the Gα(q/11-family and is a molecular pathogenetic basis for understanding the role of the toxin in bone loss during progressive atrophic rhinitis induced by Pasteurella multocida.

  7. Pasteurella multocida toxin prevents osteoblast differentiation by transactivation of the MAP-kinase cascade via the Gα(q/11)--p63RhoGEF--RhoA axis.

    Science.gov (United States)

    Siegert, Peter; Schmidt, Gudula; Papatheodorou, Panagiotis; Wieland, Thomas; Aktories, Klaus; Orth, Joachim H C

    2013-01-01

    The 146-kDa Pasteurella multocida toxin (PMT) is the main virulence factor to induce P. multocida-associated progressive atrophic rhinitis in various animals. PMT leads to a destruction of nasal turbinate bones implicating an effect of the toxin on osteoblasts and/or osteoclasts. The toxin induces constitutive activation of Gα proteins of the G(q/11)-, G₁₂/₁₃- and G(i)-family by deamidating an essential glutamine residue. To study the PMT effect on bone cells, we used primary osteoblasts derived from rat calvariae and stromal ST-2 cells as differentiation model. As marker of functional osteoblasts the expression and activity of alkaline phosphatase, formation of mineralization nodules or expression of specific transcription factors as osterix was determined. Here, we show that the toxin inhibits differentiation and/or function of osteoblasts by activation of Gα(q/11). Subsequently, Gα(q/11) activates RhoA via p63RhoGEF, which specifically interacts with Gα(q/11) but not with other G proteins like Gα₁₂/₁₃ and Gα(i). Activated RhoA transactivates the mitogen-activated protein (MAP) kinase cascade via Rho kinase, involving Ras, MEK and ERK, resulting in inhibition of osteoblast differentiation. PMT-induced inhibition of differentiation was selective for the osteoblast lineage as adipocyte-like differentiation of ST-2 cells was not hampered. The present work provides novel insights, how the bacterial toxin PMT can control osteoblastic development by activating heterotrimeric G proteins of the Gα(q/11)-family and is a molecular pathogenetic basis for understanding the role of the toxin in bone loss during progressive atrophic rhinitis induced by Pasteurella multocida.

  8. Virulence Genotyping of Pasteurella multocida Isolated from Multiple Hosts from India

    Science.gov (United States)

    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. PMID:25485303

  9. Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes.

    Directory of Open Access Journals (Sweden)

    Ahmed M Moustafa

    Full Text Available Pasteurella multocida is the primary causative agent of a range of economically important diseases in animals, including haemorrhagic septicaemia (HS, a rapidly fatal disease of ungulates. There is limited information available on the diversity of P. multocida strains that cause HS. Therefore, we determined draft genome sequences of ten disease-causing isolates and two vaccine strains and compared these genomes using a range of bioinformatic analyses. The draft genomes of the 12 HS strains were between 2,298,035 and 2,410,300 bp in length. Comparison of these genomes with the North American HS strain, M1404, and other available P. multocida genomes (Pm70, 3480, 36950 and HN06 identified a core set of 1,824 genes. A set of 96 genes was present in all HS isolates and vaccine strains examined in this study, but absent from Pm70, 3480, 36950 and HN06. Moreover, 59 genes were shared only by the Asian B:2 strains. In two Pakistani isolates, genes with high similarity to genes in the integrative and conjugative element, ICEPmu1 from strain 36950 were identified along with a range of other antimicrobial resistance genes. Phylogenetic analysis indicated that the HS strains formed clades based on their country of isolation. Future analysis of the 96 genes unique to the HS isolates will aid the identification of HS-specific virulence attributes and facilitate the development of disease-specific diagnostic tests.

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

    Science.gov (United States)

    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

  11. 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.

    Science.gov (United States)

    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.

  12. Comparison of standardised versus non-standardised methods for testing the in vitro potency of oxytetracycline against Mannheimia haemolytica and Pasteurella multocida.

    Science.gov (United States)

    Lees, P; Illambas, J; Pelligand, L; Toutain, P-L

    2016-12-01

    The in vitro pharmacodynamics of oxytetracycline was established for six isolates of each of the calf pneumonia pathogens Mannheimia haemolytica and Pasteurella multocida. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and bacterial time-kill curves were determined in two matrices, Mueller Hinton broth (MHB) and calf serum. Geometric mean MIC ratios, serum:MHB, were 25.2:1 (M. haemolytica) and 27.4:1 (P. multocida). The degree of binding of oxytetracycline to serum protein was 52.4%. Differences between serum and broth MICs could not be accounted for by oxytetracycline binding to serum protein. In vitro time-kill data suggested a co-dependent killing action of oxytetracycline. The in vitro data indicate inhibition of the killing action of oxytetracycline by serum factor(s). The nature of the inhibition requires further study. The outcome of treatment with oxytetracycline of respiratory tract infections in calves caused by M. haemolytica and P. multocida may not be related solely to a direct killing action. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. Comparative Genomic Analysis of Asian Haemorrhagic Septicaemia-Associated Strains of Pasteurella multocida Identifies More than 90 Haemorrhagic Septicaemia-Specific Genes.

    Science.gov (United States)

    Moustafa, Ahmed M; Seemann, Torsten; Gladman, Simon; Adler, Ben; Harper, Marina; Boyce, John D; Bennett, Mark D

    2015-01-01

    Pasteurella multocida is the primary causative agent of a range of economically important diseases in animals, including haemorrhagic septicaemia (HS), a rapidly fatal disease of ungulates. There is limited information available on the diversity of P. multocida strains that cause HS. Therefore, we determined draft genome sequences of ten disease-causing isolates and two vaccine strains and compared these genomes using a range of bioinformatic analyses. The draft genomes of the 12 HS strains were between 2,298,035 and 2,410,300 bp in length. Comparison of these genomes with the North American HS strain, M1404, and other available P. multocida genomes (Pm70, 3480, 36950 and HN06) identified a core set of 1,824 genes. A set of 96 genes was present in all HS isolates and vaccine strains examined in this study, but absent from Pm70, 3480, 36950 and HN06. Moreover, 59 genes were shared only by the Asian B:2 strains. In two Pakistani isolates, genes with high similarity to genes in the integrative and conjugative element, ICEPmu1 from strain 36950 were identified along with a range of other antimicrobial resistance genes. Phylogenetic analysis indicated that the HS strains formed clades based on their country of isolation. Future analysis of the 96 genes unique to the HS isolates will aid the identification of HS-specific virulence attributes and facilitate the development of disease-specific diagnostic tests.

  14. Polymerase chain reaction-restriction fragment length polymorphism analysis of the genes involved in the biosynthesis of the lipopolysaccharide of Pasteurella multocida.

    Science.gov (United States)

    Tsai, Yu-Chen; Shien, Jui-Hung; Wu, Jin-Ru; Shieh, Happy K; Chang, Poa-Chun

    2011-05-01

    The lipopolysaccharide, also known as the somatic antigen or O-antigen, is an important virulence factor of Pasteurella multocida. In the current study, the genes involved in the biosynthesis of the outer core region of the lipopolysaccharide, which were obtained from somatic type reference strains and field strains of P. multocida, were subjected to polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. The PCR-RFLP analysis classified 11 out of the 16 serotypes into 5 PCR-RFLP types (I-V). Types I and V contain strains belong to serotypes 1 and 13, respectively. The rest of the PCR-RFLP types contain strains belong to certain groups of serotypes. Typing of 38 field strains from poultry using PCR-RFLP analysis and the gel diffusion precipitation test showed consistent results. These results indicate that the PCR-RFLP analysis can be a useful tool for rapid somatic typing of some strains of P. multocida. © 2011 The Author(s)

  15. Fis Is Essential for Capsule Production in Pasteurella multocida and Regulates Expression of Other Important Virulence Factors

    OpenAIRE

    Steen, Jason A.; Steen, Jennifer A.; Paul Harrison; Torsten Seemann; Ian Wilkie; Marina Harper; Ben Adler; Boyce, John D.

    2010-01-01

    P. multocida is the causative agent of a wide range of diseases of animals, including fowl cholera in poultry and wild birds. Fowl cholera isolates of P. multocida generally express a capsular polysaccharide composed of hyaluronic acid. There have been reports of spontaneous capsule loss in P. multocida, but the mechanism by which this occurs has not been determined. In this study, we identified three independent strains that had spontaneously lost the ability to produce capsular polysacchari...

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

    Science.gov (United States)

    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 oral group. In hematology and biochemistry findings, there were significant differences (p 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 edema lesion between groups. In conclusion, this study is a proof that oral route infection of P. multocida type B:2 immunogen lipopolysaccharide can be used to stimulate host cell responses where oral vaccine through feed could be developed in the near future

  17. Regulation of Toll-like receptor 4-mediated immune responses through Pasteurella multocida toxin-induced G protein signalling

    Directory of Open Access Journals (Sweden)

    Hildebrand Dagmar

    2012-08-01

    Full Text Available Abstract Background Lipopolysaccharide (LPS-triggered Toll-like receptor (TLR 4-signalling belongs to the key innate defence mechanisms upon infection with Gram-negative bacteria and triggers the subsequent activation of adaptive immunity. There is an active crosstalk between TLR4-mediated and other signalling cascades to secure an effective immune response, but also to prevent excessive inflammation. Many pathogens induce signalling cascades via secreted factors that interfere with TLR signalling to modify and presumably escape the host response. In this context heterotrimeric G proteins and their coupled receptors have been recognized as major cellular targets. Toxigenic strains of Gram-negative Pasteurella multocida produce a toxin (PMT that constitutively activates the heterotrimeric G proteins Gαq, Gα13 and Gαi independently of G protein-coupled receptors through deamidation. PMT is known to induce signalling events involved in cell proliferation, cell survival and cytoskeleton rearrangement. Results Here we show that the activation of heterotrimeric G proteins through PMT suppresses LPS-stimulated IL-12p40 production and eventually impairs the T cell-activating ability of LPS-treated monocytes. This inhibition of TLR4-induced IL-12p40 expression is mediated by Gαi-triggered signalling as well as by Gβγ-dependent activation of PI3kinase and JNK. Taken together we propose the following model: LPS stimulates TLR4-mediated activation of the NFĸB-pathway and thereby the production of TNF-α, IL-6 and IL-12p40. PMT inhibits the production of IL-12p40 by Gαi-mediated inhibition of adenylate cyclase and cAMP accumulation and by Gβγ-mediated activation of PI3kinase and JNK activation. Conclusions On the basis of the experiments with PMT this study gives an example of a pathogen-induced interaction between G protein-mediated and TLR4-triggered signalling and illustrates how a bacterial toxin is able to interfere with the host’s immune

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

    Science.gov (United States)

    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...

  19. Pasteurella multocida cellulitis and possible septic arthritis of the knee after exposure to dog saliva after a recent total knee arthroplasty

    Directory of Open Access Journals (Sweden)

    Natallia Suvorava

    2017-01-01

    Full Text Available Prosthetic joint replacement occurs frequently. Complications of prosthetic joint replacements include bleeding, mechanical disruption, persistent pain, and infection. Infectious complications can occur early or late, measured in hours to days or months to years. The microorganism profiles vary depending on whether infection is early or late. Factors related to wound care have a significant role in the development of wound infection and coincident infection of the prosthesis. We report a wound infection and possible septic arthritis with septic shock due to Pasteurella multocida; symptoms and signs of joint involvement were noted but aspiration of the joint was declined by the orthopedic surgery service due to concerns about joint contamination from the overlying cellulitis. Blood cultures were sterile. The wound had been licked by her pet dogs on numerous occasions. Ultimately the infection was eradicated with antimicrobial agents, and the septic shock resolved.

  20. Binding of host iron-binding proteins and expression of iron-regulated membrane proteins by different serotypes of Pasteurella multocida causing haemorrhagic septicaemia.

    Science.gov (United States)

    Veken, J W; Shah, N H; Klaasen, P; Oudega, B; de Graaf, F K

    1996-07-01

    Pasteurella multocida strains of serotype B: 2,5, B: 3,4 and E: 2,5 are associated with haemorrhagic septicaemia in domestic and feral ruminants. These strains were investigated for their ability to bind transferrin, lactoferrin and haemoglobin and for their ability to use these host iron-binding proteins as a source of iron. All strains bound haemoglobin, none of the strains bound lactoferrin, whereas transferrin binding was restricted to serotype B: 2,5 strains. Growth experiments indicated that transferrin (serotype B: 2,5) and haemoglobin could restore bacterial growth under iron-depleted conditions. Two distinct serotype-independent profiles of iron-regulated membrane proteins were expressed in vitro as well as in vivo.

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

    Science.gov (United States)

    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.

  2. Estabelecimento de um índice de patogenicidade em pintos de corte de um dia de idade para amostras de Pasteurella multocida de aves e suínos

    OpenAIRE

    Roberta Marmitt Pilatti

    2014-01-01

    Pasteurella multocida, apesar de ser uma bactéria que compõe a microbiota respiratória, sob algumas circunstâncias pode manifestar-se como um agente patogênico primário ou secundário, causando doença em aves e outros animais. Como agente primário, P. multocida leva a grandes perdas econômicas, causando cólera em aves, rinite atrófica em suínos e septicemia hemorrágica em bovinos e bubalinos. P. multocida é uma espécie heterogênea, e a patogenicidade dos isolados pode ser amplamente variável. ...

  3. Fis Is Essential for Capsule Production in Pasteurella multocida and Regulates Expression of Other Important Virulence Factors

    Science.gov (United States)

    Steen, Jason A.; Steen, Jennifer A.; Harrison, Paul; Seemann, Torsten; Wilkie, Ian; Harper, Marina; Adler, Ben; Boyce, John D.

    2010-01-01

    P. multocida is the causative agent of a wide range of diseases of animals, including fowl cholera in poultry and wild birds. Fowl cholera isolates of P. multocida generally express a capsular polysaccharide composed of hyaluronic acid. There have been reports of spontaneous capsule loss in P. multocida, but the mechanism by which this occurs has not been determined. In this study, we identified three independent strains that had spontaneously lost the ability to produce capsular polysaccharide. Quantitative RT-PCR showed that these strains had significantly reduced transcription of the capsule biosynthetic genes, but DNA sequence analysis identified no mutations within the capsule biosynthetic locus. However, whole-genome sequencing of paired capsulated and acapsular strains identified a single point mutation within the fis gene in the acapsular strain. Sequencing of fis from two independently derived spontaneous acapsular strains showed that each contained a mutation within fis. Complementation of these strains with an intact copy of fis, predicted to encode a transcriptional regulator, returned capsule expression to all strains. Therefore, expression of a functional Fis protein is essential for capsule expression in P. multocida. DNA microarray analysis of one of the spontaneous fis mutants identified approximately 30 genes as down-regulated in the mutant, including pfhB_2, which encodes a filamentous hemagglutinin, a known P. multocida virulence factor, and plpE, which encodes the cross protective surface antigen PlpE. Therefore these experiments define for the first time a mechanism for spontaneous capsule loss in P. multocida and identify Fis as a critical regulator of capsule expression. Furthermore, Fis is involved in the regulation of a range of other P. multocida genes including important virulence factors. PMID:20140235

  4. Fis is essential for capsule production in Pasteurella multocida and regulates expression of other important virulence factors.

    Directory of Open Access Journals (Sweden)

    Jason A Steen

    2010-02-01

    Full Text Available P. multocida is the causative agent of a wide range of diseases of animals, including fowl cholera in poultry and wild birds. Fowl cholera isolates of P. multocida generally express a capsular polysaccharide composed of hyaluronic acid. There have been reports of spontaneous capsule loss in P. multocida, but the mechanism by which this occurs has not been determined. In this study, we identified three independent strains that had spontaneously lost the ability to produce capsular polysaccharide. Quantitative RT-PCR showed that these strains had significantly reduced transcription of the capsule biosynthetic genes, but DNA sequence analysis identified no mutations within the capsule biosynthetic locus. However, whole-genome sequencing of paired capsulated and acapsular strains identified a single point mutation within the fis gene in the acapsular strain. Sequencing of fis from two independently derived spontaneous acapsular strains showed that each contained a mutation within fis. Complementation of these strains with an intact copy of fis, predicted to encode a transcriptional regulator, returned capsule expression to all strains. Therefore, expression of a functional Fis protein is essential for capsule expression in P. multocida. DNA microarray analysis of one of the spontaneous fis mutants identified approximately 30 genes as down-regulated in the mutant, including pfhB_2, which encodes a filamentous hemagglutinin, a known P. multocida virulence factor, and plpE, which encodes the cross protective surface antigen PlpE. Therefore these experiments define for the first time a mechanism for spontaneous capsule loss in P. multocida and identify Fis as a critical regulator of capsule expression. Furthermore, Fis is involved in the regulation of a range of other P. multocida genes including important virulence factors.

  5. Fis is essential for capsule production in Pasteurella multocida and regulates expression of other important virulence factors.

    Science.gov (United States)

    Steen, Jason A; Steen, Jennifer A; Harrison, Paul; Seemann, Torsten; Wilkie, Ian; Harper, Marina; Adler, Ben; Boyce, John D

    2010-02-05

    P. multocida is the causative agent of a wide range of diseases of animals, including fowl cholera in poultry and wild birds. Fowl cholera isolates of P. multocida generally express a capsular polysaccharide composed of hyaluronic acid. There have been reports of spontaneous capsule loss in P. multocida, but the mechanism by which this occurs has not been determined. In this study, we identified three independent strains that had spontaneously lost the ability to produce capsular polysaccharide. Quantitative RT-PCR showed that these strains had significantly reduced transcription of the capsule biosynthetic genes, but DNA sequence analysis identified no mutations within the capsule biosynthetic locus. However, whole-genome sequencing of paired capsulated and acapsular strains identified a single point mutation within the fis gene in the acapsular strain. Sequencing of fis from two independently derived spontaneous acapsular strains showed that each contained a mutation within fis. Complementation of these strains with an intact copy of fis, predicted to encode a transcriptional regulator, returned capsule expression to all strains. Therefore, expression of a functional Fis protein is essential for capsule expression in P. multocida. DNA microarray analysis of one of the spontaneous fis mutants identified approximately 30 genes as down-regulated in the mutant, including pfhB_2, which encodes a filamentous hemagglutinin, a known P. multocida virulence factor, and plpE, which encodes the cross protective surface antigen PlpE. Therefore these experiments define for the first time a mechanism for spontaneous capsule loss in P. multocida and identify Fis as a critical regulator of capsule expression. Furthermore, Fis is involved in the regulation of a range of other P. multocida genes including important virulence factors.

  6. Identification of the capsule type of Pasteurella multocida isolates from cases of fowl cholera by multiplex PCR and comparison with phenotypic methods

    Directory of Open Access Journals (Sweden)

    TQ Furian

    2014-06-01

    Full Text Available The ability of Pasteurella multocida to invade and multiply in its host is enhanced by the presence of the capsule, one of the most important virulence factors for this bacterium. Capsular typing methods are often used in epidemiological and pathogenesis studies of this agent. Five different serogroups have been identified based on serological typing. However, such tests are laborious, and agglutination of homologous antiserum may fail. The aim of this study was to develop a multiplex PCR protocol for the identification of the hyaD-hyaC and dcbF genes specific to serogroups A and D, respectively, and to compare these results with those of phenotypic tests for 54 strains isolated from fowl cholera cases in southern Brazil. The kappa coefficient and chisquare statistics were calculated to assess the agreement between the diagnostic methods and to determine the significance of the results, respectively. The multiplex PCR was able to detect the evaluated genes. Forty-nine strains (90.74% were classified into serogroup A, and only two isolates (3.7% were not identified as belonging to any of the serogroups analyzed. In contrast, with the phenotypic tests, only 41 strains (75.93% were classified into serogroup A and 11 samples (20.37% were unidentifiable. Of the strains analyzed, 70.37% were classified into the same serogroup (A by both methods, and the kappa coefficient (k = 0.017 indicated poor agreement between the tests. Thus, multiplex PCR is an alternative for P. multocida capsular typing, as it allows the simultaneous and rapid detection of genes and also provides a greater strain-typing capacity.

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

    Science.gov (United States)

    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.

  8. Structural analysis of lipopolysaccharide produced by Heddleston serovars 10, 11, 12 and 15 and the identification of a new Pasteurella multocida lipopolysaccharide outer core biosynthesis locus, L6.

    Science.gov (United States)

    Harper, Marina; St Michael, Frank; John, Marietta; Steen, Jason; van Dorsten, Lieke; Parnas, Henrietta; Vinogradov, Evgeny; Adler, Ben; Cox, Andrew D; Boyce, John D

    2014-07-01

    Pasteurella multocida is a Gram-negative bacterial pathogen classified into 16 serovars based on lipopolysaccharide (LPS) antigens. Previously, we have characterized the LPS outer core biosynthesis loci L1, L2, L3, L5 and L7, and have elucidated the full range of LPS structures associated with each. In this study, we have determined the LPS structures produced by the type strains representing the serovars 10, 11, 12 and 15 and characterized a new LPS outer core biosynthesis locus, L6, common to all. The L6 outer core biosynthesis locus shares significant synteny with the L3 locus but due to nucleotide divergence, gene duplication and gene redundancy, the L6 and L3 LPS outer cores are structurally distinct. Using LPS structural and genetic differences identified in each L6 strain, we have predicted a role for most of the L6 glycosyltransferases in LPS assembly. Importantly, we have identified two glycosyltransferases, GctD and GatB, that differ by one amino acid, A162T, but use different donor sugars [uridine diphosphate (UDP)-Glc and UDP-Gal, respectively]. The longest outer core oligosaccharide, produced by the serovar 12 type strain, contained a terminal region consisting of β-Gal-(1,4)-β-GlcNAc-(1,3)-β-Gal-(1,4)-β-Glc that was identical in structure to the vertebrate glycosphingolipid, paragloboside. Mimicry of host glycosphingolipids has been observed previously in P. multocida strains belonging to L3 LPS genotype, which produce LPS similar in structure to the globo-series of glycosphingolipids. The expression of a paragloboside-like oligosaccharide on the LPS produced by the serovar 12 type strain indicates that strains belonging to the L6 LPS genotype may also engage in molecular mimicry. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  9. Pasteurella multocida CMP-sialic acid synthetase and mutants of Neisseria meningitidis CMP-sialic acid synthetase with improved substrate promiscuity.

    Science.gov (United States)

    Li, Yanhong; Yu, Hai; Cao, Hongzhi; Muthana, Saddam; Chen, Xi

    2012-03-01

    Cytidine 5'-monophosphate (CMP)-sialic acid synthetases (CSSs) catalyze the formation of CMP-sialic acid from CTP and sialic acid, a key step for sialyltransferase-catalyzed biosynthesis of sialic acid-containing oligosaccharides and glycoconjugates. More than 50 different sialic acid forms have been identified in nature. To facilitate the enzymatic synthesis of sialosides with diverse naturally occurring sialic acid forms and their non-natural derivatives, CMP-sialic acid synthetases with promiscuous substrate specificity are needed. Herein we report the cloning, characterization, and substrate specificity studies of a new CSS from Pasteurella multocida strain P-1059 (PmCSS) and a CSS from Haemophillus ducreyi (HdCSS). Based on protein sequence alignment and substrate specificity studies of these two CSSs and a Neisseria meningitidis CSS (NmCSS), as well as crystal structure modeling and analysis of NmCSS, NmCSS mutants (NmCSS_S81R and NmCSS_Q163A) with improved substrate promiscuity were generated. The strategy of combining substrate specificity studies of enzymes from different sources and protein crystal structure studies can be a general approach for designing enzyme mutants with improved activity and substrate promiscuity.

  10. Surveillance for Pasteurella multocida in Ring-Necked Pheasants (Phasianus colchicus) After an Outbreak of Avian Cholera and Apparently Successful Antibiotic Treatment.

    Science.gov (United States)

    Brown, Justin D; Dunn, Patricia; Wallner-Pendleton, Eva; Kariyawasam, Subhashinie; Schriner, Timothy; Hofacre, Charles; Johnson, Joshua; Boyd, Robert

    2016-03-01

    Avian cholera is a significant disease of domestic and wild birds caused by the bacterium Pasteurella multocida (PM). In poultry, a major source of PM infection is chronic carriers, domestic birds that have become infected and recovered or had subclinical infections. Although outbreaks of avian cholera in ring-necked pheasants (Phasianus colchicus) have been reported, the potential for chronic carriers is unknown. To address this, we conducted surveillance for PM in a flock of captive ring-necked pheasants after an outbreak of avian cholera that responded positively to antibiotic treatment based on resolution of morbidity and mortality. At approximately 1 mo after antibiotic treatment, oropharyngeal swabs were collected from 300 pheasants (out of a total population of ~2300) in a single winter holding pen. All samples were tested for PM through routine aerobic bacterial culture, but none of the samples were positive. In addition, there were no additional outbreaks within this infected pen over the subsequent months. These data provide preliminary evidence to suggest that pheasants that respond to antibiotic therapy may be less likely to become chronic carriers of PM than other poultry species, such as chickens (Gallus domesticus). However, due to marked phenotypic and biologic differences between PM strains, additional studies are needed to further support or refute these findings and better understand avian cholera in this species.

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

    DEFF Research Database (Denmark)

    Guo, Yao; Jers, Carsten; Meyer, Anne S.

    2014-01-01

    tThis study examined a recombinant Pasteurella multocida sialyltransferase exhibiting dual trans-sialidase activities. The enzyme catalyzed trans-sialylation using either 2-O-(p-nitrophenyl)--d-N-acetylneuraminic acid or casein glycomacropeptide (whey protein) as the sialyl donor and lactose asthe......, andlactose concentration) on enzymatic production of 3- and 6-sialyllactoses using 5% (w/v) casein glyco-macropeptide (equivalent to 9 mM bound sialic acid) as the donor. The maximum yield of 3-sialyllactose(2.75 ± 0.35 mM) was achieved at a reaction condition with pH 6.4, 40◦C, 100 mM lactose after 6 h......; andthe largest concentration of 6-sialyllactose (3.33 ± 0.38 mM) was achieved under a condition with pH5.4, 40◦C, 100 mM lactose after 8 h. 6-sialyllactose was presumably formed from -2,3 bound sialic acidin the casein glycomacropeptide as well as from 3-sialyllactose produced in the reaction. The kcat...

  12. Identificação de transcritos diferencialmente expressos por Pasteurella multocida em condições de privação de ferro

    Directory of Open Access Journals (Sweden)

    Mayara I.V. Silva

    Full Text Available RESUMO: Ferro (Fe é um elemento essencial e a capacidade de adquiri-lo in vivo têm sido descrita em diversos agentes patogênicos através de fatores de virulência. Análises de transcritos durante a privação de Fe tem sido descritos através da técnica de "microarray", entretanto a técnica de RNA-seq recentemente tem demonstrado resultados superiores. Neste trabalho, o isolado de Pasteurella multocida (Pm 16759 altamente patogênico em suínos foi cultivado em duas condições com diferentes concentrações de Fe (controle e privação com o objetivo de analisar transcritos diferencialmente expressos. O RNA total das duas condições foi extraído e sequenciado através da plataforma de nova geração Ion Torrent. Os dados foram analisados no Software Ion Reporter(tm e processados no programa Rockhopper. Foram obtidas 1.341.615 leituras com tamanho médio de 81pb, com 96% de alinhamento com o genoma de Pasteurella multocida subsp. multocida 3480 e 98,8% de acurácia. No mapeamento das leituras das duas condições, observou-se 2,652 transcritos e destes, 177 (6,7% foram diferencialmente expressos, sendo 93 na condição controle (Fe+ e 84 na condição de privação (Fe-. Na condição de privação de Fe, o perfil de transcritos foram associados a função de transporte celular (fbp ABC, permease de alta afinidade com Fe2+/Pb2+ e proteína periplasmática de alta afinidade com Fe2+ , reguladores transcricionais e proteínas hipotéticas. O perfil na condição controle (Fe+ apresentou transcritos diferencialmente expressos associados ao RNAs anti-sense (asRNA e genes do metabolismo energético (fructose-1,6-bisfosfatase. O estudo comprovou que a restrição de Fe aumenta a expressão de genes envolvidos no transporte celular, reguladores transcricionais, proteínas hipotéticas e desconhecidas e permitiu ainda a identificação de novos genes como a permease de alta afinidade com Fe2+/Pb2+ e proteina periplasmática de alta afinidade

  13. Efecto bactericida de la acetamida furánica bromada frente a cepas salvajes de Pasteurella multocida in vitro (Evaluation of bactericide effect of bromade furanic acetamid against to wilds strains of multocide Pasterella in vitr)

    OpenAIRE

    Estrada, C; Cadrelo, T. Surelis; Salazar, A. Ana Ivis; Almeida, S; Montero, V. R.; Peillón, V. O; Cos, D., Yusel; Bárzaga, R.; Campuzano, Y

    2007-01-01

    Resumen Se realizó una evaluación del efecto la Acetamida furánica bromada frente a cepas salvajes de Pasteurella multocida in vitro, en el Laboratorio Provincial de Diagnóstico Veterinario de Granma. Se determinó la Concentración Mínima Inhibitoria (CMI) y Concentración Máxima Bactericida (CMB), empleando el método de diluciones seriadas en tubos. La CMI y la CMB fueron iguales a 8 (µg) en una dilución de 1:4. Se realizó una prueba de hipótesis entre proporciones, con un nivel de sign...

  14. Outer Membrane Proteome Analysis of Indian Strain of Pasteurella multocida Serotype B:2 by MALDI-TOF/MS Analysis

    Directory of Open Access Journals (Sweden)

    A. Prasannavadhana

    2014-01-01

    Full Text Available Identification of outer membrane proteins (OMPs is important to understand the bacteria structure and function, host-pathogen interaction, development of novel vaccine candidates, and diagnostic antigens. But till now the key antigens of P. multocida B:2 isolate causing haemorrhagic septicaemia (HS in animals are not clearly defined. In this study, P52 strain of P. multocida serotype B:2 was grown in vitro under iron-rich and iron-limited condition. The OMPs were extracted by sarkosyl method followed by SDS-PAGE and the proteins were identified by MALDI-TOF/MS analysis. In total, 22 proteins were identified, of which 7 were observed exclusively under iron-limited condition. Most of the high molecular weight proteins (TbpA, HgbA, HgbB, HasR, IroA, and HemR identified in this study were involved in iron acquisition. Some hypothetical proteins (HP-KCU-10206, HP and AAUPMB 08244, HP AAUPMB 21592, HP AAUPMB 19766, AAUPMB 11295 were observed for the first time in this study which could be unique to serotype B:2. Further functional in vivo study of the proteins identified are required to explore the utility of these proteins in developing diagnostics and vaccine against HS.

  15. Comparative Analysis of Pasteurella multocida Isolates from Acute and Chronic Fowl Cholera Cases in Hungary During the Period 2005 Through 2010.

    Science.gov (United States)

    Sellyei, Boglárka; Thuma, Ákos; Volokhov, Dmitriy; Varga, Zsuzsanna

    2017-12-01

    Fowl cholera (FC) is a highly contagious and economically important disease of poultry worldwide. This study was performed on 218 Pasteurella multocida isolates collected from separated breeding farms or backyards with acute and chronic FC cases in multiple localities across Hungary during the period 2005-2010. All isolates were characterized by a broad range of biochemical, serological, and molecular methods, as well as their antibiotic susceptibility to aminoglycosides (A), macrolides (M), penicillins (P), quinolones (Q), cephalosporins, sulphonamides (S), and tetracyclines (T) was determined. Fifty-two percent of all isolates belonged to a well-defined type that was highly virulent, caused acute FC, and had the same character: fermented L-arabinose, possessed capsule type A, identified as Heddleston serotype 1, and possessed allele type A of the ptfA fimbrial gene. This type was widely distributed among poultry in Hungary, especially in waterfowl flocks. Isolates collected from the chronic FC cases were more diverse: none of them fermented L-arabinose; they possessed capsule type A (76%), F (9%), or was non-typeable (15%) with different Heddleston serotypes, mainly 1, 3, 4, and 5, or 7 and 16; in addition, possessed allele type B of ptfA fimbrial gene. Only 26 isolates presented characters similar to any of the chronic FC cases but caused severe disease. The antibiotic susceptibility assay presented that 80% of all isolates were resistant to 1-5 of the studied antimicrobial agents. During the survey, after two years, there was a dramatic decline both in the number of the multi-drug resistance phenotypes and the prevalence of the highly virulent type of the isolates. In the next four years, multiresistant isolates were almost completely removed, whereas the number of isolates resistant to 1 or 2 drugs was constant. Reduced frequency of antibiotic multiresistant, mostly L-arabinose-fermenting isolates, has been observed since 2007. This reduction may be a

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

    Science.gov (United States)

    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

  17. Investigation into the pathogenesis of atrophic rhinitis in pigs. I. Atrophic rhinitis caused by Bordetella bronchiseptica and Pasteurella multocida and the meaning of a thermolabile toxin of P. multocida.

    Science.gov (United States)

    de Jong, M F; Akkermans, J P

    1986-07-01

    In two groups of swine herds, herds with and without clinical AR the presence of Atrophic Rhinitis (AR) correlated with the presence of toxinogenic Pasteurella multocida (PM) and not with the Bordetella bronchiseptica (BB) infection. Six BB- and eighteen PM-strains have been investigated for AR pathogenicity. Broth cultures were injected intradermally in guinea-pigs (GPST) or intranasally in 3-week-old colostrum deprived specific pathogen free (SPF) piglets. The average atrophy of the ventral conchae (AVC) correlated with the GPST in 4 BB-and 7 PM-strains. One BB- and 2 PM-strains were qualified as doubtful, the others as non-AR pathogenic. With AR pathogenic BB-and PM-strains clinical AR could be induced in 3-and 6-week-old piglets. AVC lesions (gradation greater than 1) could be induced with BB in piglets of 6 and with pathogenic PM in 16-week-old piglets. Six of seven AR pathogenic PM-strains resembled Carter-type D and one resembled type A. No significance was found between AR pathogenicity and somatic serotypes. Intranasal instillations of cell-free broth culture filtrates of AR pathogenic PM-strains also caused AR in piglets. These filtrates also caused lethality in piglets and in mice lethalitytest (MLT) and induced a positive GPST. After heating the pathogenic effects of the filtrates disappeared. The name AR toxin has been introduced for this thermolabile, haemorrhagic dermonecrotic (HDNT) fraction of the AR inducing filtrates. The severity of the AR lesions depended on the amount of the AR toxin intranasally instilled in pigs. Cross protecting antibodies obtained in rabbits against the AR toxins of two PM strains could be demonstrated by a toxin neutralisation test in the MLT and the GPST. Broth cultures were injected intradermally in guinea-pigs (GPST) or intranasally in 3-week-old colostrum deprived specific pathogen free (SPF) piglets.

  18. Utilización de lectinas en la inhibición de la adhesión de Pasteurella multocida

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    Magda Patricia Carrillo Lamus

    2013-06-01

    Full Text Available El primer paso de la infección es la adhesión de los organismos patógenos a las células blanco. Esta característica les permite no solo penetrar y desplegar las estrategias ofensivas para iniciar la colonización, sino contribuir a la protección y el resguardo de estos a los mecanismos de defensa tanto inmunológicos como mecánicos del hospedero. Este proceso se lleva a cabo en gran medida por la interacción de lectinas, que son proteínas de origen no inmune con la capacidad de reconocer y aglutinar carbohidratos. Este mecanismo ha sido reportado para muchos microorganismos como virus y bacterias. En el caso particular de la Pasteurella multocida, que es una bacteria gramnegativa, patógena oportunista, que inicia la infección en el epitelio respiratorio de muchos animales, se han descrito sobre su superficie sustancias lectinas como la fimbria tipo IV y carbohidratos como el lipopolisacárido o la cápsula que reconocen carbohidratos y lectinas, respectivamente, sobre la superficie de las células epiteliales del tubo respiratorio, circunstancia que le permite adherirse y resguardarse del efecto mucociliar. Debido a que para la gran mayoría de estos microorganismos —incluida P. multocida— es evidente la disminución de la susceptibilidad a los antibacterianos y a la efectividad de las vacunas, se han buscado nuevas tácticas terapéuticas y profilácticas con el fin de interrumpir la infección de los patógenos por medio del bloqueo por competencia de la unión carbohidratos-lectina, donde se ve disminuida la unión del microorganismo al tejido y, por consiguiente, la infección.

  19. 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.

    Science.gov (United States)

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

    2016-05-01

    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. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  20. Pasteurella Multocida Toxin Prevents Osteoblast Differentiation by Transactivation of the MAP-Kinase Cascade via the Gαq/11 - p63RhoGEF - RhoA Axis

    Science.gov (United States)

    Siegert, Peter; Schmidt, Gudula; Papatheodorou, Panagiotis; Wieland, Thomas; Aktories, Klaus; Orth, Joachim H. C.

    2013-01-01

    The 146-kDa Pasteurella multocida toxin (PMT) is the main virulence factor to induce P. multocida-associated progressive atrophic rhinitis in various animals. PMT leads to a destruction of nasal turbinate bones implicating an effect of the toxin on osteoblasts and/or osteoclasts. The toxin induces constitutive activation of Gα proteins of the Gq/11-, G12/13- and Gi-family by deamidating an essential glutamine residue. To study the PMT effect on bone cells, we used primary osteoblasts derived from rat calvariae and stromal ST-2 cells as differentiation model. As marker of functional osteoblasts the expression and activity of alkaline phosphatase, formation of mineralization nodules or expression of specific transcription factors as osterix was determined. Here, we show that the toxin inhibits differentiation and/or function of osteoblasts by activation of Gαq/11. Subsequently, Gαq/11 activates RhoA via p63RhoGEF, which specifically interacts with Gαq/11 but not with other G proteins like Gα12/13 and Gαi. Activated RhoA transactivates the mitogen-activated protein (MAP) kinase cascade via Rho kinase, involving Ras, MEK and ERK, resulting in inhibition of osteoblast differentiation. PMT-induced inhibition of differentiation was selective for the osteoblast lineage as adipocyte-like differentiation of ST-2 cells was not hampered. The present work provides novel insights, how the bacterial toxin PMT can control osteoblastic development by activating heterotrimeric G proteins of the Gαq/11-family and is a molecular pathogenetic basis for understanding the role of the toxin in bone loss during progressive atrophic rhinitis induced by Pasteurella multocida. PMID:23696743

  1. Influence of body weight restriction in a body-weight-selected line of turkeys on response to challenge with Pasteurella multocida.

    Science.gov (United States)

    Nestor, K E; Lilburn, M S; Saif, Y M; Anderson, J W; Patterson, R A; Li, Z; Nixon, J E

    1999-09-01

    Previous research has shown that a line (F) of turkeys selected long-term for increased 16-wk BW was more susceptible to challenge with washed Pasteurella multocida (PM) than a randombred control line (RBC2), the base population of the F line. Published research indicated that the mortality of the F line following challenge with PM was similar to that of two commercial sire lines. The purpose of the present study was to determine the influence of reducing BW of the F line to that of the RBC2 line by nutrient restriction on resistance to PM. Four challenge trials were conducted over a 2-yr period. The BW of a group of F line birds was restricted to that of the RBC2 line by limiting access to feed from 1 to 6 wk of age. The F line restricted birds and full-fed RBC2 and F line birds were challenged with a field isolate of washed PM (1.2x10(7) organisms/bird of capsular serogroup A and somatic serotype 3, 4) at 6 wk of age. Birds were checked twice daily for 14 d. Resistance to PM was measured by days to death of those that died and percentage mortality. The BW of the restricted group of the F line did not differ from full-fed RBC2 birds for males or females. In males, the restricted F line birds had similar mortality (48.0%) to the full-fed RBC2 line birds (44.3%), and the mortalities in both groups were significantly lower than that observed for the full-fed F line birds (81.3%) following challenge with PM. The mortality following challenge in females did not differ significantly among groups, even though mortality of the full-fed F line birds (64.1%) and restricted F line birds (63.3%) was more than 9% higher than that (54.2%) observed for the full-fed RBC2 line birds. Days to death was not a sensitive indicator of resistance to PM, as no differences among the three groups of birds were observed for either sex.

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

    Science.gov (United States)

    Settypalli, Tirumala Bharani Kumar; 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

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

    Directory of Open Access Journals (Sweden)

    Tirumala Bharani Kumar Settypalli

    Full Text Available 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

  4. Aspects relating to use of radioactively labelled bacteria in animal experiments. 7. Intake and deposition of aerosols of /sup 59/Fe-labelled Pasteurella multocida germ suspensions in lungs of piglets and calves

    Energy Technology Data Exchange (ETDEWEB)

    Heilmann, P.; Flossmann, K.D.; Mueller, G.; Finsterbusch, L. (Akademie der Landwirtschaftswissenschaften der DDR, Jena. Inst. fuer Bakterielle Tierseuchenforschung)

    1983-03-01

    Two different types of aerosol dispensers were used in an aerosol compartment to apply /sup 59/Fe-labelled bacteria (Pasteurella multocida) to SPE Mini-LEWE piglets as well as to conventionally raised piglets and calves. Germ intake was verified by detection of radioactivity in the lungs. Antigen deposition on each lung amounted to 2-3 . 10/sup 8/ in mini-piglets, 6-8 . 10/sup 8/ in ordinary piglets, and 2 . 10/sup 9/ in conventionally raised calves, as determined by SAG-1, a Soviet model of aerosol dispenser. More or less equally high concentrations of aerosol particles were retained in the pulmonary lobes, independent of the animal species used. Antigen intake could not be influenced by addition of skim milk or by restriction of germ suspensions.

  5. Efecto bactericida de la acetamida furánica bromada frente a cepas salvajes de Pasteurella multocida in vitro (Evaluation of bactericide effect of bromade furanic acetamid against to wilds strains of multocide Pasterella in vitr)

    OpenAIRE

    Estrada, C; Cadrelo, T. Surelis; Salazar, A. Ana Ivis; Almeida, S; Montero, V. R.; Cos, D., Yusel; Bárzaga, R.; Campuzano, Y

    2007-01-01

    Se realizó una evaluación del efecto la Acetamida furánica bromada frente a cepas salvajes de Pasteurella multocida in vitro, en el Laboratorio Provincial de Diagnóstico Veterinario de Granma. Se determinó la Concentración Mínima Inhibitoria (CMI) y Concentración Máxima Bactericida (CMB), empleando el método de diluciones seriadas en tubos. La CMI y la CMB fueron iguales a 8 (µg) en una dilución de 1:4. Se realizó una prueba de hipótesis entre proporciones, con un nivel de significació...

  6. Comparison of gene expression of Toll-like receptors and cytokines between Piau and Commercial line (Landrace×Large White crossbred) pigs vaccinated against Pasteurella multocida type D.

    Science.gov (United States)

    Sousa, Katiene Régia Silva; Ribeiro, André Mauric Frossard; Dantas, Waleska de Melo Ferreira; Oliveira, Leandro Licursi de; Gasparino, Eliane; Guimarães, Simone Eliza Facioni

    2017-10-01

    We aimed to compare Toll-like receptors (TLR) and cytokines expression in local Piau breed and a Commercial line (Landrace×Large White crossbred) pigs in response to vaccination against Pasteurella multocida type D. Seronegative gilts for Pasteurella multocida type D and Mycoplasma hyopneumoniae were used, from which peripheral blood mononuclear cells (PBMC) were collected in four time points (T0, T1, T2 and T3; before and after each vaccination dose). For bronchoalveolar lavage fluid cells (BALF), we set groups of vaccinated and unvaccinated animals for both genetic groups. Gene expression was evaluated on PBMC and BALF. In PBMC, when we analyzed time points within breeds, significant differences in expression for TLRs and cytokines, except TGFβ, were observed for Commercial animals. For the Piau pigs, only TGFβ showed differential expression. Comparing the expression among genetic groups, the Commercial pigs showed higher expression for TLRs after first vaccination dose, while for IL2, IL6, IL12 and IL13, higher expression was also observed in T3 and IL8 and IL10, in T1 and T3. Still comparing the breeds, the crossbred animals showed higher expression for TNFα in T1 and T2, while for TGFβ only in T2. For gene expression in BALF, vaccinated Commercial pigs showed higher expression of TLR6, TLR10, IL6, IL8, IL10, TNFα and TGFβ genes than vaccinated Piau pigs. The Commercial line pigs showed higher sensitivity to vaccination, while in local Piau breed lower responsiveness, which may partly explain genetic variability in immune response and will let us better understand the tolerance/susceptibility for pasteurellosis. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR Pesquisa de genes associados à virulência em cepas de Pasteurella multocida isoladas em casos de cólera aviária através da técnica de multiplex-PCR

    Directory of Open Access Journals (Sweden)

    Thales Q. Furian

    2013-02-01

    Full Text Available The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25, the sodA and nanH genes were present in 96% (24/25, ptfA was present in 92% (23/25, and pfhA was present in 60% (15/25. Gene toxA was not identified in any of the samples studied (0/25. Five different genetic profiles were obtained, of which P1 (negative to toxA was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.Os atuais sistemas de criação na avicultura, baseados na alta densidade populacional, aumentam os riscos de disseminação de patógenos, especialmente das doenças respiratórias e daquelas cujos agentes etiológicos possuam mais de um hospedeiro. A Cólera Aviária (CA apresenta estas características e apesar de representar uma das patologias aviárias que deve ser considerada para o diagnóstico diferencial de

  8. Ocorrência de genes tad associados à formação de biofilme em isolados de Pasteurella multocida de pulmões de suínos com pneumonia

    Directory of Open Access Journals (Sweden)

    Danny Franciele da S.D. Moraes

    2014-12-01

    Full Text Available Os atuais sistemas de criação intensiva de suínos aumentam a pressão de seleção microbiana propiciando a disseminação de doenças respiratórias. A bactéria Pasteurella multocida é associada a diversas patologias respiratórias em animais submetidos a esse tipo de criação, causando grandes perdas econômicas. A formação de biofilme foi descrita in vitro em P. multocida e fatores analisados indicaram a facilitação na colonização dos tecidos, aumentando a resistência às defesas do hospedeiro e aos antibióticos. Os objetivos deste trabalho foram analisar a ocorrência de P. multocida em pneumonias de suínos e na microbiota de pulmões sem lesão e a ocorrência dos genes do lócus tad nestes isolados. Foram analisados 70 isolados de P. multocida de pulmões, sendo sessenta e sete com lesão e três sem lesão. Isolados do sorotipo A ocorreram principalmente em pulmões com lesões (85,71%, enquanto em pulmões sem lesão observou-se somente o sorotipo D. Os genes tadA, tadB, tadC, tadD, tadE tadF e tadG estavam presentes em 89,55% dos isolados de pulmões com lesões. Os genes tadA, tadB e tadC estavam presentes em todos os isolados de pulmões sem lesão, porém os genes tadD, tadE, tadF e tadG estavam presentes em 0%, 33,3%, 33,3% e 66,6%, dos isolados sem lesão, respectivamente. Neste trabalho observou-se a associação da ocorrência dos genes tadD, tadE e tadF em isolados de P. multocida e a presença de lesões em pulmões.

  9. C-reactive protein, haptoglobin, serum amyloid A and pig major acute phase protein response in pigs simultaneously infected with H1N1 swine influenza virus and Pasteurella multocida

    Directory of Open Access Journals (Sweden)

    Pomorska-Mól Małgorzata

    2013-01-01

    Full Text Available Abstract Background Swine influenza (SI is an acute respiratory disease caused by swine influenza virus (SIV. Swine influenza is generally characterized by acute onset of fever and respiratory symptoms. The most frequent complications of influenza are secondary bacterial pneumonia. The objective of this work was to study the acute phase proteins (APP responses after coinfection of piglets with H1N1 swine influenza virus (SwH1N1 and Pasteurella multocida (Pm in order to identify whether the individual APP response correlate with disease severity and whether APP could be used as markers of the health status of coinfected pigs. Results In all coinfected pigs clinical sings, including fever, coughing and dyspnea, were seen. Viral shedding was observed from 2 to 7 dpi. The mean level of antibodies against Pm dermonecrotoxin in infected piglets increase significantly from 7 dpi. Anti-SwH1N1 antibodies in the serum were detected from 7 dpi. The concentration of C-reactive protein (CRP increased significantly at 1 dpi as compared to control pigs, and remained significantly higher to 3 dpi. Level of serum amyloid A (SAA was significantly higher from 2 to 3 dpi. Haptoglobin (Hp was significantly elevated from 3 dpi to the end of study, while pig major acute phase protein (Pig-MAP from 3 to 7 dpi. The concentrations of CRP, Hp and SAA significantly increased before specific antibodies were detected. Positive correlations were found between serum concentration of Hp and SAA and lung scores, and between clinical score and concentrations of Pig-MAP and SAA. Conclusions The results of current study confirmed that monitoring of APP may revealed ongoing infection, and in this way may be useful in selecting clinically healthy pigs (i.e. before integration into an uninfected herd. Present results corroborated our previous findings that SAA could be a potentially useful indicator in experimental infection studies (e.g. vaccine efficiency investigations or as a

  10. Treatment history and antimicrobial susceptibility results for Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni isolates from bovine respiratory disease cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2013 to 2015.

    Science.gov (United States)

    Magstadt, Drew R; Schuler, Adlai M; Coetzee, Johann F; Krull, Adam C; O'Connor, Annette M; Cooper, Vickie L; Engelken, Terry J

    2017-10-01

    Bovine respiratory disease is the most costly disease facing the cattle industry. Increasing resistance to antimicrobial treatment has been presented as a significant contributing factor, often through summarized susceptibility testing data. We assessed the relationship between previous antimicrobial treatment and antimicrobial susceptibility results from isolates of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni cultured from bovine respiratory cases submitted to the Iowa State University Veterinary Diagnostic Laboratory from 2013 to 2015. Antimicrobial susceptibility data from 1,251 bacterial isolates were included for analysis. More bacterial isolates from cattle that received antimicrobial treatment showed resistance compared to isolates from untreated cattle, and the percentage of resistant isolates increased as the number of antimicrobial treatments increased. Resistance to enrofloxacin, spectinomycin, tilmicosin, and tulathromycin was present in >75% of M. haemolytica isolates from cattle that had received 3 or more antimicrobial treatments; resistance to each of those 4 antimicrobials was present in ≤10% of M. haemolytica isolates from untreated cattle. Similar but less dramatic trends were apparent for isolates of P. multocida and H. somni. The percentage of multi-drug resistant bacterial isolates also increased with the number of treatments. Results of our study suggest that previous antimicrobial treatment may have a profound effect on antimicrobial susceptibility testing. Summarized susceptibility results from diagnostic laboratories should not be used to make generalized statements regarding trends in antimicrobial resistance without providing context regarding antimicrobial treatment history.

  11. An outbreak of fowl cholera due to Pasteurella gallinarum in Uttar Pradesh (India).

    Science.gov (United States)

    Yadav, M P; Sharma, V D; Sethi, M S

    1977-01-01

    An acute outbreak of fowl cholera in one-month-old chickens was investigated. Pasteurella gallinarum was isolated in pure culture from the heart blood of two moribund chicks. One of the isolants on experimental inoculation was found to be nonpathogenic for rabbits, mice, and chickens. It did not provide protection in rabbits against a virulent strain of P. multocida. This seems to be the first record of the isolation of P. gallinarum in India.

  12. Taxonomy of pasteurella anatipestifer. 1. DNA base composition and DNA-DNA hybridization analysis.

    Science.gov (United States)

    Bangun, A; Johnson, J L; Tripathy, D N

    1987-01-01

    DNA was isolated from 15 strains of Pasteurella anatipestifer and from one strain each of Moraxella nonliquefaciens, M. bovis, Pasteurella multocida, P. haemolytica, P. gallinarum, P. pneumotropica, and P. ureae. The guanine-plus-cytosine contents of P. anatipestifer ranged from 32 to 35 mole %, whereas those of Moraxella and Pasteurella spp. were much higher, ranging from 40 to 45 mole %. DNA-DNA hybridization analysis revealed that homology of nine P. anatipestifer strains to strains ATCC 11845 and PA 15 was 52 to 100%, whereas homology of Moraxella and Pasteurella strains to these strains was only 3 to 17%. Similarly, homology of P. anatipestifer strains, Moraxella, and Pasteurella species other than P. multocida to P. multocida reference strain P-2192 was low. These results strongly suggest that P. anatipestifer is genetically unrelated to either Pasteurella or Moraxella.

  13. Genomic fingerprints, ARDRA profiles and quinone systems for classification of Pasteurella sensu stricto.

    Science.gov (United States)

    Kainz, A; Lubitz, W; Busse, H J

    2000-12-01

    In order to investigate the relationships between species of the genus Pasteurella sensu stricto such as Pasteurella multocida, Pasteurella canis, Pasteurella stomatis, Pasteurella dagmatis, Pasteurella avium, Pasteurella volantium, Pasteurella gallinarum, Pasteurella species A, Pasteurella species B and "Pasteurella leonis" MCCM 00659 their genomic fingerprints and ARDRA profiles were compared and their quinone systems were analysed. Visual comparison of band patterns from rep-PCR (ERIC-, REP- and BOX-PCR) and the analyses of the combined band patterns by UPGMA (unweighted pair group method with averages) dendrogram derived from the combined fingerprint profiles demonstrated that each strain displays a distinct genomic fingerprint. In members of the same species several similarities in the band patterns were observed. Combined ARDRA profiles, obtained after digestion of amplified 23S rRNA coding genes with the enzymes DdeI, MseI and RsaI, revealed a dissection of the members of the genus Pasteurella sensu stricto into two groups which was in agreement with the two groups obtained from our analyses of the quinone systems. These two groups corresponded with the two phylogenetically determined subclusters 3A and 3B described previously. The species of subcluster 3A displayed a quinone system with ubiquinone Q-7 (32-56%) and ubiquinone Q-8 (44-63%) as major compounds. Members of subcluster 3B had a quinone system with ubiquinone Q-8 (86-97%) as the major compound. Based on these results it can be suggested that the genus Pasteurella sensu stricto should be restricted to the species of subcluster 3B including the species Pasteurella multocida, Pasteurella canis, Pasteurella stomatis, Pasteurella dagmatis and Pasteurella species B. In addition, evidence was found which would indicate that: 1) Pasteurella canis MCCM 00927 is misnamed and should be reclassified with Pasteurella multocida; 2) Pasteurella multocida subsp. septica may be classified as a separate species; and

  14. 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

    National Research Council Canada - National Science Library

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

    2016-01-01

    .... 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...

  15. Indigenous Ducks are Better Reservoirs of P. multocida than ...

    African Journals Online (AJOL)

    Two experiments were performed to study cross infections from chickens to ducks and vice versa. For each experiment the source birds (chickens or ducks) were infected with Pasteurella multocida strain 10322T. The infected birds were then mixed with sentinel indigenous ducks or chickens, respectively, six hours after ...

  16. Phenotypic and Genotypic Characterization of Pasteurella Multocida ...

    African Journals Online (AJOL)

    When the 6 isolates were tested for susceptibility to 15 antimicrobial agents by the standardized single disk method and minimal inhibitory concentrations, all isolates showed multi-drug resistance, but were all found to be susceptible to ciprofloxacin, levofloxacin, cefotaxime, ampicillin-clavulanic acid and imipenem.

  17. Time-course investigation of infection with a low virulent Pasteurella multocida strain in normal and immune-suppressed 12-week-old free-range chickens

    DEFF Research Database (Denmark)

    Mbuthia, P.G.; Njagi, L.W.; Nyaga, P.N.

    2011-01-01

    in NIS birds than in IS birds and at higher frequency per organ. Many organs were positive by FISH even 14 days post infection, and it is suggested that these organs may be likely places for long-term carriage of P. multocida following infection. The present study has demonstrated the spread of P...

  18. Experimental reproduction of endocarditis with Pasteurella gallinarum in mature leghorn chickens.

    Science.gov (United States)

    Tjahjowati, G; Orr, J P; Chirino-Trejo, M; Mills, J H

    1995-01-01

    The pathogenicity of Pasteurella gallinarum for mature leghorn chickens was investigated by inoculating thirty 52-week-old chickens intravenously with live P. gallinarum. Each chicken was inoculated once daily for 5 days at one of three different dosage levels with either the type strain ATCC 13361 or a field isolate from a chicken with endocarditis. Chickens were necropsied after death or euthanasia. Valvular endocarditis was present in seven chickens given the field isolate and five chickens given the type strain. Other lesions detected were myocarditis, hepatic and splenic infarcts, nephritis, pneumonia, and encephalitis. At necropsy, P. gallinarum was reisolated from hearts, livers, spleens, lungs, kidneys, and blood. Controls injected with sterile broth had no lesions of endocarditis, nor was P. gallinarum isolated from them. The results confirm the pathogenicity of P. gallinarum for the heart valves of mature chickens.

  19. Cloning and Characterization of Sialidases with 2-6′ and 2-3′ Sialyl Lactose Specificity from Pasteurella multocida†

    OpenAIRE

    Mizan, Shaikh; Henk, Adam; Stallings, Amy; Maier, Marie; Lee, Margie D.

    2000-01-01

    Pasteurella multocida is a mucosal pathogen that colonizes the respiratory system of susceptible hosts. Most isolates of P. multocida produce sialidase activity, which may contribute to colonization of the respiratory tract or the production of lesions in an active infection. We have cloned and sequenced a sialidase gene, nanH, from a fowl cholera isolate of P. multocida. Sequence analysis of NanH revealed that it exhibited significant amino acid sequence homology with many microbial sialidas...

  20. Comparison of the Carrier Status of P. multocida Between Farm and ...

    African Journals Online (AJOL)

    A total of one hundred and seventy one indigenous birds from smallholder farms and those traded in market centers in Nairobi were examined for the presence of Pasteurella multocida. Of these, 135 were farmed and 36 were market birds. They comprised of 117 indigenous chickens and 54 ducks. Three hundred and forty ...

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

    Science.gov (United States)

    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.

  2. Mycoplasmas and acholeplasmas isolated from ducks and their possible association with pasteurellas.

    Science.gov (United States)

    Tiong, S K

    1990-07-21

    Two hundred and sixty-three cases of clinically diseased ducks of all ages were examined for the presence of mycoplasmas. Mycoplasmas and acholeplasmas belonging to more than eight serogroups were cultured from 68 of them, and comprised 12 M anatis, one M columbinasale, two M gallinaceum, two M gallinarum, nine M synoviae, three unidentified Mycoplasma species, 37 Acholeplasma laidlawii and one unclassified acholeplasma belonging to each of serogroups 7 and 8. They were identified by biochemical characterisation, disc growth inhibition and agar gel diffusion tests. Fifty-three (78 per cent) of the isolates occurred with species of Pasteurella: 33.8 per cent with Pasteurella anatipestifer, 32.4 per cent with P multocida and 11.8 per cent with both P anatipestifer and P multocida. Nine of the isolates (13.2 per cent) were in pure culture and six (8.8 per cent) with other agents. Of the ducks negative for mycoplasmas 33.3 per cent were infected with P anatipestifer, 25.1 per cent with P multocida and 14.4 per cent with both P anatipestifer and P multocida. There was no correlation between the infections with mycoplasmas and P anatipestifer but there was a weak association between the infections with mycoplasmas, especially M anatis and P multocida.

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

    Directory of Open Access Journals (Sweden)

    Thais Sebastiana Porfida Ferreira

    2015-03-01

    Full Text Available 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.

  4. Une arthrite septique sur prothèse totale de genou à Pasteurella ...

    African Journals Online (AJOL)

    La présentation clinique n'a rien de spécifique.la contamination survient après une morsure, griffure ou léchage d'un chat. L'interrogatoire et l'examen bactériologique est la clé du diagnostique. Nous rapportons le cas d'une patiente de 84 ans qui présente une infection a Pasteurella multocida suite à une morsure du chat ...

  5. Management of avian cholera Pasteurella multocida outbreaks on ...

    African Journals Online (AJOL)

    Management intervention is required to reduce the negative impacts of disturbance due to kelp gull Lasus dominicanus predation on other breeding seabirds, primarily the African penguin Spheniscus demersus, during the carcass collection process. Keywords: avian cholera; Cape cormorant; disturbance; Dyer Island; ...

  6. Taxonomy of some recently described avian Pasteurella/Actinobacillus-like organisms as indicated by deoxyribonucleic acid relatedness.

    Science.gov (United States)

    Piechulla, K; Bisgaard, M; Gerlach, H; Mannheim, W

    1985-07-01

    Members of the family Pasteurellaceae Pohl 1981 are frequently encountered in birds as parasites or pathogens, e.g. the well-known species Pasteurella multocida, Pasteurella gallinarum, Haemophilus paragallinarum, and three species containing strains that had been previously classified as "Haemophilus avium"(Pasteurella avium, Pasteurella volantium, and an unnamed Pasteurella species defined by Mutters et al., 1985). A variety of additional taxa which had been tentatively assigned to the family do not belong to recognised species and could not even be classified to the genus level. In the present investigation selected Pasteurella/Actinobacillus-like isolates from fowl and zoo birds were examined for their genetic relationships with established species of Pasteurellaceae, using the renaturation method of deoxyribonucleic acid (DNA):DNA hybridisation. Guanine plus cytosine contents and genome masses were also considered. On the basis of DNA relatedness (1) the genus Pasteurella sensu stricto was extended to include Bisgaard's taxa 1 and 4 as two new Pasteurella species. (2) Bisgaard's taxa 2 and 3 and some phenotypically similar organisms isolated from zoo birds were shown to form a large distinct group which seems to represent a new genus with several species within the family Pasteurellaceae. (3) Another large heterogeneous, genus-like group consisted of strains labelled "Actinobacillus salpingitidis" and the phenotypically similar "Avian Pasteurella haemolytica-like" organisms; this group was placed in the close vicinity of the genus Actinobacillus. (4) Finally, a group of isolates labelled "Avian haemolytic Actinobacillus-like" organisms appeared to be heterogeneous on the species level but could be included in the genus Actinobacillus on the basis of DNA binding data. By co-evaluation of genetic relatedness and phenotypic features of avian Pasteurella/Actinobacillus-like isolates some diagnostically useful criteria have been detected.

  7. A case of wound dual infection with Pasteurella dagmatis and Pasteurella Canis resulting from a dog bite - limitations of Vitek-2 system in exact identification of Pasteurella species

    Directory of Open Access Journals (Sweden)

    Akahane T

    2011-12-01

    dog-bite, followed by the first report of dual infections due to P. dagmatis and P. multocida in 1988. Our isolate finally identified as P. dagmatis was misidentified as P. pneumotripica by means of the Vitek 2 system. The species name "P. dagmatis" was not included in the database of the system. It is also important for routine clinical microbiology laboratories to know the limitation of the automated Vitek 2 system for the accurate identification of Pasteurella species especially P. dagmatis. It should be emphasized that there still exists much room for improvement in Vitek 2 system. Significant improvement of Vitek 2 system especially in the identification of Pasteurella species is urgently desired.

  8. Experimental infection with different bacterial strains in larvae and juvenile Litopenaeus vannamei reared in Santa Catarina State, Brazil - doi: 10.4025/actascibiolsci.v32i3.5471 Experimental infection with different bacterial strains in larvae and juvenile Litopenaeus vannamei reared in Santa Catarina State, Brazil - doi: 10.4025/actascibiolsci.v32i3.5471

    Directory of Open Access Journals (Sweden)

    Adolfo Jatoba

    2010-09-01

    Full Text Available This study evaluated the pathogenic characteristics of bacteria isolated from Litopenaeus vannamei during an outbreak at the Laboratory of Marine Shrimp, UFSC, Santa Catarina State, Brazil. Their virulence potential in larvae and juvenile shrimp and the effects on the total haemocyte count, phenoloxidase activity and serum agglutinate titre were examined after experimental infection. Bacterial strains were isolated from larvae and adult shrimps, identified by the AP120E biochemical system as: two strains of Vibrio alginolyticus, three of Aeromonas salmonicida and one of Pasteurella multocida sp. and Pasteurella sp. All the bacterial strains isolated in this study caused mortality in shrimp. One strain of V. alginolyticus was responsible for 97.3 and 88.7% mortality in larvae and juvenil shrimps, respectively. The shrimp immunological system was influenced by experimental infection with V. alginolyticus. Decrease in the total haemocyte count and increase in the phenoloxidase activity and the serum agglutinate titre (p V. alginolyticus isolated from larvae and juvenile reared marine shrimp.This study evaluated the pathogenic characteristics of bacteria isolated from Litopenaeus vannamei during an outbreak at the Laboratory of Marine Shrimp, UFSC, Santa Catarina State, Brazil. Their virulence potential in larvae and juvenile shrimp and the effects on the total haemocyte count, phenoloxidase activity and serum agglutinate titre were examined after experimental infection. Bacterial strains were isolated from larvae and adult shrimps, identified by the AP120E biochemical system as: two strains of Vibrio alginolyticus, three of Aeromonas salmonicida and one of Pasteurella multocida sp. and Pasteurella sp. All the bacterial strains isolated in this study caused mortality in shrimp. One strain of V. alginolyticus was responsible for 97.3 and 88.7% mortality in larvae and juvenil shrimps, respectively. The shrimp immunological system was influenced by

  9. Bibersteinia trehalosi inhibits the growth of mannheimia haemolytica by a proximity-dependent mechanism

    Science.gov (United States)

    Mannheimia (Pasteurella) haemolytica is the only pathogen that consistently causes severe bronchopneumonia and rapid death of bighorn sheep (BHS; Ovis canadensis) under experimental conditions. Paradoxically, Bibersteinia (Pasteurella) trehalosi and Pasteurella multocida have been isolated from BHS ...

  10. A comparative study of the major outer membrane proteins of the avian haemophili and Pasteurella gallinarum.

    Science.gov (United States)

    Hartmann, L; Schröder, W; Lübke-Becker, A

    1996-06-01

    A polyclonal antibody prepared against the 35 kDa outer membrane protein (a putative porin) of Pasteurella (P.) multocida revealed binding to the 36 kDa major outer membrane protein (major Omp) of Haemophilus (H.) paragallinarum, to the 38 kDa major Omp of P.gallinarum, to the 39 kDa major Omp of P.volantium and to the 38.5 kDa major Omp of P. avium in immunoblotting studies. Comparison of N-terminal amino acid sequences also confirmed the relationship between the major Omps of most of the members of the family Pasteurellaceae.

  11. [In vitro susceptibility of Pasteurella and related bacteria to five orally administered antibiotics].

    Science.gov (United States)

    Lion, C; Lozniewski, A; Conroy, M C; Weber, M

    1996-05-01

    Tetracyclines and beta-lactam antibiotics are usually recommended for the treatment of pasteurellosis following bite wounds. However other oral antimicrobial agents could be proposed. In vitro susceptibility of 94 Pasteurella strains [P. multocida (79), P. stomatis (11), P. dagmatis (2), P. canis (1), P. "SP" (1)], 20 group EF-4 strains and 28 Neisseria weaveri strains (formerly group M-5), that are bacteria often isolated after animal-inflicted wounds, was studied towards five antibiotics: clarithromycin, azithromycin, pristinamycin, trimethoprim-sulfamethoxazole and ciprofloxacin. MICs were determined by the agar dilution method using HTM medium (Oxoïd), and for pristinamycin using both HTM and Mueller-Hinton agar supplemented with 5% horse blood (BMH). Most of Pasteurella isolates showed intermediate susceptibility to clarithromycin (63%) and to azithromycin (90.5%) with lower MICs for azithromycin. Fourty-two and thirty-two percent of Pasteurella strains were susceptible to pristinamycin respectively on HTM and on BMH. EF-4 and N. weaveri were more sensitive than Pasteurella to macrolides and to pristinamycin. Trimethoprim-sulfamethoxazole was active against all isolates, with higher MICs for EF-4 and N. weaveri. On all strains tested, the lowest MICs were observed for ciprofloxacin. Trimethoprim-sulfamethoxazole and ciprofloxacin could be proposed as a therapeutic alternative in case of pasteurellosis following animal bites.

  12. Pasteurella gallinarum neonatal meningitis.

    Science.gov (United States)

    Ahmed, K; Sein, P P; Shahnawaz, M; Hoosen, A A

    2002-01-01

    A 4-day-old baby weighing 1.7 kg was admitted to the neonatal intensive care unit of Ga-Rankuwa Hospital, Pretoria, with a history of apneic attacks. On examination there was an umbilical sepsis and the neonate was septicemic. The baby had been delivered at home and the umbilical cord had been cut by the grandmother using unclean scissors and chimney soot applied to the umbilical stump. On admission, a septic screen was done and antibiotic treatment was started with penicillin and amikacin. The investigations showed that the baby was slightly anemic, with hemoglobin levels of 10.0 g/dL (14.9-23.7 g/dL), and a pure growth of a Gram-negative bacillus was obtained from the cerebrospinal fluid, blood culture and suprapubic aspirate urine specimens. The Gram-negative bacillus was catalase and oxidase positive and it was identified as Pasteurella gallinarum. Antimicrobial profiling showed the organism to be susceptible to penicillin, cefotaxime, gentamicin and amikacin. Despite having received antimicrobial agents to which the etiological agent was susceptible, the neonate died within 5 days of admission. The cause of death was postulated to be due to overwhelming sepsis which resulted in septic shock.

  13. The efficacy of valnemulin (Econor) in the control of disease caused by experimental infection of calves with Mycoplasma bovis.

    Science.gov (United States)

    Stipkovits, L; Ripley, P H; Tenk, M; Glávits, R; Molnár, T; Fodor, L

    2005-06-01

    Mycoplasma bovis infection was experimentally induced in groups of six young calves. A further group was uninfected and served as a control. Ten days after infection, medication with either enrofloxacin (Baytril, Bayer) or valnemulin (Econor, Novartis) was instituted via the milk replacer for a further 10 days, after which all calves were killed. Infection resulted in depression, pyrexia, inappetance and prominent respiratory signs. Arthritis occurred in two animals and two (unmedicated) animals died. At post-mortem examination extensive lesions were present in the lungs and M. bovis was re-isolated from infected unmedicated calves' lungs. Medication with either enrofloxacin or valnemulin resulted in a rapid diminution of clinical signs, restoration of appetite and reversal of weight loss. Isolation of Pasteurella multocida from the calves' lungs was suppressed by both medicaments. Valnemulin resulted in a more rapid reduction of clinical scores and eliminated M. bovis from the lungs more effectively than enrofloxacin.

  14. Cloning and Characterization of Sialidases with 2-6′ and 2-3′ Sialyl Lactose Specificity from Pasteurella multocida†

    Science.gov (United States)

    Mizan, Shaikh; Henk, Adam; Stallings, Amy; Maier, Marie; Lee, Margie D.

    2000-01-01

    Pasteurella multocida is a mucosal pathogen that colonizes the respiratory system of susceptible hosts. Most isolates of P. multocida produce sialidase activity, which may contribute to colonization of the respiratory tract or the production of lesions in an active infection. We have cloned and sequenced a sialidase gene, nanH, from a fowl cholera isolate of P. multocida. Sequence analysis of NanH revealed that it exhibited significant amino acid sequence homology with many microbial sialidases. Insertional inactivation of nanH resulted in a mutant strain that was not deficient in sialidase production. However, this mutant exhibited reduced enzyme activity and growth rate on 2-3′ sialyl lactose compared to the wild type. Subsequently, we demonstrated the presence of two sialidases by cloning another sialidase gene that differed from nanH in DNA sequence and substrate specificity. NanB demonstrated activity on both 2-3′ and 2-6′ sialyl lactose, while NanH demonstrated activity only on 2-3′ sialyl lactose. Neither enzyme liberated sialic acid from colominic acid (2-8′ sialyl lactose). Recombinant E. coli containing the sialidase genes were able to utilize several sialoconjugants when they were provided as sole carbon sources in minimal medium. These data suggest that sialidases have a nutritional function and may contribute to the ability of P. multocida to colonize and persist on vertebrate mucosal surfaces. PMID:11092845

  15. Experimental infection with different bacterial strains in larvae and juvenile Litopenaeus vannamei reared in Santa Catarina State, Brazil = Infecção experimental em larvas e juvenis de Litopaenaeus vannamei cultivados no Estado de Santa Catarina, Brasil

    Directory of Open Access Journals (Sweden)

    Celso Carlos Buglione

    2010-07-01

    Full Text Available This study evaluated the pathogenic characteristics of bacteria isolated from Litopenaeus vannamei during an outbreak at the Laboratory of Marine Shrimp, UFSC, Santa Catarina State, Brazil. Their virulence potential in larvae and juvenile shrimp and theeffects on the total haemocyte count, phenoloxidase activity and serum agglutinate titre were examined after experimental infection. Bacterial strains were isolated from larvae and adult shrimps, identified by the AP120E biochemical system as: two strains of Vibrioalginolyticus, three of Aeromonas salmonicida and one of Pasteurella multocida sp. and Pasteurella sp. All the bacterial strains isolated in this study caused mortality in shrimp. One strain of V. alginolyticus was responsible for 97.3 and 88.7% mortality in larvae and juvenil shrimps, respectively. The shrimp immunological system was influenced by experimental infection with V. alginolyticus. Decrease in the total haemocyte count and increase in the phenoloxidase activity and the serum agglutinate titre (p Este estudo avaliou as características patogênicas de cepas de bactérias isoladas de Litopenaeus vannamei durante surto de mortalidade no Laboratório de Camarões Marinhos, UFSC, Estado de Santa Catarina, Brasil. Seu potencial de virulência em larvas e juvenis de camarão marinho e os efeitos sobre a contagem total de hemócito, atividade de fenoloxidase e título aglutinante do soro foramavaliados após infecção experimental. As cepas bacterianas foram isoladas de larvas e de camarões adultos e identificadas bioquimicamente pelo sistema API20E como: duas cepas de Vibrio alginolyticus, três de Aeromonas salmonicida e uma de Pasteurella sp. e P. multocida. Todas as cepas isoladas provocaram mortalidade em L. vannamei, e uma de V. alginolyticus resultou em mortalidade de 97,3 e 88,7% para larvas e juvenis de camarões, respectivamente. O sistema imunológico dos camarões juvenis sofreu influência da infecção experimental

  16. An angry cat causing Pasteurella multocida endocarditis and aortic valve replacement—A case report

    Directory of Open Access Journals (Sweden)

    Anders Ahlsson

    2016-01-01

    Conclusion: Cat bites are often deep, and in rare circumstances can lead to life-threatening endocarditis. Proper surgical revision, antibiotic treatment, and patient compliance are necessary components in patient care to avoid this complication.

  17. 21 CFR 522.955 - Florfenicol.

    Science.gov (United States)

    2010-04-01

    ... treatment of bovine respiratory disease (BRD) associated with Mannheimia haemolytica, Pasteurella multocida.... For treatment of BRD associated with Mannheimia (Pasteurella) haemolytica, P. multocida, and... Mannheimia (Pasteurella) haemolytica, P. multocida, and Haemophilus somnus. (C) Limitations. Do not slaughter...

  18. 76 FR 22610 - Implantation or Injectable Dosage Form New Animal Drugs; Enrofloxacin

    Science.gov (United States)

    2011-04-22

    ... with Mannheimia haemolytica, Pasteurella multocida, Histophilus somni, and Mycoplasma bovis in beef and... (BRD) associated with Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni in beef and...

  19. Avaliação de ferramentas para monitoria da infecção por P. multocida em suínos

    OpenAIRE

    Marina Moreno

    2011-01-01

    Pasteurella multocida é um importante patógeno para suínos, causando rinite atrófica progressiva, pneumonia, pleurite, e septicemia. Para implantação de estratégias de controle e prevenção desta infecção, torna-se necessário o conhecimento a respeito do perfil de disseminação do agente em condições naturais e em diferentes tipos de sistema de produção. Tendo em vista a inexistência de técnicas sorológicas padronizadas para esta finalidade na espécie suína, o presente estudo teve por objetivos...

  20. Mannheimiose pulmonar experimental em bezerros: swab nasal e nasofaringeano como auxílio diagnóstico Experimental pneumonic mannheimiosis in calves: nasal and nasopharingeal swabs for diagnostic

    Directory of Open Access Journals (Sweden)

    Adriana de Souza Coutinho

    2009-01-01

    Full Text Available Um modelo experimental de mannheimiosepneumônica bovina (MPB foi utilizado com o objetivo de avaliar as espécies bacterianas das cavidades nasais e nasofaringeanas em diferentes momentos do curso da doença, bem como verificar a eficiência diagnóstica do exame microbiológico dos swabs nasais (SN e nasofaringeanos (SNF. Um total de 28 bezerros foi distribuído aleatoriamente em quatro grupos experimentais (G1 a G4. SN e SNF foram colhidos sete dias antes e 12 (G1, 24 (G2, 48 (G3 e 72 (G4 horas após a inoculação intrabronquial de Mannheimia haemolytica. Após a indução da MPB, a bactéria M. haemolytica biotipo A foi predominante nos SN e SNF, sendo isolada em todos os momentos avaliados, com exceção de um SN colhido 24 horas após a indução da infecção. Não houve diferença significativa nas taxas de isolamento de Pasteurella multocida nos SN ou SNF, colhidos antes e após a indução da MPB. Contudo, esta bactéria passou a ser isolada mais freqüentemente após a indução da MPB, principalmente no SNF. Portanto, pode-se concluir que o exame microbiológico de SN e SNF é um teste auxiliar no diagnóstico da MPB.An experimental model of bovine pneumonic mannheimiosis (BPM was used to evaluate the nasal and nasopharynx bacterial species of calves during the course of the disease and for checking the diagnostic efficiency of nasal swab (NS and nasopharingeal swab (NPS microbiological exams. A total of 28 calves were randomized into four experimental groups (G1-G4. NS and NPS were obtained 7 days before and 12 (G1, 24 (G2, 48 (G3 e 72 (G4 hours after intrabronchial inoculation of Mannheimia haemolytica. After the induction of BPM, M. haemolytica biotype A was the predominant isolated bacterium in NS and NPS in all evaluated sampling times, except for one NS (harvested 24 hours. There were no significant statistical differences for the rates of Pasteurella multocida isolation in NS and NPS, harvested before and after the induction

  1. Enteritis caused by Pasteurella pneumotropica infection in hamsters.

    OpenAIRE

    Lesher, R J; Jeszenka, E V; Swan, M E

    1985-01-01

    Pasteurella pneumotropica was isolated in essentially pure cultures from the bowels of hamsters with enteritis 7 days after parturition. Newly received hamsters showed presence of P. pneumotropica in their nasal cavities but not in their uteri, lungs, spleens, or bowels.

  2. 21 CFR 522.313a - Ceftiofur crystalline free acid.

    Science.gov (United States)

    2010-04-01

    ... Actinobacillus pleuropneumoniae, Pasteurella multocida, Haemophilus parasuis, and Streptococcus suis. (iii... fever, pneumonia) associated with Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni.... haemolytica, P. multocida, and H. somni. For the treatment of bovine foot rot (interdigital necrobacillosis...

  3. Macrolide resistance conferred by rRNA mutations in field isolates of Mannheimia haemolytica and Pasteurella multocida

    DEFF Research Database (Denmark)

    Olsen, Anders S; Warrass, Ralf; Douthwaite, Stephen Roger

    2014-01-01

    . haemolytica identified as being highly resistant (MICs >64 mg/L) to the macrolides erythromycin, gamithromycin, tilmicosin, tildipirosin and tulathromycin were screened by multiplex PCR for the previously identified resistance genes erm(42), msr(E) and mph(E). Strains lacking these determinants were analysed...

  4. Literatuuronderzoek naar gegevens betreffende de betekenis van een aantal verwekkers van zoonosen in verband met de vleesconsumptie. V. Pasteurella multocida

    NARCIS (Netherlands)

    Bos; J.M.; Engel; H.W.B.; Groothuis; D.G.; Knapen; F.van; Oosterom; J.; Weiss; J.W.

    1985-01-01

    Infecties bij de mens manifesteren zich voornamelijk als wondinfecties of luchtwegaandoeningen. Predisponerende (weerstandverminderende) factoren zijn in de meeste gevallen echter noodzakelijk voor het aanslaan van een infectie. Het microorganisme komt momenteel voor bij tal van zoogdieren en

  5. Complete chemoenzymatic synthesis of the Forssman antigen using novel glycosyltransferases identified in Campylobacter jejuni and Pasteurella multocida

    Science.gov (United States)

    We have identified an alpha1,4-galactosyltransferase (CgtD) and a beta1,3-N-acetylgalactosaminyltransferase (CgtE) in the lipooligosaccharide (LOS) locus of Campylobacter jejuni LIO87. Strains that carry these genes may have the capability of synthesizing mimics of the P blood group antigens of the ...

  6. Isolation and characterization of some previously unreported taxa from poultry with phenotypical characters related to Actinobacillus-an Pasteurella species.

    Science.gov (United States)

    Bisgaard, M

    1982-02-01

    Cultural, morphologic, and biochemical characteristics of previously unreported taxa isolated from poultry and tentatively assigned to genus Actinobacillus Brumpt 1910 were compared to those of Actinobacillus lignieresii, A equuli, A. seminis, A. suis, avian haemolytic Actinobacillus sp., A.salpingitidis, avian Pasteurella haemolytica-like strains, P haemolytica biovar T, P, ureae. P. multocida, P. pneumotropica, P. gallinarum and P. anatipestifer. Evidence as obtained to indicate that taxon 1--3 was closely related to genus Actinobacillus Brumpt 1910, but sufficiently different from established species within that genus to constitute new distinct species. Taxon 4 had the cultural and biochemical characters of strains previously described by Clark and Godfrey. Strains designated P haemolytica-like could not be separated from A. salpingitidis on the basis of phenotypical characters. The final taxonomical position of taxon 1-4 in addition to strains designated avian haemolytic Actinobacillus sp. and P. haemolytica-like, however, has to await further taxonomical investigations including determination of mol % G + C in DNA and DNA hybridization, for which reason species names have been omitted.

  7. Pasteurella gallinarum: Zimbabwean experience of a versatile pathogen.

    Science.gov (United States)

    Mohan, K; Dziva, F; Chitauro, D

    2000-12-01

    Pasteurella gallinarum-related outbreaks in chickens and African guinea fowls are described. Four outbreaks were recorded in chickens and one in guinea fowls. Periorbital swelling and keratoconjunctivitis were the consistently present clinical signs in all the diseased birds. In several, swollen hocks and wattles were also discerened. Birds which succumbed to the infection showed petechiation in the internal organs and evidence of airsacculitis. Pasteurella gallinarum was isolated from the lesions and also from conjunctival swabs of the apparently healthy in-contact birds. There was no evidence of concurrent infection with Haemophilus, Mycoplasma or Chlamydia. Quinolone therapy when resorted to on one of the farms resolved the clinical signs. Phenotypes of 28 isolates were studied. The results compared well with the Pasteurella gallinarum isolates reported earlier from elsewhere. It was also found that results of xylose fermentation and ONPG test appear to be a variable character. There is no earlier report of P. gallinarum infection in guinea fowls.

  8. 2 original article investigations on the carrier rate of pasteurella

    African Journals Online (AJOL)

    Dr Oboro VO

    farmhouse with recurrent fowl cholera outbreaks and also to evaluate the association between the P. multocida found in rats co-habiting quail poultry houses and isolates from outbreaks of fowl cholera. Thus 100 pharyngeal and 100 rectum swabs samples taken from rats co-habiting farmhouse were obtained and evaluated ...

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

    Science.gov (United States)

    2010-01-01

    ... established as pure, safe, and immunogenic shall be used for vaccine production. All serials of vaccine shall.... (b) Each lot of Master Seed used for vaccine production shall be tested for immunogenicity. The... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Haemolytica Vaccine...

  10. Pasteurellosis and Pasteurellae inZimbabwe: An update | Dziva ...

    African Journals Online (AJOL)

    A wide spectra of hosts affected by species of the genus Pasteurella include cattle, rabbits, chickens, sheep, pigs, goats, crocodiles, dogs and cats. In cattle, the condition has predominantly been the pneumonic form. In pigs, pneumonia has commonly been encountered. Classical progressive atrophic rhinitis was recorded ...

  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.

    Science.gov (United States)

    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. Effects of avian cholera on survival of Lesser Snow geese Anser caerulescens: An experimental approach

    Science.gov (United States)

    Samuel, Michael D.; Takekawa, John Y.; Baranyuk, Vasily V.; Orthmeyer, Dennis L.

    1999-01-01

    Avian cholera, caused by the bacterium Pasteurella multocida, is one of the most important diseases affecting waterfowl in North America but little is known about the epizootiology of this disease or its impacts on annual survival rates. We ringed Lesser Snow Geese Anser caerulescens nesting at Wrangel Island, Russia and Banks Island, Canada with metal rings and individually coded plastic neck-collars or radio-transmitters to determine survival, movement and cause of death. We vaccinated half of the neck-collared and radiotagged geese to provide protection from avian cholera for up to one year following ringing and thus experimentally determine the impacts of this disease on survival. We found that vaccination did not reduce short-term survival of the experimental birds, compared with control geese. Neck-collared geese vaccinated in 1993 at Wrangel Island had higher survival during winter 1993–94 than control birds. In contrast, we found similar survival during winter 1994–95 between vaccinated and control geese neck-collared in 1994 at Wrangel and Banks Islands. Survival of radiotagged geese on wintering areas during 1994–95 was consistent with the vaccination versus control results for neck-collared geese during the same winter. However, we found that radiotagged geese that were vaccinated had better survival than control geese during winter 1995–96. We believe that harvest and avian cholera are the two principal causes of mortality for Lesser Snow Geese wintering in the Pacific Flyway and that avian cholera may be one of the factors affecting these populations.

  13. Pasteurella Species Peritoneal Dialysis-Associated Peritonitis: Household Pets as a Risk Factor

    Directory of Open Access Journals (Sweden)

    Philippe Guillaume Poliquin

    2015-01-01

    Full Text Available 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.

  14. Role of Pasteurella granulomatis and Dermatobia hominis in the etiology of lechiguana in cattle.

    Science.gov (United States)

    Ladeira, S L; Riet-Correa, F; Pereira, D B; Carter, G R

    1996-07-23

    Attempts were made to reproduce bovine lechiguana, a disease associated with Dermatobia hominis and Pasteurella granulomatis infections. Suspensions of Pasteurella granulomatis were mixed with each of the following: saponin, oil adjuvant, ground Dermatobia hominis, or 5% mucin. Each preparation was inoculated into 6 cattle. Twelve more cattle, 6 of which received dexamethasone, were inoculated with bacterial suspension alone. Abscesses but no lechiguana was produced in all 36 cattle. After abscess regression, 12 cattle were reinoculated with a suspension of mouse-passed P. granulomatis. Only abscesses were produced. The intralymphatic inoculation of P. granulomatis in 6 cattle did not produce the disease. Eleven cattle infected naturally with D. hominis had lesions containing dead larvae. These lesions were inoculated with P. granulomatis. Nine cattle were experimentally infected with larvae of D. hominis that had been contaminated with the bacteria. No lechiguana lesions were produced in these 20 cattle. Six cattle with severe natural D. hominis infection were inoculated in the larval lesions with P. granulomatis. One developed lesions indistinguishable from those of natural lechiguana. The lesions regressed after treatment with chloramphenicol. D. hominis larvae and exudate from lesions caused by the fly were collected from 7 cattle on 3 farms and examined bacteriologically. P. granulomatis was isolated from the larvae and the exudate of a healthy calf from a farm where lechiguana had never been observed. These results suggest that P. granulomatis has a causal role in lechiguana, and that D. hominis may be a carrier of the bacterium. These observations suggest that lechiguana occurs when severe D. hominis lesions are infected with P. granulomatis. The apparent long incubation period, the negative results obtained in the other experiments, and also the infrequent occurrence of the natural disease suggest that lechiguana is a disease for which Koch's postulates are

  15. 21 CFR 520.1660c - Oxytetracycline hydrochloride tablets/boluses.

    Science.gov (United States)

    2010-04-01

    ... (shipping fever complex, pasteurellosis) caused by Pasteurella multocida. (2)(i) Amount. 500 milligrams per... Pasteurella multocida. (3) Limitations. Dosage should continue until the animal returns to normal and for 24...

  16. 77 FR 76862 - New Animal Drugs; Enrofloxacin; Melengestrol; Meloxicam; Pradofloxacin; Tylosin

    Science.gov (United States)

    2012-12-31

    ... caused by susceptible strains of Pasteurella multocida, Streptococcus canis, Staphylococcus aureus, Staphylococcus felis, and Staphylococcus pseudintermedius. PART 522--IMPLANTATION OR INJECTABLE DOSAGE FORM NEW... 66201. strains of Pasteurella multocida, Streptococcus canis, S. aureus, S. felis, and S...

  17. [Pasteurelloses].

    Science.gov (United States)

    Avril, J L; Donnio, P Y

    1995-03-18

    According to the genetic relationships among Gram-negative bacilli the genus Pasteurella is included with the genus Haemophilus and the genus Acinobacillus within the family Pasteurellacae. Pasteurella multocida, the type species, is responsible for the majority of human Pasteurella infections. P. multocida is a member of the normal flora in the upper respiratory tract of many mammals or birds. It causes sporadic or epidemic diseases among different animal species, particularly pneumonia and atrophic rhinitis in swine in intensive breeding stations. The most common human infection with P. multocida is a local cellulitis following dog or cat bites and scratches. Serious local complications are sometimes responsible for prolonged disability. The respiratory tract is the second human source of P. multocida isolates. The frequency of recovery of P. multocida from oropharynx of apparently healthy pig breeders suggests that respiratory pasteurellosis could be an occupational disease. The mechanisms of virulence of P. multocida are unclear. Several factors are involved: capsules preventing phagocytosis, a dermonecrotic toxin causing experimental atrophic rhinitis, hyaluronidase, neuraminidase and proteases. Penicillin is considered to be the drug of choice for Pasteurella infection. Tetracyclin is efficient for bites but has no bactericidal effect. Oxacillin, first-generation cephalosporins, macrolides and aminoglycosides have poor activities. In the case of beta-lactamase producing strains a bactericidal effect could be achieved with fluoroquinolones or third generation cephalosporins.

  18. NCBI nr-aa BLAST: CBRC-OANA-01-0048 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-OANA-01-0048 ref|NP_246945.1| hypothetical protein PM2006 [Pasteurella multocida subsp. multoci...da str. Pm70] gb|AAK04090.1| unknown [Pasteurella multocida subsp. multocida str. Pm70] NP_246945.1 1.3 21% ...

  19. NCBI nr-aa BLAST: CBRC-TTRU-01-0264 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-TTRU-01-0264 ref|NP_246278.1| sensory histidine kinase UhpB [Pasteurella multocida subsp. multoci...da str. Pm70] gb|AAK03423.1| UhpB [Pasteurella multocida subsp. multocida str. Pm70] NP_246278.1 0.009 30% ...

  20. NCBI nr-aa BLAST: CBRC-BTAU-01-2867 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-BTAU-01-2867 ref|NP_245635.1| hypothetical protein PM0698 [Pasteurella multocida subsp. multoci...da str. Pm70] gb|AAK02782.1| unknown [Pasteurella multocida subsp. multocida str. Pm70] NP_245635.1 5.8 33% ...

  1. NCBI nr-aa BLAST: CBRC-CREM-01-1342 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-CREM-01-1342 ref|NP_245519.1| hypothetical protein PM0582 [Pasteurella multocida subsp. multoci...da str. Pm70] gb|AAK02666.1| unknown [Pasteurella multocida subsp. multocida str. Pm70] NP_245519.1 2e-25 33% ...

  2. NCBI nr-aa BLAST: CBRC-LAFR-01-1776 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-LAFR-01-1776 ref|NP_244964.1| NrfE [Pasteurella multocida subsp. multocida str.... Pm70] gb|AAK02111.1| NrfE [Pasteurella multocida subsp. multocida str. Pm70] NP_244964.1 5.2 28% ...

  3. Genotypic heterogeneity of Pasteurella gallinarum as shown by ribotyping and 16S rRNA sequencing

    National Research Council Canada - National Science Library

    Christensen, Henrik; Dziva, Francis; Olsen, John Elmerdahl; Bisgaard, Magne

    2002-01-01

    Forty-five strains mainly isolated from chickens in Zimbabwe and Denmark, two pig and three rat isolates all identified as Pasteurella gallinarum by conventional phenotypic tests were characterized...

  4. Avaliação de bacterina e Lactobacillus plantarum frente à infecção experimental por Vibrio harveyi em pós-larvas de Litopenaeus vannamei

    Directory of Open Access Journals (Sweden)

    Celso Carlos Buglione

    2008-12-01

    Full Text Available This study aimed to verify the effect of probiotics and inactivated cells of bacterias such as Vibrio alginolyticus, Aeromonas salmonicida and Pasteurella multocida in larvae survival of Litopenaeus vannamei, in stress test and experimental infection with Vibrio harveyi. Conic tanks of 30 L, were stocked with 400 post-larvae stage five. Four experimental treatments with triplicates consisted of: 1: commercial feed (control, 2: commercial feed plus bacterin by oral administration in artemia, 3: commercial feed plus bacterin by immersion administration, 4: commercial feed with Lactobacillus plantarum inoculation. Bacterin application was conducted 6h before the infection and stress test, while probiotic administration was for 15 days before challenges. In stress test, post-larvae of treatment 4 (commercial feed supplemented with Lactobacillus plantarum with reached the highest survival rate (87,86 ± 2,35% followed by the ones of treatment 3 and 2 (bacterim by immersion and bacterim by oral administration in artemia with 81,54±1,50% and 80,16 ± 2,15%, respectively, which were superior to the control treatment (72,63 ± 3,34%. Next to V. harveyi challenge, animals from treatment 3 presented the highest survival rate (79,60 ± 7,12% followed by treatments 4 (69,60 ± 10,43%, 2 (65,60 ± 5,18% and control (56,4 ± 5,58%. All treatments were different from control. The present results demonstrate the possible use of L. plantarum and bacterin as promoters in survival rates of L. vannamei post-larvae in the stress tests and challenges with Vibrio harveyi.

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

    National Research Council Canada - National Science Library

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

    2016-01-01

    ..., 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...

  6. Pasteurella canis Isolation following Penetrating Eye Injury: A Case Report

    Directory of Open Access Journals (Sweden)

    Noor-Khairul Rashid

    2012-01-01

    Full Text Available A 3-year-old boy presented with history of trauma to the left eye after he accidentally injured his eye with a broom stick made up from coconut skewers. There was history of cats as their pets but not dogs. Ocular examination revealed left superonasal conjunctival laceration and scleral perforation with prolapsed vitreous. Fundus examination showed minimal vitreous haemorrhage and flat retina. Conjunctiva swab at the wound site was sent for gram staining, culture, and sensitivity. He underwent scleral suturing, vitreous tap, and intravitreal injection of Ceftazidime and Amikacin. Vitreous tap was sent for gram stained, culture and sensitivity. Postoperatively, he was started empirically on IV Ciprofloxacin 160 mg BD, Guttae Ciprofloxacin, and Guttae Ceftazidime. Conjunctiva swab grew Pasteurella canis which was sensitive to all Beta lactams, Ciprofloxacin, Chloramphenicol, and Aminoglycoside. Post-operative was uneventful, absent signs of endophthalmitis or orbital cellulitis.

  7. Pasteurella Pneumotropica causa la regresión de tumores humanos trasplantados en ratones inmunodeficientes Pasteurella pneumotropica produces regression of human tumors transplanted in immunodeficiency mice

    Directory of Open Access Journals (Sweden)

    Martín Carriquiriborde

    2006-06-01

    Full Text Available La técnica de trasplante de tumores humanos en ratones inmunodeficientes es muy utilizada como modelo en investigaciones sobre el cáncer. De acuerdo con las recomendaciones internacionales, los animales de experimentación deben estar libres de los micoorganismos que interfieren en los resultados finales de las investigaciones, dentro de los cuales se encuentra Pasteurella pneumotropica. En el presente trabajo se evaluó la interferencia que produceesta bacteria en el crecimiento de la línea celular A549 de adenocarcinoma humano trasplantada en ratones de la cepa nude N:NIH (S-nu. Se utilizaron 40 ratones divididos en 4 grupos de 10 animales cada uno. Grupo 1: inoculados con la línea celular; grupo 2, con la bacteria; grupo 3, con la línea celular y la bacteria y grupo 4, el control sin inoculaciones. Se observaron diferencias significativas en el crecimiento tumoral entre los animales de los grupos 1 y 3. Si bien este microorganismo es un patógeno oportunista no letal, los ratones trasplantados con la línea celular A549 e infectados con P. pneumotropica no son aptos para utilizarse como modelo animal en estudios sobre el cáncer debido a que esta bacteria interfiere en el desarrollo de la línea tumoral, con la consecuente interpretación errónea de los resultados. Pero el hecho que la bacteria haya causado la regresión de un tumor en pleno crecimiento es inesperado y el mecanismo de acción será objeto de futuros experimentos.The technique of human tumor cell line transplantation in immunodeficient mice is used worldwide as a model for cancer research. In accordance with international recommendations, animals used in biomedical research should be free of microorganisms which can interfere in experimental results; including Pasteurella pneumotropica. The object of this study was to evaluate the interference produced by P. pneumotropica in the human adenocarcinoma cell line A549 transplanted in N:NIH(S-nu mice. A total of 40 mice

  8. Clonal outbreaks of [Pasteurella] pneumotropica biovar Heyl in two mouse colonies

    DEFF Research Database (Denmark)

    Adhikary, Sadhana; Bisgaard, Magne; Dagnæs-Hansen, Frederik

    2017-01-01

    The aim of this study was to document the pathogenic role of biovar Heyl of [Pasteurella] pneumotropica in mouse colonies. Fifty-three isolates associated with mastitis and orbital, cutaneous and vaginal abscesses as well as isolates from the nose and vagina of healthy mice were investigated...

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

    Science.gov (United States)

    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. Copyright © 2014 Sasaki et al.

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

    OpenAIRE

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

    2014-01-01

    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.

  11. 21 CFR 558.415 - Novobiocin.

    Science.gov (United States)

    2010-04-01

    ... strains of Pasteurella multocida susceptible to novobiocin following initial treatment with 7-8 mgs. per... cholera in ducks caused by Pasteurella anatipestifer and P. multocida, susceptible to novobiocin. (iii... susceptible to novobiocin; treatment of acute outbreaks of fowl cholera caused by strains of Pasteurella...

  12. Genotypic heterogeneity of Pasteurella gallinarum as shown by ribotyping and 16S rRNA sequencing.

    Science.gov (United States)

    Christensen, Henrik; Dziva, Francis; Olsen, John Elmerdahl; Bisgaard, Magne

    2002-12-01

    Forty-five strains mainly isolated from chickens in Zimbabwe and Denmark, two pig and three rat isolates all identified as Pasteurella gallinarum by conventional phenotypic tests were characterized by ribotyping, and selected strains were subsequently analysed by 16S rRNA gene sequencing. High genotypic diversity was observed, the number of ribotypes totalling 24. A major group of 47 isolates including the type strain of P. gallinarum clustered at 56% similarity and included 21 ribotypes. Ribotyping showed that some genotypes of P. gallinarum seem to be globally distributed. The three isolates from rodents did not share even a single common ribotype fragment with strains from birds and the pig isolates. Two avian isolates from Denmark and Zimbabwe and the pig strain showed from 97.6 to 99.8% 16S rRNA sequence similarity with the type strain of P. gallinarum and with type strains of Pasteurella volantium and Pasteurella avium. Two rat strains showed 98.6% 16S rRNA gene sequence similarity with each other, but were only related with P. gallinarum at 93% similarity. These isolates showed the highest similarity with [Actinobacillus] muris at 96.4 to 95.0% similarity. We suggest that conventional identification of P. gallinarum consequently should consider the source of isolation to obtain a correct diagnosis, and that isolation from animals other than fowl should be confirmed by genotypic analysis such as 16S rRNA gene sequence comparison.

  13. Efforts Towards The Development Of Recombinant Vaccines Against

    African Journals Online (AJOL)

    ABSTRACT. Hemorrhagic septicemia is caused by gram-negative bacterium of Pasteurella multocida (P. multocida) strains. Most of the current vaccines against P. multocida have shortcomings. Presently, there is increasing efforts towards construction of recombinant clone for vaccine development against P. multocida.

  14. Efforts towards the development of recombinant Vaccines against ...

    African Journals Online (AJOL)

    Hemorrhagic septicemia is caused by gram-negative bacterium of Pasteurella multocida (P. multocida) strains. Most of the current vaccines against P. multocida have shortcomings. Presently, there is increasing efforts towards construction of recombinant clone for vaccine development against P. multocida. In this review an ...

  15. Evaluation of a PCR for detection of Actinobacillus pleuropneumoniae in mixed bacterial cultures from tonsils

    DEFF Research Database (Denmark)

    Gram, T.; Ahrens, Peter; Nielsen, J.P.

    1996-01-01

    . rossii, A. suis, Escherichia coli, Bordetella bronchiseptica, Streptococcus suis, Pasteurella haemolytica, Pasteurella multocida, Haemophilus parasuis, Haemophilus taxon Minor group, Haemophilus taxon D/E and Haemophilus taxon F. Amplification of a 985 bp product was, however, observed when testing...

  16. Elimination of Pasteurella pneumotropica from a Mouse Barrier Facility by Using a Modified Enrofloxacin Treatment Regimen

    OpenAIRE

    Towne, Justin W; Wagner, April M; Griffin, Kurt J; Buntzman, Adam S.; Frelinger, Jeffrey A.; Besselsen, David G.

    2014-01-01

    Multiple NOD.Cg-Prkdcscid Il2rgtm1WjlTg(HLA-A2.1)Enge/Sz (NSG/A2) transgenic mice maintained in a mouse barrier facility were submitted for necropsy to determine the cause of facial alopecia, tachypnea, dyspnea, and sudden death. Pneumonia and soft-tissue abscesses were observed, and Pasteurella pneumotropica biotype Jawetz was consistently isolated from the upper respiratory tract, lung, and abscesses. Epidemiologic investigation within the facility revealed presence of this pathogen in mice...

  17. Effect of Diluent and Relative Humidity on Apparent Viability of Airborne Pasteurella pestis

    Science.gov (United States)

    Won, William D.; Ross, Harold

    1966-01-01

    Airborne Pasteurella pestis (A-1122) at low humidities [20 to 50% relative humidity (RH)] exhibited exponential decay when either 1% peptone or Heart Infusion Broth (HIB) was used as the diluent in the viable assay system. At higher RH values (65 and 87%), however, the 1% peptone diluent adversely affected the viability assay. In contrast, HIB as diluent was remarkably effective in demonstrating a higher number of viable cells in aerosols held at high RH values. Similarly, with HIB as diluent, aerosols were shown to contain viable cells during 90 min of observation; with 1% peptone, viability was not detectable after 20 min in the airborne state. PMID:5970462

  18. Effect of diluent and relative humidity on apparent viability of airborne Pasteurella pestis.

    Science.gov (United States)

    Won, W D; Ross, H

    1966-09-01

    Airborne Pasteurella pestis (A-1122) at low humidities [20 to 50% relative humidity (RH)] exhibited exponential decay when either 1% peptone or Heart Infusion Broth (HIB) was used as the diluent in the viable assay system. At higher RH values (65 and 87%), however, the 1% peptone diluent adversely affected the viability assay. In contrast, HIB as diluent was remarkably effective in demonstrating a higher number of viable cells in aerosols held at high RH values. Similarly, with HIB as diluent, aerosols were shown to contain viable cells during 90 min of observation; with 1% peptone, viability was not detectable after 20 min in the airborne state.

  19. Phenotypic and genotypic characterization of Mannheimia (Pasteurella) haemolytica-like strains isolated from diseased animals in Denmark

    DEFF Research Database (Denmark)

    Angen, Øystein; Ahrens, Peter; Bisgaard, M.

    2002-01-01

    Trehalose-negative strains of the Pasteurella haemolytica complex have recently been transferred to a new genus, Mannheimia. This genus presently consists of five named species: M. haemolytica, M. glucosida, M. granulomatis, M. ruminalis and M. varigena. The purpose of this study was to investigate...

  20. [Pasteurella] caballi infection not limited to horses - a closer look at taxon 42 of Bisgaard

    DEFF Research Database (Denmark)

    Christensen, Henrik; Hommez, J.; Olsen, John Elmerdahl

    2006-01-01

    Aim To investigate if taxon 42 of Bisgaard isolated from pigs represents genuine [Pasteurella] caballi which has previously only been isolated from horses. Methods and Results A total of 15 field isolates from horses and pigs from 5 different countries representing three continents were to subjec......Aim To investigate if taxon 42 of Bisgaard isolated from pigs represents genuine [Pasteurella] caballi which has previously only been isolated from horses. Methods and Results A total of 15 field isolates from horses and pigs from 5 different countries representing three continents were...... to subjected extended phenotypical characterization. Although minor differences were observed between taxon 42 and [P.] caballi, these differences did not allow phenotypic separation. Ribotyping based on HindIII digestion showed five profiles based on 9 band positions. One [P.] caballi strain and two taxon 42...... strains shared the same profile. Ribotyping using HpaII gave a higher diversity with 9 profiles based on 10 band positions. While no profiles were shared between the Taxon 42 and [P.] caballi strains, pattern analysis showed that two of the Taxon 42 isolates were most similar (91 % similarity) with a [P...

  1. Studies of the binding of ficolin-2 and ficolin-3 from the complement lectin pathway to Leptospira biflexa, Pasteurella pneumotropica and Diarrheagenic Escherichia coli

    DEFF Research Database (Denmark)

    Sahagún-Ruiz, Alfredo; Breda, Leandro Carvalho Dantas; Valencia, Mónica Marcela Castiblanco

    2015-01-01

    . We show for the first time that human ficolin-2 recognizes the nonpathogenic spirochete Leptospira biflexa serovar Patoc, but not the pathogenic Leptospira interrogans serovar Kennewicki strain Fromm. Additionally, human ficolin-2 and ficolin-3 recognize pathogenic Pasteurella pneumotropica...

  2. 77 FR 4226 - Implantation or Injectable Dosage Form New Animal Drugs; Danofloxacin

    Science.gov (United States)

    2012-01-27

    ... haemolytica, and Pasteurella multocida in beef cattle. The supplemental NADA is approved as of December 16... treatment of bovine respiratory disease (BRD) associated with Mannheimia haemolytica and Pasteurella multocida. * * * * * Dated: January 23, 2012. William T. Flynn, Acting Director, Center for Veterinary...

  3. Severe mortality in broiler chickens associated with Mycoplasma synoviae and Pasteurella gallinarum.

    Science.gov (United States)

    Droual, R; Shivaprasad, H L; Meteyer, C U; Shapiro, D P; Walker, R L

    1992-01-01

    Severe economic loss due to high mortality and condemnation rates occurred on two commercial broiler facilities. Chickens had moderate-to-severe airsacculitis, pericarditis, perihepatitis, tracheitis, and synovitis. Pasteurella gallinarum was isolated from 16 of 18 pericardia, four of 14 livers, 11 of 16 air sacs, six of seven joints and one of 28 tracheas in pure culture. In addition, Mycoplasma synoviae was isolated from trachea and air sac. Lesions were suggestive of an Escherichia coli septicemia, but E. coli was isolated from only four of 28 tracheas and one of 14 livers in pure culture. A coronavirus was isolated from trachea and lung. Whether this coronavirus represented a vaccine or field strain of infectious bronchitis was not determined. These findings suggested that the severe lesions were due to a concomitant infection with an atypical strain of P. gallinarum.

  4. An atypical strain of Pasteurella gallinarum: pathogenic, phenotypic, and genotypic characteristics.

    Science.gov (United States)

    Droual, R; Walker, R L; Shivaprasad, H L; Jeffrey, J S; Meteyer, C U; Chin, R P; Shapiro, D P

    1992-01-01

    The pathogenicity of a strain of Pasteurella gallinarum isolated in Fresno County, Calif., was compared with the American Type Culture Collection (ATCC) strain. Broiler chickens were inoculated intranasally with 10(7) colony-forming units (CFU) and intramuscularly with 10(5) CFU of each strain. The only notable lesions were in chickens inoculated intramuscularly with 10(5) CFU of the Fresno strain, which developed severe myositis at the inoculation site, pericarditis, perihepatitis, airsacculitis, and synovitis. P. gallinarum was reisolated from these lesions. Phenotypic characteristics of the two strains were identical except in reactions in ONPG broth and fermentation of xylose. Protein-banding patterns for the two strains were identical except for a single band difference in the 35-kilodalton region. Restriction endonuclease analysis confirmed that the Fresno strain was a distinct one. Plasmid analysis revealed that the ATCC strain had two plasmids and the Fresno strain had none.

  5. Elimination of Pasteurella pneumotropica from a mouse barrier facility by using a modified enrofloxacin treatment regimen.

    Science.gov (United States)

    Towne, Justin W; Wagner, April M; Griffin, Kurt J; Buntzman, Adam S; Frelinger, Jeffrey A; Besselsen, David G

    2014-09-01

    Multiple NOD. Cg-Prkdc(scid)Il2rg(tm1Wjl)Tg(HLA-A2.1)Enge/Sz (NSG/A2) transgenic mice maintained in a mouse barrier facility were submitted for necropsy to determine the cause of facial alopecia, tachypnea, dyspnea, and sudden death. Pneumonia and soft-tissue abscesses were observed, and Pasteurella pneumotropica biotype Jawetz was consistently isolated from the upper respiratory tract, lung, and abscesses. Epidemiologic investigation within the facility revealed presence of this pathogen in mice generated or rederived by the intramural Genetically Engineered Mouse Model (GEMM) Core but not in mice procured from several approved commercial vendors. Epidemiologic data suggested the infection originated from female or vasectomized male ND4 mice obtained from a commercial vendor and then comingled by the GEMM Core to induce pseudopregnancy in female mice for embryo implantation. Enrofloxacin delivered in drinking water (85 mg/kg body weight daily) for 14 d was sufficient to clear bacterial infection in normal, breeding, and immune-deficient mice without the need to change the antibiotic water source. This modified treatment regimen was administered to 2400 cages of mice to eradicate Pasteurella pneumotropica from the facility. Follow-up PCR testing for P. pneumotropica biotype Jawetz remained uniformly negative at 2, 6, 12, and 52 wk after treatment in multiple strains of mice that were originally infected. Together, these data indicate that enrofloxacin can eradicate P. pneumotropica from infected mice in a less labor-intensive approach that does not require breeding cessation and that is easily adaptable to the standard biweekly cage change schedule for individually ventilated cages.

  6. Outbreak of Pasteurella pneumotropica in a closed colony of STOCK-Cd28(tm1Mak) mice

    DEFF Research Database (Denmark)

    Artwohl, J.E.; Flynn, J.C.; Bunte, R.M.

    2000-01-01

    Fifteen mice with Pasteurella pneumotropica orbital abscesses were noted in mice that were homozygous for a targeted Cd28 gene mutation. Only one mouse heterozygous for the Cd28 mutation was affected. According to phenotypic reactions and 16S rDNA sequencing, the isolates were most similar to bio...... to biotype Heyl, This article provides evidence for an immunologic basis of susceptibility to P. pneumotropica infection....

  7. DYNAMICS OF GOBLET CELLS DURING INFECTION BY P. multocida AND B. bronchiseptica IN RABBITS

    Directory of Open Access Journals (Sweden)

    B Doncel

    2007-01-01

    Full Text Available Se evaluó el número y las características histoquímicas de las células caliciformes (CC en la cavidad nasal y la nasofaringe de conejos sanos y enfermos de septicemia y rinitis inducida por P. multocida y B. bronchiseptica. Se usó la técnica histoquímica de azul de Alciano y ácido Periódico de Schiff. Los resultados mostraron que la población de CC en ambas regiones anatómicas permaneció constante durante las diferentes etapas del desarro-llo de los conejos sanos, y estuvo entre 50-57 ±25 CC/mm de lámina basal. Se determinó el número de CC en el maxilocornete (63 ±24 CC/mm, en el septo nasal (53 ±18 CC/mm y la nasofaringe (45 ±22 CC/mm. En todas las etapas del desarrollo de los conejos, regiones anatómicas y animales enfermos hubo predominancia de glicoproteínas ácidas. Los conejos septicémicos mostraron un incremento altamente signiÀcativo (P<0,01 en el número de CC comparado con los animales sanos y aquellos que sufrían de rinitis. Este estudio demostró un incremento en el número de CC y secreción de glicoproteínas ácidas durante la enferme-dad respiratoria de los conejos.

  8. Molecular and virulence characteristics of an outer membrane-associated RTX exoprotein in Pasteurella pneumotropica

    Directory of Open Access Journals (Sweden)

    Kawamoto Eiichi

    2011-03-01

    Full Text Available Abstract Background Pasteurella pneumotropica is a ubiquitous bacterium that is frequently isolated from laboratory rodents and causes various clinical symptoms in immunodeficient animals. Currently two RTX toxins, PnxIA and PnxIIA, which are similar to hemolysin-like high-molecular-weight exoproteins are known in this species. In this study, we identified and analyzed a further RTX toxin named PnxIIIA and the corresponding type I secretion system. Results The RTX exoprotein, PnxIIIA, contains only a few copies of the RTX repeat-like sequence and 3 large repeat sequences that are partially similar to the outer membrane protein found in several prokaryotes. Recombinant PnxIIIA protein (rPnxIIIA was cytotoxic toward J774A.1 mouse macrophage cells, whereas cytotoxicity was attenuated by the addition of anti-CD11a monoclonal antibody. rPnxIIIA could bind to extracellular matrices (ECMs and cause hemagglutination of sheep erythrocytes. Binding was dependent on the 3 large repeat sequences in PnxIIIA. Protein interaction analyses indicated that PnxIIIA is mainly localized in the outer membrane of P. pneumotropica ATCC 35149 in a self-assembled oligomeric form. PnxIIIA is less cytotoxic to J774A.1 cells than PnxIA and PnxIIA. Conclusions The results implicate that PnxIIIA is located on the cell surface and participates in adhesion to ECMs and enhanced hemagglutination in the rodent pathogen P. pneumotropica.

  9. Pathology of an atypical strain of Pasteurella gallinarum infection in chickens.

    Science.gov (United States)

    Shivaprasad, H L; Droual, R

    2002-08-01

    Gross and microscopic pathology caused by an atypical strain of Pasteurella gallinarum (Fresno strain) was compared in chickens with that caused by the American Type Culture Collection type strain. Ten 21-day-old broiler chickens were inoculated intranasally with 10(7) colony forming units or intramuscularly with 10(5) colony forming units of either strain. The birds were killed 7 days later, and gross and microscopic lesions were studied. Grossly, there was extensive white discoloration of pectoral muscles with mild fibrinous exudate in birds inoculated intramuscularly with the Fresno strain of P. gallinarum. Most of these birds also had severe fibrinous exudation over the heart, the capsule of the liver, the air sac, and in the hock joints. Microscopically, there was severe chronic pyogranulomatous airsacculitis, pericarditis, perihepatitis, myositis, synovitis, and granulomatous pneumonia. One bird had severe acute multifocal hepatitis. From this study, it is evident that the Fresno strain of P. gallinarum was more pathogenic than the American Type Culture Collection type strain when given intramuscularly.

  10. Assessment of rpoB and 16S rRNA Genes as Targets for PCR-Based Identification of Pasteurella pneumotropica

    OpenAIRE

    Dole, Vandana S.; Banu, Laila A; Fister, Richard D; Nicklas, Werner; Henderson, Kenneth S.

    2010-01-01

    Diagnosis of Pasteurella pneumotropica in laboratory animals relies on isolation of the organism, biochemical characterization, and, more recently, DNA-based diagnostic methods. 16S rRNA and rpoB gene sequences were examined for development of a real-time PCR assay. Partial sequencing of rpoB (456 bp) and 16S rRNA (1368 bp) of Pasteurella pneumotropica isolates identified by microbiologic and biochemical assays indicated that either gene sequence can be used to distinguish P. pneumotropica fr...

  11. Investigations on the clonality of isolates of Pasteurella gallinarum obtained from turkeys in Germany.

    Science.gov (United States)

    Bisgaard, Magne; Christensen, Henrik; Behr, Klaus-Peter; Baron, Gerhard; Christensen, Jens P

    2005-04-01

    Pasteurella gallinarum has been considered an opportunistic pathogen rather than a primary pathogen for chickens. As P. gallinarum has been found to have a high genotypic diversity, one would expect a polyclonal distribution among isolates from different farms if this organism is a secondary invader. The aims of this investigation were to genetically characterize isolates obtained from outbreaks affecting several turkey farms to confirm the existence of the infection in turkeys and to investigate the genetic relationship between isolates from affected farms. A total of 17 isolates from 14 outbreaks of respiratory disease in Germany were subjected to extended phenotypic and genotypic characterization. All isolates were of the same phenotype, typical of P. gallinarum. Ribotyping of three isolates using either HpaII or HindIII showed that they had identical profiles and indicated that the isolates all originated from the same clone. Comparison with HpaII ribotypes from a previous study showed that the pattern was identical to that obtained with isolates from a Zimbabwean outbreak in chickens during 1999 to 2000. Restriction endonuclease analysis typing of 14 isolates from all 14 farms showed that they had identical profiles but these differed from those obtained with isolates from the Zimbabwean outbreak. Sequencing of the 16S rRNA gene and sequence comparisons with other Pasteurellaceae confirmed their classification as P. gallinarum. Identification of the same clone of P. gallinarum from 14 outbreaks of acute respiratory disease in turkeys within a time period of 2 months suggests a common source of infection, and that P. gallinarum probably played a primary role rather than a secondary role in the outbreaks.

  12. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin.

    Directory of Open Access Journals (Sweden)

    Martin Sager

    Full Text Available [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

  13. Characterization of Biofilm Formation in [Pasteurella] pneumotropica and [Actinobacillus] muris Isolates of Mouse Origin.

    Science.gov (United States)

    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. First case of human infection caused by Pasteurella gallinarum causing infective endocarditis in an adolescent 10 years after surgical correction for truncus arteriosus.

    Science.gov (United States)

    al Fadel Saleh, M; al-Madan, M S; Erwa, H H; Defonseka, I; Sohel, S Z; Sanyal, S K

    1995-06-01

    To report the first case of human infection (infective endocarditis [IE]) caused by Pasteurella gallinarum and to review the literature regarding IE caused by the genus Pasteurella. University hospital based. An adolescent boy who underwent successful correction for truncus arteriosus 10 years before the present illness. Persistent fever, pallor, and a palpable spleen suggested IE clinically. Echocardiography documented vegetation in the conduit that was used for surgical correction. Blood cultures grew P. gallinarum and confirmed its role as the causative organism for IE in the patient. This case illustrates that IE may develop in a child with congenital heart disease several years after surgical intervention using material that is foreign to the body (conduit), and that such a complication may involve unusual pathogens. These observations emphasize the need for careful long-term follow-up of children with congenital heart disease even after successful surgical correction.

  15. Isolation and characterization of a bacteriocin produced by an ...

    African Journals Online (AJOL)

    involved in domestic animal diseases, including Escherichia coli, Salmonella pullorum, Pseudomonas aeruginosa, Pasteurella multocida, Clostridium perfringens, Micrococcus luteus, Streptococcus bovis and Staphylococcus aureus. Two multidrug-resistant clinical isolates and a phytopathogenic yeast strain were also ...

  16. Disease: H00306 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available 766 PMID:6371440 (drug) Weber DJ, Wolfson JS, Swartz MN, Hooper DC Pasteurella multocida infections. Report ...not susceptible to dicloxacillin, cephalexin, clindamycin, or erythromycin. ICD-10: A28.0 MeSH: D010326 D012

  17. 21 CFR 522.1660b - Oxytetracycline solution, 300 milligrams/milliliter.

    Science.gov (United States)

    2010-04-01

    ... Escherichia coli, wooden tongue caused by Actinobacillus lignieresii, leptospirosis caused by Leptospira... by Pasteurella multocida, and leptospirosis caused by Leptospira pomona. (C) 9 mg/lb BW as a single...

  18. Inhibition of protein synthesis on the ribosome by tildipirosin compared with other veterinary macrolides

    DEFF Research Database (Denmark)

    Andersen, Niels Møller; Poehlsgaard, Jacob; Warrass, Ralf

    2012-01-01

    Tildipirosin is a 16-membered-ring macrolide developed to treat bacterial pathogens, including Mannheimia haemolytica and Pasteurella multocida, that cause respiratory tract infections in cattle and swine. Here we evaluated the efficacy of tildipirosin at inhibiting protein synthesis...

  19. Disease: H00306 [KEGG MEDICUS

    Lifescience Database Archive (English)

    Full Text Available found in the animal's oral cavity. Disseminated Pasteurella infections can lead to serious diseases including septic shock and menin...gitis mostly in infants and pregnant women. Many patients with P. multocida-related

  20. Demonstration of Ornithobacterium rhinotracheale in pheasants (Phasianus colchicus) with pneumonia and airsacculitis

    DEFF Research Database (Denmark)

    Welchman, D. de B.; Ainsworth, H. L.; Jensen, Tim Kåre

    2013-01-01

    type 2, avian coronavirus, Mycoplasma gallisepticum, Mycoplasma synoviae and other Mycoplasma species, Escherichia coli, Pasteurella multocida, other Pasteurellaceae and Syngamus trachea, suggesting synergism with other agents. Exposure to other intercurrent factors, including adverse weather...

  1. Pathomorphology and aerobic bacteria associated with pneumonia ...

    African Journals Online (AJOL)

    Aerobic bacteria isolated from the pneumonic lungs were Escherichia coli, Klebsiella pneumoniae, Mannheimia haemolytica, Streptococcus pyogenes, Staphylococcus aureus and Pasteurella multocida respectively. There was no significant seasonal, species and breed associations (p>0.05) between pneumonic lesions ...

  2. Comparison of the diagnostic performance of bacterial culture of nasopharyngeal swab and bronchoalveolar lavage fluid samples obtained from calves with bovine respiratory disease

    Science.gov (United States)

    Objective: Examine the culture results, gamithromycin susceptibility, predictive values, and agreement of pooled bilateral nasopharyngeal swabs (NPS) and bronchoalveolar lavages (BAL) for identification of Mannheimia haemolytica genotypes, Pasteurella multocida, and Histophilus somni in calves treat...

  3. Observations on macrolide resistance and susceptibility testing performance in field isolates collected from clinical bovine respiratory disease cases

    Science.gov (United States)

    The objectives of this study were; first, to describe gamithromycin susceptibility of Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni isolated from cattle diagnosed with bovine respiratory disease (BRD) and previously treated with either gamithromycin for control of BRD (mass me...

  4. Assessment of rpoB and 16S rRNA genes as targets for PCR-based identification of Pasteurella pneumotropica.

    Science.gov (United States)

    Dole, Vandana S; Banu, Laila A; Fister, Richard D; Nicklas, Werner; Henderson, Kenneths S

    2010-12-01

    Diagnosis of Pasteurella pneumotropica in laboratory animals relies on isolation of the organism, biochemical characterization, and, more recently, DNA-based diagnostic methods. 16S rRNA and rpoB gene sequences were examined for development of a real-time PCR assay. Partial sequencing of rpoB (456 bp) and 16S rRNA (1368 bp) of Pasteurella pneumotropica isolates identified by microbiologic and biochemical assays indicated that either gene sequence can be used to distinguish P. pneumotropica from other members of the Pasteurellaceae family. However, alignment of rpoB sequences from the Pasteurella pneumotropica Heyl (15 sequences) and Jawetz (16 sequences) biotypes with other Pasteurellaceae sequences from GenBank indicated that although rpoB DNA sequencing could be used for diagnosis, development of diagnostic primers and probes would be difficult, because the sequence variability between Heyl and Jawetz biotypes is not clustered in any particular region of the rpoB sequence. In contrast, alignment of 16S rRNA sequences revealed a region with unique and stable nucleotide motifs sufficient to permit development of a specific fluorogenic real-time PCR assay to confirm P. pneumotropica isolated by culture and to differentiate Heyl and Jawetz biotypes.

  5. Studies on hemorrhagic pneumonia in Moschus sifanicus | Lu ...

    African Journals Online (AJOL)

    The disease is caused by single infection of Pasteurella multocida or mix of P. multocida, Escherichia coli and Pseudomonas aeruginosa, and is a contagious disease. The pathological changes were mainly manifested in the vessel wall of bronchia and bronchiole appeared congested, bleeding, edemic with infiltration of ...

  6. Mechanistic study of CMP-Neu5Ac hydrolysis by α2,3-sialyltransferase from Pasteurella dagmatis.

    Science.gov (United States)

    Schmölzer, Katharina; Luley-Goedl, Christiane; Czabany, Tibor; Ribitsch, Doris; Schwab, Helmut; Weber, Hansjörg; Nidetzky, Bernd

    2014-08-25

    Bacterial sialyltransferases of the glycosyltransferase family GT-80 exhibit pronounced hydrolase activity toward CMP-activated sialyl donor substrates. Using in situ proton NMR, we show that hydrolysis of CMP-Neu5Ac by Pasteurella dagmatis α2,3-sialyltransferase (PdST) occurs with axial-to-equatorial inversion of the configuration at the anomeric center to release the α-Neu5Ac product. We propose a catalytic reaction through a single displacement-like mechanism where water replaces the sugar substrate as a sialyl group acceptor. PdST variants having His(284) in the active site replaced by Asn, Asp or Tyr showed up to 10(4)-fold reduced activity, but catalyzed CMP-Neu5Ac hydrolysis with analogous inverting stereochemistry. The proposed catalytic role of His(284) in the PdST hydrolase mechanism is to facilitate the departure of the CMP leaving group. Copyright © 2014 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  7. Identification of a virulence determinant that is conserved in the Jawetz and Heyl biotypes of [Pasteurella] pneumotropica.

    Science.gov (United States)

    Sasaki, Hiraku; Ishikawa, Hiroki; Terayama, Hayato; Asano, Ryoki; Kawamoto, Eiichi; Ishibashi, Hidetoshi; Boot, Ron

    2016-08-01

    [Pasteurella] pneumotropica is a ubiquitous bacterium frequently isolated from laboratory rodents. Although this bacterium causes various diseases in immunosuppressed animals, little is known about major virulence factors and their roles in pathogenicity. To identify virulence factors, we sequenced the genome of [P.] pneumotropica biotype Heyl strain ATCC 12555, and compared the resulting non-contiguous draft genome sequence with the genome of biotype Jawetz strain ATCC 35149. Among a large number of genes encoding virulence-associated factors in both strains, four genes encoding for YadA-like proteins, which are known virulence factors that function in host cell adherence and invasion in many pathogens. In this study, we assessed YadA distribution and biological activity as an example of one of virulence-associated factor shared, with biotype Jawetz and Heyl. More than half of mouse isolates were found to have at least one of these genes; whereas, the majority of rat isolates did not. Autoagglutination activity, and ability to bind to mouse collagen type IV and mouse fibroblast cells, was significantly higher in YadA-positive than YadA-negative strains. To conclude, we identified a large number of candidate genes predicted to influence [P.] pneumotropica pathogenesis. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Pasteurella pneumotropica evades the human complement system by acquisition of the complement regulators factor H and C4BP.

    Directory of Open Access Journals (Sweden)

    Alfredo Sahagún-Ruiz

    Full Text Available Pasteurella pneumotropica is an opportunist Gram negative bacterium responsible for rodent pasteurellosis that affects upper respiratory, reproductive and digestive tracts of mammals. In animal care facilities the presence of P. pneumotropica causes severe to lethal infection in immunodeficient mice, being also a potential source for human contamination. Indeed, occupational exposure is one of the main causes of human infection by P. pneumotropica. The clinical presentation of the disease includes subcutaneous abscesses, respiratory tract colonization and systemic infections. Given the ability of P. pneumotropica to fully disseminate in the organism, it is quite relevant to study the role of the complement system to control the infection as well as the possible evasion mechanisms involved in bacterial survival. Here, we show for the first time that P. pneumotropica is able to survive the bactericidal activity of the human complement system. We observed that host regulatory complement C4BP and Factor H bind to the surface of P. pneumotropica, controlling the activation pathways regulating the formation and maintenance of C3-convertases. These results show that P. pneumotropica has evolved mechanisms to evade the human complement system that may increase the efficiency by which this pathogen is able to gain access to and colonize inner tissues where it may cause severe infections.

  9. 21 CFR 520.88b - Amoxicillin trihydrate for oral suspension.

    Science.gov (United States)

    2010-04-01

    ... respiratory tract due to Staphylococcus spp., Streptococcus spp., Hemophilus spp., E. coli, Pasteurella spp.... mirabilis, and Corynebacterium spp.; gastrointestinal tract due to E. coli, Proteus spp., Staphylococcus spp... Staphylococcus spp., Streptococcus spp., E. coli, and Pasteurella multocida. (C) Limitations. Use for 5 to 7 days...

  10. 21 CFR 522.88 - Sterile amoxicillin trihydrate for suspension.

    Science.gov (United States)

    2010-04-01

    ..., Staphylococcus spp., Streptococcus spp., Hemophilus spp., E. coli, Pasteurella spp., and P. mirabilis... Corynebacterium spp.; gastrointestinal infections due to E. coli, Proteus spp., Staphylococcus spp., and..., Staphylococcus spp., Streptococcus spp., E. coli, and Pasteurella multocida. (3) Limitations. For use in dogs and...

  11. Lack of Virus-Specific Bacterial Adherence to Bovine Embryonic Lung Cells Infected with Bovine Parainfluenza Virus Type 3 †

    OpenAIRE

    Toth, Thomas E.; Gates, Connie

    1983-01-01

    Infection of bovine embryonic lung cells with bovine parainfluenza virus type 3 did not induce in vitro, virus-specific, hemadsorption-related adherence of Corynebacterium pyogenes, Haemophilus somnus, Staphylococcus aureus, Streptococcus zooepidemicus, Pasteurella haemolytica, Listeria monocytogenes, Escherichia coli, Pasteurella multocida, Brucella sp., or Salmonella typhimurium.

  12. DEVELOPMENT OF A PEROXIDASE-ANTIPEROXIDASE (PAP) TECHNIQUE FOR THE IDENTIFICATION OF HAEMOPHILUS-SOMNUS IN PNEUMONIC CALF LUNGS IN DENMARK

    DEFF Research Database (Denmark)

    Tegtmeier, Conny; Jensen, N.E.; Jensen, H.E.

    1995-01-01

    for the determination of the sensitivity and specificity of antiserum that had been heterologously absorbed with antigens of cross-reacting bacteria, i.e. Pasteurella haemolytica and Pasteurella multocida. None of the antisera reacted with Actinomyces pyogenes. An antiserum raised against somatic antigens of the Danish...

  13. An ecologic study comparing distribution of Pasteurella trehalosi and Mannheimia haemolytica between Sierra Nevada bighorn sheep, White Mountain bighorn sheep, and domestic sheep.

    Science.gov (United States)

    Tomassini, Letizia; Gonzales, Ben; Weiser, Glen C; Sischo, William

    2009-10-01

    The prevalence and phenotypic variability of Pasteurella and Mannheimia isolates from Sierra Nevada bighorn sheep (Ovis canadensis sierrae), White Mountain bighorn sheep (Ovis canadensis nelsoni), and domestic sheep (Ovis aries) from California, USA, were compared. The White Mountain bighorn sheep population had a recent history of pneumonia-associated mortality, whereas the Sierra Nevada bighorn sheep population had no recent history of pneumonia-associated mortality. The domestic sheep flocks were pastured in areas geographically near both populations but were not known to have direct contact with either bighorn sheep population. Oropharyngeal swab samples were collected from healthy domestic and bighorn sheep and cultured to characterize bacterial species, hemolysis, biogroups, and biovariants. Pasteurella trehalosi and Mannheimia haemolytica were detected in all of the study populations, but the relative proportion of each bacterial species differed among sheep populations. Pasteurella trehalosi was more common than M. haemolytica in the bighorn sheep populations, whereas the opposite was true in domestic sheep. Mannheimia haemolytica was separated into 11 biogroups, and P. trehalosi was characterized into two biogroups. Biogroup distributions for M. haemolytica and P. trehalosi differed among the three populations; however, no difference was detected for the distribution of P. trehalosi biogroups between the Sierra Nevada bighorn sheep and domestic sheep. The prevalence odds ratios (pOR) for the distribution of M. haemolytica biogroups suggested little difference between White Mountain bighorn sheep and domestic sheep compared with Sierra Nevada bighorn sheep and domestic sheep, although these comparisons had relatively large confidence intervals for the point estimates. Hemolytic activity of the isolates was not different among the sheep populations for M. haemolytica but was different for P. trehalosi. No clear evidence of association was found in the

  14. Vibrio sp. strain NM 10, isolated from the intestine of a Japanese coastal fish, has an inhibitory effect against Pasteurella piscicida.

    Science.gov (United States)

    Sugita, H; Matsuo, N; Hirose, Y; Iwato, M; Deguchi, Y

    1997-01-01

    Vibrio sp. strain NM 10 with an inhibitory activity against Pasteurella piscicida K-III was isolated from the intestine of a spotnape ponyfish (Leiognathus nuchalis). This bacterium efficiently produced an antibacterial substance after growth at 20 degrees C for 24 h on 1/5 PYBG agar prepared with 50% seawater at pHs of 7.5 to 9.0. The antibacterial substance was heat labile and proteinaceous, with a molecular mass of less than 5 kDa, possibly a bacteriocin or a bacteriocin-like substance. PMID:9406423

  15. Artritis de codo por Pasteurella sp. Reporte de un caso y revisión de la bibliografía.

    OpenAIRE

    Dany Olivera; Paola Filomeno

    2014-01-01

    La mayoría de las infecciones por Pasteurella sp. se producen por la inoculación percutánea después de la mordedura de un gato o perro. El espectro de enfermedades producidas oscila entre abscesos localizados, celulitis, artritis séptica, osteomielitis, septicemia, infecciones respiratorias y del sistema nervioso central. Una inmunosupresión subyacente puede predisponer a manifestaciones sistémicas. Presentamos un caso de artritis séptica de codo tras la mordida y arañazo de gato en un pac...

  16. Molecular diagnosis of Haemorrhagic Septicaemia - A Review

    Directory of Open Access Journals (Sweden)

    Ranjan Rajeev

    2011-08-01

    Full Text Available Pasteurella multocida is associated with hemorrhagic septicaemia in cattle and buffaloes, pneumonic pasteurellosis in sheep and goats, fowl cholera in poultry, atrophic rhinitis in pigs and snuffles in rabbits. Haemorrhagic septicaemia is caused by Pasteurella multocida type B:2, B:2,5 and B:5 in Asian countries and type E:2 in African countries. Pasteurella multocida have five types of capsular serotype i.e. type A, B, D, E and F. Diagnosis of the disease is mainly based on the clinical sign and symptom, post mortem findings. Confirmatory diagnosis is done by isolation and identification of causative agent. A variety of laboratory diagnostic techniques have been developed over the years for pasteurellosis and used routinely in the laboratory. Among these techniques molecular techniques of diagnosis is most important. This technique not only gives diagnosis but it also provides information regarding capsular type of Pasteurella multocida. Techniques which are used for molecular diagnosis of haemorrhagic septicaemia are PCR based diagnosis, Restriction endonuclease analysis (REA, Ribotyping, Colony hybridization assay, Filled alternation gel electrophoresis (FAGE, Detection of Pasteurella multocida by Real Time PCR. Among these techniques real time PCR is most sensitive and specific. [Vet. World 2011; 4(4.000: 189-192

  17. A specific polymerase chain reaction based on the gyrB gene sequence and subsequent restriction fragment length polymorphism analysis of Pasteurella pneumotropica isolates from laboratory mice.

    Science.gov (United States)

    Hayashimoto, Nobuhito; Ueno, Masami; Takakura, Akira; Itoh, Toshio

    2007-03-01

    For a molecular identification and typing tool, we developed a specific polymerase chain reaction (PCR) based on the gyrB gene sequence and a subsequent restriction fragment length polymorphism (RFLP) analysis using the products amplified from the specific PCR to facilitate discrimination of biotypes of Pasteurella pneumotropica from laboratory mice. Appropriate PCR products, a 1039-basepair fragment for biotype Jawetz and a 1033-basepair fragment for biotype Heyl, were amplified by use of the primers CZO-1 and NJO-2 from all 105 P. pneumotropica isolates tested and the 2 reference strains but not from other bacterial species tested. MspI digestion of PCR-generated products showed 3 RFLP patterns among the 105 isolates, and these patterns correlated with the biotype of the isolate (RFLP pattern 1, biotype Jawetz; RFLP pattern 2, biotype Heyl; and RFLP pattern 3, biotype Jawetz with Beta-hemolytic activity). Our procedure identifies and biotypes isolates of P. pneumotropica from laboratory mice, using simple PCR and enzymatic restriction techniques. Therefore, this procedure is useful as a rapid identification and biotyping tool for isolates of P. pneumotropica from laboratory mice.

  18. Identification by 16S rDNA fragment amplification and determination of genetic diversity by random amplified polymorphic DNA analysis of Pasteurella pneumotropica isolated from laboratory rodents.

    Science.gov (United States)

    Kodjo, A; Villard, L; Veillet, F; Escande, F; Borges, E; Maurin, F; Bonnod, J; Richard, Y

    1999-02-01

    Pasteurella pneumotropica is an opportunistic bacterium frequently isolated from colonies of various laboratory rodents. Identification of this species, including its differentiation into two distinct biotypes (Jawetz and Heyl), is usually based on the use of conventional bacteriologic methods. In this study, a 16S rDNA fragment amplification procedure was developed for use as an alternative method for identification and differentiation of P. pneumotropica. Polymerase chain reaction (PCR) products were two distinctive fragments of 937 and 564 bp specific for biotypes Jawetz and Heyl, respectively. Specificity of PCR products could be achieved by EcoRI cleavage, leading to 596 plus 341-bp and 346 plus 218-bp fragments for each of the amplification products. Use of this procedure confirmed identification of 34 field isolates and allowed definitive identification of some strains that could not have been done by use of bacteriologic examinations. Field isolates subjected to random amplified polymorphic DNA (RAPD) analysis had high genetic diversity among biotype Jawetz strains in contrast to biotype Heyl strains. In conclusion, RAPD could represent an additional means for identification of ambiguous strains of biotype Heyl and a valuable epidemiologic tool for identification of biotype Jawetz strains of P. pneumotropica.

  19. Active-Site His85 of Pasteurella dagmatis Sialyltransferase Facilitates Productive Sialyl Transfer and So Prevents Futile Hydrolysis of CMP-Neu5Ac.

    Science.gov (United States)

    Schmölzer, Katharina; Eibinger, Manuel; Nidetzky, Bernd

    2017-08-04

    Sialyltransferases of the GT-80 glycosyltransferase family are considered multifunctional because of the array of activities detected. They exhibit glycosyl transfer, trans-sialylation, and hydrolysis activities. How these enzymes utilize their active-site residues in balancing the different enzymatic activities is not well understood. In this study of Pasteurella dagmatis α2,3sialyltransferase, we show that the conserved His85 controls efficiency and selectivity of the sialyl transfer. A His85→Asn variant was 200 times less efficient than wild-type for sialylation of lactose, and exhibited relaxed site selectivity to form not only the α2,3- but also the α2,6-sialylated product (21 %). The H85N variant was virtually inactive in trans-sialylation but showed almost the same CMP-Neu5Ac hydrolase activity as wild-type. The competition between sialyl transfer and hydrolysis in the conversion of CMP-Neu5Ac was dependent on the lactose concentration; this was characterized by a kinetic partition ratio of 85 m-1 for the H85N variant, compared to 17 000 m-1 for the wild-type enzyme. His85 promotes the productive sialyl transfer to lactose and so prevents hydrolysis of CMP-Neu5Ac in the reaction. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. FOWL CHOLERA IN A BREEDER FLOCK

    Directory of Open Access Journals (Sweden)

    Z. Parveen, A. A. Nasir, K.Tasneem and A. Shah

    2003-12-01

    Full Text Available During January, 2003 Pasteurella multocida the causative agent of fowl cholera was isolated from a breeder flock in Lahore District. The age of the flock was 245 days. Increased mortality, swollen wattles and lameness were the clinical findings present in almost all the affected birds, while gross lesions were typical of fowl cholera. To prove the virulence of the organism, mice and six-week old cockerals were infected and P. multocida was reisolated.