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

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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.

Restriction endonuclease analysis of Pasteurella multocida isolates from three California turkey premises.

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Christiansen, K H; Carpenter, T E; Snipes, K P; Hird, D W; Ghazikhanian, G Y

1992-01-01

Three California turkey premises that had repeated outbreaks of fowl cholera were studied for periods of 2 to 4 years. Using biochemical, serologic, plasmid DNA, and restriction endonuclease analyses of isolates of Pasteurella multocida from turkeys and wildlife on the premises, strains of the organism were found to be enzootic on two of the premises. On the third, a variety of strains of P. multocida were isolated from fowl cholera outbreak flocks.

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

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

2004-01-01

Strains deviating in key phenotypic characters, mainly isolated from cases of bovine pneumonia in five European countries, were genotyped in order to examine their genotypic relationship with Pasteurella multocida. Twenty-two strains of Pasteurella avium biovar 2, including variants in indole, xy...

Transmission of Pasteurella multocida on California turkey premises in 1988-89.

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Christiansen, K H; Carpenter, T E; Snipes, K P; Hird, D W

1992-01-01

Restriction endonuclease analysis (REA) of whole-cell DNA was used to determine possible sources of Pasteurella multocida for each outbreak of fowl cholera occurring in turkey flocks in eight commercial poultry companies in California from October 1988 to September 1989. Over this period, 179 isolates of P. multocida were obtained from dead turkeys in 80 meat and breeder flocks on 43 premises. P. multocida was isolated from wildlife on five premises. Isolates were characterized by subspecies, serotype, presence of plasmid DNA, and REA type. In 52 (65%) flocks, all isolates of P. multocida had the same REA pattern as the M9 live vaccine strain following digestion of DNA with the restriction enzyme SmaI. Field strains of P. multocida were obtained from 27 (34%) flocks, and one flock (1%) yielded both M9 and a field strain of the organism. REA of field strains of P. multocida revealed 17 different SmaI REA types. Based on matching SmaI REA types, potential sources of P. multocida were identified for 15 of the 28 flocks infected with field strains of the organism, and transmission between turkey premises was a possibility in only seven flocks.

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

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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 ...

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

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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.

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

International Nuclear Information System (INIS)

Hoefer, M.; Flossmann, K.D.

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 14 C- and 3 H-labelled substrates, the degradation of these substances proceeds analogously via the citric acid cycle. (author)

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.

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

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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.

Pneumonia in calves produced with aerosols of Pasteurella multocida alone and in combination with bovine herpesvirus 1.

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Jericho, K W; Carter, G R

1985-01-01

Pathological changes in respiratory tracts were studied in 30 calves following exposure to aerosols of Pasteurella multocida or to bovine herpesvirus 1 and P. multocida. Two groups of five calves were exposed to aerosols of one of two types of P. multocida only, which produced lobar pneumonia in one calf of each group. Another five groups of four calves were exposed to aerosols of bovine herpesvirus 1 and four to seven days later to one of the two types or one sub-type of P. multocida. Extens...

Pasteurella multocida Involved in Respiratory Disease of Wild Chimpanzees

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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

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

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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...

Antimicrobial susceptibility of Pasteurella multocida and Haemophilus parasuis isolates associated with porcine pneumonia

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Kateřina Nedbalcová

2013-01-01

Full Text Available Pasteurella multocida and Haemophilus parasuis pig isolates obtained in the Czech Republic were tested for their susceptibility against selected antimicrobial agents by broth microdilution method between 2008 and 2011. A low degree of resistance was observed for ampicillin, amoxicillin/clavulanic acid, ceftiofur, tulathromycin, tilmicosin, florfenicol and enrofloxacin in 20 (6.0%, 15 (4.5 %, 2 (0.6%, 8 (2.4%, 13 (3.9%, 5 (1.5% and 5 (1.5% P. multocida isolates as well as for tiamulin, gentamicin, tulathromycin, tilmicosin and ampicillin in 2 (2.4%, 2 (2.4%, 3 (3.6%, 3 (3.6% and 6 (7.2% H. parasuis isolates. In addition, moderate level of resistance to tiamulin was found in 60 (18.1% P. multocida isolates and high level of resistance for tetracycline was detected in 107 (32.2 % P. multocida isolates and in 23 (27.7 % H. parasuis isolates. Differences between resistance rates of P. multocida and H. parasuis were significant (P ≤ 0.5 only for tiamulin. These data confirmed that antimicrobial resistance is not very widespread among current porcine P. multocida and H. parasuis isolates in the Czech Republic.

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

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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...... showed the monophyly of the strains characterized and the reference strains of P. multocida. The sucrose-negative strains formed two groups, one related to reference strains of P. multocida and the other related to a separate species-like group (taxon 45 of Bisgaard). DNA-DNA hybridization further...... 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-negative...

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

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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...

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

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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.

The Capsule Is a Virulence Determinant in the Pathogenesis of Pasteurella multocida M1404 (B:2)

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Boyce, John D.; Adler, Ben

2000-01-01

Capsules from a range of pathogenic bacteria are key virulence determinants, and the capsule has been implicated in virulence in Pasteurella multocida. We have previously identified and determined the nucleotide sequence of the P. multocida M1404 (B:2) capsule biosynthetic locus (J. D. Boyce, J. Y. Chung, and B. Adler, Vet. Microbiol. 72:121–134, 2000). The cap locus consists of 15 genes, which can be grouped into three functional regions. Regions 1 and 3 contain genes proposed to encode prot...

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

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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.

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...... with inoculations with M. hyopneumoniae and toxigenic P. multocida had no significant effect on disease development, but may have enhanced colonization by toxigenic P. multocida on the nasal turbinates....

Anatomopathological pneumonic aspects associated with highly pathogenic Pasteurella multocida in finishing pigs

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Eliana S. Paladino

Full Text Available ABSTRACT: The bacterium Pasteurella multocida is a frequent cause of porcine respiratory disease complex in finishing pigs. Historically, the bacterium is recognized as an opportunistic agent, causing secondary bacterial pneumonia in pigs. Several Brazilian reports have suggested the ability of P. multocida to cause primary pulmonary infection that leads to the death of finishing pigs prior to slaughter. The aim of this study was to evaluate anatomopathological pulmonary findings associated with P. multocida infection that were obtained from animals with clinical respiratory disease and from animals at slaughter. Twenty-five lung samples from 14 herds of finishing pigs with acute clinical respiratory disease and 19 lungs collected at slaughter from a different set of 14 herds were studied. In all lung samples, bacterial isolation was performed, and only samples with pure P. multocida growth were included in the study. Gross and histopathological lesions were evaluated, as well as Influenza A, porcine circovirus type 2 (PCV2 and Mycoplasma hyopneumoniae co-infections. Pleuritis and pericarditis were more often observed in clinical samples (P<0.05. Moreover, there was a numerical trend indicating that pericarditis, lymphadenomegaly and cavity exudates were more often present in clinical samples. Thirteen lung samples were negative to M. hyopneumoniae, Influenza A and PCV2 by immunohistochemistry (IHC, with only P. multocida identified. In these cases, gross lesions such as pleuritis, pericarditis and lymphadenomegaly were always present, and no histologic lesions indicative of other agents such as Actinobacillus pleuropneumoniae, Actinobacillus suis or Haemophilus parasuis were observed. These findings suggest the ability of some P. multocida isolates to cause primary respiratory and systemic infection. However, in this study, it was not possible to determine specific virulence markers to explain these findings.

Capsular Polysaccharide Interferes with Biofilm Formation by Pasteurella multocida Serogroup A

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Briana Petruzzi

2017-11-01

Full Text Available 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. multocida.

Serotypes and DNA fingerprint profiles of Pasteurella multocida isolated from raptors

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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.

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...

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

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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.

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

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...

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

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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 (P<0.05, two-tailed Mann-Whitney U test). The results suggest that there may be at least two groups of P. multocida in sheep - a genetically homogeneous group that resides in the respiratory tract and a genetically heterogeneous group that is the predominant cause of disease.

Comparison of standardised versus non-standardised methods for testing the in vitro potency of oxytetracycline against mannheimia haemolytica and pasteurella multocida

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

2016-01-01

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...

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...... without any dimethyltransferase activity. Erm(42) is a novel addition to the Erm family: it is phylogenetically distant from the other Erm family members and it is unique in being a bona fide monomethyltransferase that is disseminated between bacterial pathogens....

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

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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 LD 50 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.

Isolation and Identification of Pasteurella multocida from Sheep & Goat in Iran

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Valadan, M.,

2014-05-01

Full Text Available This study has been carried out with the objective of isolation and identification of agent(s of pasteurella pneumonia in sheep and goat in Iran using bacteriological and biochemical assays to be identified in the pursuant researches to be used in pasteurellosis vaccine production. To accomplish this objective, samples were gathered from areas suspicious to pasteurellosis infection and industrial abbatoirs according to clinical and autopsy symptoms from eight provinces of Bushehr, Esfahan, Kerman, Kohgilooyeh & Boyr Ahmad, Fars, Qom, Tehran and Qazvin in a period from spring 2008 to spring 2011. Samples were different in sort due to the existent condition but generally were comprised of palatine tonsil swabs or blood samples taken from jugular vein in live animals and lungs or upper respiratory tract lymph glands in dead or slaughtered animals. Totally, 1454 samples (1120 samples of sheep, 334 samples of goat composed if 1084 samples of live animals and 370 samples of dead or slaughterd animals were tested. Considering results obtained from assays, only 54 samples (3.71% were assessed as being pasteurella, genus of which was totally identified as multocida.

Why your housecat's trite little bite could cause you quite a fright: a study of domestic felines on the occurrence and antibiotic susceptibility of Pasteurella multocida.

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Freshwater, A

2008-10-01

Approximately four to five million animal bite wounds are reported in the USA each year. Domestic companion animals inflict the majority of these wounds. Although canine bites far outnumber feline bites, unlike the dog, the cat's bite is worse than its bark; 20-80% of all cat bites will become infected, compared with only 3-18% of dog bite wounds. Pasteurella multocida is the most commonly cultured bacterium from infected cat bite wounds. Anyone seeking medical attention for a cat-inflicted bite wound is given prophylactic/empiric penicillin or a derivative to prevent Pasteurella infection (provided they are not allergic to penicillins). In an effort to establish a carriage rate of P. multocida in the domestic feline, bacterial samples from the gingival margins of domestic northern Ohio cats (n=409) were cultured. Isolates were tested for antibiotic sensitivity as prophylactic/empiric use of penicillin and its derivatives could potentially give rise to antibiotic resistance in P. multocida. The high carriage rate (approximately 90%) of P. multocida observed was found to be independent of physiological and behavioural variables including age, breed, food type, gingival scale, lifestyle and sex. High antibiotic susceptibility percentages were observed for benzylpenicillin, amoxicillin-clavulanate, cefazolin, and azithromycin (100%, 100%, 98.37% and 94.02%, respectively) in P. multocida isolates. The high prevalence of P. multocida in the feline oral cavity indicates that prophylactic/empiric antibiotic therapy is still an appropriate response to cat bite wounds. Additionally, the susceptibility of P. multocida to penicillin and its derivatives indicates that they remain reliable choices for preventing and treating P. multocida infections.

Multiorgan Failure and Refractory Lactic Acidosis due to Pasteurella multocida Septicemia in a Patient with No Animal Exposure

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Damaris Pena

2018-01-01

Full Text Available Introduction. Pasteurella multocida is a gram-negative coccobacillus pathogenic to animals. It can cause infection in humans by a bite, scratch, or lick from a cat or dog. P. multocida can cause a variety of infections in humans, including cellulitis, osteomyelitis, endocarditis, peritonitis, and septic shock. Case Presentation. A 56-year-old male presented to our hospital with a 2-day history of fever, abdominal pain, nausea, and vomiting. He denied exposure to cats, dogs or other pets. He had severe respiratory distress requiring ventilator support, profound septic shock requiring multiple vasopressors, severe lactic acidosis, and renal failure requiring emergent hemodialysis. Blood cultures confirmed the presence of P. multocida. The patient subsequently died of cardiopulmonary arrest due to multiorgan failure with refractory shock. Conclusion. P. multocida septicemia can lead to septic shock. Early identification of this organism may decrease mortality. Although our patient had no known cat or dog exposure, physicians should enquire about a history of animal exposure when a patient presents with an infection with no obvious cause.

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.

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

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...

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.

Structure and function of C-terminal catalytic region of pasteurella multocida toxin

International Nuclear Information System (INIS)

Kitadokoro, Kengo; Kamitami, Shigeki; Horiguchi, Yasuhiko

2008-01-01

Pasteurella multocida toxin (PMT) is one of virulence factors responsible for the pathogenesis in some Pasteurellosis. We determined the crystal structure of the C-terminal region of PMT (C-PMT), which carries an intracellularly active moiety. The overall structure of C-PMT displays three different domains designated C1, C2 and C3. We found in the C3 domain 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 the Cys-His-Asp triad. Our results demonstrate that PMT is an enzymatic toxin carrying the cysteine-protease like catalytic triad, which is organized only under reducing conditions. (author)

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

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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.

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

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

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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.

Pharmacokinetic/pharmacodynamic integration and modelling of oxytetracycline for the porcine pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida.

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Dorey, L; Pelligand, L; Cheng, Z; Lees, P

2017-10-01

Pharmacokinetic-pharmacodynamic (PK/PD) integration and modelling were used to predict dosage schedules of oxytetracycline for two pig pneumonia pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida. Minimum inhibitory concentration (MIC) and mutant prevention concentration (MPC) were determined in broth and porcine serum. PK/PD integration established ratios of average concentration over 48 h (C av0-48 h )/MIC of 5.87 and 0.27 μg/mL (P. multocida) and 0.70 and 0.85 μg/mL (A. pleuropneumoniae) for broth and serum MICs, respectively. PK/PD modelling of in vitro time-kill curves established broth and serum breakpoint values for area under curve (AUC 0-24 h )/MIC for three levels of inhibition of growth, bacteriostasis and 3 and 4 log 10 reductions in bacterial count. Doses were then predicted for each pathogen, based on Monte Carlo simulations, for: (i) bacteriostatic and bactericidal levels of kill; (ii) 50% and 90% target attainment rates (TAR); and (iii) single dosing and daily dosing at steady-state. For 90% TAR, predicted daily doses at steady-state for bactericidal actions were 1123 mg/kg (P. multocida) and 43 mg/kg (A. pleuropneumoniae) based on serum MICs. Lower TARs were predicted from broth MIC data; corresponding dose estimates were 95 mg/kg (P. multocida) and 34 mg/kg (A. pleuropneumoniae). © 2017 The Authors. Journal of Veterinary Pharmacology and Therapeutics Published by John Wiley & Sons Ltd.

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.

What is the true in vitro potency of oxytetracycline for the pig pneumonia pathogens Actinobacillus pleuropneumoniae and Pasteurella multocida?

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Dorey, L; Hobson, S; Lees, P

2017-10-01

The pharmacodynamics of oxytetracycline was determined for pig respiratory tract pathogens, Actinobacillus pleuropneumoniae and Pasteurella multocida. Indices of potency were determined for the following: (i) two matrices, broth and pig serum; (ii) five overlapping sets of twofold dilutions; and (iii) a high strength starting culture. For A. pleuropneumoniae, minimum inhibitory concentration (MIC) was similar for the two matrices, but for P. multocida, differences were marked and significantly different. MIC and minimum bactericidal concentration (MBC) serum: broth ratios for A. pleuropneumoniae were 0.83:1 and 1.22:1, respectively, and corresponding values for P. multocida were 22.0:1 and 7.34:1. For mutant prevention concentration (MPC) serum: broth ratios were 0.79:1 (A. pleuropneumoniae) and 20.9:1 (P. multocida). These ratios were corrected for serum protein binding to yield fraction unbound (fu) serum: broth MIC ratios of 0.24:1 (A. pleuropneumoniae) and 6.30:1 (P. multocida). Corresponding fu serum: broth ratios for MPC were almost identical, 0.23:1 and 6.08:1. These corrections for protein binding did not account for potency differences between serum and broth for either species; based on fu serum MICs, potency in serum was approximately fourfold greater than predicted for A. pleuropneumoniae and sixfold smaller than predicted for P. multocida. For both broth and serum and both bacterial species, MICs were also dependent on initial inoculum strength. The killing action of oxytetracycline had the characteristics of codependency for both A. pleuropneumoniae and P. multocida in both growth media. The in vitro potency of oxytetracycline in pig serum is likely to be closer to the in vivo plasma/serum concentration required for efficacy than potency estimated in broths. © 2017 The Authors. Journal of Veterinary Pharmacology and Therapeutics Published by John Wiley & Sons Ltd.

Oral exposure to culture material extract containing fumonisins predisposes swine to the development of pneumonitis caused by Pasteurella multocida

International Nuclear Information System (INIS)

Halloy, David J.; Gustin, Pascal G.; Bouhet, Sandrine; Oswald, Isabelle P.

2005-01-01

Fumonisin B 1 (FB 1 ) is a mycotoxin produced by Fusarium verticillioides and F. proliferatum that commonly occurs in maize. In swine, consumption of contaminated feed induces liver damage and pulmonary edema. Pasteurella multocida is a secondary pathogen, which can generate a respiratory disorder in predisposed pigs. In this study, we examined the effect of oral exposure to fumonisin-containing culture material on lung inflammation caused by P. multocida. Piglets received by gavage a crude extract of fumonisin, 0.5 mg FB 1 /kg body weight/day, for 7 days. One day later, the animals were instilled intratracheally with a non toxin producing type A strain of P. multocida and followed up for 13 additional days. Pig weight and cough frequency were measured throughout the experiment. Lung lesions, bronchoalveolar lavage fluid (BALF) cell composition and the expression of inflammatory cytokines were evaluated at the autopsy. Ingestion of fumonisin culture material or infection with P. multocida did not affect weight gain, induced no clinical sign or lung lesion, and only had minimal effect on BALF cell composition. Ingestion of mycotoxin extract increased the expression of IL-8, IL-18 and IFN-γ mRNA compared with P. multocida infection that increased the expression of TNF-α. The combined treatment with fumonisin culture material and P. multocida delayed growth, induced cough, and increased BALF total cells, macrophages and lymphocytes. Lung lesions were significantly enhanced in these animals and consisted of subacute interstitial pneumonia. TNF-α, IFN-γ and IL-18 mRNA expression was also increased. Taken together, our data showed that fumonisin culture material is a predisposing factor to lung inflammation. These results may have implications for humans and animals consuming FB 1 contaminated food or feed

Oral exposure to culture material extract containing fumonisins predisposes swine to the development of pneumonitis caused by Pasteurella multocida

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Halloy, David J [Department of Functional Sciences, Unit of Pharmacology, Pharmacotherapy and Toxicology, Faculty of Veterinary Medicine, University of Liege, Liege (Belgium); Gustin, Pascal G [Department of Functional Sciences, Unit of Pharmacology, Pharmacotherapy and Toxicology, Faculty of Veterinary Medicine, University of Liege, Liege (Belgium); Bouhet, Sandrine [INRA, UR66, Laboratory of Pharmacology and Toxicology, 180 Chemin de Tournefeuille, BP3, 31931 Toulouse (France); Oswald, Isabelle P [INRA, UR66, Laboratory of Pharmacology and Toxicology, 180 Chemin de Tournefeuille, BP3, 31931 Toulouse (France)

2005-09-15

Fumonisin B{sub 1} (FB{sub 1}) is a mycotoxin produced by Fusarium verticillioides and F. proliferatum that commonly occurs in maize. In swine, consumption of contaminated feed induces liver damage and pulmonary edema. Pasteurella multocida is a secondary pathogen, which can generate a respiratory disorder in predisposed pigs. In this study, we examined the effect of oral exposure to fumonisin-containing culture material on lung inflammation caused by P. multocida. Piglets received by gavage a crude extract of fumonisin, 0.5 mg FB{sub 1}/kg body weight/day, for 7 days. One day later, the animals were instilled intratracheally with a non toxin producing type A strain of P. multocida and followed up for 13 additional days. Pig weight and cough frequency were measured throughout the experiment. Lung lesions, bronchoalveolar lavage fluid (BALF) cell composition and the expression of inflammatory cytokines were evaluated at the autopsy. Ingestion of fumonisin culture material or infection with P. multocida did not affect weight gain, induced no clinical sign or lung lesion, and only had minimal effect on BALF cell composition. Ingestion of mycotoxin extract increased the expression of IL-8, IL-18 and IFN-{gamma} mRNA compared with P. multocida infection that increased the expression of TNF-{alpha}. The combined treatment with fumonisin culture material and P. multocida delayed growth, induced cough, and increased BALF total cells, macrophages and lymphocytes. Lung lesions were significantly enhanced in these animals and consisted of subacute interstitial pneumonia. TNF-{alpha}, IFN-{gamma} and IL-18 mRNA expression was also increased. Taken together, our data showed that fumonisin culture material is a predisposing factor to lung inflammation. These results may have implications for humans and animals consuming FB{sub 1} contaminated food or feed.

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

Science.gov (United States)

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

2011-05-01

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

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.

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

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...... 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...

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

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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

Isolation, identification, and monitoring of antibiotic resistance in Pasteurella multocida and Mannheimia haemolytica isolated from sheep in East Azerbaijan province, Iran

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I. Khalili

2016-09-01

Full Text Available The present study was carried out in order to isolate, identify, and assess the antimicrobial susceptibility of the causative agent(s of pneumonic pasteurellosis in sheep in East Azerbaijan province, northwest of Iran. Pneumonia was detected in 320 cases, and the affected lungs were sampled in the slaughterhouse. The samples were investigated bacteriologically for the isolation of two microorganisms from the Pasteurellaceae family. Pasteurella multocida was isolated from six (1.87% samples, while none of the lung tissues were positive for Mannheimia haemolytica. After the isolation and detection of microorganisms via cultural and morphological tests, the bacteria were identified on the basis of biochemical criteria and polymerase chain reaction (PCR technique. Antimicrobial susceptibility testing was performed on all P. multocida isolates, using broth microdilution method. Evaluation of the minimum inhibitory concentration (MIC of eight antimicrobial agents against the tested isolates showed that all the organisms were resistant to amoxicillin and relatively susceptible to ceftiofur. In conclusion, P. multocida was introduced as the main cause of ovine pneumonic pasteurellosis in the studied district, and the outbreak frequency significantly varied in different seasons of the year (P

Ocorrência de genes tad associados à formação de biofilme em isolados de Pasteurella multocida de pulmões de suínos com pneumonia

OpenAIRE

Moraes, Danny Franciele da S.D.; Brandão, Laila Natasha S.; Pitchenin, Leticia C.; O. Filho, João Xavier; Morés, Nelson; Nakazato, Luciano; Dutra, Valéria

2014-01-01

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 ...

21 CFR 558.630 - Tylosin and sulfamethazine.

Science.gov (United States)

2010-04-01

... grams sulfamethazine. (2) Indications for use-(i) Maintaining weight gains and feed efficiency in the presence of atrophic rhinitis; lowering the incidence and severity of Bordetella bronchiseptica rhinitis... (Pasteurella multocida and/or Corynebacterium pyogenes); for reducing the incidence of cervical lymphadenitis...

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

[Effect of lead microparticles introduced into the respiratory system of the sensitivity of mice to Pasteurella multocida infection via aerosol].

Science.gov (United States)

Bouley, G; Dubreuil, A; Arsac, F; Boudène, C

1977-12-19

Lead microparticles, resulting from the pyrolysis of organic lead used as an anti-knock agent in gasoline, were introduced into the lungs of Mice, during a short single exposure. When 6 microgram of lead were retained in the lungs (mean value per Mouse), the phagocytic ability of the pulmonary alveolar macrophages harvested 6 and 18 hrs. later, was significantly reduced. It was observed, in the same conditions, that the resistance of Mice to experimental infection by aerosolized Pasteurella multocida, was significantly reduced. When 3 microgram of lead were retained in the lungs, there was no significant difference between control and intoxicated Mice.

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

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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.

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.

Efeito do extrato alcoólico de própolis sobre a Pasteurella multocida “in vitro” e em coelhos - DOI: 10.4025/actascianimsci.v26i1.1952 Effect of alcohol extract of propolis on the Pasteurella multocida in vitro and in the rabbits - DOI: 10.4025/actascianimsci.v26i1.1952

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Hélio Langoni

2004-04-01

Full Text Available O ensaio in vitro objetivou avaliar o efeito do álcool PA e de diferentes concentrações (5%, 10% e 15% no Agar, do Extrato Alcoólico de Própolis (EAP, de três regiões do Estado de São Paulo, sobre a Pasteurella multocida, obtida a partir de swab nasal de coelhos. Por meio de outro ensaio, analisou-se o comportamento dessas bactérias em lavado tráqueo-brônquico de coelhos, alimentados com rações contendo álcool PA e diferentes concentrações de EAP (0,1%; 0,2% e 0,3%. O álcool só foi efetivo na inibição do desenvolvimento bacteriano in vitro na concentração mais elevada, enquanto o EAP demonstrou efetividade em todas as concentrações. As amostras de própolis das três regiões não diferiram entre si. No lavado tráqueo-brônquico, o número de Unidades Formadoras de Colônias por mililitro não diferiu entre os tratamentos, porém houve uma tendência de maior efetividade nos tratamentos com EAP em relação ao álcool, acompanhando a ação in vitro.The essay in vitro aimed to evaluate the effect of alcohol and different concentrations (5, 10 and 15% in the agar, of Alcoholic Extract of Propolis (AEP, from three regions of São Paulo State, Brazil, on Pasteurella multocida, obtained from rabbits’ nasal swabs. In other essay the reaction of this bacteria was observed. In tracheal regions of rabbits, fed on diets with alcohol and AEP in different concentrations (0.1; 0.2 and 0.3%. The alcohol was only effective on the inhibition of the bacteria in vitro on highest concentration, while the AEP was effective in all concentrations. The propolis samples from three regions did not differ. In the other assay, there wasn’t any difference between treatments, but the AEP showed to be more active than alcohol against Pasteurella multocida, in the tracheal region of the rabbits, as to the action in vitro.

Field study of pneumonia in vaccinated cattle associated with incorrect vaccination and Pasteurella multocida infection.

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Crawshaw, W M; Caldow, G L

2015-04-25

This field study used data on the vaccine courses against bovine respiratory disease sold by one pharmaceutical company in conjunction with pharmacovigilance data to explore reported suspected lack of expected efficacy and the reasons for this. The study ran from May 1, 2007, to April 30, 2010, and covered vaccines sold in Scotland and part of Northumberland. In total, 83 groups of cattle reported suspected lack of expected efficacy, representing 1.6 per cent of the 804,618 vaccine courses sold. It was possible to investigate 45 of these outbreaks in depth using a standard questionnaire and diagnostic protocol. Vaccine usage outwith the specific product characteristics (SPC) occurred in 47 per cent of cases (21/45). The proportion of vaccination courses used where a pathogen contained in the vaccine was detected in the diseased cattle and vaccine use was consistent with the SPC was estimated at 0.12 per cent of the courses sold. Pasteurella multocida was the most common pathogen detected and was found in 21 of the outbreaks. For outbreaks where a pathogen contained in the vaccine was detected, P. multocida was found at a significantly greater frequency (P=0.03) where vaccine use was compliant with the SPC (five of six outbreaks) compared with outbreaks where vaccine use had not been compliant with the SPC (one of seven outbreaks). The limitations of the study, including the diagnostic tests employed and definition of vaccination outwith the SPC, are discussed. British Veterinary Association.

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.

Investigation of genetic diversity and epidemiological characteristics of Pasteurella multocida isolates from poultry in southwest China by population structure, multi-locus sequence typing and virulence-associated gene profile analysis.

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Li, Zhangcheng; Cheng, Fangjun; Lan, Shimei; Guo, Jianhua; Liu, Wei; Li, Xiaoyan; Luo, Zeli; Zhang, Manli; Wu, Juan; Shi, Yang

2018-04-25

Fowl cholera caused by Pasteurella multocida has always been a disease of global importance for poultry production. The aim of this study was to obtain more information about the epidemiology of avian P. multocida infection in southwest China and the genetic characteristics of clinical isolates. P. multocida isolates were characterized by biochemical and molecular-biological methods. The distributions of the capsular serogroups, the phenotypic antimicrobial resistance profiles, lipopolysaccharide (LPS) genotyping and the presence of 19 virulence genes were investigated in 45 isolates of P. multocida that were associated with clinical disease in poultry. The genetic diversity of P. multocida strains was performed by 16S rRNA and rpoB gene sequence analysis as well as multilocus sequence typing (MLST). The results showed that most (80.0%) of the P. multocida isolates in this study represented special P. multocida subspecies, and 71.1% of the isolates showed multiple-drug resistance. 45 isolates belonged to capsular types: A (100%) and two LPS genotypes: L1 (95.6%) and L3 (4.4%). MLST revealed two new alleles (pmi77 and gdh57) and one new sequence type (ST342). ST129 types dominated in 45 P. multocida isolates. Isolates belonging to ST129 were with the genes ompH+plpB+ptfA+tonB, whereas ST342 included isolates with fur+hgbA+tonB genes. Population genetic analysis and the MLST results revealed that at least one new ST genotype was present in the avian P. multocida in China. These findings provide novel insights into the epidemiological characteristics of avian P. multocida isolates in southwest China.

Serological patterns of Actinobacillus pleuropneumoniae, Mycoplasma hyopneumoniae, Pasteurella multocida and Streptococcus suis in pig herds affected by pleuritis.

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Wallgren, Per; Nörregård, Erik; Molander, Benedicta; Persson, Maria; Ehlorsson, Carl-Johan

2016-10-04

Respiratory illness is traditionally regarded as the disease of the growing pig, and has historically mainly been associated to bacterial infections with focus on Mycoplasma hyopneumoniae and Actinobacillus pleuropneumoniae. These bacteria still are of great importance, but continuously increasing herd sizes have complicated the scenario and the influence of secondary invaders may have been increased. The aim of this study was to evaluate the presence of A. pleuropneumoniae and M. hyopneumoniae, as well as that of the secondary invaders Pasteurella multocida and Streptococcus suis by serology in four pig herds (A-D) using age segregated rearing systems with high incidences of pleuritic lesions at slaughter. Pleuritic lesions registered at slaughter ranged from 20.5 to 33.1 % in the four herds. In herd A, the levels of serum antibodies to A. pleuropneumoniae exceeded A 450  > 1.5, but not to any other microbe searched for. The seroconversion took place early during the fattening period. Similar levels of serum antibodies to A. pleuropneumoniae were also recorded in herd B, with a subsequent increase in levels of antibodies to P. multocida. Pigs seroconverted to both agents during the early phase of the fattening period. In herd C, pigs seroconverted to P. multocida during the early phase of the fattening period and thereafter to A. pleuropneumoniae. In herd D, the levels of antibodies to P. multocida exceeded A 450  > 1.0 in absence (A 450  hyopneumoniae and to S. suis remained below A 450  hyopneumoniae late during the rearing period (herd B-D), or not at all (herd A). Different serological patterns were found in the four herds with high levels of serum antibodies to A. pleuropneumoniae and P. multocida, either alone or in combination with each other. Seroconversion to M. hyopneumoniae late during the rearing period or not at all, confirmed the positive effect of age segregated rearing in preventing or delaying infections with M

Rapid diagnosis of virulent Pasteurella multocida isolated from farm animals with clinical manifestation of pneumonia respiratory infection using 16S rDNA and KMT1 gene

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Gamal Mohamedin Hassan

2016-01-01

Full Text Available Objective: To characterize intra-isolates variation between clinical isolates of Pasteurella multocida (P. multocida isolated from sheep, cattle and buffalo at molecular level to check the distribution of pneumonia and hemorrhagic septicemia in some regions of Fayoum, Egypt. Methods: These isolates were obtained from various locations in the Fayoum Governorate, Egypt and they were identified by amplifying 16S rDNA and KMT1 genes using their DNA as a template in PCR reaction. Results: The results demonstrated that the five selective isolates of P. multocida had similar size of PCR products that generated one band of 16S rDNA having 1 471 bp and KMT1 gene having 460 bp. The phylogenetic tree and similarity of the five selective isolates of P. multocida which were collected from GenBank database were calculated and analyzed for the nucleotide sequence of 16S rDNA and KMT1 genes. The sequencing result of 16S rRNA gene product (1 471 bp for the five selective isolates of P. multocida showed that the isolates of sheep (FUP2 shared 94.08%, 88.10% homology with the buffalo isolate (FUP8 and cattle isolate (FUP9 respectively, whereas, the buffalo isolate (FUP5 shared 98.18% and 94.40% homology with the cattle isolates (FUP12 and FUP9. Conclusions: The results indicated the relationships of P. multocida isolated from buffalo and cattle rather than the close relationships between P. multocida isolated from cattle and sheep. Diagnosis of P. multocida by 16S rDNA and KMT1 gene sequences was important to determine the antigen that is responsible for protective cover within the same group of animals and to help for the production of new vaccines for the control of microbial infection for domestic animals.

Forekomst af resistente bakterier og forbrug af antibiotika til hunde

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Pedersen, Karl; Pedersen, Kristina; Jensen, Helene

2007-01-01

), Pasteurella multocida (n=25), Bordetella bronchiseptica (n=14), Proteus spp. (n=29), og E. coli (n=449). I undersøgelsen anvendtes data fra VetStat databasen. Størstedelen af de antibiotika, der bruges til hunde er bredspektrede. Penicilliner med udvidet spektrum, cephalosporiner samt sulphonamider...

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

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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.

Mutant prevention concentration and PK-PD relationships of enrofloxacin for Pasteurella multocida in buffalo calves.

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Balaje, R M; Sidhu, P K; Kaur, G; Rampal, S

2013-12-01

This study validated the use of mutant prevention concentration (MPC) and pharmacokinetic and pharmacodynamic (PK-PD) modeling approach for optimization of dose regimen of enrofloxacin to contain the emergence of Pasteurella multocida resistance. The PK and PD characteristics of enrofloxacin were investigated in buffalo calves after intramuscular administration at a dose rate of 12 mg/kg. The concentration of enrofloxacin and ciprofloxacin in serum were determined by high-performance liquid chromatography. The serum peak concentration (Cmax), terminal half-life (t1/2K10), volume of distribution (Vd(area)/F) and mean residence time (MRT) of enrofloxacin were 1.89 ± 0.35 μg/ml, 5.14 ± 0.66 h, 5.59 ± 0.99 l/kg/h and 8.52 ± 1.29 h, respectively. The percent metabolite conversion ratio of ciprofloxacin to enrofloxacin was 79. The binding of enrofloxacin to plasma proteins was 11%. The MIC, MBC and MPC for enrofloxacin against P. multocida were 0.05, 0.06 μg/ml and 1.50 μg/ml.In vitro and ex-vivo bactericidal activity of enrofloxacin was concentration dependent. Modeling of ex-vivo growth inhibition data to the sigmoid Emax equation provided AUC24h/MIC values to produce bacteriostatic (19 h), bactericidal (43 h) and bacterial eradication (64 h). PK-PD data in conjunction with MPC and MIC90 data predicted dosage schedules for enrofloxacin that may achieve optimum efficacy in respect of bacteriological and clinical cure and minimize the risk of emergence of resistance. Copyright © 2013 Elsevier Ltd. All rights reserved.

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.

A capsule/lipopolysaccharide/MLST genotype D/L6/ST11 of Pasteurella multocida is likely to be strongly associated with swine respiratory disease in China.

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Peng, Zhong; Wang, Haonan; Liang, Wan; Chen, Yibao; Tang, Xibiao; Chen, Huanchun; Wu, Bin

2018-01-01

Pasteurella multocida is a leading cause of respiratory disease in pigs worldwide. In this study, we determined the genetic characteristics of 115 P. multocida isolates from the lungs of pigs with respiratory disease in China in 2015 using capsular typing, lipopolysaccharide (LPS) genotyping, and virulence genotyping based on the detection of virulence-associated genes. The results showed that the isolates belonged to three capsular types: A (49.6%), D (46.1%), and nontypable (4.3%); and two LPS genotypes: L3 (22.6%) and L6 (77.4%). When combining the capsular types with the LPS genotypes, a genotype group D: L6 (46.1%) was the most prevalent among the strains. Among the 23 virulence-associated genes detected in this study, a small number of them displayed a certain level of "genotype-preference". We found that pfhA, hgbA, and hgbB had a close association with P. multocida LPS genotypes, while tadD was more associated with P. multocida capsular types. In addition, multilocus sequence typing (MLST) on 40 P. multocida isolates identified four sequence types: ST3, ST10, ST11, and ST16, and the distribution of ST11 was significantly higher than the other MLST genotypes. Interestingly, all of the ST11 isolates detected in this study were genotype D: L6 strains and they were 100% positive for hgbB. Our data suggest that a capsule/LPS/MLST genotype D/L6/ST11 is likely to be strongly associated with respiratory clinical manifestation of the disease in pigs.

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

2012-01-01

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

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

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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.

The capsule is a virulence determinant in the pathogenesis of Pasteurella multocida M1404 (B:2).

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Boyce, J D; Adler, B

2000-06-01

Capsules from a range of pathogenic bacteria are key virulence determinants, and the capsule has been implicated in virulence in Pasteurella multocida. We have previously identified and determined the nucleotide sequence of the P. multocida M1404 (B:2) capsule biosynthetic locus (J. D. Boyce, J. Y. Chung, and B. Adler, Vet. Microbiol. 72:121-134, 2000). The cap locus consists of 15 genes, which can be grouped into three functional regions. Regions 1 and 3 contain genes proposed to encode proteins involved in capsule export, and region 2 contains genes proposed to encode proteins involved in polysaccharide biosynthesis. In order to construct a mutant impaired in capsule export, the final gene of region 1, cexA, was disrupted by insertion of a tetracycline resistance cassette by allelic replacement. The genotype of the tet(M) OmegacexA mutant was confirmed by Southern hybridization and PCR. The acapsular phenotype was confirmed by immunofluorescence, and the strain could be complemented and returned to capsule production by the presence of a cloned uninterrupted copy of cexA. Wild-type, mutant, and complemented strains were tested for virulence by intraperitoneal challenge of mice; the presence of the capsule was shown to be a crucial virulence determinant. Following intraperitoneal challenge of mice, the acapsular bacteria were removed efficiently from the blood, spleen, and liver, while wild-type bacteria multiplied rapidly. Acapsular bacteria were readily taken up by murine peritoneal macrophages, but wild-type bacteria were significantly resistant to phagocytosis. Both wild-type and acapsular bacteria were resistant to complement in bovine and murine serum.

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.

Characterization of Pasteurella multocida involved in rabbit infections

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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...

Determination of the minimum inhibitory concentration (MIC and mutant prevention concentration (MPC of selected antimicrobials in bovine and swine Pasteurella multocida, Escherichia coli, and Staphylococcus aureus isolates

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Kateřina Nedbalcová

2015-01-01

Full Text Available We compared the values of the minimum inhibitory concentration (MIC and mutant prevention concentration (MPC values ​​of three antimicrobial agents for 72 bovine isolates of Pasteurella multocida, 80 swine isolates of P. multocida, 80 bovine isolates of Escherichia coli, 80 swine isolates of E. coli, and 80 isolates of Staphylococcus aureus from bovine mastitis. The ratio of MIC90​​/MPC90 which limited mutant selection window (MSW was ≤ 0.12/4 mg/l for enrofloxacin, 0.5/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in bovine P. multocida isolates, ≤ 0.12/2 mg/l for enrofloxacin, 0.5/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in swine P. multocida isolates, 1/16 mg/l for enrofloxacin, 8/≥ 64 mg/l for florfenicol and 8/≥ 128 mg/l for tulathromycin in bovine E. coli isolates, 0.5/16 mg/l for enrofloxacin, ≥ 64/≥ 64 mg/l for florfenicol and 8/≥ 128 mg/l for tulathromycin in swine E. coli isolates, and 0.25/16 mg/l for enrofloxacin, 4/≥ 64 mg/l for florfenicol and 4/≥ 128 mg/l for tulathromycin in S. aureus isolates. These findings indicate that the dosage of antimicrobial agents to achieve serum concentration equal to or higher than MPC could reduce selection of resistant bacterial subpopulation.

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

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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 hematology and biochemistry findings, there were 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. Copyright © 2015 Elsevier Ltd. All rights reserved.

Identificação de transcritos diferencialmente expressos por Pasteurella multocida em condições de privação de ferro

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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

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

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

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.

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

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Glass-Kaastra, Shiona K.; 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 trends in 12-month rolling averages, logistic-regression modeling, temporal-scan statistics, and a scan with the “What’s strange about recent events?” (WSARE) algorithm. The AMR trends varied among the antimicrobial drugs for a single pathogen and between pathogens for a single antimicrobial, suggesting that pathogen-specific AMR surveillance may be preferable to indicator data. The 4 methods provided complementary and, at times, redundant results. The most appropriate combination of analysis methods for surveillance using these data included temporal-scan statistics with a visualization method (rolling-average or predicted-probability plots following logistic-regression models). The WSARE algorithm provided interesting results for quality control and has the potential to detect new resistance patterns; however, missing data created problems for displaying the results in a way that would be meaningful to all surveillance stakeholders. PMID:25355992

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.

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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.

[In vitro activity of 12 antibiotics used in veterinary medicine against Mannheimia haemolytica and Pasteurella multocida isolated from calves in the Netherlands].

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Mevius, D J; Hartman, E G

2000-03-01

Results of susceptibility tests of clinical isolates of animal pathogens are periodically summarized and reported by the Animal Health Service. However, these results are based upon qualitative test methods. In the present paper results of quantitative susceptibility tests of twelve antibacterial agents against Mannheimia haemolytica (MHA) and Pasteurella multocida (PMU) isolated from Dutch calves in 1996 and 1997 are presented. Minimum inhibitory concentrations of amoxicillin, ceftiofur, tetracycline, trimethoprim-sulphamethoxazole, tilmicosin, neomycin, gentamicin, spectinomycin, flumequine, enrofloxacin, chloramphenicol and florfenicol were determined. No resistance was detected for ceftiofur and florfenicol. Three strains had an intermediate susceptibility to tilmicosin. The resistance percentages of MHA and PMU for neomycin, gentamicin, spectinomycin, flumequine, enrofloxacin, and chloramphenicol varied from 2% to 16%. Higher resistance percentages (16%-53%) were observed for amoxicillin, tetracycline, and trimethoprim-sulphamethoxazole. The MIC breakpoints used to determine whether a strain is susceptible, intermediate, or resistant are arbitrary and discussed in this paper.

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

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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.

Cellular defense of the avian respiratory system: effects of Pasteurella multocida on respiratory burst activity of avian respiratory tract phagocytes.

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

1988-12-01

The respiratory tract of healthy chickens contain few free-residing phagocytic cells. Intratracheal inoculation with Pasteurella multocida stimulated a significant (P less than 0.05) migration of cells to the lungs and air sacs of White Rock chickens within 2 hours after inoculation. We found the maximal number of avian respiratory tract phagocytes (22.9 +/- 14.0 x 10(6] at 8 hours after inoculation. Flow cytometric analysis of these cells revealed 2 populations on the basis of cell-size and cellular granularity. One of these was similar in size and granularity to those of blood heterophils. Only this population was capable of generating oxidative metabolites in response to phorbol myristate acetate. The ability of the heterophils to produce hydrogen peroxide, measured as the oxidation of intracellularly loaded 2',7'-dichlorofluorescein, decreased with time after inoculation. These results suggest that the migration of heterophils, which are capable of high levels of oxidative metabolism, to the lungs and air sacs may be an important defense mechanism of poultry against bacterial infections of the respiratory tract.

Demonstration that Australian Pasteurella multocida isolates from sporadic outbreaks of porcine pneumonia are non-toxigenic (toxA-) and display heterogeneous DNA restriction endonuclease profiles compared with toxigenic isolates from herds with progressive atrophic rhinitis.

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Djordjevic, S P; Eamens, G J; Ha, H; Walker, M J; Chin, J C

1998-08-01

Capsular types A and D of Pasteurella multocida cause economic losses in swine because of their association with progressive atrophic rhinitis (PAR) and enzootic pneumonia. There have been no studies comparing whole-cell DNA profiles of isolates associated with these two porcine respiratory diseases. Twenty-two isolates of P. multocida from diseased pigs in different geographic localities within Australia were characterised genotypically by restriction endonuclease analysis (REA) with the enzyme CfoI. Seven of 12 P. multocida isolates from nasal swabs from pigs in herds where PAR was either present or suspected displayed a capsular type D phenotype. These were shown to possess the toxA gene by polymerase chain reaction (PCR) and Southern hybridisation, and further substantiated by production of cytotoxin in vitro. The CfoI profile of one of these seven isolates, which was from the initial outbreak of PAR in Australia (in Western Australia, WA), was identical with profiles of all six other toxigenic isolates from sporadic episodes in New South Wales (NSW). The evidence suggests that the strain involved in the initial outbreak was responsible for the spread of PAR to the eastern states of Australia. Another 10 isolates, representing both capsular types A and D, were isolated exclusively from porcine lung lesions after sporadic outbreaks of enzootic pneumonia in NSW and WA. CfoI restriction endonuclease profiles of these isolates revealed considerable genomic heterogeneity. Furthermore, none of these possessed the toxA gene. This suggests that P. multocida strains with the toxA gene do not have a competitive survival advantage in the lower respiratory tract or that toxin production does not play a role in the pathology of pneumonic lesions, or both. REA with polyacrylamide gel electrophoresis and silver staining was found to be a practical and discriminatory tool for epidemiological tracing of P. multocida outbreaks associated with PAR or pneumonia in pigs.

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

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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

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.

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

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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 Apa I 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.

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

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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...

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.

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

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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

Characterization of Pasteurella multocida associated with ovine pneumonia using multi-locus sequence typing (MLST) and virulence-associated gene profile analysis and comparison with porcine isolates.

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García-Alvarez, Andrés; Vela, Ana Isabel; San Martín, Elvira; Chaves, Fernando; Fernández-Garayzábal, José Francisco; Lucas, Domínguez; Cid, Dolores

2017-05-01

Pasteurella multocida is a pathogen causing disease in a wide range of hosts including sheep and pigs. Isolates from ovine pneumonia were characterized by MLST (Multi-host and RIRDC databases) and virulence-associated gene (VAG) typing and compared with porcine isolates. Ovine and porcine isolates did not share any STs as determined by both schemes and exhibited different VAG profiles. With the Multi-host database, sixteen STs were identified among 43 sheep isolates with two STs (ST50 and ST19) comprising 53.5% of the isolates, and seven MLST genotypes (ST3, ST11 and ST62 included 75% of the isolates) among the 48 pig isolates. The most frequent VAG profile among sheep isolates was tbpA+/toxA+ (69.8% of isolates) and pfhA+ (62.5%) and hgbB+ (33.3%) among pig isolates. Representative ovine and porcine isolates of those STs identified by the Multi-host scheme were further typed using the RIRDC scheme. Seven STs were identified among the ovine isolates (ST95 RIRDC , ST131 RIRDC , ST203 RIRDC , ST320 RIRDC , ST324 RIRDC , ST321 RIRDC , and ST323 RIRDC ), with the latter four sequence types being new STs identified in this study, and six STs (ST9 RIRDC , ST13 RIRDC , ST27 RIRDC , ST50 RIRDC , and ST74 RIRDC and a new sequence type ST322 RIRDC ) among the porcine isolates. STs identified among ovine isolates have been detected exclusively in small ruminants, suggesting an adaptation to these hosts, while the genotypes identified among pig isolates have been previously identified in multiple hosts and therefore they are not restricted to pigs. The differences in genotypes and VAG profiles between ovine and pig isolates suggest they could represent different subpopulations of P. multocida. Copyright © 2017 Elsevier B.V. All rights reserved.

ORF Alignment: NC_002663 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002663 gi|15602008 >1p3dA 1 463 17 479 e-156 ... ref|NP_245080.1| MurC [Pasteurell...a multocida subsp. multocida str. Pm70] ... gb|AAK02227.1| MurC [Pasteurella multocida subsp. ...

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.

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....

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

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...

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

Science.gov (United States)

2010-04-01

...) As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to... days. If disease recurs, repeat treatment. (ii) Limitations. Make fresh solution daily. Do not treat... acute fowl cholera caused by Pasteurella multocida susceptible to sulfamerazine, sulfamethazine, and...

Molecular diagnosis of Haemorrhagic Septicaemia - A Review

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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

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...

Virulence Genotyping of Pasteurella multocida Isolated from Multiple Hosts from India

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Laxmi Narayan Sarangi

2014-01-01

Full Text Available 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.

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.

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.

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

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......) 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...

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 ...

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

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Sandra Prüller

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

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.

In vitro activity of flumequine in comparison with several other antimicrobial agents against five pathogens isolated in calves in The Netherlands.

Science.gov (United States)

Mevius, D J; Breukink, H J; van Miert, A S

1990-10-01

The in vitro activity of flumequine in comparison with several other drugs was tested against 17 P. multocida, 16 P. haemolytica, 21 S. dublin, 21 S. typhimurium and 21 E. coli strains, isolated in (veal) calves in the Netherlands. The MIC50 of flumequine for the respective pasteurellas was 0.25 and 1 microgram/ml, for the salmonellas and E. coli 0.5 micrograms/ml. In comparison with flumequine, enrofloxacin and ciprofloxacin showed higher in vitro activity, with MIC50 less than or equal to 0.008 micrograms/ml for ciprofloxacin. Decreased susceptibility of the pasteurellas was found for kanamycin, neomycin, streptomycin, gentamicin, oxytetracycline and doxycycline. The MIC50 of minocycline for P. multocida was 0.5 micrograms/ml and there was no cross resistance with the other tetracyclines. P. multocida was very susceptible to ampicillin (MIC50 less than or equal to 0.03 micrograms/ml), P. haemolytica, however, was 100% resistant to this drug. Both pasteurellas were susceptible to cephalothin and approximately 50% of the strains of both bacteria were resistant to chloramphenicol. The MIC50 of either spiramycin or tylosin was greater than or equal to their respective breakpoint-MIC values. Both pasteurellas were susceptible to the combination of trimethoprim and sulphamethoxazole. However, for P. multocida, the addition of sulphamethoxazole to trimethoprim had no synergistic effect on its MIC. In comparison with trimethorpim, aditoprim was less potent. Therefore only P. multocida was susceptible to aditoprim.

Bordetella pertussis transmission

Science.gov (United States)

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

Characterization of fimbrial subunits from Bordetella species

NARCIS (Netherlands)

Mooi, F.R.; Heide, H.G.J. van der; Avest, A.R. ter; Welinder, K.G.; Livey, I.; Zeijst, B.A.M. van der; Gaastra, W.

Using antisera raised against serotype 2 and 3 fimbrial subunits from Bordetella pertussis, serologically related polypeptides were detected in Bordetella bronchiseptica, Bordetella parapertussis and Bordetella avium strains. The two B. pertussis fimbrial subunits, and three of the serologically

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

Science.gov (United States)

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

2011-10-01

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

21 CFR 522.2630 - Tulathromycin.

Science.gov (United States)

2010-04-01

.... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for the control of SRD associated with A. pleuropneumoniae, P. multocida, and M. hyopneumoniae in groups of pigs..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk of...

Aspectos patológicos e microbiológicos das doenças respiratórias em suínos de terminação no Brasil

Directory of Open Access Journals (Sweden)

Marcos A.Z. Morés

2015-08-01

Full Text Available Resumo: Para avaliação dos aspectos patológicos e microbiológicos de casos clínicos de doenças respiratórias em suínos de terminação foram analisados 75 suínos doentes oriundos de 36 lotes. Suínos que apresentavam sinais clínicos respiratórios evidentes foram necropsiados para avaliação macroscópica e colheita de amostras para análise histopatológica e microbiológica. Foram realizados testes de isolamento bacteriano para as principais bactérias do sistema respiratório dos suínos, PCR para Mycoplasma hyorhinis, imuno-histoquímica para Influenza A, Circovirus suíno tipo 2 e Mycoplasma hyopneumoniae. A sensibilidade antimicrobiana de 24 amostras de Pasteurella multocida tipo A foi avaliada por testes de concentração inibitória mínima para os principais antimicrobianos utilizados em suinocultura. Mycoplasma hyopneumoniae e Pasteurella multocida tipo A foram os agentes infecciosos mais prevalentes. Broncopneumonia supurativa e pleurite foram as principais lesões respiratórias encontradas. Pasteurella multocida tipo A, quando presente, aumentou a extensão das lesões pulmonares. Todas as amostras de Pasteurella multocida testadas foram sensíveis aos antimicrobianos Doxiciclina, Enrofloxacina e Tilmicosina. Em 58% das amostras foi identificado mais de um agente infeccioso, evidenciando a alta prevalência da associação de agentes nas doenças respiratórias de suínos em terminação.

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

International Nuclear Information System (INIS)

Settypalli, T.B.K.; Lamien, C.; Spergser, J.; Lelenta, M.; Wade, A.; Gelaye, E.; Loitsch, A.; Minoungou, G.; Thiaucourt, F.; Diallo, A.

2016-01-01

Full text: 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

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

Evaluation of the Specificity of BP3385 for Bordetella pertussis

Science.gov (United States)

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

[Features of Bordetella pertussis, Bordetella spp. infection and whopping cough in Córdoba province, Argentina].

Science.gov (United States)

Giayetto, Víctor O; Blanco, Sebastián; Mangeaud, Arnaldo; Barbás, María G; Cudolá, Analía; Gallego, Sandra V

2017-04-01

Whooping cough is a re-emerging infection in the world and Latin America. It was considered relevant to investigate the clinical and epidemiological profile of Bordetella spp. and Bordetella pertussis infection in Córdoba province, Argentina; evaluating, at the same time, the co-infection with virus producing respiratory infections that may be confused with whooping cough. All whooping cough suspected cases were studied by Polimerase Chain Reaction, amplifying the repeated insertion sequence (IS) 481 and the promoter gene encoding pertussis toxin, between 2011 and 2013. The data were obtained from the clinical and epidemiological records. From 2,588 whooping cough suspected cases, 11.59% was infected by Bordetella spp. and 9.16% was confirmed as Bordetella pertussis infection. The rate of infection was 7.22 and 1.84 per 100,000 for 2011 and 2012, respectively. The infection presented a seasonal tendency and it was mainly found on the group of children between 13 and 24 months old. The co-infection with virus producing respiratory infections, were uncommon. Paroxysmal cough, cyanosis and/or vomiting were predictors of the infection for Bordetella pertussis. To deal with the re-emergence of whooping cough is important the knowledge of the regional epidemiological situation. This paper shows the situation of these infections in the regional clinical and epidemiological context, and makes the information available for health decision-making.

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

Science.gov (United States)

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

2014-01-01

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

Effect of radiation on bacterial cells pathogenesis

International Nuclear Information System (INIS)

Szulc, M.; Tropilo, J.; Zajaczkowska, E.

1983-01-01

In order to determine the effect of X rays on the pathogeny of Erysipelothrix rhusiopathiae and Pasteurella multocida 334 mice were infected subcutaneously with the germs exposed to 10 Gy and D 10 . It was found that a single exposition of E. rhusiopathiae and P. multocida to 10 Gy and D 10 did not change pathogenic properties of these cells. P. multocida showed higher sensitivity to X rays: D 10 for that species was 94 Gy and for E. rhusiopathiae - 156 Gy. (author)

Bacteremia and Peritonitis in a Patient With Cirrhosis: A Life-Threatening Case From a Prick of a Cactus

Directory of Open Access Journals (Sweden)

Jodi-Anne Wallace MD

2017-08-01

Full Text Available A 58-year-old male with nonalcoholic steatohepatitis cirrhosis presents with right lower extremity cellulitis, abdominal tenderness, and severe sepsis after sustaining puncture injury from a cactus on a property with feral cats. Blood cultures and diagnostic paracentesis were consistent with spontaneous bacterial peritonitis due to Pasteurella multocida , a gram-negative coccobacillus found in the respiratory tract of domestic animals. The patient received timely antibiotic coverage with resolution of spontaneous bacterial peritonitis and sepsis after 14-day treatment. This case emphasizes the life-threatening nature of systemic Pasteurella multocida infection as well as an indirect way of acquiring a zoonotic infection in a patient with end-stage liver disease.

21 CFR 866.3065 - Bordetella spp. serological reagents.

Science.gov (United States)

2010-04-01

... (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3065 Bordetella... serological tests to identify Bordetella spp. from cultured isolates or directly from clinical specimens. The...

Monitoring of antimicrobial susceptibility of respiratory tract pathogens isolated from diseased cattle and pigs across Europe, 2009-2012: VetPath results.

Science.gov (United States)

El Garch, Farid; de Jong, Anno; Simjee, Shabbir; Moyaert, Hilde; Klein, Ulrich; Ludwig, Carolin; Marion, Hervé; Haag-Diergarten, Silke; Richard-Mazet, Alexandra; Thomas, Valérie; Siegwart, Ed

2016-10-15

VetPath is an ongoing pan-European antibiotic susceptibility monitoring programme that collects pathogens from diseased cattle, pigs and poultry. In the current study, 996 isolates from cattle and pig respiratory tract infections were tested for their antimicrobial susceptibilities. Non-replicate lung samples or nasopharyngeal/nasal swabs were collected from animals with acute clinical signs in 10 countries during 2009-2012. Pasteurella multocida, Mannheimia haemolytica and Histophilus somni from cattle and P. multocida, Actinobacillus pleuropneumoniae, Haemophilus parasuis, Bordetella bronchiseptica and Streptococcus suis from pigs were isolated by standard methods. S. suis was also isolated from meningitis cases. MIC values of 16 or 17 antibiotics were assessed centrally by broth microdilution following CLSI standards. Results were interpreted using CLSI breakpoints where available. Cattle isolates were generally highly susceptible to most antibiotics, except to tetracycline (3.0-12.0% resistance). Low levels of resistance (0-4.0%) were observed for the macrolide antibiotics. Resistance to spectinomycin varied from 0 to 6.0%. In pig isolates similar observations were made. Resistance to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin, florfenicol, tulathromycin, tiamulin and tilmicosin was absent or <2%. Trimethoprim/sulfamethoxazole resistance varied from 1.9 to 5.3%, but tetracycline resistance varied from 20.4% in P. multocida to 88.1% in S. suis. For most antibiotics and pathogens the percentage resistance remained unchanged or only increased numerically as compared to that of the period 2002-2006. In conclusion, absence or low resistance to antibiotics with defined clinical breakpoints, except for tetracycline, was observed among the major respiratory tract pathogens recovered from livestock. Comparison of all antibiotics and organisms was hampered since for almost half of the antibiotics no CLSI-defined breakpoints were available. Copyright © 2016

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...

The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae

Directory of Open Access Journals (Sweden)

Schneiker-Bekel Susanne

2008-09-01

Full Text Available Abstract Background Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. Results In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. Conclusion The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches.

Indução de empiema em ratos através da inoculação pleural de bactérias Experimental empyema in rats through intrapleural injection of bacteria

Directory of Open Access Journals (Sweden)

José Carlos Fraga

2001-12-01

Full Text Available OBJETIVOS: avaliar a indução experimental de empiema em ratos, através da inoculação intrapleural de duas bactérias (Pasteurella multocida e Staphylococcus aureus, utilizando técnica cirúrgica simples e de fácil execução. MÉTODOS: foram utilizados 24 ratos albinos da raça Wistar, de ambos os sexos, pesando entre 250 e 300g, que, após a anestesia geral, foram submetidos à toracotomia anterior direita, afastamento da musculatura e inoculação de 0,2ml de solução, conforme descrição a seguir: grupo I (n=12, inoculação de Pasteurella multocida, 10(10 unidades formadoras de colônia/ml cultivados em caldo cérebro-coração; grupo II (n=8, inoculação de Staphylococcus aureus, 10(10 unidades formadoras de colônia/ml cultivados em caldo cérebro-coração; e grupo III (n=4, inoculação de caldo cérebro-coração estéril (controle. Os animais foram sacrificados em até 7 dias e a intensidade da reação pleural, analisada macroscopicamente conforme escala padronizada. Também foram avaliados a mortalidade, o volume de líquido na cavidade pleural e o exame bacteriológico (animais mortos e líquido pleural. RESULTADOS: no grupo I (Pasteurella multocida, sete ratos morreram nas primeiras 48 horas de experimento. Cinco ratos foram sacrificados no período programado, mas nenhum deles apresentava empiema. No grupo II (Staphylococcus aureus, somente um animal morreu nas primeiras 24 horas, os outros 7 (88% foram sacrificados e apresentavam empiema. No grupo III, considerados controles, todos os animais sobreviveram, não se observando nenhuma anormalidade torácica ao sacrifício. Analisando conjuntamente os grupos, a indução de empiema esteve associada de maneira significativa à inoculação de Staphylococcus aureus no espaço pleural (p OBJECTIVE: to evaluate empyema formation in rats through the injection of two bacteria (Pasteurella multocida and Staphylococcus aureus, using a simple, easy-to-use surgical technique

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

21 CFR 558.586 - Sulfaquinoxaline.

Science.gov (United States)

2010-04-01

... use. As an aid in the control of acute fowl cholera caused by Pasteurella multocida susceptible to.... Treatment to be started after weaning. Feed continuously for 30 days or feed medicated feed for 2 days out...

Avian cholera

Science.gov (United States)

Friend, Milton

1999-01-01

Avian cholera is a contagious disease resulting from infection by the bacterium Pasteurella multocida. Several subspecies of bacteria have been proposed for P. multocida, and at least 16 different P. multocida serotypes or characteristics of antigens in bacterial cells that differentiate bacterial variants from each other have been recognized. The serotypes are further differentiated by other methods, including DNA fingerprinting. These evaluations are useful for studying the ecology of avian cholera (Fig. 7.1), because different serotypes are generally found in poultry and free-ranging migratory birds. These evaluations also show that different P. multocida serotypes are found in wild birds in the eastern United States than those that are found in the birds in the rest of the Nation (Fig. 7.2).

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...

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

Science.gov (United States)

Heininger, Ulrich; Schlassa, Detlef

2016-11-01

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

The isolation and antibiogram of aerobic nasal bacterial flora of ...

African Journals Online (AJOL)

The following microorganisms were identified from the normal flora of the grasscutters, they are; Staphylococcus epidermidis, Staphylococcus aureus, Enterococcus sp, Micrococcus sp, Bacillus cerus , E. coli, Serratia sp, Streptococcus sp, Pasteurella multocida, Streptacoccus, sp., Mannheimia heamolytica, Klebsiella sp., ...

Quantification and characterization of enzymatically produced hyaluronan with fluorophore-assisted carbohydrate electrophoresis

NARCIS (Netherlands)

Kooy, F.K.; Muyuan Ma,; Beeftink, H.H.; Eggink, G.; Tramper, J.; Boeriu, C.G.

2009-01-01

Hyaluronan (HA) is a polysaccharide with high-potential medical applications, depending on the chain length and the chain length distribution. Special interest goes to homogeneous HA oligosaccharides, which can be enzymatically produced using Pasteurella multocida hyaluronan synthase (PmHAS). We

Bordetella bronchiseptica and fatal pneumonia of dogs and cats

Science.gov (United States)

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

75 FR 1274 - Implantation or Injectable Dosage Form New Animal Drugs; Florfenicol and Flunixin

Science.gov (United States)

2010-01-11

... flunixin meglumine), a combination injectable solution, for treatment of bovine respiratory disease (BRD... treatment of bovine respiratory disease (BRD) associated with Mannheimia haemolytica, Pasteurella multocida... female dairy cattle 20 months of age or older. Use of florfenicol in this class of cattle may cause milk...

21 CFR 522.955 - Florfenicol.

Science.gov (United States)

2010-04-01

... treatment of bovine respiratory disease (BRD) associated with Mannheimia haemolytica, Pasteurella multocida... cause milk residues. A withdrawal period has not been established in preruminating calves. Do not use in... use. For control of respiratory disease in cattle at high risk of developing BRD associated with...

21 CFR 520.2260b - Sulfamethazine sustained-release boluses.

Science.gov (United States)

2010-04-01

... pneumonia caused or complicated by Pasteurella multocida; as an aid in the treatment of foot rot, mastitis... sulfamethazine-sensitive organisms as follows: bacterial pneumonia and bovine respiratory disease complex... mastitis and acute metritis caused by Streptococcus spp.)1 (iii) Limitations. After 72 hours, all animals...

Fulminant infection by uncommon organisms in animal bite wounds.

Science.gov (United States)

Dutta, J K

1998-10-01

In 1995 and 1996, 215 patients exposed to different species of animals were treated at the Amarnath Polyclinic, Balasore, in India. Among them were two children infected by uncommon organisms, i.e., Capnocytophaga canimorsus and Pasteurella multocida; the patients recovered with appropriate antibiotic therapy.

Fulminant infection by uncommon organisms in animal bite wounds.

OpenAIRE

Dutta, J. K.

1998-01-01

In 1995 and 1996, 215 patients exposed to different species of animals were treated at the Amarnath Polyclinic, Balasore, in India. Among them were two children infected by uncommon organisms, i.e., Capnocytophaga canimorsus and Pasteurella multocida; the patients recovered with appropriate antibiotic therapy.

Management of common animal bites in the emergency centre

African Journals Online (AJOL)

Professor Engelbrecht's current fields of interest are bites, stings and poisonous plants. Correspondence to: ... Cross War Memorial Children's Hospital in. Cape Town ... infections. Wound infection with Pasteurella multocida usually occurs early (within 12 ..... Dog bite prevention: an assessment of child knowledge. J Pediatr ...

Improving specificity of Bordetella pertussis detection using a four target real-time PCR.

Directory of Open Access Journals (Sweden)

Helena Martini

Full Text Available The incidence of whooping cough, a contagious respiratory disease caused by Bordetella pertussis, is on the rise despite existing vaccination programmes. Similar, though usually milder, respiratory symptoms may be caused by other members of the Bordetella genus: B. parapertussis, B. holmesii, and B. bronchiseptica. Pertussis diagnosis is mostly done using PCR, but the use of multiple targets is necessary in order to differentiate the different Bordetella spp. with sufficient sensitivity and specificity. In this study we evaluate a multiplex PCR assay for the differentiation of B. pertussis from other Bordetella spp., using the targets IS481, IS1001, IS1002, and recA. Moreover, we retrospectively explore the epidemiology of Bordetella spp. infections in Belgium, using the aforementioned assay over a three-year period, from 2013 until 2015.

Improving specificity of Bordetella pertussis detection using a four target real-time PCR

Science.gov (United States)

Detemmerman, Liselot; Soetens, Oriane; Yusuf, Erlangga; Piérard, Denis

2017-01-01

The incidence of whooping cough, a contagious respiratory disease caused by Bordetella pertussis, is on the rise despite existing vaccination programmes. Similar, though usually milder, respiratory symptoms may be caused by other members of the Bordetella genus: B. parapertussis, B. holmesii, and B. bronchiseptica. Pertussis diagnosis is mostly done using PCR, but the use of multiple targets is necessary in order to differentiate the different Bordetella spp. with sufficient sensitivity and specificity. In this study we evaluate a multiplex PCR assay for the differentiation of B. pertussis from other Bordetella spp., using the targets IS481, IS1001, IS1002, and recA. Moreover, we retrospectively explore the epidemiology of Bordetella spp. infections in Belgium, using the aforementioned assay over a three-year period, from 2013 until 2015. PMID:28403204

Essential role of Bordetella NadC in a quinolinate salvage pathway for NAD biosynthesis.

Science.gov (United States)

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

2017-02-01

Nicotinamide adenine dinucleotide (NAD) is produced via de novo biosynthesis pathways and by salvage or recycling routes. The classical Bordetella bacterial species are known to be auxotrophic for nicotinamide or nicotinic acid. This study confirmed that Bordetella bronchiseptica, Bordetella pertussis and Bordetella parapertussis have the recycling/salvage pathway genes pncA and pncB, for use of nicotinamide or nicotinic acid, respectively, for NAD synthesis. Although these Bordetellae lack the nadA and nadB genes needed for de novo NAD biosynthesis, remarkably, they have one de novo pathway gene, nadC, encoding quinolinate phosphoribosyltransferase. Genomic analyses of taxonomically related Bordetella and Achromobacter species also indicated the presence of an 'orphan' nadC and the absence of nadA and nadB. When supplied as the sole NAD precursor, quinolinate promoted B. bronchiseptica growth, and the ability to use it required nadC. Co-expression of Bordetella nadC with the nadB and nadA genes of Paraburkholderia phytofirmans allowed B. bronchiseptica to grow in the absence of supplied pyridines, indicative of de novo NAD synthesis and functional confirmation of Bordetella NadC activity. Expression of nadC in B. bronchiseptica was influenced by nicotinic acid and by a NadQ family transcriptional repressor, indicating that these organisms prioritize their use of pyridines for NAD biosynthesis. © 2016 John Wiley & Sons Ltd.

(PCR) for the identification of Mycoplasma capricolum subsp.

African Journals Online (AJOL)

ajl yemi

2011-10-07

Oct 7, 2011 ... lactalbumin hydrolysate, 1% MEM, 20% decomplemented horse serum, 5% fresh yeast extract, 1% thallium acetate and 0.4% sodium pyruvate) in a high security laboratory. The DNA of. Pasteurella multocida and Mannheimia haemolytica was maintained in the State Key Laboratory of Veterinary Etiological ...

In vitro screening of methanol plant extracts for their antibacterial activity

International Nuclear Information System (INIS)

Hussain, T.; Arshad, M.; Khan, S.; Sattar, H.

2011-01-01

The purpose of this study was to observe the antibacterial activity of aqueous methanolic extracts of 10 plants against 2-gram negative bacteria (Pasteurella multocida, Escherichia coli) and 3-gram positive bacteria (Bacillus cereus, Staphylococcus aureus, Corynebacterium bovis) by using disc diffusion method. The minimum inhibitory concentration (MIC) was determined by agar well diffusion method and agar dilution method. All the bacteria were susceptible to different plant extracts. Lawsonia inermis, Embellia ribes and Santalum album showed antibacterial activity against all the tested bacteria. The extract of Santalum album showed maximum antibacterial activity of the 10 plant extracts used. Bacillus cereus and Pasteurella multocida were the most sensitive bacteria against most of the plant extracts. It is clear from the results of the present studies that the plant extracts have great potential as antimicrobial compounds against bacteria. However, there is a need of further research to isolate the active ingredients for further pharmacological evaluation. (author)

Prevalence of Bordetella pertussis and Bordetella parapertussis in Samples Submitted for RSV Screening

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Walsh, Paul

2008-08-01

Full Text Available BACKGROUND: The clinical presentation of Bordetella pertussis can overlap with that of respiratory syncytial virus (RSV; however, management differs.HYPOTHESIS: First, the prevalence of B. pertussis is less than 2% among patients screened for RSV, and second the prevalence of B. parapertussis is also less than 2% among these patients.METHODS: Nasal washings submitted to a clinical laboratory for RSV screening were tested for B. pertussis and B. parapertussis, using species-specific real-time polymerase chain reaction (PCR assays. These were optimized to target conserved regions within a complement gene and the CarB gene, respectively. A Bordetella spp. genus-specific real-time PCR assay was designed to detect the Bhur gene of B. pertussis, B. parapertussis, and B. bronchiseptica. RSV A and B subtypes were tested by reverse transcription-PCR.RESULTS: Four hundred and eighty-nine clinical samples were tested. There was insufficient material to complete testing for one B. pertussis, 10 RSV subtype A, and four RSV subtype B assays. Bordetella pertussis was detected in 3/488 (0.6% (95% CI 0.1% to 1.8%, while B. parapertussis was detected in 5/489 (1.0% (95% CI 0.3% to 2.4%. Dual infection of B. pertussis with RSV and of B. parapertussis with RSV occurred in two and in three cases respectively. RSV was detected by PCR in 127 (26.5%.CONCLUSION: The prevalence of B. pertussis in nasal washings submitted for RSV screening was less than 2%. The prevalence of parapertussis may be higher than 2%. RSV with B. pertussis and RSV with B. parapertussis coinfection do occur.

Bordetella pertussis diagnosed by polymerase chain reaction

DEFF Research Database (Denmark)

Birkebaek, N H; Heron, I; Skjødt, K

1994-01-01

The object of this work was to test the polymerase chain reaction (PCR) for demonstration of Bordetella pertussis (BP) in nasopharyngeal secretions. The method was applied to patients with recently diagnosed pertussis, as verified by BP culture. In order to test the sensitivity and specificity...... in 25 patients in whose nasopharyngeal secretions BP had been demonstrated after 4-7 days of culture. The detection limit of PCR in aqueous solution was 1-2 BP bacteria per reaction tube. PCR was 100% specific for BP, showing no response with other Bordetella species or other bacteria known to colonize...

21 CFR 522.2471 - Tilmicosin.

Science.gov (United States)

2010-04-01

...) Indications for use. For the treatment of bovine respiratory disease (BRD) associated with Mannheimia haemolytica, Pasteurella multocida, and Histophilus somni. For the control of respiratory disease in cattle at... dairy cattle 20 months of age or older. Use of this antibiotic in this class of cattle may cause milk...

Whole Genome Sequence Analysis of Pig Respiratory Bacterial Pathogens with Elevated Minimum Inhibitory Concentrations for Macrolides.

Science.gov (United States)

Dayao, Denise Ann Estarez; Seddon, Jennifer M; Gibson, Justine S; Blackall, Patrick J; Turni, Conny

2016-10-01

Macrolides are often used to treat and control bacterial pathogens causing respiratory disease in pigs. This study analyzed the whole genome sequences of one clinical isolate of Actinobacillus pleuropneumoniae, Haemophilus parasuis, Pasteurella multocida, and Bordetella bronchiseptica, all isolated from Australian pigs to identify the mechanism underlying the elevated minimum inhibitory concentrations (MICs) for erythromycin, tilmicosin, or tulathromycin. The H. parasuis assembled genome had a nucleotide transition at position 2059 (A to G) in the six copies of the 23S rRNA gene. This mutation has previously been associated with macrolide resistance but this is the first reported mechanism associated with elevated macrolide MICs in H. parasuis. There was no known macrolide resistance mechanism identified in the other three bacterial genomes. However, strA and sul2, aminoglycoside and sulfonamide resistance genes, respectively, were detected in one contiguous sequence (contig 1) of A. pleuropneumoniae assembled genome. This contig was identical to plasmids previously identified in Pasteurellaceae. This study has provided one possible explanation of elevated MICs to macrolides in H. parasuis. Further studies are necessary to clarify the mechanism causing the unexplained macrolide resistance in other Australian pig respiratory pathogens including the role of efflux systems, which were detected in all analyzed genomes.

Bordetella pertussis en estudiantes adolescentes de la Ciudad de México Bordetella pertussis em estudantes adolescentes da Cidade do México Bordetella pertussis in adolescents students in Mexico City

Directory of Open Access Journals (Sweden)

Patricia Tomé Sandoval

2008-08-01

Full Text Available OBJETIVO: Estimar la seroprevalencia a Bordetella pertussis en escolares y sus contactos escolares y familiares. MÉTODOS: Un total de 12.273 estudiantes de 12 a 15 años de edad, de 14 escuelas secundarias públicas de la Ciudad de México fueron estudiados durante los meses de Septiembre 2002 a Marzo 2003. Se tomó muestra de exudado nasofaríngeo en adolescentes con tos de más de 14 días de evolución. La infección fue confirmada por la técnica de reacción en cadena de polimerasa. Se realizó estudio de contactos escolares y familiares. RESULTADOS: La incidencia de tos fue de 5 para 1.000 estudiantes. De los 61 estudiantes con tos incluidos en la muestra, 20 (32,8% fueron positivos para Bordetella. De los 152 contactos escolares, 16 (10,6% resultaron positivos, y ocho tenían tos. Uno de esos contactos fue el director de una de las escuelas responsable de más del 60% de los casos positivos (12/20, quien también dio lecciones a diez de los estudiantes infectados. De los 29 familiares, ocho (27,6% fueron positivos, pertenecientes a tres familias. CONCLUSIONES: Los resultados muestran que la frecuencia de la enfermedad fue similar al comunicado en la población adolescente de otros países. Sin embargo, este trastorno no tiene necesariamente signos clínicos de la tos persistente y está sujeto a la existencia de infectados asintomáticos con Bordetella.OBJETIVO: Estimar a soroprevalência a Bordetella pertussis em escolares e seus contatos. MÉTODOS: Foram examinados 12.273 alunos entre 12 e 15 anos de idade, de 14 escolas secundárias públicas da Cidade do México, de setembro de 2002 a março de 2003. Amostras de exudado nasofaríngeo foram coletadas de adolescentes com tosse por mais de 14 dias. A infecção foi confirmada por reação em cadeia da polimerase. Todos os alunos e funcionários dos colégios dos casos confirmados por reação em cadeia da polimerase e seus familiares foram testados. RESULTADOS: A incidência de tosse

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

Science.gov (United States)

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

Gamma-Irradiated Mannheimia (Pasteurella) Haemolytica Identified by rRNA Gene Sequencing as a Potential Vaccine in Mice

International Nuclear Information System (INIS)

Araby, E.

2014-01-01

Pneumonic pasteurellosis is a significant disease in beef production medicine. The most information suggests that this disease is a $700 million dollar per year economic burden in bovine food animal production. The current study was designed to assess the immune efficacy of whole cell killed of M. haemolytica strain from satisfactory cases (infected lung from sheep). The efficacy of gamma- irradiated M. haemolytica vaccine (GIV) was evaluated in mice in comparison to the classical aqueous formalized (AFV) one. The bacteria under study were cultivation on blood agar, purification and genetically identified. Then the bacterial cells were exposed to different doses of gamma radiation (2- 20 kGy) with 2 kGy intervals and the dose response curve of the survivors was plotted and 20 kGy was selected as the dose for the preparation of the vaccine. A total of 30 male mice (two weeks – old) were used for the further experimental investigations. Animals were divided into three equal groups each of 10 animals. The first group (group A) was given GIV . The second group (group B) received AFV. The third group (group C) was injected with sterile saline solution and represents the control. Animals were vaccinated via intraperitoneal (i.p) injection with 1x10 8 CFU per treated mouse. After vaccination, the immuno response was determined by cellular surface antigens-reactive antibodies using a modified protein- electrophoresis procedure. Antibody-antigen hybrids was visualized at molecular weight more than 225 KDa in samples represented M. haemolytica antibodies group (A, B) against both bacterial samples (M. haemolytica and Pasteurella multocida ) , while non-treated bacterial cells in which cells incubated with serum of mice group (C) revealed no hybridization reaction, this results verify that, there is shared cellular surface antigens among the two Pasteurella species. Also, the bacterial distribution with (LD 50 ) 2x10 7 CFU of a live M. heamolytica into vaccinated and non

Pasteurella aerogenes as an Asymptomatic Bacteriuria Agent.

Science.gov (United States)

Alaygut, Demet; Engin, Aynur

2018-02-01

'Asymptomatic bacteriuria' (ASB) is isolation of a specified quantitative count of bacteria in an appropriately collected urine specimen obtained from a person without symptoms or signs referable to urinary infection. Catheterized specimens are less likely to be contaminated compared with voided specimens; therefore, positive cultures of catheterized specimens are more likely to reflect true bladder bacteriuria even with low colony counts. The common pathogens for ASB are Escherichia coli, Klebsiella and Streptococcus spp. Pasteurella spp. was not previously reported as an ASB agent. ASB is important for pregnant women, children, individuals with obstructive uropathy, chronic renal failure and neutropenia, before the urologic procedures and after renal transplantation. Treatment of ASB is required for above situations. We report an 11-year-old-girl with neurogenic bladder who made clean intermittent catheterization and had Pasteurella aerogenes as an ASB agent. © The Author [2017]. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Application of Enrofloxacin and Orbifloxacin Disks Approved in Japan for Susceptibility Testing of Representative Veterinary Respiratory Pathogens

Science.gov (United States)

HARADA, Kazuki; USUI, Masaru; ASAI, Tetsuo

2014-01-01

ABSTRACT In this study, susceptibilities of Pasteurella multocida, Mannheimia haemolytica and Actinobacillus pleuropneumoniae to enrofloxacin and orbifloxacin were tested using an agar diffusion method with the commercial disks and a broth microdilution method. Good correlation between the 2 methods for enrofloxacin and orbifloxacin was observed for P. multocida (r = −0.743 and −0.818, respectively), M. haemolytica (r = −0.739 and −0.800, respectively) and A. pleuropneumoniae (r = −0.785 and −0.809, respectively). Based on the Clinical and Laboratory Standards Institute interpretive criteria for enrofloxacin, high-level categorical agreement between the 2 methods was found for P. multocida (97.9%), M. haemolytica (93.8%) and A. pleuropneumoniae (92.0%). Our findings indicate that the tested commercial disks can be applied for susceptibility testing of veterinary respiratory pathogens. PMID:25008965

Phenotypic variability among strains of Pasteurella multocida ...

African Journals Online (AJOL)

STORAGESEVER

2008-05-02

May 2, 2008 ... Available online at http://www.academicjournals.org/AJB. ISSN 1684–5315 ... extended phenotypic characterization methods supported by DNA ... septicaemia African (Obudu) strain (E:2) which are currently employed as ...

9 CFR 113.68 - Pasteurella Haemolytica Vaccine, Bovine.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Pasteurella Haemolytica Vaccine, Bovine. 113.68 Section 113.68 Animals and Animal Products ANIMAL AND PLANT HEALTH INSPECTION SERVICE... Service. (4) A satisfactory challenge shall be evidenced in the controls by progression of clinical signs...

Epidemiology of whooping cough & typing of Bordetella pertussis.

Science.gov (United States)

Hegerle, Nicolas; Guiso, Nicole

2013-11-01

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

[Infection by Bordetella pertussis and bordetella parapertussis in cases of suspected whooping cough (2011-2015) in Mar del Plata, Argentina].

Science.gov (United States)

Lavayén, Silvina; Zotta, Claudia; Cepeda, Marcela; Lara, Claudia; Rearte, Analia; Regueira, Mabel

2017-01-01

. To classify the study population in the Argentinian National Health Surveillance System framework, determine the proportion of infection by Bordetella pertussis and Bordetella parapertussis, and identify factors associated with the cases of suspected whooping cough attended to in the city of Mar del Plata and its outskirts during the period 2011- 2015. An observational and descriptive study was carried out. Clinical cases with suspicion of whooping cough were diagnosed by laboratory. The laboratory studies consisted of culture, PCR, and serology using the ELISA technique. A total of 572 cases were evaluated. The female sex was the most frequent (51.9%). The most frequent age range was 2 to 17 moths (51.1%; 290/568), which was also the group with the most confirmed cases. Only 47.8% (155/324) of the population studied had complete vaccination for their age. Whooping cough due to B. pertussis was confirmed in 15.5% (89/572) of cases and one case with B. parapertussis. Those cases that had contact with a coughing relative were significantly associated with the confirmation of Bordetella spp. by the laboratory (odds ratio: 3.3; 95% confidence interval: 1.9-5.4). The results show the need to suspect whooping cough and diagnose it early in children, adolescents, and adults in order to better control the disease. Likewise, continuing prevention and containment measures are fundamental in decreasing the circulation of the causal agent.

Genomic island excisions in Bordetella petrii

Directory of Open Access Journals (Sweden)

Levillain Erwan

2009-07-01

Full Text Available Abstract Background Among the members of the genus Bordetella B. petrii is unique, since it is the only species isolated from the environment, while the pathogenic Bordetellae are obligately associated with host organisms. Another feature distinguishing B. petrii from the other sequenced Bordetellae is the presence of a large number of mobile genetic elements including several large genomic regions with typical characteristics of genomic islands collectively known as integrative and conjugative elements (ICEs. These elements mainly encode accessory metabolic factors enabling this bacterium to grow on a large repertoire of aromatic compounds. Results During in vitro culture of Bordetella petrii colony variants appear frequently. We show that this variability can be attributed to the presence of a large number of metastable mobile genetic elements on its chromosome. In fact, the genome sequence of B. petrii revealed the presence of at least seven large genomic islands mostly encoding accessory metabolic functions involved in the degradation of aromatic compounds and detoxification of heavy metals. Four of these islands (termed GI1 to GI3 and GI6 are highly related to ICEclc of Pseudomonas knackmussii sp. strain B13. Here we present first data about the molecular characterization of these islands. We defined the exact borders of each island and we show that during standard culture of the bacteria these islands get excised from the chromosome. For all but one of these islands (GI5 we could detect circular intermediates. For the clc-like elements GI1 to GI3 of B. petrii we provide evidence that tandem insertion of these islands which all encode highly related integrases and attachment sites may also lead to incorporation of genomic DNA which originally was not part of the island and to the formation of huge composite islands. By integration of a tetracycline resistance cassette into GI3 we found this island to be rather unstable and to be lost from

Avian cholera causes marine bird mortality in the Bering Sea of Alaska

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Bodenstein, Barbara L.; Kimberlee Beckmen,; Gay Sheffield,; Kathy Kuletz,; Van Hemert, Caroline R.; Berlowski-Zier, Brenda M.; Shearn-Bochsler, Valerie I.

2015-01-01

The first known avian cholera outbreak among wild birds in Alaska occurred during November 2013. Liver, intestinal, and splenic necrosis consistent with avian cholera was noted, and Pasteurella multocida serotype 1 was isolated from liver and lung or spleen in Crested Auklets (Aethia cristatella), Thick-billed Murres (Uria lomvia), Common Eider (Somateria mollissima), Northern Fulmars (Fulmarus glacialis), and Glaucous-winged Gulls (Larus glaucescens).

Septicemic pasteurellosis in free-ranging neonatal pronghorn in Oregon

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Dunbar, Michael R.; Wolcott, Mark J.; Rimler, R.B.; Berlowski, Brenda M.

2000-01-01

As part of a study to determine the cause(s) of population decline and low survival of pronghorn (Antilocapra americana) neonates on Hart Mountain National Antelope Refuge (HMNAR), Oregon (USA), 55 of 104 neonates captured during May 1996 and 1997 were necropsied (n = 28, 1996; n = 27, 1997) to determine cause of death. Necropsies were conducted on fawns that died during May, June, or July of each year. The objectives of this study were to report the occurrence and pathology of pasteurellosis in neonates and determine if the isolated strain of Pasteurella multocida was unique. Septicemic pasteurellosis, caused by P. multocida, was diagnosed as the cause of death for two neonates in May and June 1997. Necropsy findings included widely scattered petechial and ecchymotic hemorrhages found over a large portion of the subcutaneous tissue, meninges of the brain, epicardium, skeletal muscle, and serosal surface of the thorasic and abdominal cavities. Histological examination of lung tissues revealed diffuse congestion and edema and moderate to marked multifocal infiltrate of macrophages, neutrophils, and numerous bacteria within many terminal bronchioles and alveoli. Pasteurella multocida serotypes A:3,4, and B:1 were isolated from several tissues including lung, intestinal, thorasic fluid, and heart blood. Each B:1 isolate had DNA restriction endonuclease fingerprint profiles distinct from isolates previously characterized from domestic cattle, swan (Olor spp.), moose (Alces alces), and pronghorn from Montana (USA). This is the first report of pasteurellosis in pronghorn from Oregon and the B:1 isolates appear to be unique in comparison to DNA fingerprint profiles from selected domestic and wild species.

Use of radioactively labelled bacteria in animal experiments. 4

International Nuclear Information System (INIS)

Flossmann, K.D.; Mueller, G.; Heilmann, P.; Finsterbusch, L.; Hubald, J.

1981-01-01

Intratracheal administration of 3 H-, 14 C-, 59 Fe- or 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 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. (author)

Survey of susceptibility to marbofloxacin in bacteria isolated from diseased pigs in Europe.

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El Garch, F; Kroemer, S; Galland, D; Morrissey, I; Woehrle, F

2017-06-17

A monitoring programme of marbofloxacin susceptibility of bacteria from Europe causing respiratory tract infection and meningitis in pigs has been active since 1994 and 2002, respectively. Monitoring digestive, metritis and urinary tract infection (UTI) in pigs has been active since 2005 and susceptibility results until 2013 are presented. Minimum inhibitory concentration (MIC) was determined by broth microdilution. For MIC interpretation, Vétoquinol-evaluated breakpoints were applied. For digestive pathogens, Escherichia coli and Salmonella species (1717 and 300 isolates, respectively) exhibited 7.5 per cent resistance in E coli and no resistance in Salmonella species. Similarly, E coli from metritis (369 isolates) had 7.0 per cent resistance to marbofloxacin. However, E coli from UTI (633 isolates) had higher resistance (10.4 per cent). For Streptococcus suis causing meningitis (585 isolates), marbofloxacin susceptibility was very high with only 0.5 per cent resistance and 0.4 per cent resistance was observed with S suis causing respiratory disease (729 isolates). Other respiratory pathogens were also highly susceptible to marbofloxacin with no resistance in Actinobacillus pleuropneumoniae (647 isolates) or Bordetella bronchiseptica (504 isolates), 0.1 per cent resistance in Pasteurella multocida (1373 isolates) and 1.4 per cent resistance in Haemophilus parasuis (145 isolates). There was no apparent change in marbofloxacin MIC over time for any bacterial pathogen based on MIC 50/90 These data confirm previously published MIC results from porcine and other animal infections. British Veterinary Association.

Isolation of obligate and facultative anaerobic bacteria from feline subcutaneous abscesses.

Science.gov (United States)

Hoshuyama, S; Kanoe, M; Amimoto, A

1996-03-01

A total of 113 specimens collected from purulent skin lesions of household cats was examined bacteriologically. Ninety seven isolates obtained from 74 specimens (65.5%). Of these, 11 specimens (9.7%) contained obligate anaerobes only, 18 specimens (15.9%) yielded both obligate and facultative anaerobes. In the obligate anaerobes detected, genus Fusobacterium was the most frequently observed and F. nucleatum was most common species. Pasteurella multocida was the facultative anaerobe which was most frequently detected.

Proteome analysis of Bordetella pertussis isolated from human macrophages

Czech Academy of Sciences Publication Activity Database

Lamberti, Y.; Cafiero, J.H.; Surmann, K.; Valdez, H.; Holubová, Jana; Večerek, Branislav; Šebo, Peter; Schmidt, F.; Völker, U.; Rodriguez, M.E.

2016-01-01

Roč. 136, MAY16 (2016), s. 55-67 ISSN 1874-3919 R&D Projects: GA MŠk(CZ) 7AMB14AR028 Institutional support: RVO:61388971 Keywords : Bordetella pertussis * Intracellular survival * Proteomics Subject RIV: EE - Microbiology, Virology Impact factor: 3.914, year: 2016

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

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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

Induction and maintenance of Bordetella pertussis specific immune responses

NARCIS (Netherlands)

Stenger, R.M.

2010-01-01

Pertussis, also referred to as whooping cough, is a serious respiratory disease mainly caused by the gram-negative bacterium Bordetella pertussis. The disease is most severe in neonates and children under the age of 1. Before childhood vaccination was introduced in the 1950s, pertussis was an

Occurrence of 3 Bordetella species during an outbreak of cough illness in Ohio: epidemiology, clinical features, laboratory findings and antimicrobial susceptibility.

Science.gov (United States)

Spicer, Kevin B; Salamon, Doug; Cummins, Carol; Leber, Amy; Rodgers, Loren E; Marcon, Mario J

2014-07-01

An increase in laboratory diagnosis of pertussis was noted in central Ohio during 2010. Diagnosis was made using a polymerase chain reaction assay targeting the multicopy insertion sequence IS481, which is found in both Bordetella pertussis (Bp) and Bordetella holmesii (Bh). An increase in specimens testing positive for Bordetella parapertussis (Bpp) using insertion sequence IS1001 was also noted. Nasopharyngeal swab specimens submitted April 1, 2010, to March 31, 2011, were tested using a multiplex polymerase chain reaction assay for Bp/Bh (IS481) and Bpp followed by singleplex assays for Bp and Bh. A subgroup of specimens was also cultured for Bordetella species, and antimicrobial susceptibility testing was performed on recovered organisms. Demographic and clinical features were compared for patients with Bp, Bh and Bpp. Of 520 IS481-positive specimens, 214 (41.1%) were positive for Bp, 79 (15.2%) were positive for Bh and 5 (1.0%) were positive for both Bp and Bh; 222 (42.7%) were negative for both targets. An additional 220 specimens were positive for Bpp. Among a sample of 155 IS481-positive specimens, 40, 15 and 0 were culture positive for Bp, Bh and Bpp, respectively. Among a sample of 55 BparaIS1001-positive (Bpp) specimens, 22, 0 and 0 were culture positive for Bpp, Bp and Bh, respectively. All Bordetella species were susceptible to macrolide antibiotics. Patients with Bh were older than patients with Bp, who were older than those positive for Bpp (mean ages: 12.0, 8.0 and 4.2 years, respectively; P Bpp and 100 negative for Bordetella species), but did not differ statistically among the groups (χ = 5.1, P = 0.17). All 3 Bordetella species, Bp, Bh and Bpp, were detected during on outbreak of pertussis-like cough illness. There were noted differences in age and seasonality, but clinical features at the time of presentation did not allow clear differentiation of these infections. All Bordetella species recovered from culture and tested were susceptible in

Antibacterial Activity of the Hydro-Alcoholic Extract of Juglans regia L. Stem Bark on Human Bacterial Infection

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Moori Bakhtiari N.* PhD,

2015-12-01

Full Text Available Aims Bovine mastitis continues to be the most costly disease to the dairy farmers. It dominates in Iran as one of the most prevalent diseases in dairy cattle among the dairy farms. Mastitis treatment with antibiotics leads to the development of antibiotic resistant strains and consumer health problem.This study was performed for the first time to analyze in vitro effects of hydro-alcoholic extract of Juglans regia L. stem bark on 6 mastitis pathogens. Materials & Methods the susceptibility of 6 strains of bacteria (Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Streptococcus spp., Pasteurella multocida and Mannheimia haemolytica were analyzed against hydro-alcoholic extract of Juglans regia L. stem bark with minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC methods. Findings Hydro-alcoholic extract did not have antibacterial effects on E. coli and K. pneumoniae. Minimum inhibitory concentration for S. aureus, P. multocida, M. haemolytica and Streptococcus spp. was 62.5mg/ml of hydro-alcoholic extract. There was not any significant response with concentrations below 100mg/disc on S. aureus, Streptococcus species, P. multocida and M. haemolytica. Minimum bactericidal concentration of this extract was 100mg/ml in all isolates. Conclusion Juglans regia L. have some antibacterial effects on S. aureus, P. multocida, M. haemolytica and Streptococcus species.

Birds in Human Modified Environments and Bird Damage Control: Social, Economic, and Health Implications

Science.gov (United States)

1989-12-01

Paratyphoid (Salmonella typhimurium) x x x + + .+ + + . + + + + Pasteurellasis (Fowl cholera ) (Pasteurella multocida) x x...enteritis (Clostridlum colinum) x 0 0 Vibriosis ( Vibrio fetu) x 30 Table 4 (ContWd Carriers r. .14 0 C c to to z ca -W V 0 4 w .,.4 ". Ca 0 3d ca DI .C...endemic vector of EEE in North America (Reeves et al. 1958, Howard and Wallis 1974). This species breeds in freshwater swamps from the Gulf of Mexico

Crucial role of antibodies to pertactin in Bordetella pertussis immunity

NARCIS (Netherlands)

Hellwig, SMM; Rodriguez, ME; Berbers, GAM; de Winkel, JGJV; Mooi, FR

2003-01-01

Pertussis, a serious infectious disease of the respiratory tract caused by Bordetella pertussis, is reemerging in vaccinated populations. Efforts to curtail this disease are hampered by limited insight into the basis of protective immunity. Opsonophagocytosis was recently found to play a central

Extracellular DNA is essential for maintaining Bordetella biofilm integrity on abiotic surfaces and in the upper respiratory tract of mice.

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Matt S Conover

2011-02-01

Full Text Available Bacteria form complex and highly elaborate surface adherent communities known as biofilms which are held together by a self-produced extracellular matrix. We have previously shown that by adopting a biofilm mode of existence in vivo, the gram negative bacterial pathogens Bordetella bronchiseptica and Bordetella pertussis are able to efficiently colonize and persist in the mammalian respiratory tract. In general, the bacterial biofilm matrix includes polysaccharides, proteins and extracellular DNA (eDNA. In this report, we investigated the function of DNA in Bordetella biofilm development. We show that DNA is a significant component of Bordetella biofilm matrix. Addition of DNase I at the initiation of biofilm growth inhibited biofilm formation. Treatment of pre-established mature biofilms formed under both static and flow conditions with DNase I led to a disruption of the biofilm biomass. We next investigated whether eDNA played a role in biofilms formed in the mouse respiratory tract. DNase I treatment of nasal biofilms caused considerable dissolution of the biofilm biomass. In conclusion, these results suggest that eDNA is a crucial structural matrix component of both in vitro and in vivo formed Bordetella biofilms. This is the first evidence for the ability of DNase I to disrupt bacterial biofilms formed on host organs.

A longitudinal study of serological patterns of respiratory infections in nine infected Danish swine herds

DEFF Research Database (Denmark)

Andreasen, Margit; Nielsen, Jens; Bækbo, Poul

2000-01-01

Sixteen litters of seven pigs from each of nine Danish farrow-to-finish herds were followed to investigate the serological patterns caused by natural infection with Mycoplasma hyponeumoniae, Pasteurella multocida toxin and Actinobacillus pleuroneumoniae serotypes 2, 5-7, 12. In seven of the herds......, pigs were followed as two separate cohorts started 4 weeks apart, and in two herds only one cohort was followed. A total of 999 pigs were included in the study. The pigs were blood sampled at weaning and subsequently every fourth week until slaughter. All pigs were examined for antibodies against M....... hyopneumoniae (enzyme-linked immunosorbent assay), P. multocida toxin (enzyme-linked immunosorbent assay) and A. pleuropneumoniae serotypes 2, 5-7, 12 (complement-fixation tests). The most-common pattern (28%) of seroconversion was that of pigs first seroconverting to A. pleuropneumoniae serotype 2, followed...

Occurrence of antimicrobial resistance in bacteria from diagnostic samples from dogs

DEFF Research Database (Denmark)

Pedersen, Karl; Pedersen, Kristina; Jensen, Helene

2007-01-01

Pseudomonas aeruginosa, 25 Pasteurella multocida, 29 Proteus spp. and 449 Escherichia coli isolates from clinical submissions from dogs were determined by a broth-dilution method for determination of minimal inhibitory concentration. Data for consumption of antimicrobials were retrieved from Vet....... intermedius and Proteus isolates. Conclusions: This investigation provided data on occurrence of antimicrobial resistance in important pathogenic bacteria from dogs, which may be useful for the small animal practitioner. Resistance was low to the compounds that were most often used, but unfortunately...

Significant Decrease in Pertactin-Deficient Bordetella pertussis Isolates, Japan

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

2017-01-01

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

Identification of Bordetella bronchseptica in fatal pneumonia of dogs and cats

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Infection with Bordetella bronchiseptica is a common cause of tracheobronchitis and upper respiratory disease in dogs and cats, but it can also lead to fatal pneumonia. Identification of this pathogen is important due the risk of transmission to other animals, availability of vaccines and potential...

Avian cholera in waterfowl: the role of lesser snow and Ross's geese as carriers of avian cholera in the Playa Lakes region

Science.gov (United States)

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

2005-01-01

We collected samples from apparently healthy geese in the Playa Lakes Region (USA) during the winters of 2000a??01 and 2001a??02 to determine whether carriers of Pasteurella multocida, the bacterium that causes avian cholera, were present in wild populations. With the use of methods developed in laboratory challenge trials (Samuel et al., 2003a) and a serotype-specific polymerase chain reaction method for identification of P. multocida serotype 1, we found that a small proportion of 322 wild birds (cholera infection. Our results confirm the hypothesis that wild waterfowl are carriers of avian cholera and add support for the hypothesis that wild birds are a reservoir for this disease. In concert with other research, this work indicates that enzootic infection with avian cholera occurs in lesser snow goose (Chen caerulescens caerulescens) populations throughout their annual cycle. Although fewer Rossa??s geese (Chen rossii) were sampled, we also found these birds were carriers of P. multocida. Even in the absence of disease outbreaks, serologic evidence indicates that chronic disease transmission and recent infection are apparently occurring year-round in these highly gregarious birds and that a small portion of these populations are potential carriers with active infection.

Polymorphisms influencing expression of dermonecrotic toxin in Bordetella bronchiseptica.

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Keisuke Okada

Full Text Available Bordetella bronchiseptica is a pathogenic bacterium causing respiratory infections in a broad range of mammals. Recently, we determined the whole genome sequence of B. bronchiseptica S798 strain isolated from a pig infected with atrophic rhinitis and found four single-nucleotide polymorphisms (SNPs at positions -129, -72, +22, and +38 in the region upstream of dnt encoding dermonecrotic toxin (DNT, when compared with a rabbit isolate, RB50. DNT is known to be involved in turbinate atrophy observed in atrophic rhinitis. Immunoblotting, quantitative real-time PCR, and β-galactosidase reporter assay revealed that these SNPs resulted in the increased promoter activity of dnt and conferred the increased ability to produce DNT on the bacteria. Similar or identical SNPs were also found in other pig isolates kept in our laboratory, all of which produce a larger amount of DNT than RB50. Our analysis revealed that substitution of at least two of the four bases, at positions -72 and +22, influenced the promoter activity for dnt. These results imply that these SNPs are involved in the pathogenicity of bordetellae specific to pig diseases.

An Investigation of the Pathology and Pathogens Associated with Porcine Respiratory Disease Complex in Denmark

DEFF Research Database (Denmark)

Hansen, Mette Sif; Pors, S. E.; Jensen, H. E.

2010-01-01

), porcine reproductive and respiratory syndrome virus (both European and US type), porcine circovirus type 2 (PCV2), porcine respiratory coronavirus, porcine cytomegalovirus, Mycoplasma hyopneumoniae and Mycoplasma hyorhinis. All cases had cranioventral lobular bronchopneumonia consistent with PRDC....... There was a broad range of microscopical lesions and the cases were characterized as acute (n=10), subacute (n=24) or chronic (n=114) bronchopneumonia. Five bacterial species, five viruses and two Mycoplasma spp. were detected in different combinations. PCV2, M. hyopneumoniae, M. hyorhinis and Pasteurella multocida...

Fatal pneumonia of bighorn sheep following association with domestic sheep.

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Foreyt, W J; Jessup, D A

1982-04-01

During 1979-1980 acute fibrinopurulent bronchopneumonia resulted in high mortality or total loss of herds of bighorn sheep (Ovis canadensis) in California and Washington. Contact with domestic sheep occurred shortly before the onset of disease in each case. Circumstantial evidence indicated that the apparently healthy domestic sheep transmitted pathogenic bacteria to the bighorns, resulting in mortality. Pasteurella multocida and Corynebacterium pyogenes were isolated from pulmonary tissue of dead bighorns. The presence of domestic sheep may have been an important stress which initiated or compounded the disease.

Biased distribution of DNA uptake sequences towards genome maintenance genes

DEFF Research Database (Denmark)

Davidsen, T.; Rodland, E.A.; Lagesen, K.

2004-01-01

Repeated sequence signatures are characteristic features of all genomic DNA. We have made a rigorous search for repeat genomic sequences in the human pathogens Neisseria meningitidis, Neisseria gonorrhoeae and Haemophilus influenzae and found that by far the most frequent 9-10mers residing within...... in these organisms. Pasteurella multocida also displayed high frequencies of a putative DUS identical to that previously identified in H. influenzae and with a skewed distribution towards genome maintenance genes, indicating that this bacterium might be transformation competent under certain conditions....

Bordetella pertussis pathogenesis: current and future challenges

Science.gov (United States)

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

2014-01-01

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

Antimicrobial susceptibility monitoring of respiratory tract pathogens isolated from diseased cattle and pigs across Europe: the VetPath study.

Science.gov (United States)

de Jong, Anno; Thomas, Valérie; Simjee, Shabbir; Moyaert, Hilde; El Garch, Farid; Maher, Kirsty; Morrissey, Ian; Butty, Pascal; Klein, Ulrich; Marion, Hervé; Rigaut, Delphine; Vallé, Michel

2014-08-06

VetPath is an ongoing pan-European antibiotic susceptibility monitoring programme collecting pathogens from diseased antimicrobial non-treated cattle, pigs and poultry. In the current study, 1001 isolates from cattle and pig respiratory tract infections were tested for their antimicrobial susceptibilities. Non-replicate lung samples or nasopharyngeal/nasal swabs were collected from animals with acute clinical signs in 11 countries during 2002-2006. Pasteurella multocida and Mannheimia haemolytica from cattle and P. multocida, Actinobacillus pleuropneumoniae and Streptococcus suis from pigs were isolated by standard methods. S. suis was also isolated from meningitis cases. MICs of 16 antibiotics were assessed centrally by broth microdilution following CLSI recommendations. Results were interpreted using CLSI breakpoints where available. P. multocida (231) and M. haemolytica (138) isolates were all susceptible to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin and trimethoprim/sulfamethoxazole. Resistance to florfenicol and spectinomycin was 0.4% and 3.5% in P. multocida, respectively, and absent in M. haemolytica isolates. Tetracycline resistance was 5.7% and 14.6% for P. multocida and M. haemolytica. In pigs, 230 P. multocida, 220 A. pleuropneumoniae and 182 S. suis isolates were recovered. Resistance to amoxicillin/clavulanic acid, ceftiofur, enrofloxacin, florfenicol, tiamulin and tilmicosin was absent or <1%. Trimethoprim/sulfamethoxazole resistance was 3-6% and tetracycline resistance varied from 14.7% in A. pleuropneumoniae to 81.8% in S. suis. In conclusion, low resistance to antibiotics with defined clinical breakpoints, except for tetracycline, was observed among the major respiratory tract pathogens recovered from cattle and pigs. Since for approximately half of the antibiotics in this panel no CLSI-defined breakpoints were available, setting of the missing veterinary breakpoints is important. Copyright © 2014 Elsevier B.V. All rights reserved.

Microbiological and serological monitoring in hooded crow (Corvus corone cornix in the Region Lombardia, Italy

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Guido Grilli

2010-01-01

Full Text Available The health status of 276 hooded crows (Corvus corone cornix from various provinces of Lombardy was monitored for three years. Bacteriological examination detected E. coli (76%, Campylobacter jejuni (17%, Salmonella typhimurium (11.6%, Yersinia spp. (6.5%, Clamydophila abortus and C. psittaci (2.6%; from six birds showing severe prostration Pasteurella multocida was isolated. Virological and serological tests were negative for Avian Influenza virus (AIV, West Nile virus (WNV and only three samples were positive for Newcastle disease virus (NDV but only at serology (titre 1:16.

Filamentous hemagglutinin of Bordetella pertussis: a key adhesin with immunomodulatory properties?

Czech Academy of Sciences Publication Activity Database

Romero, Rodrigo, Villarino; Osička, Radim; Šebo, Peter

2014-01-01

Roč. 9, č. 12 (2014), s. 1339-1360 ISSN 1746-0913 R&D Projects: GA ČR(CZ) P302/11/0580; GA ČR(CZ) GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella * adhesion * integrins * filamentous hemagglutinin Subject RIV: EE - Microbiology, Virology Impact factor: 4.275, year: 2014

A real-time PCR assay with improved specificity for detection and discrimination of all clinically relevant Bordetella species by the presence and distribution of three Insertion Sequence elements

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Ossewaarde Jacobus M

2011-01-01

Full Text Available Abstract Background In Dutch laboratories molecular detection of B. pertussis and B. parapertussis is commonly based on insertion sequences IS481 and IS1001, respectively. Both IS elements are more widely spread among Bordetella species. Both Bordetella holmesii, and B. bronchiseptica can harbour IS481. Also, IS1001 is found among B. bronchiseptica. IS481, and IS1001 based PCR thus lacks specificity when used for detection of specific Bordetella spp. Findings We designed a PCR based on IS1002, another IS element that is present among Bordetella species, and exploited it as a template in combination with PCR for IS481, and IS1001. In combining the PCRs for IS481, IS1001, and IS1002, and including an inhibition control, we were able to detect and discriminate all clinically relevant Bordetella species. Conclusions We developed an improved PCR method for specific detection of B. pertussis, B. parapertussis, B. holmesii, and B. bronchiseptica.

Bordetella petrii recovered from chronic pansinusitis in an adult with cystic fibrosis

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Laura Biederman

2015-01-01

Full Text Available To date Bordetella petrii has infrequently been identified within the clinical setting likely due to the asaccharolytic nature of this organism. We present a case of B. petrii recovered on two separate events in a patient with adult cystic fibrosis experiencing chronic pansinusitis.

Bordetella pertussis pertactin knock-out strains reveal immunomodulatory properties of this virulence factor.

NARCIS (Netherlands)

Hovingh, Elise Sofie; Mariman, Rob; Solans, Luis; Hijdra, Daniëlle; Hamstra, Hendrik-Jan; Jongerius, Ilse; van Gent, Marjolein; Mooi, Frits; Locht, Camille; Pinelli, Elena

2018-01-01

Whooping cough, caused by Bordetella pertussis, has resurged and presents a global health burden worldwide. B. pertussis strains unable to produce the acellular pertussis vaccine component pertactin (Prn), have been emerging and in some countries represent up to 95% of recent clinical isolates.

Real-time PCR-based detection of Bordetella pertussis and Bordetella parapertussis in an Irish paediatric population.

LENUS (Irish Health Repository)

Grogan, Juanita A

2011-06-01

Novel real-time PCR assays targeting the Bordetella pertussis insertion sequence IS481, the toxin promoter region and Bordetella parapertussis insertion sequence IS1001 were designed. PCR assays were capable of detecting ≤10 copies of target DNA per reaction, with an amplification efficiency of ≥90 %. From September 2003 to December 2009, per-nasal swabs and nasopharyngeal aspirates submitted for B. pertussis culture from patients ≤1 month to >15 years of age were examined by real-time PCR. Among 1324 patients, 76 (5.7 %) were B. pertussis culture positive and 145 (10.95 %) were B. pertussis PCR positive. Of the B. pertussis PCR-positive patients, 117 (81 %) were aged 6 months or less. A total of 1548 samples were examined, of which 87 (5.6 %) were culture positive for B. pertussis and 169 (10.92 %) were B. pertussis PCR positive. All culture-positive samples were PCR positive. Seven specimens (0.5 %) were B. parapertussis culture positive and 10 (0.8 %) were B. parapertussis PCR positive, with all culture-positive samples yielding PCR-positive results. A review of patient laboratory records showed that of the 1324 patients tested for pertussis 555 (42 %) had samples referred for respiratory syncytial virus (RSV) testing and 165 (30 %) were positive, as compared to 19.4 % of the total 5719 patients tested for RSV in this period. Analysis of the age distribution of RSV-positive patients identified that 129 (78 %) were aged 6 months or less, similar to the incidence observed for pertussis in that patient age group. In conclusion, the introduction of the real-time PCR assays for the routine detection of B. pertussis resulted in a 91 % increase in the detection of the organism as compared to microbiological culture. The incidence of infection with B. parapertussis is low while the incidence of RSV infection in infants suspected of having pertussis is high, with a similar age distribution to B. pertussis infection.

Molecular evolution of the two-component system BvgAS involved in virulence regulation in Bordetella.

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Julien Herrou

Full Text Available The whooping cough agent Bordetella pertussis is closely related to Bordetella bronchiseptica, which is responsible for chronic respiratory infections in various mammals and is occasionally found in humans, and to Bordetella parapertussis, one lineage of which causes mild whooping cough in humans and the other ovine respiratory infections. All three species produce similar sets of virulence factors that are co-regulated by the two-component system BvgAS. We characterized the molecular diversity of BvgAS in Bordetella by sequencing the two genes from a large number of diverse isolates. The response regulator BvgA is virtually invariant, indicating strong functional constraints. In contrast, the multi-domain sensor kinase BvgS has evolved into two different types. The pertussis type is found in B. pertussis and in a lineage of essentially human-associated B. bronchiseptica, while the bronchiseptica type is associated with the majority of B. bronchiseptica and both ovine and human B. parapertussis. BvgS is monomorphic in B. pertussis, suggesting optimal adaptation or a recent population bottleneck. The degree of diversity of the bronchiseptica type BvgS is markedly different between domains, indicating distinct evolutionary pressures. Thus, absolute conservation of the putative solute-binding cavities of the two periplasmic Venus Fly Trap (VFT domains suggests that common signals are perceived in all three species, while the external surfaces of these domains vary more extensively. Co-evolution of the surfaces of the two VFT domains in each type and domain swapping experiments indicate that signal transduction in the periplasmic region may be type-specific. The two distinct evolutionary solutions for BvgS confirm that B. pertussis has emerged from a specific B. bronchiseptica lineage. The invariant regions of BvgS point to essential parts for its molecular mechanism, while the variable regions may indicate adaptations to different lifestyles. The

Development of pneumonia in desert bighorn sheep after exposure to a flock of exotic wild and domestic sheep.

Science.gov (United States)

Callan, R J; Bunch, T D; Workman, G W; Mock, R E

1991-03-15

From 1986 to 1989, 5 desert bighorn sheep (3 Ovis canadensis mexicana and 2 O c nelsoni), ranging in age from 2 to 3 years, were exposed to a flock of exotic wild and domestic sheep to potentially achieve naturally acquired pneumonia. Pasteurella multocida was isolated from nasal samples from 4 of 6 sheep randomly sampled from the flock. Bighorn sheep were exposed individually and each exposure period was a trial. Treatment before and after exposure varied and included combinations of alpha interferon, antibiotics, anti-inflammatory drugs, and vaccines. Treatments were chosen on the basis of recommendations of others for treating pneumonia in desert bighorn sheep as well as our own experience in sheep and cattle. Regardless of treatment used, bighorn sheep in trials 1 to 4 developed signs of pneumonia within 10 to 14 days of exposure. Bighorn sheep in trials 1 to 3 died within 11 to 17 days of initial exposure. In trial 4, the bighorn sheep was isolated from the carrier sheep for treatment of pneumonia on day 14 and died on day 30. Pasteurella multocida was isolated from lung tissue in 3 of the 4 bighorn sheep. On the basis of results of trials 1 to 4, a more in depth clinical study was conducted in trial 5. Nasal and blood specimens were collected prior to and during trial 5 for bacteriologic culturing and serologic testing for bovine viral diarrhea virus, infectious bovine rhinotracheitis, parainfluenza-3 virus, and respiratory syncytial virus.(ABSTRACT TRUNCATED AT 250 WORDS)

[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...

Pasteurella testudinis associated with respiratory disease and septicaemia in leopard (Geochelone pardalis and other tortoises in South Africa

Directory of Open Access Journals (Sweden)

M.M. Henton

2003-07-01

Full Text Available The first recorded isolates of Pasteurella testudinis from South African tortoises kept in captivity is presented. P. testudinis was found in association with respiratory disease in affected animals.

RESPIRATORY SYNDROME: A MAJOR THREAT TO THE LIVESTOCK FARMERS AND ITS ECONOMIC IMPACT

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A. B. ZAHUR, U. FAROOQ, M. HUSSAIN1, S. H. HASHMI2 AND R. MUNEER

2007-10-01

Full Text Available Epidemiology of a respiratory syndrome was studied at Landhi Dairy Colony (LDC, Karachi, Pakistan and its economic impact was estimated. Among 5889 buffaloes examined, 2.3% animals were suffering from this syndrome. From some of the sick animals, Pasteurella multocida, the causative agent of haemorrhagic septicaemia, was isolated. In the present study, an average loss of Rs. 0.2 million per farm was calculated and the extrapolated values for 0.2 and 0.8 million animals present in LDC and other dairy colonies in Karachi were Rs. 225.6 and Rs. 1128.1 million, respectively.

Seroprevalence of Bordetella pertussis among vaccinated and unvaccinated pregnant women and newborn infants in a university hospital of Buenos Aires.

Science.gov (United States)

Bosch, Juan J; Fernández, Hilaria; Polak, Fernando P; Musante, Gabriel; Libster, Romina; Rocca Rivarola, Manuel

2017-08-01

Pertussis is a highly contagious disease caused by Bordetella pertussis. It poses a high morbidity and mortality rate, especially among infants younger than 6 months old. In Argentina, pertussis incidence and mortality have increased over the past three decades. To establish Bordetella pertussisantibody titers among pregnant women in their third trimester and among newborn infants, as measured in cord blood. This was an observational, cross-sectional study. The study started in 2011; at that time, pertussis vaccination was not mandatory for pregnant women as per the national immunization schedule, only optional. Maternal antibodies were measured in the last trimester of pregnancy for women and in cord blood for newborn infants. Antibody titers were determined using Abcam's anti-Bordetella pertussis toxin (PT) IgG in vitro ELISA kit. The χ² test was used to compare prevalence rates. The study included 111 mother-newborn infant dyads; 35 infants from unvaccinated mothers (before the introduction of the vaccine) and 76 from vaccinated mothers. Positive IgG antibodies were found in 92% (70/76) of infants born from vaccinated mothers whereas 100% (35/35) of infants born from unvaccinated mothers had negative results for antibodies; p < 0.001. In the vaccinated population of this study, 92% of infants had positive IgG antibodies. This study supports the need for maternal immunization against Bordetella pertussis to provide protection to newborn infants. Sociedad Argentina de Pediatría

Association of Bordetella dermonecrotic toxin with the extracellular matrix

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Miyake Masami

2010-09-01

Full Text Available Abstract Background Bordetella dermonecrotic toxin (DNT causes the turbinate atrophy in swine atrophic rhinitis, caused by a Bordetella bronchiseptica infection of pigs, by inhibiting osteoblastic differentiation. The toxin is not actively secreted from the bacteria, and is presumed to be present in only small amounts in infected areas. How such small amounts can affect target tissues is unknown. Results Fluorescence microscopy revealed that DNT associated with a fibrillar structure developed on cultured cells. A cellular component cross-linked with DNT conjugated with a cross-linker was identified as fibronectin by mass spectrometry. Colocalization of the fibronectin network on the cells with DNT was also observed by fluorescence microscope. Several lines of evidence suggested that DNT interacts with fibronectin not directly, but through another cellular component that remains to be identified. The colocalization was observed in not only DNT-sensitive cells but also insensitive cells, indicating that the fibronectin network neither serves as a receptor for the toxin nor is involved in the intoxicating procedures. The fibronectin network-associated toxin was easily liberated when the concentration of toxin in the local environment decreased, and was still active. Conclusions Components in the extracellular matrix are known to regulate activities of various growth factors by binding and liberating them in response to alterations in the extracellular environment. Similarly, the fibronectin-based extracellular matrix may function as a temporary storage system for DNT, enabling small amounts of the toxin to efficiently affect target tissues or cells.

Effects of Enrofloxacin on Porcine Phagocytic Function

Science.gov (United States)

Schoevers, E. J.; van Leengoed, L. A. M. G.; Verheijden, J. H. M.; Niewold, T. A.

1999-01-01

The interaction between enrofloxacin and porcine phagocytes was studied with clinically relevant concentrations of enrofloxacin. Enrofloxacin accumulated in phagocytes, with cellular concentration/extracellular concentration ratios of 9 for polymorphonuclear leukocytes (PMNs) and 5 for alveolar macrophages (AMs). Cells with accumulated enrofloxacin brought into enrofloxacin-free medium released approximately 80% (AMs) to 90% (PMNs) of their enrofloxacin within the first 10 min, after which no further release was seen. Enrofloxacin affected neither the viability of PMNs and AMs nor the chemotaxis of PMNs at concentrations ranging from 0 to 10 μg/ml. Enrofloxacin (0.5 μg/ml) did not alter the capability of PMNs and AMs to phagocytize fluorescent microparticles or Actinobacillus pleuropneumoniae, Pasteurella multocida, and Staphylococcus aureus. Significant differences in intracellular killing were seen with enrofloxacin at 5× the MIC compared with that for controls not treated with enrofloxacin. PMNs killed all S. aureus isolates in 3 h with or without enrofloxacin. Intracellular S. aureus isolates in AMs were less susceptible than extracellular S. aureus isolates to the bactericidal effect of enrofloxacin. P. multocida was not phagocytosed by PMNs. AMs did not kill P. multocida, and similar intra- and extracellular reductions of P. multocida isolates by enrofloxacin were found. Intraphagocytic killing of A. pleuropneumoniae was significantly enhanced by enrofloxacin at 5× the MIC in both PMNs and AMs. AMs are very susceptible to the A. pleuropneumoniae cytotoxin. This suggests that in serologically naive pigs the enhancing effect of enrofloxacin on the bactericidal action of PMNs may have clinical relevance. PMID:10471554

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...

Third Acivity of Bordetella Adenylate Cyclase (AC) Toxin-Hemolysin

Czech Academy of Sciences Publication Activity Database

Fišer, Radovan; Mašín, Jiří; Basler, Marek; Krůšek, Jan; Špuláková, V.; Konopásek, Ivo; Šebo, Peter

2007-01-01

Roč. 282, č. 5 (2007), s. 2808-2820 ISSN 0021-9258 R&D Projects: GA MŠk 1M0506; GA AV ČR IAA5020406 Grant - others:XE(XE) LSHB-CT-2003-503582; Univerzita Karlova(CZ) 146/2005/B-BIO Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50110509 Source of funding: R - rámcový projekt EK ; V - iné verejné zdroje Keywords : bordetella * adenylate cyclase toxin * enzymatic aktivity Subject RIV: EE - Microbiology, Virology Impact factor: 5.581, year: 2007

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...... strains with the same rpoB sequence type, as shown by the PFGE profiles. The investigation documented that members of biovar Heyl of [P.] pneumotropica caused disease outbreaks in mouse colonies since the clonality indicated a primary role of [P.] pneumotropica biovar Heyl in the infections observed....

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...

Bordetella pertussis Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers

Czech Academy of Sciences Publication Activity Database

Hasan, Shakir; Kulkarni, N.N.; Asbjarnarson, A.; Linhartová, Irena; Osička, Radim; Šebo, Peter; Gudmundsson, H.

2018-01-01

Roč. 86, č. 3 (2018), č. článku e00445-17. ISSN 0019-9567 R&D Projects: GA ČR GA15-09157S; GA MZd(CZ) NV16-28126A; GA MŠk(CZ) LM2015064 Institutional support: RVO:61388971 Keywords : Bordetella pertussis * airway epithelia * CyaA Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 3.593, year: 2016

Standard PK/PD concepts can be applied to determine a dosage regimen for a macrolide: the case of tulathromycin in the calf.

Science.gov (United States)

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

2017-01-01

The pharmacokinetic (PK) profile of tulathromycin, administered to calves subcutaneously at the dosage of 2.5 mg/kg, was established in serum, inflamed (exudate), and noninflamed (transudate) fluids in a tissue cage model. The PK profile of tulathromycin was also established in pneumonic calves. For Mannheimia haemolytica and Pasteurella multocida, tulathromycin minimum inhibitory concentrations (MIC) were approximately 50 times lower in calf serum than in Mueller-Hinton broth. The breakpoint value of the PK/pharmacodynamic (PD) index (AUC (0-24 h) /MIC) to achieve a bactericidal effect was estimated from in vitro time-kill studies to be approximately 24 h for M. haemolytica and P. multocida. A population model was developed from healthy and pneumonic calves and, using Monte Carlo simulations, PK/PD cutoffs required for the development of antimicrobial susceptibility testing (AST) were determined. The population distributions of tulathromycin doses were established by Monte Carlo computation (MCC). The computation predicted a target attainment rate (TAR) for a tulathromycin dosage of 2.5 mg/kg of 66% for M. haemolytica and 87% for P. multocida. The findings indicate that free tulathromycin concentrations in serum suffice to explain the efficacy of single-dose tulathromycin in clinical use, and that a dosage regimen can be computed for tulathromycin using classical PK/PD concepts. © 2016 John Wiley & Sons Ltd.

Bordetella Pertussis virulence factors in the continuing evolution of whooping cough vaccines for improved performance.

NARCIS (Netherlands)

Dorji, Dorji; Mooi, Frits; Yantorno, Osvaldo; Deora, Rajendar; Graham, Ross M; Mukkur, Trilochan K

Despite high vaccine coverage, whooping cough caused by Bordetella pertussis remains one of the most common vaccine-preventable diseases worldwide. Introduction of whole-cell pertussis (wP) vaccines in the 1940s and acellular pertussis (aP) vaccines in 1990s reduced the mortality due to pertussis.

Evidence for an intracellular niche for Bordetella pertussis in broncho-alveolar lavage cells of mice

NARCIS (Netherlands)

Hellwig, SMM; Hazenbos, WLW; van de Winkel, JGJ; Mooi, FR

1999-01-01

Bordetella pertussis can attach, invade and survive intracellularly in human macrophages in vitro. To study the significance of this bacterial feature in vivo, we analyzed the presence of viable bacteria in broncho-alveolar lavage (BAL) cells of mice infected with B, pertussis. We found B. pertussis

Uptake of [14C]nicotinic acid and [14C]nicotinamide by Bordetella pertussis

International Nuclear Information System (INIS)

McPheat, W.L.; Wardlaw, A.C.

1980-01-01

Bordetella pertussis has an absolute requirement for either nicotinic acid (NA) or nicotinamide (ND), both being equally effective in supporting growth. The results of an investigation to compare the rates of uptake of NA and ND, to determine the influence of factors such as energy source and temperature on uptake, and to measure the Ksub(m) and Vsub(max) values are reported. (Auth.)

A Multiplex PCR for Detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in Clinical Specimens

National Research Council Canada - National Science Library

McDonough, E. A; Barrozo, C. P; Russell, K. L; Metzgar, D

2005-01-01

..., and Bordetella pertussis in uncultured patient specimens. These organisms cause similar symptomologies and are often not diagnosed because they are difficult to identify with classical methods such as culture and serology...

Dicty_cDB: AFK338 [Dicty_cDB

Lifescience Database Archive (English)

Full Text Available AF (Link to library) AFK338 (Link to dictyBase) - - - - AFK338P (Link to Original s...ite) AFK338F 659 AFK338Z 221 AFK338P 880 - - Show AFK338 Library AF (Link to library) Clone ID AFK338 (Link to dic...01175_2502( CP001175 |pid:none) Listeria monocytogenes HCC23, c... 40 0.099 AY714838_16( AY714838 |pid:none) Unculture...A sequence. 44 3.0 1 AE006080 |AE006080.1 Pasteurella multocida PM70 section 47 o... 0.003 BX294148_242( BX294148 |pid:none) Rhodopirellula baltica SH 1 comp... 44 0.005 CP000910_1528( CP00091

Retrospective Analysis of Bacterial and Viral Co-Infections in Pneumocystis spp. Positive Lung Samples of Austrian Pigs with Pneumonia.

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Christiane Weissenbacher-Lang

Full Text Available Aim of this study was the retrospective investigation of viral (porcine circovirus type 2 (PCV2, porcine reproductive and respiratory syndrome virus (PRRSV, torque teno sus virus type 1 and 2 (TTSuV1, TTSuV2 and bacterial (Bordetella bronchiseptica (B. b., Mycoplasma hyopneumoniae (M. h., and Pasteurella multocida (P. m. co-infections in 110 Pneumocystis spp. positive lung samples of Austrian pigs with pneumonia. Fifty-one % were positive for PCV2, 7% for PRRSV, 22% for TTSuV1, 48% for TTSuV2, 6% for B. b., 29% for M. h., and 21% for P. m. In 38.2% only viral, in 3.6% only bacterial and in 40.0% both, viral and bacterial pathogens were detected. In 29.1% of the cases a co-infection with 1 pathogen, in 28.2% with 2, in 17.3% with 3, and in 7.3% with 4 different infectious agents were observed. The exposure to Pneumocystis significantly decreased the risk of a co-infection with PRRSV in weaning piglets; all other odds ratios were not significant. Four categories of results were compared: I = P. spp. + only viral co-infectants, II = P. spp. + both viral and bacterial co-infectants, III = P. spp. + only bacterial co-infectants, and IV = P. spp. single infection. The evaluation of all samples and the age class of the weaning piglets resulted in a predomination of the categories I and II. In contrast, the suckling piglets showed more samples of category I and IV. In the group of fattening pigs, category II predominated. Suckling piglets can be infected with P. spp. early in life. With increasing age this single infections can be complicated by co-infections with other respiratory diseases.

Channel formation in model membranes by the adenylate cyclase toxin of Bordetella pertussis: Effect if Calcium

Czech Academy of Sciences Publication Activity Database

Knapp, O.; Maier, E.; Polleichtner, G.; Mašín, Jiří; Šebo, Peter; Benz, R.

2003-01-01

Roč. 42, - (2003), s. 8077-8084 ISSN 0006-2960 R&D Projects: GA AV ČR IPP1050128 Institutional research plan: CEZ:AV0Z5020903 Keywords : bordetella pertussis * calcium Subject RIV: EE - Microbiology, Virology Impact factor: 3.922, year: 2003

Bordetella pertussis evolution in the (functional) genomics era

Science.gov (United States)

Belcher, Thomas; Preston, Andrew

2015-01-01

The incidence of whooping cough caused by Bordetella pertussis in many developed countries has risen dramatically in recent years. This has been linked to the use of an acellular pertussis vaccine. In addition, it is thought that B. pertussis is adapting under acellular vaccine mediated immune selection pressure, towards vaccine escape. Genomics-based approaches have revolutionized the ability to resolve the fine structure of the global B. pertussis population and its evolution during the era of vaccination. Here, we discuss the current picture of B. pertussis evolution and diversity in the light of the current resurgence, highlight import questions raised by recent studies in this area and discuss the role that functional genomics can play in addressing current knowledge gaps. PMID:26297914

Experimental contact transmission of Pasteurella haemolytica from clinically normal domestic sheep causing pneumonia in Rocky Mountain bighorn sheep.

Science.gov (United States)

Onderka, D K; Wishart, W D

1988-10-01

Two Rocky Mountain bighorn lambs (Ovis canadensis canadensis) were held in captivity for 120 days before being housed with two domestic sheep. The lambs were clinically normal and had no Pasteurella spp. on nasal swab cultures. The domestic sheep were known to carry Pasteurella haemolytica biotype A in the nasal passages. After being in close contact for 19 days. P. haemolytica biotype A was cultured from nasal swabs of one of the bighorn lambs. By 26 days, both bighorn sheep developed coughs, were anorectic and became lethargic and nasal swabs yielded P. haemolytica biotype T, serotype 10. Twenty-nine days after contact, the lambs were necropsied and found to have extensive fibrinous bronchopneumonia. From affected tissues pure cultures of beta-hemolytic P. haemolytica biotype T, serotype 10 were grown. Both domestic sheep remained clinically normal and had no gross or microscopic lesions, but they carried the same P. haemolytica serotype in their tonsils. Behavioural observations gave no indication of stress in the bighorn lambs.

sigE facilitates the adaptation of Bordetella bronchiseptica to stress conditions and lethal infection in immunocompromised mice

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Barchinger Sarah E

2012-08-01

Full Text Available Abstract Background The cell envelope of a bacterial pathogen can be damaged by harsh conditions in the environment outside a host and by immune factors during infection. Cell envelope stress responses preserve the integrity of this essential compartment and are often required for virulence. Bordetella species are important respiratory pathogens that possess a large number of putative transcription factors. However, no cell envelope stress responses have been described in these species. Among the putative Bordetella transcription factors are a number of genes belonging to the extracytoplasmic function (ECF group of alternative sigma factors, some of which are known to mediate cell envelope stress responses in other bacteria. Here we investigate the role of one such gene, sigE, in stress survival and pathogenesis of Bordetella bronchiseptica. Results We demonstrate that sigE encodes a functional sigma factor that mediates a cell envelope stress response. Mutants of B. bronchiseptica strain RB50 lacking sigE are more sensitive to high temperature, ethanol, and perturbation of the envelope by SDS-EDTA and certain β-lactam antibiotics. Using a series of immunocompromised mice deficient in different components of the innate and adaptive immune responses, we show that SigE plays an important role in evading the innate immune response during lethal infections of mice lacking B cells and T cells. SigE is not required, however, for colonization of the respiratory tract of immunocompetent mice. The sigE mutant is more efficiently phagocytosed and killed by peripheral blood polymorphonuclear leukocytes (PMNs than RB50, and exhibits decreased cytotoxicity toward macrophages. These altered interactions with phagocytes could contribute to the defects observed during lethal infection. Conclusions Much of the work on transcriptional regulation during infection in B. bronchiseptica has focused on the BvgAS two-component system. This study reveals that the Sig

Seroprevalence of pertussis in the Gambia : evidence for continued circulation of bordetella pertussis despite high vaccination rates

NARCIS (Netherlands)

Scott, Susana; van der Sande, Marianne; Faye-Joof, Tisbeh; Mendy, Maimuna; Sanneh, Bakary; Barry Jallow, Fatou; de Melker, Hester; van der Klis, Fiona; van Gageldonk, Pieter; Mooi, Frits; Kampmann, Beate

BACKGROUND: Bordetella pertussis can cause severe respiratory disease and death in children. In recent years, large outbreaks have occurred in high-income countries; however, little is known about pertussis incidence in sub-Saharan Africa. METHODS: We evaluated antibody responses to pertussis toxin

Evaluation of Amplification Targets for the Specific Detection of Bordetella pertussis Using Real-Time Polymerase Chain Reaction

Directory of Open Access Journals (Sweden)

Mohammad Rubayet Hasan

2014-01-01

Full Text Available BACKGROUND: Bordetella pertussis infections continue to be a major public health challenge in Canada. Polymerase chain reaction (PCR assays to detect B pertussis are typically based on the multicopy insertion sequence IS481, which offers high sensitivity but lacks species specificity.

The Bordetella Bps polysaccharide is required for biofilm formation and persistence in the lower respiratory tract of swine

Science.gov (United States)

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

Fatal pneumonia following inoculation of healthy bighorn sheep with Pasteurella haemolytica from healthy domestic sheep.

Science.gov (United States)

Foreyt, W J; Snipes, K P; Kasten, R W

1994-04-01

In a series of three experiments, isolates of Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1, from healthy domestic sheep, were inoculated intratracheally into eight bighorn sheep (Ovis canadensis canadensis) and seven domestic sheep with doses of bacteria ranging from 5.3 x 10(8) to 8.6 x 10(11) colony forming units. Seven of eight inoculated bighorn sheep died from acute pneumonia within 48 hr of inoculation, whereas all seven domestic sheep inoculated with comparable or greater doses of bacteria remained healthy. One contact control bighorn sheep also died 6 days after its penmates received P. haemolytica. Three other noncontact control bighorn sheep remained healthy during the experiments. Pasteurella haemolytica biotype A, serotype 2, ribotype reference WSU-1 in the inocula was recovered from one or more tissues from all bighorns that died; whereas, it was not detected in any bighorn sheep before inoculation. Three different ribotypes of P. haemolytica A2 were recovered from bighorn sheep; however, only the ribotype reference WSU-1 in the domestic sheep-origin inoculum was recovered from all dead bighorn sheep, and was not recovered from bighorn sheep that survived the experiments. Thus, a relatively nonpathogenic and common isolate of P. haemolytica from healthy domestic sheep was lethal in bighorn sheep under experimental conditions.

Identifying long-term memory B-cells in vaccinated children despite waning antibody levels specific for Bordetella pertussis proteins

NARCIS (Netherlands)

Hendrikx, Lotte H.; Ozturk, Kemal; de Rond, Lia G. H.; Veenhoven, Reinier H.; Sanders, Elisabeth A. M.; Berbers, Guy A. M.; Buisman, Anne-Marie

2011-01-01

Whooping cough is a respiratory disease caused by Bordetella pertussis. Since the 1950s in developed countries pertussis vaccinations are included in the national immunization program. However, antibody levels rapidly wane after both whole cell and acellular pertussis vaccination. Therefore

Ectopic Expression of O Antigen in Bordetella pertussis by a Novel Genomic Integration System.

Science.gov (United States)

Ishigaki, Keisuke; Shinzawa, Naoaki; Nishikawa, Sayaka; Suzuki, Koichiro; Fukui-Miyazaki, Aya; Horiguchi, Yasuhiko

2018-01-01

We describe a novel genome integration system that enables the introduction of DNA fragments as large as 50 kbp into the chromosomes of recipient bacteria. This system, named BPI, comprises a bacterial artificial chromosome vector and phage-derived gene integration machinery. We introduced the wbm locus of Bordetella bronchiseptica , which is required for O antigen biosynthesis, into the chromosome of B. pertussis , which intrinsically lacks O antigen, using the BPI system. After the introduction of the wbm locus, B. pertussis presented an additional substance in the lipooligosaccharide fraction that was specifically recognized by the anti- B. bronchiseptica antibody but not the anti- B. pertussis antibody, indicating that B. pertussis expressed O antigen corresponding to that of B. bronchiseptica . O antigen-expressing B. pertussis was less sensitive to the bactericidal effects of serum and polymyxin B than the isogenic parental strain. In addition, an in vivo competitive infection assay showed that O antigen-expressing B. pertussis dominantly colonized the mouse respiratory tract over the parental strain. These results indicate that the BPI system provides a means to alter the phenotypes of bacteria by introducing large exogenous DNA fragments. IMPORTANCE Some bacterial phenotypes emerge through the cooperative functions of a number of genes residing within a large genetic locus. To transfer the phenotype of one bacterium to another, a means to introduce the large genetic locus into the recipient bacterium is needed. Therefore, we developed a novel system by combining the advantages of a bacterial artificial chromosome vector and phage-derived gene integration machinery. In this study, we succeeded for the first time in introducing a gene locus involved in O antigen biosynthesis of Bordetella bronchiseptica into the chromosome of B. pertussis , which intrinsically lacks O antigen, and using this system we analyzed phenotypic alterations in the resultant

Comparative studies on [Pasteurella] testudinis and [P.] testudinis-like bacteria and proposal of Chelonobacter oris gen. nov., sp. nov. as a new member of the family Pasteurellaceae

DEFF Research Database (Denmark)

Gregersen, Rikke Heidemann; Neubauer, Claudia; Christensen, Henrik

2009-01-01

Abstract A collection of 12 strains isolated from diseased tortoises tentatively identified as [Pasteurella] testudinis-like based on phenotypical characters were compared with three reference strains of [Pasteurella] testudinis. All strains could be separated from the reference strains with resp...

Laboratory-based surveillance of pertussis using multitarget real-time PCR in Japan: evidence for Bordetella pertussis infection in preteens and teens

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K. Kamachi

2015-11-01

Full Text Available Between January 2013 and December 2014, we conducted laboratory-based surveillance of pertussis using multitarget real-time PCR, which discriminates among Bordetella pertussis, Bordetella parapertussis, Bordetella holmesii and Mycoplasma pneumoniae. Of 355 patients clinically diagnosed with pertussis in Japan, B. pertussis, B. parapertussis and M. pneumoniae were detected in 26% (n = 94, 1.1% (n = 4 and 0.6% (n = 2, respectively, whereas B. holmesii was not detected. It was confirmed that B. parapertussis and M. pneumoniae are also responsible for causing pertussis-like illness. The positive rates for B. pertussis ranged from 16% to 49%, depending on age. Infants aged ≤ 3 months had the highest rate (49%, and children aged 1 to 4 years had the lowest rate (16%, p < 0.01 vs. infants aged ≤ 3 months. Persons aged 10 to 14 and 15 to 19 years also showed high positive rates (29% each; the positive rates were not statistically significant compared with that of infants aged ≤ 3 months (p ≥ 0.06. Our observations indicate that similar to infants, preteens and teens are at high risk of B. pertussis infection.

Mycoplasma ovipneumoniae - A Primary Cause of Severe Pneumonia Epizootics in the Norwegian Muskox (Ovibos moschatus) Population

Science.gov (United States)

Handeland, Kjell; Tengs, Torstein; Kokotovic, Branko; Vikøren, Turid; Ayling, Roger D.; Bergsjø, Bjarne; Sigurðardóttir, Ólöf G.; Bretten, Tord

2014-01-01

The Norwegian muskox (Ovibos moschatus) population lives on the high mountain plateau of Dovre and originates from animals introduced from Greenland. In the late summers of 2006 and 2012, severe outbreaks of pneumonia with mortality rates of 25-30% occurred. During the 2012 epidemic high quality samples from culled sick animals were obtained for microbiological and pathological examinations. High throughput sequencing (pyrosequencing) of pneumonic lung tissue revealed high concentrations of Mycoplasma ovipneumoniae in all six animals examined by this method and Pasteurella multocida subsp. multocida in four animals, whereas no virus sequences could be identified. Mycoplasma ovipneumoniae and P. multocida multocida were also isolated by culture. Using real time PCR on lung swabs, M. ovipneumoniae was detected in all of the 19 pneumonic lungs examined. Gross pathological examination revealed heavy consolidations primarily in the cranial parts of the lungs and it also identified one case of otitis media. Histologically, lung lesions were characterized as acute to subacute mixed exudative and moderately proliferative bronchoalveolar pneumonia. Immunohistochemical (IHC) examination revealed high load of M. ovipneumoniae antigens within lung lesions, with particularly intensive staining in the neutrophils. Similar IHC finding were observed in archived lung tissue blocks from animals examined during the 2006 epidemic. An M. ovipneumoniae specific ELISA was applied on bio-banked muskox sera from stray muskoxen killed in the period 2004–2013 and sick muskoxen culled, as well as sera from wild reindeer (Rangifer tarandus tarandus) on Dovre and muskoxen from Greenland. Serology and mycoplasma culturing was also carried out on sheep that had been on pasture in the muskox area during the outbreak in 2012. Our findings indicated separate introductions of M. ovipneumoniae infection in 2006 and 2012 from infected co-grazing sheep. Salt licks shared by the two species were a

Mycoplasma ovipneumoniae--a primary cause of severe pneumonia epizootics in the Norwegian Muskox (Ovibos moschatus population.

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Kjell Handeland

Full Text Available The Norwegian muskox (Ovibos moschatus population lives on the high mountain plateau of Dovre and originates from animals introduced from Greenland. In the late summers of 2006 and 2012, severe outbreaks of pneumonia with mortality rates of 25-30% occurred. During the 2012 epidemic high quality samples from culled sick animals were obtained for microbiological and pathological examinations. High throughput sequencing (pyrosequencing of pneumonic lung tissue revealed high concentrations of Mycoplasma ovipneumoniae in all six animals examined by this method and Pasteurella multocida subsp. multocida in four animals, whereas no virus sequences could be identified. Mycoplasma ovipneumoniae and P. multocida multocida were also isolated by culture. Using real time PCR on lung swabs, M. ovipneumoniae was detected in all of the 19 pneumonic lungs examined. Gross pathological examination revealed heavy consolidations primarily in the cranial parts of the lungs and it also identified one case of otitis media. Histologically, lung lesions were characterized as acute to subacute mixed exudative and moderately proliferative bronchoalveolar pneumonia. Immunohistochemical (IHC examination revealed high load of M. ovipneumoniae antigens within lung lesions, with particularly intensive staining in the neutrophils. Similar IHC finding were observed in archived lung tissue blocks from animals examined during the 2006 epidemic. An M. ovipneumoniae specific ELISA was applied on bio-banked muskox sera from stray muskoxen killed in the period 2004-2013 and sick muskoxen culled, as well as sera from wild reindeer (Rangifer tarandus tarandus on Dovre and muskoxen from Greenland. Serology and mycoplasma culturing was also carried out on sheep that had been on pasture in the muskox area during the outbreak in 2012. Our findings indicated separate introductions of M. ovipneumoniae infection in 2006 and 2012 from infected co-grazing sheep. Salt licks shared by the two

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

Science.gov (United States)

Bouchez, Valérie; Guiso, Nicole

2015-10-01

Whooping cough is a vaccine-preventable disease due to Bordetella pertussis and B. parapertussis. This highly contagious respiratory disease occurs through epidemic cycles every 3-5 years and vaccination did not change this frequency. Models suggest that the cyclic increase of susceptibles is linked to demographic differences and different vaccine coverage. However, differences in surveillance of the disease as well as adaptation of the agents of the disease to their human hosts and to vaccine pressure might also play an important role. These parameters are discussed in this review. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease.

Science.gov (United States)

Ivanov, Yury V; Shariat, Nikki; Register, Karen B; Linz, Bodo; Rivera, Israel; Hu, Kai; Dudley, Edward G; Harvill, Eric T

2015-10-26

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR. The CRISPR-Cas system has been identified using Basic Local Alignment Search Tool (BLAST) against other sequenced and annotated genomes and confirmed via CRISPRfinder program. Using Polymerase Chain Reactions (PCR) and Sanger DNA sequencing, we discovered CRISPRs in additional bacterial isolates of the same species of Bordetella. Transcriptional activity and processing of the CRISPR have been assessed via RT-PCR. Here we describe a novel Type II-C CRISPR and its associated genes-cas1, cas2, and cas9-in several isolates of a newly discovered Bordetella species. The CRISPR-cas locus, which is absent in all other Bordetella species, has a significantly lower GC-content than the genome-wide average, suggesting acquisition of this locus via horizontal gene transfer from a currently unknown source. The CRISPR array is transcribed and processed into mature CRISPR RNAs (crRNA), some of which have homology to prophages found in closely related species B. hinzii. Expression of the CRISPR-Cas system and processing of crRNAs with perfect homology to prophages present in closely related species, but absent in that containing this CRISPR-Cas system, suggest it provides protection against phage predation. The 3,117-bp cas9 endonuclease gene from this novel CRISPR-Cas system is 990 bp smaller than that of Streptococcus pyogenes, the 4,017-bp allele currently used for genome editing, and which may make it a useful tool in various CRISPR-Cas technologies.

Detection of IgG antibodies against Bordetella pertussis with 125I-protein A

International Nuclear Information System (INIS)

Wirsing von Koenig, C.H.; Finger, H.

1981-01-01

A method for the detection of IgG antibodies against Bordetella pertussis is described, based on the principle of 'sandwich' radioimmunoassay. 125 I protein A is used as radioactive tracer. The influence of amounts of antigen, antibody, radioactive tracer, incubation time and temperature were tested and the optimal conditions for the assay are described. The procedure offers a simple, quick, and sensitive method for detecting antibodies against B. pertussis. Application and limitation of the test are discussed. (orig.)

Immunization status of Iranian military recruits against Bordetella pertussis infection (whooping cough).

Science.gov (United States)

Izadi, Morteza; Afsharpaiman, Shahla; Jonaidi Jafari, Nematollah; Ranjbar, Reza; Gooya, Mohammad Mahdi; Robat Sarpooshi, Javad; Esfahani, Ali Akbar; Soheylipoor, Hamid

2011-03-21

Military recruits are susceptible to respiratory pathogens because of increased antibiotic resistance and the lack of an effective vaccine. The goal of the current study was to determine the immunological status of the Bordetella pertussis among conscripts in Iranian military garrisons. The study population consisted of 424 conscripts aged 18 to 21 years who enrolled for military service. They were selected using cluster stratified sampling from all military garrisons in Tehra, Iran. To determine the seroprevalence of infection, blood specimens from all recruits were collected and stored at - 20 °C until assayed. All serum samples were screened for immunoglobulin G (IgG) antibodies against Bordetella pertussis toxin (PT) and by using enzyme-linked immunosorbent assay (ELISA). The overall prevalence of B. pertussis seropositivity in military recruits was 60.6. Only 55.0% of the recruits had low awareness about the record of vaccination against B. pertussis during childhood. Among 424 studied individuals, 48 recruits (11.3%) had a positive history of whooping cough; prevalence of seropositivity in these recruits was 70.0%. Among these subjects, 61.7% were referred to a physician for treatment and only 39.6% of them were administered anti-pertussis therapy. Our study showed that military conscripts in Tehran garrisons were not serologically immune to pertussis and also confirmed the low awareness about vaccination and medical history related to pertussis infection in this high-risk subgroup of the Iranian population. Routine acellular booster vaccination, particularly before 18 years of age, is recommended.

Bordetella pertussis filamentous hemagglutinin itself does not trigger anti-inflammatory interleukin-10 production by human dendritic cells

Czech Academy of Sciences Publication Activity Database

Romero, Rodrigo, Villarino; Hasan, Shakir; Faé, K.; Holubová, Jana; Geurtsen, J.; Schwarzer, Martin; Wiertsema, S.; Osička, Radim; Poolman, J.; Šebo, Peter

2016-01-01

Roč. 306, č. 1 (2016), s. 38-47 ISSN 1438-4221 R&D Projects: GA ČR GA15-09157S; GA ČR GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella pertussis * Cytokine * Dendritic cell Subject RIV: EE - Microbiology, Virology Impact factor: 3.391, year: 2016

Pore-Forming and Enzymatic Activities of Bordetella pertussis Adenylate Cyclase Toxin Synergize in Promoting Lysis of Monocytes

Czech Academy of Sciences Publication Activity Database

Basler, Marek; Mašín, Jiří; Osička, Radim; Šebo, Peter

2006-01-01

Roč. 74, č. 5 (2006), s. 2207-2214 ISSN 0019-9567 R&D Projects: GA AV ČR IAA5020406; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50200510 Keywords : bordetella pertussis * cyaa * cytotoxicity Subject RIV: EE - Microbiology, Virology Impact factor: 4.004, year: 2006

Delivery of multiple epitopes by recombinant detoxified adenylate cyclase of Bordetella pertussis induces protective antiviral immunity

Czech Academy of Sciences Publication Activity Database

Fayolle, C.; Osičková, Adriana; Osička, Radim; Henry, T.; Rojas, M. J.; Saron, M. F.; Šebo, Peter; Leclers, C.

2001-01-01

Roč. 75, č. 16 (2001), s. 7330-7338 ISSN 0022-538X R&D Projects: GA ČR GA310/98/0432; GA MŠk ME 167; GA MŠk VS96149 Institutional research plan: CEZ:AV0Z5020903 Keywords : antiviral immunity * Bordetella pertusis Subject RIV: EE - Microbiology, Virology Impact factor: 5.622, year: 2001

Structure of Bordetella pertussis peptidoglycan

International Nuclear Information System (INIS)

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

1987-01-01

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

Segments Crucial for Membrane Translocation and Pore-forming Activity of Bordetella Adenylate Cyclase Toxin

Czech Academy of Sciences Publication Activity Database

Basler, Marek; Knapp, O.; Mašín, Jiří; Fišer, R.; Maier, E.; Benz, R.; Šebo, Peter; Osička, Radim

2007-01-01

Roč. 282, č. 17 (2007), s. 12419-12429 ISSN 0021-9258 R&D Projects: GA MŠk 1M0506; GA AV ČR IAA5020406 Grant - others:XE(XE) European Union 6th FP contract LSHB-CT-2003-503582 THERAVAC Institutional research plan: CEZ:AV0Z50200510 Source of funding: R - rámcový projekt EK Keywords : bordetella * adenylate cyclase toxin * ac membrane translocation Subject RIV: EE - Microbiology, Virology Impact factor: 5.581, year: 2007

Amidate prodrugs of 9-[2-(Phosphonomethoxy)ethyl]adenine (PMEA) as inhibitors of adenylate cyclase toxin from Bordetella pertussis

Czech Academy of Sciences Publication Activity Database

Šmídková, Markéta; Dvořáková, Alexandra; Tloušťová, Eva; Česnek, Michal; Janeba, Zlatko; Mertlíková-Kaiserová, Helena

2014-01-01

Roč. 281, Suppl S1 (2014), s. 729 ISSN 1742-464X. [FEBS EMBO 2014 Conference. 30.08.2014-04.09.2014, Paris] R&D Projects: GA MŠk LO1302; GA MV VG20102015046 Institutional support: RVO:61388963 Keywords : Bordetella pertussis * adenylyl cyclase toxin * inhibitors Subject RIV: CE - Biochemistry

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)

Capik, Sarah F; White, Brad J; Lubbers, Brian V; Apley, Michael D; DeDonder, Keith D; Larson, Robert L; Harhay, Greg P; Chitko-McKown, Carol G; Harhay, Dayna M; Kalbfleisch, Ted S; Schuller, Gennie; Clawson, Michael L

2017-03-01

OBJECTIVE To compare predictive values, extent of agreement, and gamithromycin susceptibility between bacterial culture results of nasopharyngeal swab (NPS) and bronchoalveolar lavage fluid (BALF) samples obtained from calves with bovine respiratory disease (BRD). ANIMALS 28 beef calves with clinical BRD. PROCEDURES Pooled bilateral NPS samples and BALF samples were obtained for bacterial culture from calves immediately before and at various times during the 5 days after gamithromycin (6 mg/kg, SC, once) administration. For each culture-positive sample, up to 12 Mannheimia haemolytica, 6 Pasteurella multocida, and 6 Histophilus somni colonies underwent gamithromycin susceptibility testing. Whole-genome sequencing was performed on all M haemolytica isolates. For paired NPS and BALF samples collected 5 days after gamithromycin administration, the positive and negative predictive values for culture results of NPS samples relative to those of BALF samples and the extent of agreement between the sampling methods were determined. RESULTS Positive and negative predictive values of NPS samples were 67% and 100% for M haemolytica, 75% and 100% for P multocida, and 100% and 96% for H somni. Extent of agreement between results for NPS and BALF samples was substantial for M haemolytica (κ, 0.71) and H somni (κ, 0.78) and almost perfect for P multocida (κ, 0.81). Gamithromycin susceptibility varied within the same sample and between paired NPS and BALF samples. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated culture results of NPS and BALF samples from calves with BRD should be interpreted cautiously considering disease prevalence within the population, sample collection relative to antimicrobial administration, and limitations of diagnostic testing methods.

Amidate Prodrugs of 9-[2-(Phosphonomethoxy)Ethyl]Adenine as Inhibitors of Adenylate Cyclase Toxin from Bordetella pertussis

Czech Academy of Sciences Publication Activity Database

Šmídková, Markéta; Dvořáková, Alexandra; Tloušťová, Eva; Česnek, Michal; Janeba, Zlatko; Mertlíková-Kaiserová, Helena

2014-01-01

Roč. 58, č. 2 (2014), s. 664-671 ISSN 0066-4804 R&D Projects: GA MV VG20102015046 Grant - others:OPPC(XE) CZ.2.16/3.1.00/24016 Institutional support: RVO:61388963 Keywords : Bordetella pertussis * adenylate cyclase toxin * ACT * inhibitors * PMEA * amidate prodrugs Subject RIV: CC - Organic Chemistry Impact factor: 4.476, year: 2014

Evidence of Bordetella pertussis infection in vaccinated 1-year-old Danish children

DEFF Research Database (Denmark)

von Linstow, Marie-Louise; Pontoppidan, Peter Lotko; von König, Carl-Heinz Wirsing

2010-01-01

We measured IgA and IgG antibodies to pertussis toxin (PT) and filamentous hemagglutinin (FHA) in sera from 203 1-year-old children who had received one to three doses of a monocomponent PT toxoid vaccine. Ten children (5%) had IgA antibody to PT indicating recent infection; seven of these children......%. The apparent high Bordetella pertussis infection rate in Danish infants suggests that the monocomponent PT toxoid vaccine used in Denmark has limited efficacy against B. pertussis infection. A prospective immunization study comparing a multi-component vaccine with the present monocomponent PT toxoid vaccine...

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

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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

Mouse lung adhesion assay for Bordetella pertussis

Energy Technology Data Exchange (ETDEWEB)

Burns, K A; Freer, J H [Department of Microbiology, Alexander Stone Building, Bearsden, Glasgow, Scotland

1982-03-01

The ability of Bordetella pertussis to adhere to cell surfaces has been demonstrated by adhesion to tissue culture cells and adhesion to chicken, hamster or rabbit trachea in organ culture. In this report a mouse lung assay for adhesion is described and the results obtained using two virulent strains of B. pertussis and their avirulent counterparts. These were a C modulation of one of the original virulent strains and a phase IV variant of the other virulent strain. Organisms were radiolabelled by adding 1 ..mu..Ci (37 K Bq) of (/sup 14/C)glutamic acid per 10 ml of culture medium before inoculation and incubation for 5 days. The lungs were washed by perfusion in situ with at least two volumes (1 ml) of sterile 1% (w/v) casamino acids. The percentage of the inoculated organisms retained in the lungs was determined, after removal of the lungs, by one of the following two methods: viable count or radioactive count. Results for both methods were expressed as the percentage of the inoculum retained in the lungs plus or minus one standard deviation.

Mouse lung adhesion assay for Bordetella pertussis

International Nuclear Information System (INIS)

Burns, K.A.; Freer, J.H.

1982-01-01

The ability of Bordetella pertussis to adhere to cell surfaces has been demonstrated by adhesion to tissue culture cells and adhesion to chicken, hamster or rabbit trachea in organ culture. In this report a mouse lung assay for adhesion is described and the results obtained using two virulent strains of B. pertussis and their avirulent counterparts. These were a C modulation of one of the original virulent strains and a phase IV variant of the other virulent strain. Organisms were radiolabelled by adding 1 μCi (37 K Bq) of [ 14 C]glutamic acid per 10 ml of culture medium before inoculation and incubation for 5 days. The lungs were washed by perfusion in situ with at least two volumes (1 ml) of sterile 1% (w/v) casamino acids. The percentage of the inoculated organisms retained in the lungs was determined, after removal of the lungs, by one of the following two methods: viable count or radioactive count. Results for both methods were expressed as the percentage of the inoculum retained in the lungs plus or minus one standard deviation. (Auth.)

Short communication: Interaction of the isomers carvacrol and thymol with the antibiotics doxycycline and tilmicosin: In vitro effects against pathogenic bacteria commonly found in the respiratory tract of calves.

Science.gov (United States)

Kissels, W; Wu, X; Santos, R R

2017-02-01

Bovine respiratory disease is the major problem faced by cattle, specially calves, leading to reduced animal performance and increased mortality, consequently causing important economic losses. Hence, calves must be submitted to antibiotic therapy to counteract this infection usually initiated by the combination of environmental stress factors and viral infection, altering the animal's defense mechanism, and thus allowing lung colonization by the opportunistic bacteria Mannheimia haemolytica and Pasteurella multocida. Essential oils appear to be candidates to replace antibiotics or to act as antibiotic adjuvants due to their antimicrobial properties. In the present study, we aimed to evaluate the 4 essential oil components carvacrol, thymol, trans-anethole, and 1,8 cineole as antibacterial agents or as adjuvants for the antibiotics doxycycline and tilmicosin against M. haemolytica and P. multocida. Bacteria were cultured according to standard protocols, followed by the determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration. A checkerboard assay was applied to detect possible interactions between components, between antibiotics, and between components and antibiotics. Doxycycline at 0.25 and 0.125 μg/mL inhibited the growth of P. multocida and M. haemolytica, respectively, whereas tilmicosin MIC values were 1.0 and 4.0 μg/mL for P. multocida and M. haemolytica, respectively. Carvacrol MIC values were 2.5 and 1.25 mM for P. multocida and M. haemolytica, respectively, whereas thymol MIC values were 1.25 and 0.625 mM for P. multocida and M. haemolytica, respectively. Trans-anethole and 1,8 cineole did not present any antibacterial effect even at 40 mM against the investigated pathogens. All minimum bactericidal concentration values were the same as MIC, except when thymol was tested against M. haemolytica, being twice the MIC data (i.e., 1.25 mM thymol). Based on fractional inhibitory concentration checkerboard assay, no

Investigation of genome sequences within the family Pasteurellaceae

DEFF Research Database (Denmark)

Angen, Øystein; Ussery, David

Introduction The bacterial genome sequences are now available for an increasing number of strains within the family Pasteurellaceae. At present, 24 Pasteurellaceae genomes are publicly available through internet databases, and another 40 genomes are being sequenced. This investigation will describe...... the core genome for both the family Pasteurellaceae and for the species Haemophilus influenzae. Methods Twenty genome sequences from the following species were included: Haemophilus influenzae (11 strains), Haemophilus ducreyi (1 strain), Histophilus somni (2 strains), Haemophilus parasuis (1 strain......), Actinobacillus pleuropneumoniae (2 strains), Actinobacillus succinogenes (1 strain), Mannheimia succiniciproducens (1 strain), and Pasteurella multocida (1 strain). The predicted proteins for each genome were BLASTed against each other, and a set of conserved core gene families was determined as described...

Aspects relating to use of radioactively labelled bacteria in animal experiments. 7

International Nuclear Information System (INIS)

Heilmann, P.; Flossmann, K.D.; Mueller, G.; Finsterbusch, L.

1983-01-01

Two different types of aerosol dispensers were used in an aerosol compartment to apply 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 8 in mini-piglets, 6-8 . 10 8 in ordinary piglets, and 2 . 10 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. (author)

Rapport for Center for Vildtsundhed 1. halvår 2012

DEFF Research Database (Denmark)

Chriél, Mariann; Enemark, Heidi L.; Therkildsen, Ole Roland

for vurderingen af parasitbelastningen hos raske ederfugle. Redemateriale til undersøgelse af Pasteurella multocida, der forårsager udbrud af fjerkrækolera, fortsættes i et specialestudie af de dyrkbare bakterier. Med henblik på kortlægning af rævens dværgbændelorm (Echinococcus multilocularis) hos mårhund og ræv...... Danmark siden 2000, og det er første gang bændelormen er påvist i jyske ræve. Echinococcus multilocularis kan smitte til mennesker, hvor de i larvestadiet angriber leveren, hvilket kan have fatale følger, så det er vigtigt, at folk er opmærksomme på risikoen og tager deres forbehold. Mennesker smittes...

Structure-Function Relationships Underlying the Capacity of Bordetella Adenylate Cyclase Toxin to Disarm Host Phagocytes

Czech Academy of Sciences Publication Activity Database

Novák, Jakub; Černý, Ondřej; Osičková, Adriana; Linhartová, Irena; Mašín, Jiří; Bumba, Ladislav; Šebo, Peter; Osička, Radim

2017-01-01

Roč. 9, č. 10 (2017), s. 1-28, č. článku 300. E-ISSN 2072-6651 R&D Projects: GA ČR GA15-09157S; GA ČR(CZ) GA16-05919S; GA MŠk(CZ) LM2015064; GA MZd(CZ) NV16-28126A Institutional support: RVO:61388971 Keywords : adenylate cyclase toxin * Bordetella * cAMP Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 3.030, year: 2016

Cyclic di-GMP regulation of the bvg-repressed genes and the orphan response regulator RisA in Bordetella pertussis

Science.gov (United States)

Expression of Bordetella pertussis virulence factors is activated by the BvgAS two-component system. Under modulating growth conditions BvgAS indirectly represses another set of genes through the action of BvgR, a bvg-activated protein. BvgR blocks activation of the response regulator RisA which is ...

Cell envelope of Bordetella pertussis: immunological and biochemical analyses and characterization of a major outer membrane porin protein

International Nuclear Information System (INIS)

Armstrong, S.K.

1986-01-01

Surface molecules of Bordetella pertussis which may be important in metabolism, pathogenesis, and immunity to whooping cough were examined using cell fractionation and 125 I cell surface labeling. Antigenic envelope proteins were examined by immunofluorescence microscopy and Western blotting procedures using monoclonal antibodies and convalescent sera. A surface protein with a high M/sub r/, missing in a mutant lacking the filamentous hemagglutinin, was identified in virulent Bordetella pertussis but was absent in virulent B. pertussis strains. At least three envelope proteins were found only in virulent B. pertussis strains and were absent or diminished in avirulent and most phenotypically modulated strains. Transposon-induced mutants unable to produce hemolysin, dermonecrotic toxin, pertussis toxin, and filamentous hemagglutinin also lacked these three envelope proteins, confirming that virulence-associated envelope proteins were genetically regulated with other virulence-associated traits. Two dimensional gel electrophoresis revealed at least five heat modifiable proteins which migrated as higher or lower M/sub r/ moieties if solubilized at 25 0 C instead of 100 0 C

Toll-like receptor 4-positive macrophages protect mice from Pasteurella pneumotropica-induced pneumonia

Science.gov (United States)

Hart, Marcia L.; Mosier, Derek A.; Chapes, Stephen K.

2003-01-01

This study investigates Toll-like receptor 4 (TLR4)-positive macrophages in early recognition and clearance of pulmonary bacteria. TLR4 is a trans-membrane receptor that is the primary recognition molecule for lipopolysaccharide of gram-negative bacteria. The TLR4(Lps-del) mouse strains C57BL10/ScN (B10) and STOCK Abb(tm1) TLR4(Lps-del) Slc11a1(s)(B10 x C2D) are susceptible to pulmonary infections and develop pneumonia when naturally or experimentally infected by the opportunistic bacterium Pasteurella pneumotropica. Since these mice have the TLR4(Lps-del) genotype, we hypothesized that reconstitution of mice with TLR4-positive macrophages would provide resistance to this bacterium. A cultured macrophage cell line (C2D macrophages) and bone marrow cells from C2D mice were adoptively transferred to B10 and B10 x C2D mice by intraperitoneal injection. C2D macrophages increased B10 and B10 x C2D mouse resistance to P. pneumotropica. In C2D-recipient mice there was earlier transcription of tumor necrosis factor alpha and chemokines JE and macrophage inflammatory protein 2 (MIP-2) in the lungs of B10 and B10 x C2D mice, and there was earlier transcription of KC and MIP-1alpha in B10 x C2D mice. In addition, the course of inflammation following experimental Pasteurella challenge was altered in C2D recipients. C2D macrophages also protected B10 x C2D mice, which lack CD4(+) T cells. These data indicate that macrophages are critical for pulmonary immunity and can provide host resistance to P. pneumotropica. This study indicates that TLR4-positive macrophages are important for early recognition and clearance of pulmonary bacterial infections.

Pathological and molecular based study of pneumonic pasteurellosis in cattle and buffalo (bubalus bubalis)

International Nuclear Information System (INIS)

Hussain, R.

2014-01-01

In present study the clinico-pathological findings were recorded in naturally infected cattle and buffaloes due to pasteurella multocida during an outbreak at different livestock herds. There was no significant difference in mortality among various groups of buffaloes (p>0.78) and cattle (p>0.49). The infected animals showed clinical signs of moderate to acute anorexia, salivation, fever, depression, dysponea, submandibular edema, mucopurulent nasal discharge and respiratory grunts. Few of infected animals died due to septicemia. the necropsy of dead animals was performed and visceral organs lungs, kidneys, heart and lymph nodes were observed for gross and histopathological lesions. The tissue samples from these organs were fixed in formalin for pathological changes. Necropsy of dead animals revealed severe pneumonia, consolidation of lungs and intense pleural adhesions. serosanguinous fluid was accumulated in pericardium and peritoneal cavities. Histopathologically affected lungs exhibited severe congestion, mononuclear cell infiltration, thick interlobular septae punctuated with macrophages, plasma cells and peri-vascular cuffing. Liver, kidneys and lymph nodes had degenerative changes in histological sections. The specificity of p. multocida was determined by colony characteristics on macconkey's agar and morphological features with gram's iodine. The pcr product size approximately 511bp from lung tissues confirmed a total of 82% (19/23) bacterial isolates from dead animals. (author)

Acylation of Lysine 860 Allows Tight Binding and Cytotoxicity of Bordetella Adenylate Cyclase on CD1 1b-Expressing Cells

Czech Academy of Sciences Publication Activity Database

Mašín, Jiří; Basler, Marek; Knapp, O.; El-Azami-El-Idrissi, M.; Maier, E.; Konopásek, I.; Benz, R.; Leclerc, C.; Šebo, Peter

2005-01-01

Roč. 44, - (2005), s. 12766-12759 ISSN 0006-2960 R&D Projects: GA AV ČR IAA5020406; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50200510 Keywords : lysine 860 * bordetella Subject RIV: EE - Microbiology, Virology Impact factor: 3.848, year: 2005

Fatal Pasteurella haemolytica pneumonia in bighorn sheep after direct contact with clinically normal domestic sheep.

Science.gov (United States)

Foreyt, W J

1989-03-01

Six Rocky Mountain bighorn sheep were raised in captivity from birth (n = 5) or taken from the wild as a lamb (n = 1). After the bighorn sheep were in captivity for over a year, 6 clinically normal domestic sheep were placed on the 2 ha of pasture on which the bighorn sheep were kept. Nasal swab specimens were obtained from all sheep at the time the domestic sheep were introduced. Pasteurella haemolytica was isolated from swab specimens obtained from 4 of 6 domestic sheep, but not from specimens obtained from the bighorn sheep. All 6 bighorn sheep died of acute hemorrhagic pneumonia after exposure to domestic sheep. Death in the bighorn sheep occurred on days 4, 27, 27, 29, 36, or 71 after initial exposure to domestic sheep. Pasteurella haemolytica was isolated from respiratory tract tissue specimens of all bighorn sheep at the time of death. None of the domestic sheep were clinically ill during the study. At the end of the study, 3 of 6 domestic sheep were euthanatized, and at necropsy, P haemolytica was isolated from 2 of them. The most common serotypes in bighorn and domestic sheep were P haemolytica T-3 and A-2. Other serotypes isolated included P haemolytica A-1, A-9, and A-11 in bighorn sheep and A-1 in domestic sheep. On the basis of results of this study and of other reports, domestic sheep and bighorn sheep should not be managed in proximity to each other because of the potential fatal consequences in bighorn sheep.

Phylogeny of 54 representative strains of species in the family Pasteurellaceae as determined by comparison of 16S rRNA sequences.

Science.gov (United States)

Dewhirst, F E; Paster, B J; Olsen, I; Fraser, G J

1992-03-01

Virtually complete 16S rRNA sequences were determined for 54 representative strains of species in the family Pasteurellaceae. Of these strains, 15 were Pasteurella, 16 were Actinobacillus, and 23 were Haemophilus. A phylogenetic tree was constructed based on sequence similarity, using the Neighbor-Joining method. Fifty-three of the strains fell within four large clusters. The first cluster included the type strains of Haemophilus influenzae, H. aegyptius, H. aphrophilus, H. haemolyticus, H. paraphrophilus, H. segnis, and Actinobacillus actinomycetemcomitans. This cluster also contained A. actinomycetemcomitans FDC Y4, ATCC 29522, ATCC 29523, and ATCC 29524 and H. aphrophilus NCTC 7901. The second cluster included the type strains of A. seminis and Pasteurella aerogenes and H. somnus OVCG 43826. The third cluster was composed of the type strains of Pasteurella multocida, P. anatis, P. avium, P. canis, P. dagmatis, P. gallinarum, P. langaa, P. stomatis, P. volantium, H. haemoglobinophilus, H. parasuis, H. paracuniculus, H. paragallinarum, and A. capsulatus. This cluster also contained Pasteurella species A CCUG 18782, Pasteurella species B CCUG 19974, Haemophilus taxon C CAPM 5111, H. parasuis type 5 Nagasaki, P. volantium (H. parainfluenzae) NCTC 4101, and P. trehalosi NCTC 10624. The fourth cluster included the type strains of Actinobacillus lignieresii, A. equuli, A. pleuropneumoniae, A. suis, A. ureae, H. parahaemolyticus, H. parainfluenzae, H. paraphrohaemolyticus, H. ducreyi, and P. haemolytica. This cluster also contained Actinobacillus species strain CCUG 19799 (Bisgaard taxon 11), A. suis ATCC 15557, H. ducreyi ATCC 27722 and HD 35000, Haemophilus minor group strain 202, and H. parainfluenzae ATCC 29242. The type strain of P. pneumotropica branched alone to form a fifth group. The branching of the Pasteurellaceae family tree was quite complex. The four major clusters contained multiple subclusters. The clusters contained both rapidly and slowly evolving

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

International Nuclear Information System (INIS)

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

1985-01-01

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

Bisamidate Prodrugs of 2-Substituted 9-[2-(Phosphonomethoxy)ethyl]adenine (PMEA, adefovir) as Selective Inhibitors of Adenylate Cyclase Toxin from Bordetella pertussis

Czech Academy of Sciences Publication Activity Database

Česnek, Michal; Jansa, Petr; Šmídková, Markéta; Mertlíková-Kaiserová, Helena; Dračínský, Martin; Brust, T. F.; Pávek, P.; Trejtnar, F.; Watts, V. J.; Janeba, Zlatko

2015-01-01

Roč. 10, č. 8 (2015), s. 1351-1364 ISSN 1860-7179 R&D Projects: GA MV VG20102015046 Institutional support: RVO:61388963 Keywords : adenylate cyclase toxin * bisamidates * Bordetella pertussis * nucleosides * phosphonates Subject RIV: CC - Organic Chemistry Impact factor: 2.980, year: 2015

Real-Time Polymerase Chain Reaction-Based Detection of Bordetella pertussis in Mexican Infants and Their Contacts: A 3-Year Multicenter Study.

Science.gov (United States)

Aquino-Andrade, Alejandra; Martínez-Leyva, Gabriel; Mérida-Vieyra, Jocelin; Saltigeral, Patricia; Lara, Antonino; Domínguez, Wendy; García de la Puente, Silvestre; De Colsa, Agustín

2017-09-01

To evaluate the usefulness of real-time polymerase chain reaction (RT-PCR) as a diagnostic method for the detection of Bordetella pertussis in hospitalized patients aged pertussis detection and symptoms in household contacts of patients diagnosed with pertussis were studied. A total of 286 patients were included; of these, 67.1% had B pertussis and 4.5% had Bordetella spp. Complications occurred in 20% of patients, and the mortality rate was 6.7%. Of 434 contacts studied, 111 were mothers of study infants, representing the most frequently B pertussis-infected group and the main symptomatic contact. The use of RT-PCR permits improved detection and diagnosis of pertussis and a better understanding of the epidemiology of sources of infection. The complications and mortality rate of pertussis continue to be high. Household contacts are confirmed as a frequent source of infection of B pertussis in young children. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

Antimicrobial Susceptibility of Bacteria That Cause Bovine Respiratory Disease Complex in Alberta, Canada

Directory of Open Access Journals (Sweden)

R. Michele Anholt

2017-12-01

Full Text Available Bovine respiratory disease (BRD is the most important illness of feedlot cattle. Disease management targets the associated bacterial pathogens, Mannheimia haemolytica, Mycoplasma bovis, Pasteurella multocida, Histophilus somni, and Trueperella pyogenes. We conducted a cross-sectional study to measure the frequencies of antimicrobial-resistant BRD pathogens using a collaborative network of veterinarians, industry, government, and a diagnostic laboratory. Seven private veterinary practices in southern Alberta collected samples from both living and dead BRD-affected animals at commercial feedlots. Susceptibility testing of 745 isolates showed that 100% of the M. haemolytica, M. bovis, P. multocida, and T. pyogenes isolates and 66.7% of the H. somni isolates were resistant to at least one antimicrobial class. Resistance to macrolide antimicrobials (90.2% of all isolates was notable for their importance to beef production and human medicine. Multidrug resistance (MDR was high in all target pathogens with 47.2% of the isolates resistant to four or five antimicrobial classes and 24.0% resistance to six to nine classes. We compared the MDR profiles of isolates from two feedlots serviced by different veterinary practices. Differences in the average number of resistant classes were found for M. haemolytica (p < 0.001 and P. multocida (p = 0.002. Compared to previous studies, this study suggests an increasing trend of resistance in BRD pathogens against the antimicrobials used to manage the disease in Alberta. For the veterinary clinician, the results emphasize the importance of ongoing susceptibility testing of BRD pathogens to inform treatment protocols. Surveillance studies that collect additional epidemiological information and manage sampling bias will be necessary to develop strategies to limit the spread of resistance.

Bordetella bronchiseptica Pneumonia in an Extremely-Low-Birth-Weight Neonate

Directory of Open Access Journals (Sweden)

Yuk Joseph Ting

2011-12-01

Full Text Available Bordetella bronchiseptica, a gram-negative coccobacillus, is a common veterinary pathogen. In both domestic and wild animals, this bacterium causes respiratory infections including infectious tracheobronchitis in dogs and atrophic rhinitis in swine. Human infections are rare and have been documented in immunocompromised hosts. Here, we describe an extremely-low-birth-weight infant with B. bronchiseptica pneumonia. This is the first report that describes the microorganism's responsibility in causing nosocomial infection in a preterm neonate. He recovered uneventfully after a course of meropenem. It is possible that the bacteria colonize the respiratory tracts of our health care workers or parents who may have had contact with pets and then transmitted the bacterium to our patient. Follow-up until 21 months of age showed normal growth and development. He did not suffer from any significant residual respiratory disease.

Aqueous Humor Antimicrobial Activity: In Vitro Analysis after Topical 0.5% Chloramphenicol Application.

Science.gov (United States)

Cagini, Carlo; Dragoni, Annalisa; Orsolini, Giampaolo; Fiore, Tito; Beccasio, Alfredo; Spadea, Leopoldo; Moretti, Amedeo; Mencacci, Antonella

2017-06-01

To assess aqueous humor antimicrobial activity in vitro after topical 0.5% chloramphenicol application. This investigation included 63 eyes from 65 cataract surgery patients. The study group of 48 eyes received preoperatively four topical applications of 0.5% chloramphenicol. The control group of 15 eyes was given no topical applications. Aqueous humor samples were collected for in vitro antimicrobial analysis using Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, and Pasteurella multocida organisms by means of disk diffusion test. No inhibition halo was observed around all aqueous humor samples from all chloramphenicol-treated patients, irrespective of the sample quantity added to the paper disks, with no significant difference from aqueous humor from untreated control patients. Aqueous humor displayed no bactericidal effect against any of the microorganisms evaluated after topical 0.5% chloramphenicol application.

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

2014-01-01

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 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. PMID:25255075

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.

Complete genome sequence of a clinical Bordetella pertussis isolate from Brazil

Directory of Open Access Journals (Sweden)

Bruno Gabriel N Andrade

2014-11-01

Full Text Available There has been a resurgence in the number of pertussis cases in Brazil and around the world. Here, the genome of a clinical Bordetella pertussis strain (Bz181 that was recently isolated in Brazil is reported. Analysis of the virulence-associated genes defining the pre- and post-vaccination lineages revealed the presence of the prn2-ptxS1A-fim3B-ptxP3 allelic profile in Bz181, which is characteristic of the current pandemic lineage. A putative metallo-β-lactamase gene presenting all of the conserved zinc-binding motifs that characterise the catalytic site was identified, in addition to a multidrug efflux pump of the RND family that could confer resistance to erythromycin, which is the antibiotic of choice for treating pertussis disease.

Expression of bvg-repressed genes in Bordetella pertussis is controlled by RisA through a novel c-di-GMP signaling pathway

Science.gov (United States)

The BvgAS two component system of Bordetella pertussis controls virulence factor expression. In addition, BvgAS controls expression of the bvg-repressed genes through the action of the repressor, BvgR. The transcription factor RisA is inhibited by BvgR, and when BvgR is not expressed RisA induces th...

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

The Bordetella pertussis Type III Secretion System Tip Complex Protein Bsp22 Is Not a Protective Antigen and Fails To Elicit Serum Antibody Responses during Infection of Humans and Mice

Science.gov (United States)

Villarino Romero, Rodrigo; Bibova, Ilona; Cerny, Ondrej; Vecerek, Branislav; Wald, Tomas; Benada, Oldrich; Zavadilova, Jana; Sebo, Peter

2013-01-01

The type III secretion system (T3SS) of pathogenic bordetellae employs a self-associating tip complex protein Bsp22. This protein is immunogenic during infections by Bordetella bronchiseptica and could be used as a protective antigen to immunize mice against B. bronchiseptica challenge. Since low-passage clinical isolates of the human pathogen Bordetella pertussis produce a highly homologous Bsp22 protein (97% homology), we examined its vaccine and diagnostic potential. No Bsp22-specific antibodies were, however, detected in serum samples from 36 patients with clinically and serologically confirmed whooping cough disease (pertussis syndrome). Moreover, although the induction of Bsp22 secretion by the laboratory-adapted 18323 strain in the course of mice lung infection was observed, the B. pertussis 18323-infected mice did not mount any detectable serum antibody response against Bsp22. Furthermore, immunization with recombinant Bsp22 protein yielded induction of high Bsp22-specific serum antibody titers but did not protect mice against an intranasal challenge with B. pertussis 18323. Unlike for B. bronchiseptica, hence, the Bsp22 protein is nonimmunogenic, and/or the serum antibody response to it is suppressed, during B. pertussis infections of humans and mice. PMID:23690400

Crystallographic characterization of the passenger domain of the Bordetella autotransporter BrkA

International Nuclear Information System (INIS)

Zhao, Li; Nguyen, Nham T.; Fernandez, Rachel C.; Murphy, Michael E. P.

2009-01-01

A secreted bacterial protein from a human pathogen that mediates serum resistance and adherence was overexpressed, purified, refolded and crystallized. Preliminary X-ray diffraction data are presented. Autotransporters (ATs) are proteins that deliver effectors (the passenger domain) to the surface of Gram-negative bacteria by the type V secretion pathway. The passenger domain of BrkA, a Bordetella pertussis autotransporter mediating serum resistance and adherence, was cloned in a pET expression system and overexpressed in Escherichia coli. The gene product was correctly refolded, purified to homogeneity and crystallized. The crystals diffracted to 2.8 Å resolution. The space group was assumed to be P4 1 2 1 2, with unit-cell parameters a = b = 108.19, c = 115.35 Å

Susceptibility of Rocky Mountain bighorn sheep and domestic sheep to pneumonia induced by bighorn and domestic livestock strains of Pasteurella haemolytica.

OpenAIRE

Onderka, D K; Rawluk, S A; Wishart, W D

1988-01-01

Bighorn sheep were inoculated intratracheally with suspensions of nonhemolytic Pasteurella haemolytica biotype T (10(12) organisms) unique to wild bighorns, with beta-hemolytic P. haemolytica biotype T (10(12) organisms) isolated from clinically normal domestic sheep or intradermally with half a dose of a cattle vaccine containing P. haemolytica biotype A (10(5) organisms). The bighorn strain caused lobar necrotizing bronchopneumonia whereas both domestic livestock strains precipitated fatal ...

Albumin, in the Presence of Calcium, Elicits a Massive Increase in Extracellular Bordetella Adenylate Cyclase Toxin.

Science.gov (United States)

Gonyar, Laura A; Gray, Mary C; Christianson, Gregory J; Mehrad, Borna; Hewlett, Erik L

2017-06-01

Pertussis (whooping cough), caused by Bordetella pertussis , is resurging in the United States and worldwide. Adenylate cyclase toxin (ACT) is a critical factor in establishing infection with B. pertussis and acts by specifically inhibiting the response of myeloid leukocytes to the pathogen. We report here that serum components, as discovered during growth in fetal bovine serum (FBS), elicit a robust increase in the amount of ACT, and ≥90% of this ACT is localized to the supernatant, unlike growth without FBS, in which ≥90% is associated with the bacterium. We have found that albumin, in the presence of physiological concentrations of calcium, acts specifically to enhance the amount of ACT and its localization to the supernatant. Respiratory secretions, which contain albumin, promote an increase in amount and localization of active ACT that is comparable to that elicited by serum and albumin. The response to albumin is not mediated through regulation of ACT at the transcriptional level or activation of the Bvg two-component system. As further illustration of the specificity of this phenomenon, serum collected from mice that lack albumin does not stimulate an increase in ACT. These data, demonstrating that albumin and calcium act synergistically in the host environment to increase production and release of ACT, strongly suggest that this phenomenon reflects a novel host-pathogen interaction that is central to infection with B. pertussis and other Bordetella species. Copyright © 2017 American Society for Microbiology.

The host defense peptide beta-defensin 1 confers protection against Bordetella pertussis in newborn piglets.

Science.gov (United States)

Elahi, Shokrollah; Buchanan, Rachelle M; Attah-Poku, Sam; Townsend, Hugh G G; Babiuk, Lorne A; Gerdts, Volker

2006-04-01

Innate immunity plays an important role in protection against respiratory infections in humans and animals. Host defense peptides such as beta-defensins represent major components of innate immunity. We recently developed a novel porcine model of pertussis, an important respiratory disease of young children and infants worldwide. Here, we investigated the role of porcine beta-defensin 1 (pBD-1), a porcine defensin homologue of human beta-defensin 2, in conferring protection against respiratory infection with Bordetella pertussis. In this model, newborn piglets were fully susceptible to infection and developed severe bronchopneumonia. In contrast, piglets older than 4 weeks of age were protected against infection with B. pertussis. Protection was associated with the expression of pBD-1 in the upper respiratory tract. In fact, pBD-1 expression was developmentally regulated, and the absence of pBD-1 was thought to contribute to the increased susceptibility of newborn piglets to infection with B. pertussis. Bronchoalveolar lavage specimens collected from older animals as well as chemically synthesized pBD-1 displayed strong antimicrobial activity against B. pertussis in vitro. Furthermore, in vivo treatment of newborn piglets with only 500 mug pBD-1 at the time of challenge conferred protection against infection with B. pertussis. Interestingly, pBD-1 displayed no bactericidal activity in vitro against Bordetella bronchiseptica, a closely related natural pathogen of pigs. Our results demonstrate that host defense peptides play an important role in protection against pertussis and are essential in modulating innate immune responses against respiratory infections.

مطالعه بر روی ایمنی زایی یک واکسن آزمایشی سپتی سمی هموراژیک با ادجوان روغنی در گاو

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ا. ستوده نیا

2005-01-01

Full Text Available An oil adjuvant vaccine (OAV was prepared from a local strain of Pasteurella multocida. Strain 6:B was cultured and inactivated by formalin. Bacterial pellet was prepared by centrifugation and subsequently adjuvanted by Montannide oil ISA-70. A dose of prepared vaccine containing 3ml (2mg dry weight/ml was injected into five calves by IM route. Animals were bled before and at 24, 90, 150, and 200 days post-vaccination. Collected sera were used in passive mouse protection test (PMPT. Active mouse protection test (AMPT was carried out for OAV according to standard method. Results of PMPT showed 100% protection up to 150 days and 66-83% up to 200 days post-vaccination. In AMPT, 4 log of protection was gained. In this experiment the immunity induced by OAV adjuvanted by ISA-70 could protect the calves.

Bordetella Bronchiseptica in the Immunosuppressed Population – A Case Series and Review.

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Abraham Tareq Yacoub

2014-04-01

Full Text Available Organisms that are not known to cause serious infection in the immunocompetent population can in fact cause devastating illness in immunosupressed neutropenic populations especially those who are undergoing hematopoietic stem cell transplantation (HSCT, and solid organ transplantation or a history of malignancy.  One organism of interest isolated from immunosupressed patients at our institution was Bordetella bronchiseptica. B. bronchiseptica is a Gram-negative, rod shaped bacterium, primary respiratory tract organism known to cause respiratory tract disease in the animal population which include dogs, cats, and rabbits. This organism rarely causes serious infection in the immunocompetent population. However; in immunosupressed patients, it can cause serious pulmonary disease. We present three cases of B. bronchiseptica isolated from patients with a history of malignancy a serious pulmonary infection.

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

of these species exhibit resistance to veterinary macrolides with phenotypes that fall into three distinct classes. The first class has type I macrolide, lincosamide, and streptogramin B antibiotic resistance and, consistent with this, the 23S rRNA nucleotide A2058 is monomethylated by the enzyme product......(E) genes within an isogenic Escherichia coli background to assess their individually contributions to resistance. Our findings indicate what types of compounds might have driven the selection for these resistance determinants....

Pathogenical Studies And Preliminary Prevention on Bordetella Avium%禽波氏杆菌病原性研究及防治初探

Institute of Scientific and Technical Information of China (English)

王玉燕; 朱瑞良

2001-01-01

Straining characters, antibiotic sensibilities and biochemical characters of 8 strains of Bordetella avium have been experimented. Their pathogenicities to chicks were concentrated on , and proved they caused chicks acute death. The main pathogenic changes were haemorrhage on lung, liver, kidney and serous membrane of glandular stomach and muscular stomach. The resistance of the tested individuals was positively correlated with their age. The older, the resistance of chicks to bordetella avium is stronger. We also developed a preprolis vaccine with 8 strains of bordetella avium coming from different area. Used the vaccine to vaccinate 1-day-old and 11-day-old chick groups, the preventive rate is low.%本文介绍了禽波氏杆菌的培养特性、药物敏感性及生化特性，重点进行了禽波氏杆菌对雏鸡的致病性研究，结果证明本菌主要引起雏鸡的急性死亡，主要病变为肺脏出血，肝脏出血、边缘坏死，肾脏出血，腺胃、肌胃浆膜出血，肠道出血。不同日龄的雏鸡对禽波氏杆菌的抵抗力不同，与日龄成正相关。同时还用不同来源的8株禽波氏杆菌制成蜂胶灭活苗，并分别免疫1日龄和11日龄雏鸡，结果发现本疫苗对雏鸡的保护率低。

Evidence for an intact polysaccharide capsule in Bordetella pertussis.

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Neo, YiLin; Li, Rui; Howe, Josephine; Hoo, Regina; Pant, Aakanksha; Ho, SiYing; Alonso, Sylvie

2010-03-01

Polysaccharide capsules contribute to the pathogenesis of many bacteria species by providing resistance against various defense mechanisms. The production of a capsule in Bordetella pertussis, the etiologic agent of whooping cough, has remained controversial; earlier studies reported this pathogen as a capsulated microorganism whereas the recent B. pertussis genome analysis revealed the presence of a truncated capsule locus. In this work, using transmission electron microscopy and immunostaining approaches, we provide a formal evidence for the presence of an intact microcapsule produced at the surface of both laboratory strain and clinical isolates of B. pertussis. In agreement with previous studies, we found that the capsule is optimally produced in avirulent phase. Unexpectedly, the presence of the capsule was also detected at the surface of virulent B. pertussis bacteria. Consistently, a substantial transcriptional activity of the capsule operon was detected in virulent phase, suggesting that the capsular polysaccharide may play a role during pertussis pathogenesis. In vitro assays indicated that the presence of the capsule does not affect B. pertussis adherence to mammalian cells and does not further protect the bacterium from phagocytosis, complement-mediated killing or antimicrobial peptide attack. Copyright 2009. Published by Elsevier SAS.

Complement evasion by Bordetella pertussis: implications for improving current vaccines.

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Jongerius, Ilse; Schuijt, Tim J; Mooi, Frits R; Pinelli, Elena

2015-04-01

Bordetella pertussis causes whooping cough or pertussis, a highly contagious disease of the respiratory tract. Despite high vaccination coverage, reported cases of pertussis are rising worldwide and it has become clear that the current vaccines must be improved. In addition to the well-known protective role of antibodies and T cells during B. pertussis infection, innate immune responses such as the complement system play an essential role in B. pertussis killing. In order to evade this complement activation and colonize the human host, B. pertussis expresses several molecules that inhibit complement activation. Interestingly, one of the known complement evasion proteins, autotransporter Vag8, is highly expressed in the recently emerged B. pertussis isolates. Here, we describe the current knowledge on how B. pertussis evades complement-mediated killing. In addition, we compare this to complement evasion strategies used by other bacterial species. Finally, we discuss the consequences of complement evasion by B. pertussis on adaptive immunity and how identification of the bacterial molecules and the mechanisms involved in complement evasion might help improve pertussis vaccines.

Purification, crystallization and preliminary crystallographic characterization of the α2,6-sialyltransferase from Photobacterium sp. JT-ISH-224

International Nuclear Information System (INIS)

Okino, Nozomu; Kakuta, Yoshimitsu; Kajiwara, Hitomi; Ichikawa, Masako; Takakura, Yoshimitsu; Ito, Makoto; Yamamoto, Takeshi

2007-01-01

Crystallization of the α2,6-sialyltransferase from Photobacterium. Sialyltransferases transfer sialic acid from cytidine-5-monophospho-N-acetylneuraminic acid (CMP-NeuAc) to the nonreducing termini of the oligosaccharyl structures of various glycoproteins and glycolipids. The newly cloned α2,6-sialyltransferase from Photobacterium sp. JT-ISH-224 (from the Vibrionaceae family) is composed of two domains: an unknown N-terminal domain and a catalytic C-terminal domain which shares significant homology with the Pasteurella multocida multifunctional sialyltransferase. The putative mature form of JT-ISH-224 α2,6-sialyltransferase was overproduced in Escherichia coli, purified and crystallized using the hanging-drop vapour-diffusion method at 293 K. The crystal belonged to space group P3 1 21 or P3 2 21, with unit-cell parameters a = b = 90.29, c = 204.33 Å. X-ray diffraction data were collected to 2.5 Å resolution

Pneumonia in slaughtered sheep in south-western Iran: pathological characteristics and aerobic bacterial aetiology.

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Azizi, Shahrzad; Korani, Farzad Shahrani; Oryan, Ahmad

2013-01-01

In this study, the lungs of 1,000 sheep carcasses were subjected to gross examination and those suspected to be infected with pneumonia were studied at histopathological level as well as examined for presence of bacteria. Pneumonia was detected in 42 (4.2%) carcasses. Based on histopathological lesions, 45.24% were affected with suppurative bronchopneumonia, 20.93% with interstitial pneumonia, 11.9% bronchointerstitial pneumonia, 7.14% with fibrinous bronchopneumonia and 2.38% with embolic pneumonia. In addition, 11.9% of the lungs showed lung abscesses and 2.33% were affected with pleuritis without involving pulmonary parenchyma. Bacteriological examination revealed presence of ovine pathogens, such as Pasteurella multocida (24.53%), Staphylococcus aureus (20.75%), Klebsiella pneumoniae (15.09%), Corynebacterium pseudotuberculosis (7.55%) and Actinomyces pyogenes (1.89%). The most common form of pneumonia was suppurative bronchopneumonia with moderate amounts of fibrin deposits on the pleural surface and inside the bronchioles and alveoli.

Evidence of Bordetella pertussis infection in vaccinated 1-year-old Danish children

DEFF Research Database (Denmark)

von Linstow, Marie-Louise; Pontoppidan, Peter Lotko; von König, Carl-Heinz Wirsing

2010-01-01

We measured IgA and IgG antibodies to pertussis toxin (PT) and filamentous hemagglutinin (FHA) in sera from 203 1-year-old children who had received one to three doses of a monocomponent PT toxoid vaccine. Ten children (5%) had IgA antibody to PT indicating recent infection; seven of these children...... had received three doses of vaccine. PT IgA responders did not have significantly longer coughing episodes than PT IgA non-responders. Since an IgA antibody response occurs in only approximately 50% of infected children, the actual infection rate in our cohort is estimated to approximately 10......%. The apparent high Bordetella pertussis infection rate in Danish infants suggests that the monocomponent PT toxoid vaccine used in Denmark has limited efficacy against B. pertussis infection. A prospective immunization study comparing a multi-component vaccine with the present monocomponent PT toxoid vaccine...

Enhanced Ex Vivo Stimulation of Mycobacterium tuberculosis-Specific T Cells in Human Immunodeficiency Virus-Infected Persons via Antigen Delivery by the Bordetella pertusis Adenylate Cyclase Vector

Czech Academy of Sciences Publication Activity Database

Connell, T. G.; Shey, M. S.; Seldon, R.; Rangaka, M. X.; van Cutsem, G.; Šimšová, Marcela; Marčeková, Zuzana; Šebo, Peter; Curtis, N.; Diwakar, L.; Meintjes, G. A.; Leclerc, C.; Wilkinson, R. J.; Wilkinson, K. A.

2007-01-01

Roč. 14, č. 7 (2007), s. 847-854 ISSN 1556-6811 R&D Projects: GA MŠk 2B06161 Institutional research plan: CEZ:AV0Z50200510 Keywords : mycobacterium tuberculosis * bordetella pertusis * human immunodeficiency virus Subject RIV: EE - Microbiology, Virology Impact factor: 1.995, year: 2007

Development of colloidal gold immunochromatographic strips for detection of Riemerella anatipestifer.

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Wanwan Hou

Full Text Available Riemerella anatipestifer is one of the most important bacterial pathogen of ducks and causes a contagious septicemia. R. anatipestifer infection causes serositis syndromes similar to other bacterial infections in ducks, including infection by Escherichia coli, Salmonella enterica and Pasteurella multocida. Clinically differentiating R. anatipestifer infections from other bacterial pathogen infections is usually difficult. In this study, MAb 1G2F10, a monoclonal antibody against R. anatipestifer GroEL, was used to develop a colloidal gold immunochromatographic strip. Colloidal gold particles were prepared by chemical synthesis to an average diameter of 20 ± 5.26 nm by transmission electron microscope imaging. MAb 1G2F10 was conjugated to colloidal gold particles and the formation of antibody-colloidal gold conjugates was monitored by UV/Vis spectroscopy. Immunochromatographic strips were assembled in regular sequence through different accessories sticked on PVC plate. Strips specifically detected R. anatipestifer within 10 min, but did not detect E. coli, S. enterica and P. multocida. The detection limit for R. anatipestifer was 1 × 10(6 colony forming units, which was 500 times higher than a conventional agglutination test. Accuracy was 100% match to multiplex PCR. Assay stability and reproducibility were excellent after storage at 4°C for 6 months. The immunochromatographic strips prepared in this study offer a specific, sensitive, and rapid detection method for R. anatipestifer, which is of great importance for the prevention and control of R. anatipestifer infections.

Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

Energy Technology Data Exchange (ETDEWEB)

Gangi Setty, Thanuja [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Cho, Christine [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Govindappa, Sowmya [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India); Apicella, Michael A. [Carver College of Medicine, University of Iowa, Iowa City, IA 52242-1109 (United States); Ramaswamy, S., E-mail: ramas@instem.res.in [Institute for Stem Cell Biology and Regenerative Medicine, NCBS Campus, GKVK Post, Bangalore, Karnataka 560 065 (India)

2014-07-01

Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states.

Bacterial periplasmic sialic acid-binding proteins exhibit a conserved binding site

International Nuclear Information System (INIS)

Gangi Setty, Thanuja; Cho, Christine; Govindappa, Sowmya; Apicella, Michael A.; Ramaswamy, S.

2014-01-01

Structure–function studies of sialic acid-binding proteins from F. nucleatum, P. multocida, V. cholerae and H. influenzae reveal a conserved network of hydrogen bonds involved in conformational change on ligand binding. Sialic acids are a family of related nine-carbon sugar acids that play important roles in both eukaryotes and prokaryotes. These sialic acids are incorporated/decorated onto lipooligosaccharides as terminal sugars in multiple bacteria to evade the host immune system. Many pathogenic bacteria scavenge sialic acids from their host and use them for molecular mimicry. The first step of this process is the transport of sialic acid to the cytoplasm, which often takes place using a tripartite ATP-independent transport system consisting of a periplasmic binding protein and a membrane transporter. In this paper, the structural characterization of periplasmic binding proteins from the pathogenic bacteria Fusobacterium nucleatum, Pasteurella multocida and Vibrio cholerae and their thermodynamic characterization are reported. The binding affinities of several mutations in the Neu5Ac binding site of the Haemophilus influenzae protein are also reported. The structure and the thermodynamics of the binding of sugars suggest that all of these proteins have a very well conserved binding pocket and similar binding affinities. A significant conformational change occurs when these proteins bind the sugar. While the C1 carboxylate has been identified as the primary binding site, a second conserved hydrogen-bonding network is involved in the initiation and stabilization of the conformational states

Seroprevalence of Bordetella pertussis in the Mexican population: a cross-sectional study.

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Conde-Glez, C; Lazcano-Ponce, E; Rojas, R; DeAntonio, R; Romano-Mazzotti, L; Cervantes, Y; Ortega-Barría, E

2014-04-01

SUMMARY Serum samples collected during the National Health and Nutrition survey (ENSANUT 2006) were obtained from subjects aged 1-95 years (January-October 2010) and analysed to assess the seroprevalence of Bordetella pertussis (BP) in Mexico. Subjects' gender, age, geographical region and socioeconomic status were extracted from the survey and compiled into a subset database. A total of 3344 subjects (median age 29 years, range 1-95 years) were included in the analysis. Overall, BP seroprevalence was 47.4%. BP seroprevalence was significantly higher in males (53.4%, P = 0.0007) and highest in children (59.3%) decreasing with advancing age (P = 0.0008). BP seroprevalence was not significantly different between regions (P = 0.1918) and between subjects of socioeconomic status (P = 0.0808). Women, adolescents and young adults were identified as potential sources of infection to infants. Booster vaccination for adolescents and primary contacts (including mothers) for newborns and infants may provide an important public health intervention to reduce the disease burden.

Bordetella pertussis diagnosis in children under five years of age in the Regional Hospital of Cajamarca, Northern Peru.

Science.gov (United States)

Del Valle-Mendoza, Juana; Casabona-Oré, Veronica; Petrozzi-Helasvuo, Veronica; Cornejo-Tapia, Angela; Weilg, Pablo; Pons, Maria J; Cieza-Mora, Erico; Bazán-Mayra, Jorge; Cornejo-Pacherres, Hernan; Ruiz, Joaquin

2015-11-30

Bordetella pertussis is an important human pathogen that causes whooping cough (pertussis), an endemic illness responsible of significant morbidity and mortality, especially in infants and children. Worldwide, there are an estimated of 16 million cases of pertussis, resulting in about 195,000 child deaths per year. In Peru, pertussis is a major health problem that has been on the increase despite immunization efforts. The objective of this study was to determine the prevalence of B. pertussis among children under five years of age suspected to have whopping cough in Cajamarca, Peru. Children diagnosed with whooping cough admitted to the Hospital Regional de Cajamarca from August 2010 to July 2013 were included. Nasopharyngeal samples were obtained for B. pertussis culture and polymerase chain reaction (PCR) detection. In 133 children, the pertussis toxin and IS481 gene were detected in 38.35% (51/133) of the cases by PCR, while only 9.02% (12/133) of the Bordetella cultures were positive. The most frequent symptoms in patients with positive B. pertussis were paroxysm of coughing 68.63% (35/51), cyanosis 56.86% (29/51), respiratory distress 43.14% (22/51), and fever 39.22% (20/51). Pneumonia and acute bronchial obstructive syndrome were present in 17.65% (9/51) and 13.72% (7/51) of the cases, respectively. B. pertussis is responsible for an important proportion of whooping cough in hospitalized children in Cajamarca. Epidemiologic surveillance programs for B. pertussis are essential in Peru, especially in children who could most benefit from the vaccine.

Pneumonia caused by Bordetella bronchiseptica in two HIV-positive patients

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Roberta Filipini Rampelotto

Full Text Available ABSTRACT: CONTEXT AND OBJECTIVE: Bordetella bronchiseptica (BB is a Gram-negative coccobacillus responsible for respiratory diseases in dogs, cats and rabbits. Reports on its development in humans are rare. However, in immunosuppressed patients, especially in those with the immunodeficiency virus (HIV, BB can cause severe pulmonary infections. We report on two cases of pneumonia caused by BB in HIV-positive male patients in a university hospital. CASE REPORT: The first case comprised a 43-year-old patient who was admitted presenting chronic leg pain and coughing, with suspected pneumonia. BB was isolated from sputum culture and was successfully treated with trimethoprim/sulfamethoxazole in association with levofloxacin. The second case comprised a 49-year-old patient who was admitted presenting fever, nausea, sweating and a dry cough, also with suspected pneumonia. BB was isolated from sputum culture, tracheal secretions and bronchoalveolar lavage. The disease was treated with ciprofloxacin but the patient died. CONCLUSION: BB should be included in the etiology of pneumonia in immunodeficient HIV patients. As far as we know, these two were the first cases of pneumonia due to BB to occur in this university hospital.

Monospecific antibody against Bordetella pertussis Adenylate Cyclase protects from Pertussis

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Yasmeen Faiz Kazi

2012-06-01

Full Text Available Objectives: Acellular pertussis vaccines has been largely accepted world-wide however, there are reports about limitedantibody response against these vaccines suggesting that multiple antigens should be included in acellular vaccinesto attain full protection. The aim of present study was to evaluate the role of Bordetella pertussis adenylate cyclase as aprotective antigen.Materials and methods: Highly mono-specific antibody against adenylate cyclase (AC was raised in rabbits usingnitrocellulose bound adenylate cyclase and the specificity was assessed by immuoblotting. B.pertussis 18-323, wasincubated with the mono-specific serum and without serum as a control. Mice were challenged intra-nasally and pathophysiolgicalresponses were recorded.Results: The production of B.pertussis adenylate cyclase monospecific antibody that successfully recognized on immunoblotand gave protection against fatality (p< 0.01 and lung consolidation (p <0.01. Mouse weight gain showedsignificant difference (p< 0.05.Conclusion: These preliminary results highlight the role of the B.pertussis adenylate cyclase as a potential pertussisvaccine candidate. B.pertussis AC exhibited significant protection against pertussis in murine model. J Microbiol InfectDis 2012; 2(2: 36-43Key words: Pertussis; monospecific; antibody; passive-protection

[Transformation from chronic subdural hematoma into subdural empyema following cat bites: a case report].

Science.gov (United States)

Konno, Takuya; Yamada, Kei; Kasahara, Sou; Umeda, Yoshitaka; Oyake, Mutsuo; Fujita, Nobuya

2015-01-01

A 69-year-old man developed motor aphasia and right hemiparesis with severe headache, during the treatment of cellulitis and sepsis due to cat bites. Brain CT showed a low density, crescent-shaped lesion in the left subdural space, which was hypointense on brain diffusion-weighted imaging (DWI). One week later, when his neurological symptoms had worsened, the signal of the subdural lesion had changed to hyperintense on DWI. The lesion was capsule-shaped when enhanced by Gadolinium. The signal changes on DWI of the lesion indicated the existing hematoma had changed to an empyema, or so-called infected subdural hematoma, due to a hematogenous bacterial infection. Pasteurella multocida, a resident microbe in the oral cavity of cats, could be the responsible pathogen in this case. The patient recovered completely after treatment with intravenous high dose antibiotics. This is an important case report describing the transformation from a chronic subdural hematoma into a subdural empyema by DWI.

Clinical and microbiological study of otitis externa in sheep

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M. I. Al-Farwachi

2008-01-01

Full Text Available In this study one hundred Awassi sheep were examined clinically and bacteriologically for isolation and identification of the bacterial agents of otitis externa in sheep. The main clinical signs appeared included weakness, pale mucus membrane, auricular discharge, cough, anorexia, emaciation, and nasal discharge. Results revealed isolation of bacteria from (45% examined swabs. The most being from right ear. Younger animals were more frequently infected than older animals. Pseudomonas aeruginosa, Staphylococcus aureus, Mannheimia haemolytica, Staph. epidermidis, Pasteurella multocida, Streptococcus spp., Acintobacter spp., Escherichia coli and Klebsiella pneumonia were isolated. The results revealed that the most bacterial isolates were resistance to the bactericidal effect of the normal serum included Streptococcus pneumonia, Staphylococcus aureus, Mannheimia haemolytica. While the most bacterial isolates were produced hydroxymate siderophore included Staphylococcus aureus, Mannheimia haemolytica, Streptococcus pneumonia. The obtained results indicated to the importance of determination of serum resistance as a bacterial virulence factor in otitis externa in sheep.

Enzymatic production of human milk oligosaccharides

DEFF Research Database (Denmark)

Guo, Yao

. A recombinant Pasteurella multocida sialyltransferase (EC 2.4.99.-), namely PmST, exhibiting promiscuous trans-sialidase activities was examined. The enzyme catalysed α-2,3- and α-2,6- sialylation of lactose using either 2- O -(p-nitrophenyl)-α- D - N -acetylneuraminic acid or casein glycomacropeptide...... galactooligosaccharides with use of galactooligosaccharides as acceptors. Secondly, we examined the regioselectivity of five designed mutants of PmST catalysing synthesis of 3'- and 6'-sialyllactoses using casein glycomacropeptide and lactose as substrates. The mutants PmST E271F , PmST R313Y and PmST E271F/R313Y...... was almost abolished. The k cat / K m value for PmST P34H catalysing 6'-sialyllactose synthesis using 3'-sialyllactose as donor was 31.2 M -1 s -1 . Moreover, both the wild type enzyme and PmST P34H were capable of catalysing the hydrolysis and transfer of α-2,6 bound sialic acid....

New derivatives of salicylamides: Preparation and antimicrobial activity against various bacterial species.

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Pauk, Karel; Zadražilová, Iveta; Imramovský, Aleš; Vinšová, Jarmila; Pokorná, Michaela; Masaříková, Martina; Cížek, Alois; Jampílek, Josef

2013-11-01

Three series of salicylanilides, esters of N-phenylsalicylamides and 2-hydroxy-N-[1-(2-hydroxyphenylamino)-1-oxoalkan-2-yl]benzamides, in total thirty target compounds were synthesized and characterized. The compounds were evaluated against seven bacterial and three mycobacterial strains. The antimicrobial activities of some compounds were comparable or higher than the standards ampicillin, ciprofloxacin or isoniazid. Derivatives 3f demonstrated high biological activity against Staphylococcus aureus (⩽0.03μmol/L), Mycobacterium marinum (⩽0.40μmol/L) and Mycobacterium kansasii (1.58μmol/L), 3g shows activity against Clostridium perfringens (⩽0.03μmol/L) and Bacillus cereus (0.09μmol/L), 3h against Pasteurella multocida (⩽0.03μmol/L) and M. kansasii (⩽0.43μmol/L), 3i against methicillin-resistant S. aureus and B. cereus (⩽0.03μmol/L). The structure-activity relationships are discussed for all the compounds. Copyright © 2013 Elsevier Ltd. All rights reserved.

Aerobic bacteria occurring in the vagina of bitches with reproductive disorders.

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Bjurström, L

1993-01-01

A retrospective survey was performed of aerobic bacterial species found in the vagina of 203 bitches with genital disorders, e.g. infertility, vaginitis, pyometra and puppy death. Escherichia coli, beta-hemolytic streptococci, Staphylococcus intermedius and Pasteurella multocida were the species most often isolated. From bitches with pyometra E. coli in pure culture was the most frequent isolate. In contrast, the majority of infertile bitches gave rise to mixed cultures, and no specific bacterial species was consistently associated with infertility. Thus, bacterial sampling from infertile bitches was concluded to be of low diagnostic value. Bacterial species isolated from the bitches having vaginitis were present in pure culture in 26.9% of the samples while nonspecific mixed cultures were obtained from 34.6% of the samples from these bitches. E. coli was the most frequently isolated bacterial species from bitches with dead puppies. However, in such cases it is important to relate the vaginal bacterial findings to autopsy findings and the results of bacteriological cultures of the pups.

Causes of Pneumonia Epizootics among Bighorn Sheep, Western United States, 2008–2010

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Highland, Margaret A.; Baker, Katherine; Cassirer, E. Frances; Anderson, Neil J.; Ramsey, Jennifer M.; Mansfield, Kristin; Bruning, Darren L.; Wolff, Peregrine; Smith, Joshua B.; Jenks, Jonathan A.

2012-01-01

Epizootic pneumonia of bighorn sheep is a devastating disease of uncertain etiology. To help clarify the etiology, we used culture and culture-independent methods to compare the prevalence of the bacterial respiratory pathogens Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae in lung tissue from 44 bighorn sheep from herds affected by 8 outbreaks in the western United States. M. ovipneumoniae, the only agent detected at significantly higher prevalence in animals from outbreaks (95%) than in animals from unaffected healthy populations (0%), was the most consistently detected agent and the only agent that exhibited single strain types within each outbreak. The other respiratory pathogens were frequently but inconsistently detected, as were several obligate anaerobic bacterial species, all of which might represent secondary or opportunistic infections that could contribute to disease severity. These data provide evidence that M. ovipneumoniae plays a primary role in the etiology of epizootic pneumonia of bighorn sheep. PMID:22377321

Causes of pneumonia epizootics among bighorn sheep, Western United States, 2008-2010.

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Besser, Thomas E; Highland, Margaret A; Baker, Katherine; Cassirer, E Frances; Anderson, Neil J; Ramsey, Jennifer M; Mansfield, Kristin; Bruning, Darren L; Wolff, Peregrine; Smith, Joshua B; Jenks, Jonathan A

2012-03-01

Epizootic pneumonia of bighorn sheep is a devastating disease of uncertain etiology. To help clarify the etiology, we used culture and culture-independent methods to compare the prevalence of the bacterial respiratory pathogens Mannheimia haemolytica, Bibersteinia trehalosi, Pasteurella multocida, and Mycoplasma ovipneumoniae in lung tissue from 44 bighorn sheep from herds affected by 8 outbreaks in the western United States. M. ovipneumoniae, the only agent detected at significantly higher prevalence in animals from outbreaks (95%) than in animals from unaffected healthy populations (0%), was the most consistently detected agent and the only agent that exhibited single strain types within each outbreak. The other respiratory pathogens were frequently but inconsistently detected, as were several obligate anaerobic bacterial species, all of which might represent secondary or opportunistic infections that could contribute to disease severity. These data provide evidence that M. ovipneumoniae plays a primary role in the etiology of epizootic pneumonia of bighorn sheep.

Pneumonia in slaughtered sheep in south-western Iran: pathological characteristics and aerobic bacterial aetiology

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Shahrzad Azizi

2013-03-01

Full Text Available In this study, the lungs of 1,000 sheep carcasses were subjected to gross examination and those suspected to be infected with pneumonia were studied at histopathological level as well as examined for presence of bacteria. Pneumonia was detected in 42 (4.2% carcasses. Based on histopathological lesions, 45.24% were affected with suppurative bronchopneumonia, 20.93% with interstitial pneumonia, 11.9% bronchointerstitial pneumonia, 7.14% with fibrinous bronchopneumonia and 2.38% with embolic pneumonia. In addition, 11.9% of the lungs showed lung abscesses and 2.33% were affected with pleuritis without involving pulmonary parenchyma. Bacteriological examination revealed presence of ovine pathogens, such as Pasteurella multocida (24.53%, Staphylococcus aureus (20.75%, Klebsiella pneumoniae (15.09%, Corynebacterium pseudotuberculosis (7.55% and Actinomyces pyogenes (1.89%. The most common form of pneumonia was suppurative bronchopneumonia with moderate amounts of fibrin deposits on the pleural surface and inside the bronchioles and alveoli.

Bovine mastitis caused by gram negative bacteria in Mosul

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S. Y. A. Al-Dabbagh

2012-01-01

Full Text Available A total of 90 milk samples were collected from cows with clinical and subclinical mastitis from different areas in Mosul city, in a period from October 2009 to June 2010, for the detection of gram negative bacteriological causative agents. The bacteria were identified using morphological, cultural and biochemical characteristics. thirty tow (35.3% gram negative bacterial isolates were obtained from the total count which included 14 isolates (15.5% for Escherichia coli, 7 isolates (7.7% for Klebsiella spp, 4 isolates (4.4% for Pseudomonas aeruginosa, 3 isolates (3.3% for Enterobacter aerogenes ,2 isolates for Serratia marcescens and one isolates (1.1% for each of Aeromonas hydrophila and Pasteurella multocida. Results of antibiotic sensitivity test indicated that most of these isolates were sensitive to Ciprofloxacin following by Gentamycin and Cotrimoxazole, while most of these organisms were resistant to Ampicillin, the isolates showed different percentages of sensitivity to Doxycycline, Tetracycline, Neomycin and Chloramphenicol.

Mutant prevention concentration, pharmacokinetic-pharmacodynamic integration, and modeling of enrofloxacin data established in diseased buffalo calves.

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Ramalingam, B; Sidhu, P K; Kaur, G; Venkatachalam, D; Rampal, S

2015-12-01

The pharmacokinetic-pharmacodynamic (PK/PD) modeling of enrofloxacin data using mutant prevention concentration (MPC) of enrofloxacin was conducted in febrile buffalo calves to optimize dosage regimen and to prevent the emergence of antimicrobial resistance. The serum peak concentration (Cmax ), terminal half-life (t1/2 K10) , apparent volume of distribution (Vd(area) /F), and mean residence time (MRT) of enrofloxacin were 1.40 ± 0.27 μg/mL, 7.96 ± 0.86 h, 7.74 ± 1.26 L/kg, and 11.57 ± 1.01 h, respectively, following drug administration at dosage 12 mg/kg by intramuscular route. The minimum inhibitory concentration (MIC), minimum bactericidal concentration, and MPC of enrofloxacin against Pasteurella multocida were 0.055, 0.060, and 1.45 μg/mL, respectively. Modeling of ex vivo growth inhibition data to the sigmoid Emax equation provided AUC24 h /MIC values to produce effects of bacteriostatic (33 h), bactericidal (39 h), and bacterial eradication (41 h). The estimated daily dosage of enrofloxacin in febrile buffalo calves was 3.5 and 8.4 mg/kg against P. multocida/pathogens having MIC90 ≤0.125 and 0.30 μg/mL, respectively, based on the determined AUC24 h /MIC values by modeling PK/PD data. The lipopolysaccharide-induced fever had no direct effect on the antibacterial activity of the enrofloxacin and alterations in PK of the drug, and its metabolite will be beneficial for its use to treat infectious diseases caused by sensitive pathogens in buffalo species. In addition, in vitro MPC data in conjunction with in vivo PK data indicated that clinically it would be easier to eradicate less susceptible strains of P. multocida in diseased calves. © 2015 John Wiley & Sons Ltd.

Mortalidad por meningitis por Pasteurella canis. Oportunidades de aprendizaje

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Ana Rosa Ropero Vera

2016-01-01

Full Text Available La meningitis bacteriana es una enfermedad importante de distribución mundial, causa mayor y sustancial de mortalidad y morbilidad en países en desarrollo. La Organización Mundial de la Salud (OMS sostiene que la meningitis es una de las diez afecciones principales del ser humano y debe ser considerada como una emergencia infectológica; por eso es fundamental reconocer que esta enfermedad es causa de muerte en niños de todo el mundo, sin distinción de raza, nivel económico o sociocultural. Se realizó una investigación de caso en menor de 53 días de nacido, que cumplía con los criterios clínicos y de laboratorio compatible con meningitis bacteriana, con el propósito de analizar y fortalecer la toma de decisiones en salud pública por parte de la secretaría local de salud del municipio de Valledupar (Colombia. Entre los hallazgos se encontró antecedentes infecciosos en el menor, coloración de Gram y cultivo de LCR, en el que se identificó cocobacilos Gram negativos, que fueron aislados como agente causal Pasteurella canis. Este estudio pretende sensibilizar a los prestadores de salud para que cuenten con personal altamente capacitado para brindar tratamientos adecuados y prevenir complicaciones en la meningitis bacteriana en niños, y así disminuir la posibilidad de secuelas o muerte, tanto en pacientes con compromiso inmunológico o sin este.

New Data on Vaccine Antigen Deficient Bordetella pertussis Isolates

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Valérie Bouchez

2015-09-01

Full Text Available Evolution of Bordetella pertussis is driven by natural and vaccine pressures. Isolates circulating in regions with high vaccination coverage present multiple allelic and antigenic variations as compared to isolates collected before introduction of vaccination. Furthermore, during the last epidemics reported in regions using pertussis acellular vaccines, isolates deficient for vaccine antigens, such as pertactin (PRN, were reported to reach high proportions of circulating isolates. More sporadic filamentous hemagglutinin (FHA or pertussis toxin (PT deficient isolates were also collected. The whole genome of some recent French isolates, deficient or non-deficient in vaccine antigens, were analyzed. Transcription profiles of the expression of the main virulence factors were also compared. The invasive phenotype in an in vitro human tracheal epithelial (HTE cell model of infection was evaluated. Our genomic analysis focused on SNPs related to virulence genes known to be more likely to present allelic polymorphism. Transcriptomic data indicated that isolates circulating since the introduction of pertussis vaccines present lower transcription levels of the main virulence genes than the isolates of the pre-vaccine era. Furthermore, isolates not producing FHA present significantly higher expression levels of the entire set of genes tested. Finally, we observed that recent isolates are more invasive in HTE cells when compared to the reference strain, but no multiplication occurs within cells.

Widespread Bordetella parapertussis Infections-Wisconsin, 2011-2012: Clinical and Epidemiologic Features and Antibiotic Use for Treatment and Prevention.

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Koepke, Ruth; Bartholomew, Michael L; Eickhoff, Jens C; Ayele, Roman A; Rodd, Diane; Kuennen, Joan; Rosekrans, Jean; Warshauer, David M; Conway, James H; Davis, Jeffrey P

2015-11-01

During October 2011-December 2012, concurrent with a statewide pertussis outbreak, 443 Bordetella parapertussis infections were reported among Wisconsin residents. We examined clinical features of patients with parapertussis and the effect of antibiotic use for treatment and prevention. Patients with polymerase chain reaction results positive for B. parapertussis reported during October 2011-May 2012 were interviewed regarding presence and durations of pertussis-like symptoms and receipt of azithromycin treatment. Data regarding acute cough illnesses and receipt of azithromycin prophylaxis among parapertussis patient household members (HHMs) were also collected. Using multivariate repeated measures log-binomial regression analysis, we examined associations of treatment receipt by the HHM with the earliest illness onset and prophylaxis receipt among other HHMs with the presence of any secondary cough illnesses in the household. Among 218 patients with parapertussis, pertussis-like symptoms were frequently reported. Illness durations were significantly shorter among patients with treatment initiated 0-6 days after cough onset, compared with nonrecipients (median durations: 10 vs 19 days, P = .002). Among 361 HHMs from 120 households, compared with nonrecipients, prompt prophylaxis of HHMs was associated with no secondary cough illnesses (relative risk: 0.16; 95% confidence interval, .04-.69). Bordetella parapertussis infection causes pertussis-like illness that might be misclassified as pertussis if B. parapertussis testing is not performed. Prompt treatment might shorten illness duration, and prompt HHM prophylaxis might prevent secondary illnesses. Further study is needed to evaluate antibiotic effectiveness for preventing parapertussis and to determine risks and benefits of antibiotic use. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Inhibition of PCR-based assay for Bordetella pertussis by using calcium alginate fiber and aluminum shaft components of a nasopharyngeal swab.

OpenAIRE

Wadowsky, R M; Laus, S; Libert, T; States, S J; Ehrlich, G D

1994-01-01

A PCR-based assay for Bordetella pertussis was inhibited by using a calcium alginate fiber-tipped swab with an aluminum shaft but not by using a Dacron fiber-tipped swab with a plastic shaft. The calcium alginate fiber component inhibited the assay following storage for less than 1 min in a suspension of 10(3) CFU of B. pertussis per ml, whereas the aluminum shaft component required storage for at least 48 h in order to cause inhibition. We recommend the Dacron swab over the calcium alginate ...

Survival of bacteria of laboratory animal origin on cage bedding and inactivation by hydrogen peroxide vapour.

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Benga, Laurentiu; Benten, W Peter M; Engelhardt, Eva; Gougoula, Christina; Schulze-Röbbecke, Roland; Sager, Martin

2017-08-01

This study aims to determine the ability of laboratory animal bacteria to resist desiccation and inactivation by hydrogen peroxide vapour (HPV) on paper bedding pieces. Bedding pieces were saturated with bacterial suspensions in water or 2% (w/v) bovine serum albumin (BSA) in water, and held in a mouse facility. Viable counts showed variable survival rates over time for the bacterial species used ([ Pasteurella] pneumotropica, Muribacter muris, Pseudomonas aeruginosa, Acinetobacter redioresistens, Escherichia coli, Klebsiella oxytoca, Bordetella bronchiseptica, Bordetella hinzii, Enterococcus faecalis, β-haemolytic Streptococcus spp., Staphylococcus aureus and Staphylococcus xylosus). Overall, BSA increased bacterial survival in the bedding pieces. The survival rates of Bacillus safensis were not influenced by BSA but depended on sporulation. When bedding pieces and Petri dishes inoculated with E. coli, P. aeruginosa and S. aureus were subjected to HPV disinfection, all bacterial species on the bedding pieces inoculated with bacterial suspensions in water were readily inactivated. By contrast, S. aureus and P. aeruginosa, but not E. coli cells survived HPV treatment in high numbers when inoculated on bedding pieces as a BSA suspension. Notably, all three bacterial species were readily inactivated by HPV even in the presence of BSA when smeared on smooth surfaces. In conclusion, the suspension medium and the carrier can influence the environmental survival and susceptibility of bacterial species to HPV. Our results may help to develop standard protocols that can be used to ensure the microbiological quality of experimental rodent housing.

Anti-BACE1 and Antimicrobial Activities of Steroidal Compounds Isolated from Marine Urechis unicinctus

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Yong-Zhe Zhu

2018-03-01

Full Text Available The human β-site amyloid cleaving enzyme (BACE1 has been considered as an effective drug target for treatment of Alzheimer’s disease (AD. In this study, Urechis unicinctus (U. unicinctus, which is a Far East specialty food known as innkeeper worm, ethanol extract was studied by bioassay-directed fractionation and isolation to examine its potential β-site amyloid cleaving enzyme inhibitory and antimicrobial activity. The following compounds were characterized: hecogenin, cholest-4-en-3-one, cholesta-4,6-dien-3-ol, and hurgadacin. These compounds were identified by their mass spectrometry, 1H, and 13C NMR spectral data, comparing those data with NIST/EPA/NIH Mass spectral database (NIST11 and published values. Hecogenin and cholest-4-en-3-one showed significant inhibitory activity against BACE1 with EC50 values of 116.3 and 390.6 µM, respectively. Cholesta-4,6-dien-3-ol and hurgadacin showed broad spectrum antimicrobial activity, particularly strongly against Escherichia coli (E. coli, Salmonella enterica (S. enterica, Pasteurella multocida (P. multocida, and Physalospora piricola (P. piricola, with minimal inhibitory concentration (MIC ranging from 0.46 to 0.94 mg/mL. This is the first report regarding those four known compounds that were isolated from U. unicinctus and their anti-BACE1 and antimicrobial activity, highlighting the fact that known natural compounds may be a critical source of new medicine leads. These findings provide scientific evidence for potential application of those bioactive compounds for the development of AD drugs and antimicrobial agents.

Anti-BACE1 and Antimicrobial Activities of Steroidal Compounds Isolated from Marine Urechis unicinctus.

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Zhu, Yong-Zhe; Liu, Jing-Wen; Wang, Xue; Jeong, In-Hong; Ahn, Young-Joon; Zhang, Chuan-Jie

2018-03-14

The human β-site amyloid cleaving enzyme (BACE1) has been considered as an effective drug target for treatment of Alzheimer's disease (AD). In this study, Urechis unicinctus (U. unicinctus) , which is a Far East specialty food known as innkeeper worm, ethanol extract was studied by bioassay-directed fractionation and isolation to examine its potential β-site amyloid cleaving enzyme inhibitory and antimicrobial activity. The following compounds were characterized: hecogenin, cholest-4- en -3-one, cholesta-4,6- dien -3-ol, and hurgadacin. These compounds were identified by their mass spectrometry, ¹H, and 13 C NMR spectral data, comparing those data with NIST/EPA/NIH Mass spectral database (NIST11) and published values. Hecogenin and cholest-4- en -3-one showed significant inhibitory activity against BACE1 with EC 50 values of 116.3 and 390.6 µM, respectively. Cholesta-4,6- dien -3-ol and hurgadacin showed broad spectrum antimicrobial activity, particularly strongly against Escherichia coli (E. coli) , Salmonella enterica (S. enterica) , Pasteurella multocida (P. multocida) , and Physalospora piricola (P. piricola) , with minimal inhibitory concentration (MIC) ranging from 0.46 to 0.94 mg/mL. This is the first report regarding those four known compounds that were isolated from U. unicinctus and their anti-BACE1 and antimicrobial activity, highlighting the fact that known natural compounds may be a critical source of new medicine leads. These findings provide scientific evidence for potential application of those bioactive compounds for the development of AD drugs and antimicrobial agents.

Aerobic bacteria from mucous membranes, ear canals, and skin wounds of feral cats in Grenada, and the antimicrobial drug susceptibility of major isolates.

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Hariharan, Harry; Matthew, Vanessa; Fountain, Jacqueline; Snell, Alicia; Doherty, Devin; King, Brittany; Shemer, Eran; Oliveira, Simone; Sharma, Ravindra N

2011-03-01

In a 2-year period 54 feral cats were captured in Grenada, West Indies, and a total of 383 samples consisting of swabs from rectum, vagina, ears, eyes, mouth, nose and wounds/abscesses, were cultured for aerobic bacteria and campylobacters. A total of 251 bacterial isolates were obtained, of which 205 were identified to species level and 46 to genus level. A commercial bacterial identification system (API/Biomerieux), was used for this purpose. The most common species was Escherichia coli (N=60), followed by Staphylococcus felis/simulans (40), S. hominis (16), S. haemolyticus (12), Streptococcus canis (9), Proteus mirabilis (8), Pasteurella multocida (7), Streptococcus mitis (7), Staphylococcus xylosus (7), S. capitis (6), S. chromogenes (4), S. sciuri (3), S. auricularis (2), S. lentus (2), S. hyicus (2), Streptococcus suis (2) and Pseudomonas argentinensis (2). Sixteen other isolates were identified to species level. A molecular method using 16S rRNA sequencing was used to confirm/identify 22 isolates. Salmonella or campylobacters were not isolated from rectal swabs. E. coli and S. felis/simulans together constituted 50% of isolates from vagina. S. felis/simulans was the most common species from culture positive ear and eye samples. P. multocida was isolated from 15% of mouth samples. Coagulase-negative staphylococci were the most common isolates from nose and wound swabs. Staphylococcus aureus, or S. intemedius/S. pseudintermedius were not isolated from any sample. Antimicrobial drug resistance was minimal, most isolates being susceptible to all drugs tested against, including tetracycline. Copyright © 2010 Elsevier Ltd. All rights reserved.

Demonstration of 1-year duration of immunity for attenuated Bordetella bronchiseptica vaccines in dogs.

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Lehar, Craig; Jayappa, Huchappa; Erskine, Jason; Brown, Alicia; Sweeney, Diane; Wassmoen, Terri

2008-01-01

Three groups of healthy dogs with low antibody titers to Bordetella bronchiseptica (Bb), canine parainfluenza virus (CPI), and canine adenovirus type 2 (CAV-2) were used in this study. One group was vaccinated with a single dose of monovalent attenuated Bb vaccine and one group with a trivalent vaccine containing attenuated Bb, CPI, and CAV-2; dogs were vaccinated intranasally with a single dose of the respective vaccines. The third group served as unvaccinated controls. All vaccinated dogs subsequently developed serum antibody titers to Bb that persisted for at least 1 year. Following Bb challenge 1 year after vaccination, all vaccinated dogs, regardless of group, showed significantly fewer clinical signs and shed significantly fewer challenge organisms than unvaccinated controls. These results demonstrate that intranasal administration of a single dose of monovalent attenuated Bb vaccine or trivalent vaccine containing attenuated Bb, CPI, and CAV-2 provides 1 year of protection against Bb.

The BvgAS Regulon of Bordetella pertussis

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Kyung Moon

2017-10-01

Full Text Available Nearly all virulence factors in Bordetella pertussis are activated by a master two-component system, BvgAS, composed of the sensor kinase BvgS and the response regulator BvgA. When BvgS is active, BvgA is phosphorylated (BvgA~P, and virulence-activated genes (vags are expressed [Bvg(+ mode]. When BvgS is inactive and BvgA is not phosphorylated, virulence-repressed genes (vrgs are induced [Bvg(− mode]. Here, we have used transcriptome sequencing (RNA-seq and reverse transcription-quantitative PCR (RT-qPCR to define the BvgAS-dependent regulon of B. pertussis Tohama I. Our analyses reveal more than 550 BvgA-regulated genes, of which 353 are newly identified. BvgA-activated genes include those encoding two-component systems (such as kdpED, multiple other transcriptional regulators, and the extracytoplasmic function (ECF sigma factor brpL, which is needed for type 3 secretion system (T3SS expression, further establishing the importance of BvgA~P as an apex regulator of transcriptional networks promoting virulence. Using in vitro transcription, we demonstrate that the promoter for brpL is directly activated by BvgA~P. BvgA-FeBABE cleavage reactions identify BvgA~P binding sites centered at positions −41.5 and −63.5 in bprL. Most importantly, we show for the first time that genes for multiple and varied metabolic pathways are significantly upregulated in the B. pertussis Bvg(− mode. These include genes for fatty acid and lipid metabolism, sugar and amino acid transporters, pyruvate dehydrogenase, phenylacetic acid degradation, and the glycolate/glyoxylate utilization pathway. Our results suggest that metabolic changes in the Bvg(− mode may be participating in bacterial survival, transmission, and/or persistence and identify over 200 new vrgs that can be tested for function.

Aerobic salivary bacteria in wild and captive Komodo dragons.

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Montgomery, Joel M; Gillespie, Don; Sastrawan, Putra; Fredeking, Terry M; Stewart, George L

2002-07-01

During the months of November 1996, August 1997, and March 1998, saliva and plasma samples were collected for isolation of aerobic bacteria from 26 wild and 13 captive Komodo dragons (Varanus komodoensis). Twenty-eight Gram-negative and 29 Gram-positive species of bacteria were isolated from the saliva of the 39 Komodo dragons. A greater number of wild than captive dragons were positive for both Gram-negative and Gram-positive bacteria. The average number of bacterial species within the saliva of wild dragons was 46% greater than for captive dragons. While Escherichia coli was the most common bacterium isolated from the saliva of wild dragons, this species was not present in captive dragons. The most common bacteria isolated from the saliva of captive dragons were Staphylococcus capitis and Staphylococcus capitis and Staphylococcus caseolyticus, neither of which were found in wild dragons. High mortality was seen among mice injected with saliva from wild dragons and the only bacterium isolated from the blood of dying mice was Pasteurella multocida. A competitive inhibition enzyme-linked immunosorbent assay revealed the presence of anti-Pasteurella antibody in the plasma of Komodo dragons. Four species of bacteria isolated from dragon saliva showed resistance to one or more of 16 antimicrobics tested. The wide variety of bacteria demonstrated in the saliva of the Komodo dragon in this study, at least one species of which was highly lethal in mice and 54 species of which are known pathogens, support the observation that wounds inflicted by this animal are often associated with sepsis and subsequent bacteremia in prey animals.

Detection of Bordetella pertussis from Clinical Samples by Culture and End-Point PCR in Malaysian Patients.

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Ting, Tan Xue; Hashim, Rohaidah; Ahmad, Norazah; Abdullah, Khairul Hafizi

2013-01-01

Pertussis or whooping cough is a highly infectious respiratory disease caused by Bordetella pertussis. In vaccinating countries, infants, adolescents, and adults are relevant patients groups. A total of 707 clinical specimens were received from major hospitals in Malaysia in year 2011. These specimens were cultured on Regan-Lowe charcoal agar and subjected to end-point PCR, which amplified the repetitive insertion sequence IS481 and pertussis toxin promoter gene. Out of these specimens, 275 were positive: 4 by culture only, 6 by both end-point PCR and culture, and 265 by end-point PCR only. The majority of the positive cases were from ≤3 months old patients (77.1%) (P 0.05). Our study showed that the end-point PCR technique was able to pick up more positive cases compared to culture method.

Characterization of a Bordetella pertussis Diaminopimelate (DAP) Biosynthesis Locus Identifies dapC, a Novel Gene Coding for an N-Succinyl-l,l-DAP Aminotransferase

OpenAIRE

Fuchs, Thilo M.; Schneider, Boris; Krumbach, Karin; Eggeling, Lothar; Gross, Roy

2000-01-01

The functional complementation of two Escherichia coli strains defective in the succinylase pathway of meso-diaminopimelate (meso-DAP) biosynthesis with a Bordetella pertussis gene library resulted in the isolation of a putative dap operon containing three open reading frames (ORFs). In line with the successful complementation of the E. coli dapD and dapE mutants, the deduced amino acid sequences of two ORFs revealed significant sequence similarities with the DapD and DapE proteins of E. coli...

Characterization of a Bordetella pertussis diaminopimelate (DAP) biosynthesis locus identifies dapC, a novel gene coding for an N-succinyl-L, L-DAP aminotransferase

OpenAIRE

Fuchs, T. M.; Schneider, B.; Krumbach, K.; Eggeling, L.; Gross, S. M.

2000-01-01

The functional complementation of two Escherichia coli strains defective in the succinylase pathway of meso-diaminopimelate (meso DAP) biosynthesis with a Bordetella pertussis gene library resulted in the isolation of a putative dap operon containing three open reading frames (ORFs), In line with the successful complementation of the E, coli dapD and dapE mutants, the deduced amino acid sequences of two ORFs revealed significant sequence similarities with the DapD and DapE proteins of E, coli...

Live Attenuated Pertussis Vaccine BPZE1 Protects Baboons Against Bordetella pertussis Disease and Infection

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Papin, James F.; Lecher, Sophie; Debrie, Anne-Sophie; Thalen, Marcel; Solovay, Ken; Rubin, Keith; Mielcarek, Nathalie

2017-01-01

Abstract Evidence suggests that the resurgence of pertussis in many industrialized countries may result from the failure of current vaccines to prevent nasopharyngeal colonization by Bordetella pertussis, the principal causative agent of whooping cough. Here, we used a baboon model to test the protective potential of the novel, live attenuated pertussis vaccine candidate BPZE1. A single intranasal/intratracheal inoculation of juvenile baboons with BPZE1 resulted in transient nasopharyngeal colonization and induction of immunoglobulin G and immunoglobulin A to all antigens tested, while causing no adverse symptoms or leukocytosis. When BPZE1-vaccinated baboons were challenged with a high dose of a highly virulent B. pertussis isolate, they were fully protected against disease, whereas naive baboons developed illness (with 1 death) and leukocytosis. Total postchallenge nasopharyngeal virulent bacterial burden of vaccinated animals was substantially reduced (0.002%) compared to naive controls, providing promising evidence in nonhuman primates that BPZE1 protects against both pertussis disease and B. pertussis infection. PMID:28535276

Infecção por Bordetella pertussis com hipertensão pulmonar grave num recém-nascido com boa evolução clínica – Caso clínico

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Sérgia Soares

2008-09-01

Full Text Available Resumo: Apesar da ampla cobertura vacinal, a infecção por Bordetella pertussis está longe de estar controlada. Os recém-nascidos e lactentes ainda sem imunização completa e filhos de mães com baixos títulos de anticorpos para a Bordetella pertussis são altamente susceptíveis à infecção e têm maior risco de doença grave e morte. A infecção por Bordetella pertussis associada a hipertensão pulmonar no recém-nascido é frequentemente fatal. Os autores descrevem um caso clínico de doença grave num recém-nascido com insuficiência respiratória aguda e hipertensão pulmonar grave, tratado com sucesso com sildenafil e óxido nítrico inalado.Rev Port Pneumol 2008; XIV (5: 687-692 Abstract: In spite of the availability and widespread use of vaccines, pertussis is far from controlled. Newborns and infants too young to be fully vaccinated, born from mothers with low antibody titers to Bordetella pertussis, are highly susceptible to infection and at risk of severe disease and death. Pertussis associated with pulmonary hypertension in the newborn is often fatal. The authors report a clinical case of severe pertussis-induced respiratory failure associated to severe pulmonary hypertension in a neonate successfully treated with sildenafil and inhaled nitric oxide.Rev Port Pneumol 2008; XIV (5: 687-692 Palavras-chave: Recém-nascido, oxido nítrico, pertussis, hipertensão pulmonar, sildenafil, Key-words: Neonate, nitric oxide, pertussis, pulmonary hypertension, sildenafil

Inhibition of PCR-based assay for Bordetella pertussis by using calcium alginate fiber and aluminum shaft components of a nasopharyngeal swab.

Science.gov (United States)

Wadowsky, R M; Laus, S; Libert, T; States, S J; Ehrlich, G D

1994-04-01

A PCR-based assay for Bordetella pertussis was inhibited by using a calcium alginate fiber-tipped swab with an aluminum shaft but not by using a Dacron fiber-tipped swab with a plastic shaft. The calcium alginate fiber component inhibited the assay following storage for less than 1 min in a suspension of 10(3) CFU of B. pertussis per ml, whereas the aluminum shaft component required storage for at least 48 h in order to cause inhibition. We recommend the Dacron swab over the calcium alginate swab for collecting specimens for testing in PCR-based assays.

Clinical Diagnosis of Bordetella Pertussis Infection: A Systematic Review.

Science.gov (United States)

Ebell, Mark H; Marchello, Christian; Callahan, Maria

2017-01-01

Bordetella pertussis (BP) is a common cause of prolonged cough. Our objective was to perform an updated systematic review of the clinical diagnosis of BP without restriction by patient age. We identified prospective cohort studies of patients with cough or suspected pertussis and assessed study quality using QUADAS-2. We performed bivariate meta-analysis to calculate summary estimates of accuracy and created summary receiver operating characteristic curves to explore heterogeneity by vaccination status and age. Of 381 studies initially identified, 22 met our inclusion criteria, of which 14 had a low risk of bias. The overall clinical impression was the most accurate predictor of BP (positive likelihood ratio [LR+], 3.3; negative likelihood ratio [LR-], 0.63). The presence of whooping cough (LR+, 2.1) and posttussive vomiting (LR+, 1.7) somewhat increased the likelihood of BP, whereas the absence of paroxysmal cough (LR-, 0.58) and the absence of sputum (LR-, 0.63) decreased it. Whooping cough and posttussive vomiting have lower sensitivity in adults. Clinical criteria defined by the Centers for Disease Control and Prevention were sensitive (0.90) but nonspecific. Typical signs and symptoms of BP may be more sensitive but less specific in vaccinated patients. The clinician's overall impression was the most accurate way to determine the likelihood of BP infection when a patient initially presented. Clinical decision rules that combine signs, symptoms, and point-of-care tests have not yet been developed or validated. © Copyright 2017 by the American Board of Family Medicine.

Activity Enhancement Based on the Chemical Equilibrium of Multiple-Subunit Nitrile Hydratase from Bordetella petrii.

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Liu, Yi; Liu, Ping; Lin, Lu; Zhao, Yueqin; Zhong, Wenjuan; Wu, Lunjie; Zhou, Zhemin; Sun, Weifeng

2016-09-01

The maturation mechanism of nitrile hydratase (NHase) of Pseudomonas putida NRRL-18668 was discovered and named as "self-subunit swapping." Since the NHase of Bordetella petrii DSM 12804 is similar to that of P. putida, the NHase maturation of B. petrii is proposed to be the same as that of P. putida. However, there is no further information on the application of NHase according to these findings. We successfully rapidly purified NHase and its activator through affinity his tag, and found that the cell extracts of NHase possessed multiple types of protein ingredients including α, β, α2β2, and α(P14K)2 who were in a state of chemical equilibrium. Furthermore, the activity was significantly enhanced through adding extra α(P14K)2 to the cell extracts of NHase according to the chemical equilibrium. Our findings are useful for the activity enhancement of multiple-subunit enzyme and for the first time significantly increased the NHase activity according to the chemical equilibrium.

Preliminary clinical pharmacological investigations of tylosin and tiamulin in chickens.

Science.gov (United States)

Ziv, G

1980-10-15

The minimal inhibitory concentrations (MIC) of tiamulin and tylosin for mycoplasma, Gram-positive, and Gram-negative micro-organisms isolated from chickens were determinated by the agar dilution method. Median MIC values for tiamulin against Mycoplasma gallisepticum (0.05 microgram/ml) and Mycoplasma synoviae (0.10 microgram/ml) were 2 to 4 times lower than the corresponding values for tylosin. Tiamulin was also slightly more effective in vitro in inhibiting Escherichia coli, Pasteurella multocida, and beta-haemolytic streptococci than was tylosin. Groups of chicken were offered tiamulin medicated drinking water at rates of 125 and 250 mg/litre for 48 hours. Average serum tiamulin concentrations were 0.38 and 0.78 microgram/ml, respectively. When tylosin tartrate was added to the drinking water at 500 and 700 mg/litre, average serum drug levels were 0.12 and 0.17 microgram/ml, respectively. Tiamulin was 45% bound in chicken serum, as against 30% serum protein binding for tylosin. Correlations were made between free (non protein bound) serum drug levels and the MIC values of the two drugs. Such comparisons suggest that when tiamulin is given in the drinking water at rates of 125 to 250 mg/litre, better antimycoplasmal activity is to be expected in vivo than by giving tylosin tartrate in the drinking water at 500 to 700 mg/litre. Based on these data, no clinical efficacy of these dose rates can be expected in flocks infected by gram-negative micro-organisms such as E. coli or P. multocida. The tylosin tartrate rate of 500 to 700 mg/litre, may be clinical ineffective the treatment of Staphylococcus aureus infections.

Dose dependency of outcomes of intrapleural fibrinolytic therapy in new rabbit empyema models

Science.gov (United States)

Florova, Galina; Azghani, Ali O.; Buchanan, Ann; Boren, Jake; Allen, Timothy; Rahman, Najib M.; Koenig, Kathleen; Chamiso, Mignote; Karandashova, Sophia; Henry, James; Idell, Steven

2016-01-01

The incidence of empyema (EMP) is increasing worldwide; EMP generally occurs with pleural loculation and impaired drainage is often treated with intrapleural fibrinolytic therapy (IPFT) or surgery. A number of IPFT options are used clinically with empiric dosing and variable outcomes in adults. To evaluate mechanisms governing intrapleural fibrinolysis and disease outcomes, models of Pasteurella multocida and Streptococcus pneumoniae were generated in rabbits and the animals were treated with either human tissue (tPA) plasminogen activator or prourokinase (scuPA). Rabbit EMP was characterized by the development of pleural adhesions detectable by chest ultrasonography and fibrinous coating of the pleura. Similar to human EMP, rabbits with EMP accumulated sizable, 20- to 40-ml fibrinopurulent pleural effusions associated with extensive intrapleural organization, significantly increased pleural thickness, suppression of fibrinolytic and plasminogen-activating activities, and accumulation of high levels of plasminogen activator inhibitor 1, plasminogen, and extracellular DNA. IPFT with tPA (0.145 mg/kg) or scuPA (0.5 mg/kg) was ineffective in rabbit EMP (n = 9 and 3 for P. multocida and S. pneumoniae, respectively); 2 mg/kg tPA or scuPA IPFT (n = 5) effectively cleared S. pneumoniae-induced EMP collections in 24 h with no bleeding observed. Although intrapleural fibrinolytic activity for up to 40 min after IPFT was similar for effective and ineffective doses of fibrinolysin, it was lower for tPA than for scuPA treatments. These results demonstrate similarities between rabbit and human EMP, the importance of pleural fluid PAI-1 activity, and levels of plasminogen in the regulation of intrapleural fibrinolysis and illustrate the dose dependency of IPFT outcomes in EMP. PMID:27343192

Ceftiofur sodium, a broad-spectrum cephalosporin: evaluation in vitro and in vivo in mice.

Science.gov (United States)

Yancey, R J; Kinney, M L; Roberts, B J; Goodenough, K R; Hamel, J C; Ford, C W

1987-07-01

Ceftiofur sodium, a broad-spectrum beta-lactamase-resistant cephalosporin, was evaluated in vitro and in vivo in mice. Ceftiofur is the sodium salt of (6R, 7R)-7[( 2-amino-4-thiazolyl)-Z- (methoxyimino)acetyl]amino)-3-[( (2-furanylcarbonyl)thio]methyl)-8-oxo-5- thia-1-azabicyclo-[4.2.0]oct-2-ene-2-carboxylate. Minimal inhibitory concentration values were obtained with 264 strains representing 9 genera and 17 species of bacterial pathogens from cattle, swine, sheep, horses, poultry, dogs, cats, and human beings. Ceftiofur was more active than was ampicillin against all strains tested including beta-lactamase-producing organisms. In mice with systemic infections, ceftiofur was more active than or equivalent to ampicillin, cephalothin, cefamandole, cloxacillin, cefoperazone, or pirlimycin. These protection tests included infections with Escherichia coli, Haemophilus pleuropneumoniae, H somnus, Pasteurella haemolytica, P multocida, Salmonella typhimurium, or Staphylococcus aureus. In infant mice with E coli-induced lethal diarrhea and in mice with S aureus and E coli-induced mastitis, ceftiofur was comparable or more active than was ampicillin.

Granzyme A Produces Bioactive IL-1β through a Nonapoptotic Inflammasome-Independent Pathway

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Dagmar Hildebrand

2014-11-01

Full Text Available Bacterial components are recognized by the immune system through activation of the inflammasome, eventually causing processing of the proinflammatory cytokine interleukin-1β (IL-1β, a pleiotropic cytokine and one of the most important mediators of inflammation, through the protease caspase-1. Synthesis of the precursor protein and processing into its bioactive form are tightly regulated, given that disturbed control of IL-1β release can cause severe autoinflammatory diseases or contribute to cancer development. We show that the bacterial Pasteurella multocida toxin (PMT triggers Il1b gene transcription in macrophages independently of Toll-like receptor signaling through RhoA/Rho-kinase-mediated NF-κΒ activation. Furthermore, PMT mediates signal transducer and activator of transcription (STAT protein-controlled granzyme A (a serine protease expression in macrophages. The exocytosed granzyme A enters target cells and mediates IL-1β maturation independently of caspase-1 and without inducing cytotoxicity. These findings show that macrophages can induce an IL-1β-initiated immune response independently of inflammasome activity.

Development and characterization of attenuated metabolic mutants of Bordetella bronchiseptica for applications in vaccinology.

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Yevsa, Tetyana; Ebensen, Thomas; Fuchs, Barbara; Zygmunt, Beata; Libanova, Rimma; Gross, Roy; Schulze, Kai; Guzmán, Carlos A

2013-01-01

Bordetella bronchiseptica is an important pathogen causing a number of veterinary respiratory syndromes in agriculturally important and food-producing confinement-reared animals, resulting in great economic losses annually amounting to billions of euros worldwide. Currently available live vaccines are incompletely satisfactory in terms of efficacy and safety. An efficient vaccine for livestock animals would allow reducing the application of antibiotics, thereby preventing the massive release of pharmaceuticals into the environment. Here, we describe two new potential vaccine strains based on the BB7865 strain. Two independent attenuating mutations were incorporated by homologous recombination in order to make negligible the risk of recombination and subsequent reversion to the virulent phenotype. The mutations are critical for bacterial metabolism, resistance to oxidative stress, intracellular survival and in vivo persistence. The resulting double mutants BB7865 risA aroA and BB7865 risA dapE were characterized as promising vaccine candidates, which are able to confer protection against colonization of the lower respiratory tract after sublethal challenge with the wild-type strain. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP

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Hui Zhang

2013-10-01

Full Text Available Bordetella bronchiseptica causes acute and chronic respiratory infections in diverse animal species and occasionally in humans. In this study, we described the establishment of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP assay for the detection of B. bronchiseptica. A set of primers towards a 235 bp region within the flagellum gene of B. bronchiseptica was designed with online software.. The specificity of the LAMP assay was examined by using 6 porcine pathogens and 100 nasal swabs collected from healthy pigs and suspect infected pigs. The results indicated that positive reactions were confirmed for all B. bronchiseptica and no cross-reactivity was observed from other non-B. bronchiseptica. In sensitivity evaluations, the technique successfully detected a serial dilutions of extracted B. bronchiseptica DNA with a detection limit of 9 copies, which was 10 times more sensitive than that of PCR. Compared with conventional PCR, the higher sensitivity of LAMP method and no need for the complex instrumentation make this LAMP assay a promising alternative for the diagnosis of B. bronchiseptica in rural areas and developing countries where there lacks of complex laboratory services.

Differences in Purinergic Amplification of Osmotic Cell Lysis by the Pore-Forming RTX Toxins Bordetella pertussis CyaA and Actinobacillus pleuropneumoniae ApxIA: the Role of Pore Size

Czech Academy of Sciences Publication Activity Database

Mašín, Jiří; Fišer, Radovan; Linhartová, Irena; Osička, Radim; Bumba, Ladislav; Hewlett, E. L.; Benz, R.; Šebo, Peter

2013-01-01

Roč. 81, č. 12 (2013), s. 4571-4582 ISSN 0019-9567 R&D Projects: GA ČR GAP302/12/0460; GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GAP207/11/0717; GA AV ČR IAA500200914; GA ČR GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella pertussis * Actinobacillus pleuropneumoniae * E- coli Subject RIV: EE - Microbiology, Virology Impact factor: 4.156, year: 2013

Cyclic AMP-Elevating Capacity of Adenylate Cyclase Toxin-Hemolysin Is Sufficient for Lung Infection but Not for Full Virulence of Bordetella pertussis

Czech Academy of Sciences Publication Activity Database

Škopová, Karolína; Tomalová, Barbora; Kanchev, Ivan; Rossmann, Pavel; Švédová, Martina; Adkins, Irena; Bíbová, Ilona; Tomala, Jakub; Mašín, Jiří; Guiso, N.; Osička, Radim; Sedláček, Radislav; Kovář, Marek; Šebo, Peter

2017-01-01

Roč. 85, č. 6 (2017), s. 1-22, č. článku e00937-16. ISSN 0019-9567 R&D Projects: GA MZd(CZ) NV16-28126A; GA ČR(CZ) GA13-14547S; GA ČR GA13-12885S; GA ČR GA15-09157S; GA ČR(CZ) GAP302/12/0460; GA MŠk(CZ) LM2015064; GA MŠk(CZ) LM2015040 Institutional support: RVO:61388971 ; RVO:68378050 Keywords : Bordetella pertussis * adenylate cyclase toxin-hemolysin * cAMP intoxication Subject RIV: EE - Microbiology, Virology; EE - Microbiology, Virology (UMG-J) OBOR OECD: Microbiology; Microbiology (UMG-J) Impact factor: 3.593, year: 2016

Crystallization and preliminary X-ray diffraction analysis of P30, the transmembrane domain of pertactin, an autotransporter from Bordetella pertussis

International Nuclear Information System (INIS)

Zhu, Yanshi; Black, Isobel; Roszak, Aleksander W.; Isaacs, Neil W.

2007-01-01

P30, the transmembrane C-terminal domain of pertactin from B. pertussis has been crystallized after refolding in vitro. Preliminary X-ray crystallographic data are reported. P30, the 32 kDa transmembrane C-terminal domain of pertactin from Bordetella pertussis, is supposed to form a β-barrel inserted into the outer membrane for the translocation of the passenger domain. P30 was cloned and expressed in inclusion bodies in Escherichia coli. After refolding and purification, the protein was crystallized using the sitting-drop vapour-diffusion method at 292 K. The crystals diffract to a resolution limit of 3.5 Å using synchrotron radiation and belong to the hexagonal space group P6 1 22, with unit-cell parameters a = b = 123.27, c = 134.43 Å

Modulation of the NF-kappaB pathway by Bordetella pertussis filamentous hemagglutinin.

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Tzvia Abramson

Full Text Available Filamentous hemagglutinin (FHA is a cell-associated and secreted adhesin produced by Bordetella pertussis with pro-apoptotic and pro-inflammatory activity in host cells. Given the importance of the NF-kappaB transcription factor family in these host cell responses, we examined the effect of FHA on NF-kappaB activation in macrophages and bronchial epithelial cells, both of which are relevant cell types during natural infection.Exposure to FHA of primary human monocytes and transformed U-937 macrophages, but not BEAS-2B epithelial cells, resulted in early activation of the NF-kappaB pathway, as manifested by the degradation of cytosolic IkappaB alpha, by NF-kappaB DNA binding, and by the subsequent secretion of NF-kappaB-regulated inflammatory cytokines. However, exposure of macrophages and human monocytes to FHA for two hours or more resulted in the accumulation of cytosolic IkappaB alpha, and the failure of TNF-alpha to activate NF-kappaB. Proteasome activity was attenuated following exposure of cells to FHA for 2 hours, as was the nuclear translocation of RelA in BEAS-2B cells.These results reveal a complex temporal dynamic, and suggest that despite short term effects to the contrary, longer exposures of host cells to this secreted adhesin may block NF-kappaB activation, and perhaps lead to a compromised immune response to this bacterial pathogen.

Pharmacological Cyclophilin Inhibitors Prevent Intoxication of Mammalian Cells with Bordetella pertussis Toxin.

Science.gov (United States)

Ernst, Katharina; Eberhardt, Nina; Mittler, Ann-Katrin; Sonnabend, Michael; Anastasia, Anna; Freisinger, Simon; Schiene-Fischer, Cordelia; Malešević, Miroslav; Barth, Holger

2018-05-01

The Bordetella pertussis toxin (PT) is one important virulence factor causing the severe childhood disease whooping cough which still accounted for approximately 63,000 deaths worldwide in children in 2013. PT consists of PTS1, the enzymatically active (A) subunit and a non-covalently linked pentameric binding/transport (B) subunit. After endocytosis, PT takes a retrograde route to the endoplasmic reticulum (ER), where PTS1 is released into the cytosol. In the cytosol, PTS1 ADP-ribosylates inhibitory alpha subunits of trimeric GTP-binding proteins (Giα) leading to increased cAMP levels and disturbed signalling. Here, we show that the cyclophilin (Cyp) isoforms CypA and Cyp40 directly interact with PTS1 in vitro and that Cyp inhibitors cyclosporine A (CsA) and its tailored non-immunosuppressive derivative VK112 both inhibit intoxication of CHO-K1 cells with PT, as analysed in a morphology-based assay. Moreover, in cells treated with PT in the presence of CsA, the amount of ADP-ribosylated Giα was significantly reduced and less PTS1 was detected in the cytosol compared to cells treated with PT only. The results suggest that the uptake of PTS1 into the cytosol requires Cyps. Therefore, CsA/VK112 represent promising candidates for novel therapeutic strategies acting on the toxin level to prevent the severe, life-threatening symptoms caused by PT.

Probing the genome-scale metabolic landscape of Bordetella pertussis, the causative agent of whooping cough.

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Branco Dos Santos, Filipe; Olivier, Brett G; Boele, Joost; Smessaert, Vincent; De Rop, Philippe; Krumpochova, Petra; Klau, Gunnar W; Giera, Martin; Dehottay, Philippe; Teusink, Bas; Goffin, Philippe

2017-08-25

Whooping cough is a highly-contagious respiratory disease caused by Bordetella pertussi s. Despite vaccination, its incidence has been rising alarmingly, and yet, the physiology of B. pertussis remains poorly understood. We combined genome-scale metabolic reconstruction, a novel optimization algorithm and experimental data to probe the full metabolic potential of this pathogen, using strain Tohama I as a reference. Experimental validation showed that B. pertussis secretes a significant proportion of nitrogen as arginine and purine nucleosides, which may contribute to modulation of the host response. We also found that B. pertussis can be unexpectedly versatile, being able to metabolize many compounds while displaying minimal nutrient requirements. It can grow without cysteine - using inorganic sulfur sources such as thiosulfate - and it can grow on organic acids such as citrate or lactate as sole carbon sources, providing in vivo demonstration that its TCA cycle is functional. Although the metabolic reconstruction of eight additional strains indicates that the structural genes underlying this metabolic flexibility are widespread, experimental validation suggests a role of strain-specific regulatory mechanisms in shaping metabolic capabilities. Among five alternative strains tested, three were shown to grow on substrate combinations requiring a functional TCA cycle, but only one could use thiosulfate. Finally, the metabolic model was used to rationally design growth media with over two-fold improvements in pertussis toxin production. This study thus provides novel insights into B. pertussis physiology, and highlights the potential, but also limitations of models solely based on metabolic gene content. IMPORTANCE The metabolic capabilities of Bordetella pertussis - the causative agent of whooping cough - were investigated from a systems-level perspective. We constructed a comprehensive genome-scale metabolic model for B. pertussis , and challenged its predictions

Mutation in the β-hairpin of the Bordetella pertussis adenylate cyclase toxin modulates N-lobe conformation in calmodulin

International Nuclear Information System (INIS)

Springer, Tzvia I.; Goebel, Erich; Hariraju, Dinesh; Finley, Natosha L.

2014-01-01

Highlights: • Bordetella pertussis adenylate cyclase toxin modulates bi-lobal structure of CaM. • The structure and stability of the complex rely on intermolecular associations. • A novel mode of CaM-dependent activation of the adenylate cyclase toxin is proposed. - Abstract: Bordetella pertussis, causative agent of whooping cough, produces an adenylate cyclase toxin (CyaA) that is an important virulence factor. In the host cell, the adenylate cyclase domain of CyaA (CyaA-ACD) is activated upon association with calmodulin (CaM), an EF-hand protein comprised of N- and C-lobes (N-CaM and C-CaM, respectively) connected by a flexible tether. Maximal CyaA-ACD activation is achieved through its binding to both lobes of intact CaM, but the structural mechanisms remain unclear. No high-resolution structure of the intact CaM/CyaA-ACD complex is available, but crystal structures of isolated C-CaM bound to CyaA-ACD shed light on the molecular mechanism by which this lobe activates the toxin. Previous studies using molecular modeling, biochemical, and biophysical experiments demonstrate that CyaA-ACD’s β-hairpin participates in site-specific interactions with N-CaM. In this study, we utilize nuclear magnetic resonance (NMR) spectroscopy to probe the molecular association between intact CaM and CyaA-ACD. Our results indicate binding of CyaA-ACD to CaM induces large conformational perturbations mapping to C-CaM, while substantially smaller structural changes are localized primarily to helices I, II, and IV, and the metal-binding sites in N-CaM. Site-specific mutations in CyaA-ACD’s β-hairpin structurally modulate N-CaM, resulting in conformational perturbations in metal binding sites I and II, while no significant structural modifications are observed in C-CaM. Moreover, dynamic light scattering (DLS) analysis reveals that mutation of the β-hairpin results in a decreased hydrodynamic radius (R h ) and reduced thermal stability in the mutant complex. Taken together

Mutation in the β-hairpin of the Bordetella pertussis adenylate cyclase toxin modulates N-lobe conformation in calmodulin

Energy Technology Data Exchange (ETDEWEB)

Springer, Tzvia I.; Goebel, Erich; Hariraju, Dinesh [Department of Microbiology, Miami University, Oxford, OH 45056 (United States); Finley, Natosha L., E-mail: finleynl@miamioh.edu [Department of Microbiology, Miami University, Oxford, OH 45056 (United States); Cell, Molecular, and Structural Biology Program, Miami University, Oxford, OH 45056 (United States)

2014-10-10

Highlights: • Bordetella pertussis adenylate cyclase toxin modulates bi-lobal structure of CaM. • The structure and stability of the complex rely on intermolecular associations. • A novel mode of CaM-dependent activation of the adenylate cyclase toxin is proposed. - Abstract: Bordetella pertussis, causative agent of whooping cough, produces an adenylate cyclase toxin (CyaA) that is an important virulence factor. In the host cell, the adenylate cyclase domain of CyaA (CyaA-ACD) is activated upon association with calmodulin (CaM), an EF-hand protein comprised of N- and C-lobes (N-CaM and C-CaM, respectively) connected by a flexible tether. Maximal CyaA-ACD activation is achieved through its binding to both lobes of intact CaM, but the structural mechanisms remain unclear. No high-resolution structure of the intact CaM/CyaA-ACD complex is available, but crystal structures of isolated C-CaM bound to CyaA-ACD shed light on the molecular mechanism by which this lobe activates the toxin. Previous studies using molecular modeling, biochemical, and biophysical experiments demonstrate that CyaA-ACD’s β-hairpin participates in site-specific interactions with N-CaM. In this study, we utilize nuclear magnetic resonance (NMR) spectroscopy to probe the molecular association between intact CaM and CyaA-ACD. Our results indicate binding of CyaA-ACD to CaM induces large conformational perturbations mapping to C-CaM, while substantially smaller structural changes are localized primarily to helices I, II, and IV, and the metal-binding sites in N-CaM. Site-specific mutations in CyaA-ACD’s β-hairpin structurally modulate N-CaM, resulting in conformational perturbations in metal binding sites I and II, while no significant structural modifications are observed in C-CaM. Moreover, dynamic light scattering (DLS) analysis reveals that mutation of the β-hairpin results in a decreased hydrodynamic radius (R{sub h}) and reduced thermal stability in the mutant complex. Taken

Absence of antibodies against Bordetella pertussis in pregnant women and newborns in the state of Nuevo Leon.

Science.gov (United States)

Villarreal-Pérez, Jesús Zacarías; Ramírez-Aranda, José Manuel; Rodríguez-Rodríguez, Irasema; Perales-Dávila, José; García-Elizondo, Francisco Javier; Gómez-Gómez, Celina; Galindo-Galindo, Edgar; Rodríguez-Moreno, Marco Antonio; Ballesteros-Elizondo, Romelia

2014-09-01

To assess placental transfer of antibodies to the child at birth and at 2 months of age. For the quantification of anti-PT IgG antibodies, we used an indirect enzyme-linked immunosorbent assay standardized by The National Institute of Epidemiologic Diagnosis and Reference (InDRE). Samples were considered negative from 0 to 48 IU/mL, indeterminate from 49 to 93 IU/mL and positive at ≥94 IU/mL. We performed a cross-sectional assessment of anti-PT IgG antibody levels in the mother, umbilical cord, and child. There was a higher concentration of IgG anti-TP in the umbilical cord (4.3%) and in the mother (1.4%), but a total absence was observed in the child (0%). The vulnerability of children to Bordetella pertussis shows the need to implement effective immunization strategies, whether actively, in the child, or passively through the mother, adolescents, and adults who are in contact with the child.

Crystallization and preliminary X-ray diffraction analysis of P30, the transmembrane domain of pertactin, an autotransporter from Bordetella pertussis

Energy Technology Data Exchange (ETDEWEB)

Zhu, Yanshi; Black, Isobel; Roszak, Aleksander W.; Isaacs, Neil W., E-mail: n.isaacs@chem.gla.ac.uk [Department of Chemistry and WestChem, Glasgow Biomedical Research Centre, University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom)

2007-07-01

P30, the transmembrane C-terminal domain of pertactin from B. pertussis has been crystallized after refolding in vitro. Preliminary X-ray crystallographic data are reported. P30, the 32 kDa transmembrane C-terminal domain of pertactin from Bordetella pertussis, is supposed to form a β-barrel inserted into the outer membrane for the translocation of the passenger domain. P30 was cloned and expressed in inclusion bodies in Escherichia coli. After refolding and purification, the protein was crystallized using the sitting-drop vapour-diffusion method at 292 K. The crystals diffract to a resolution limit of 3.5 Å using synchrotron radiation and belong to the hexagonal space group P6{sub 1}22, with unit-cell parameters a = b = 123.27, c = 134.43 Å.

Comparative investigation on the efficacy of tulathromycin and florfenicol in the treatment of bronchopneumonia in feedlot calves

Directory of Open Access Journals (Sweden)

Jezdimirović Milanka

2011-01-01

Full Text Available The clinical efficacy of tulathromycin (TU and florfenicol (FL in the treatment of bronchopneumonia (BP caused by Pasteurella multocida which was isolated from nose swabs of diseased calves has been examined. The symptoms of bronchopneumonia (BP were quantified by means of the clinical score (CS with a maximum of 47 points. In the current investigation the average CS in diseased calves was 23.5± 0.15. The clinical efficacy of TU and FL was assessed every day in the first week after the administration of the drugs and was based on the decrease in CS and on microbiological findings on days 7, 28 and 35 after the completion of therapy. Tulathromycin was administered s.c., in the prescribed therapeutic dose (2.5 mg/kg BW, and florfenicol s.c., twice at a 48 h interval, in its respective therapeutic dose (40 mg/kg BW. In spite of the repeated administration of FL, TU was significantly more rapid to decrease the major clinical symptoms in the first four days following the application, in comparison with FL (P<0.05. On the fourth day after the administration, the clinical efficacy of TU in the therapy of BP in calves was 43.4±1.5 %, and of florfenicol 27.2±1.6 %. However, five days after the application of TU and two days after the repeated application of FL the assessed clinical efficacy of the two antibiotics was roughly the same. The average efficacy of TU was 57.1±0.0%, and of florfenicol 58.5±0.0%, both the individual and mean CS in the treated calves was 10 points, due to hyperthermia, which remained the only symptom. Six days after the administration of TU and three days after the repeated application of FL both antibiotics had equal maximum efficacy (100% in the treatment of BP. The clinical efficacy remained unchanged on day seven. The recovery was confirmed by the absence of P. multocida in nose swabs sampled on the seventh day after the initial treatment. However, in 4 calves (21.05 % of the 19 treated Streptococcus alpha haemolyticus

Thermodynamics of ligand binding to histone deacetylase like amidohydrolase from Bordetella/Alcaligenes.

Science.gov (United States)

Meyners, Christian; Baud, Matthias G J; Fuchter, Matthew J; Meyer-Almes, Franz-Josef

2014-03-01

Thermodynamic studies on ligand-protein binding have become increasingly important in the process of drug design. In combination with structural data and molecular dynamics simulations, thermodynamic studies provide relevant information about the mode of interaction between compounds and their target proteins and therefore build a sound basis for further drug optimization. Using the example of histone deacetylases (HDACs), particularly the histone deacetylase like amidohydrolase (HDAH) from Bordetella/Alcaligenes, a novel sensitive competitive fluorescence resonance energy transfer-based binding assay was developed and the thermodynamics of interaction of both fluorescent ligands and inhibitors to histone deacetylase like amidohydrolase were investigated. The assay consumes only small amounts of valuable target proteins and is suitable for fast kinetic and mechanistic studies as well as high throughput screening applications. Binding affinity increased with increasing length of aliphatic spacers (n = 4-7) between the hydroxamate moiety and the dansyl head group of ligand probes. Van't Hoff plots revealed an optimum in enthalpy contribution to the free energy of binding for the dansyl-ligand with hexyl spacer. The selectivity in the series of dansyl-ligands against human class I HDAC1 but not class II HDACs 4 and 6 increased with the ratio of ΔH(0)/ΔG(0). The data clearly emphasize the importance of thermodynamic signatures as useful general guidance for the optimization of ligands or rational drug design. Copyright © 2014 John Wiley & Sons, Ltd.

Cloning of Bordetella pertussis putative outer protein D (BopD) and Leucin/Isoleucine/Valin binding protein (LivJ)

Science.gov (United States)

Öztürk, Burcu Emine Tefon

2017-04-01

Whooping cough also known as pertussis is a contagious acute upper respiratory disease primarily caused by Bordetella pertussis. It is known that this disease may be fatal especially in infants and recently, the number of pertussis cases has been increased. Despite the fact that there are numbers of acellular vaccines on the market, the current acellular vaccine compositions are inadequate for providing sustainable immunity and avoiding subclinical disease cases. Hence, exploring novel proteins with high immune protective capacities is essential to enhance the clinical efficacy of current vaccines. In this study, genes of selected immunogenic proteins via -omics studies, namely Putative outer protein D (BopD) and Leucin/Isoleucine/Valin Binding Protein (LivJ) were first cloned into pGEM-T Easy vector and transformed to into E. coli DH5α cells and then cloned into the expression vector pET-28a(+) and transformed into E. coli BL21 (DE3) cells to express the proteins.

[Production of pertussis toxin by Bordetella pertussis strains isolated from patients with whooping cough].

Science.gov (United States)

Zaĭtsev, E M; Mertsalova, N U; Shinkarev, A S; Mazurova, I K; Zakharova, N S

2011-01-01

To assess level of pertussin toxin (PT) production by vaccine strains of Bordetella pertussis and strains isolated from patients with whooping cough. Concentration of PT in supernatants of microbial cultures of 3 vaccine strains and 25 strains of B. pertussis isolated from patients with pertussis in 2001 - 2005 was measured with enzyme immunoassay using gamma-globulin fractions of rabbit antiserum to PT as immunosorbent or included in peroxidase conjugates. Level of PT production by strains isolated from infected persons varied from 3 +/- 0.5 to 64.8 +/- 12.2 ng/MFU/ml: in 9 strains--from 3 +/- 0.5 to 9.4 +/- 2.1 ng/MFU/ml, in 7--10.5 +/- 1.8 to 18.4 +/- 2.6 ng/MFU/ml, and in 9--23.6 +/- 4.5 to 64.8 +/- 12.2 ng/MFU/ml. B. pertussis strains isolated from patients were heterogeneous on level of PT production. Difference in expression of PT between strains were as high as 20-fold. Conditionally low, moderate and high levels of PT production had 9 (36%), 7 (28%), and 9 (36%) of 25 studied strains. Three vaccine strains had levels of toxin production similar to recently isolated strains with moderate level of its production.

Immunomodulatory activity of methanolic extract of Morus alba Linn. (mulberry) leaves.

Science.gov (United States)

Bharani, Shendige Eswara Rao; Asad, Mohammed; Dhamanigi, Sunil Samson; Chandrakala, Gowda Kallenahalli

2010-01-01

The leaves of Morus alba Linn. (Family: Moraceae) commonly known as mulberry are mainly used as food for the silkworms and they are sometimes eaten as vegetable or used as cattle fodder in different parts of the world. The effect of Morus alba on the immune system was evaluated by using different experimental models such as carbon clearance test, cyclophosphamide induced neutropenia, neutrophil adhesion test, effect on serum immunoglobulins, mice lethality test and indirect haemagglutination test. Methanolic extract of Morus alba was administered orally at low dose and high dose of 100 mg/kg and 1 g/kg respectively and Ocimum sanctum (100 mg/kg, po) was used as standard drug. Morus alba extract in both doses increased the levels of serum immunoglobulins and prevented the mortality induced by bovine Pasteurella multocida in mice. It also increased the circulating antibody titre in indirect haemagglutination test. On the other hand, it showed significant increase in the phagocytic index in carbon clearance assay, a significant protection against cyclophosphamide induced neutropenia and increased the adhesion of neutrophils in the neutrophil adhesion test. Hence, it was concluded that Morus alba increases both humoral immunity and cell mediated immunity.

How Respiratory Pathogens Contribute to Lamb Mortality in a Poorly Performing Bighorn Sheep ( Ovis canadensis ) Herd.

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Wood, Mary E; Fox, Karen A; Jennings-Gaines, Jessica; Killion, Halcyon J; Amundson, Sierra; Miller, Michael W; Edwards, William H

2017-01-01

We evaluated bighorn sheep ( Ovis canadensis ) ewes and their lambs in captivity to examine the sources and roles of respiratory pathogens causing lamb mortality in a poorly performing herd. After seven consecutive years of observed December recruitments of sheep from the remnant Gribbles Park herd in Colorado, US were captured and transported to the Thorne-Williams Wildlife Research Center in Wyoming in March 2013. Ewes were sampled repeatedly over 16 mo. In April 2014, ewes were separated into individual pens prior to lambing. Upon death, lambs were necropsied and tested for respiratory pathogens. Six lambs developed clinical respiratory disease and one lamb was abandoned. Pathology from an additional six lambs born in 2013 was also evaluated. Mycoplasma ovipneumoniae , leukotoxigenic Mannheimia spp., leukotoxigenic Bibersteinia trehalosi , and Pasteurella multocida all contributed to lamb pneumonia. Histopathology suggested a continuum of disease, with lesions typical of pasteurellosis predominating in younger lambs and lesions typical of mycoplasmosis predominating in older lambs. Mixed pathology was observed in lambs dying between these timeframes. We suspected that all the ewes in our study were persistently infected and chronically shedding the bacteria that contributed to summer lamb mortality.

A Functional Tricarboxylic Acid Cycle Operates during Growth of Bordetella pertussis on Amino Acid Mixtures as Sole Carbon Substrates.

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Marie Izac

Full Text Available It has been claimed that citrate synthase, aconitase and isocitrate dehydrogenase activities are non-functional in Bordetella pertussis and that this might explain why this bacterium's growth is sometimes associated with accumulation of polyhydroxybutyrate (PHB and/or free fatty acids. However, the sequenced genome includes the entire citric acid pathway genes. Furthermore, these genes were expressed and the corresponding enzyme activities detected at high levels for the pathway when grown on a defined medium imitating the amino acid content of complex media often used for growth of this pathogenic microorganism. In addition, no significant PHB or fatty acids could be detected. Analysis of the carbon balance and stoichiometric flux analysis based on specific rates of amino acid consumption, and estimated biomass requirements coherent with the observed growth rate, clearly indicate that a fully functional tricarboxylic acid cycle operates in contrast to previous reports.

Influence of vaccination with Bordetella pertussis cells on haemopoiesis in sublethally irradiated mice and their radiation lethality

International Nuclear Information System (INIS)

Kwiek, S.; Bitny-Szlachto, S.

1978-01-01

Post-irradiation lethality of CFW mice has turned out to be enhanced by vaccination with Bordetella pertussis cells 10 min., 48 hrs. prior or 48 hrs. after the exposure to X-rays. The sensitization factor was found to be 1.23, as it revealed by decrease of radiation LD 50 . Granulopoiesis and erythropoiesis proved to be stimulated by vaccination, in mice irradiated with 200 or 400 R but not in those after 600 R. Direct radiosensitivity of CFU was not altered by vaccination, but the subsequent loss of bone marrow stem cells was enhanced in vaccinated mice. On the other hand, endocolonization of spleens with bone marrow stem cells has turned out to be highly enhanced by the vaccine, resulting in confluent growth of colonies. This effect of the vaccine was not abolished by hydroxyurea given 15 min. or 1 hr. after vaccination. Enhanced post-irradiation lethality is considered to result from fall of the bone marrow stem cell pool below the level indispensable to ensure the post-irradiation recovery of the haemopoietic system. (author)

Serodiagnosis of whooping cough in Belgium: results of the National Reference Centre for Bordetella pertussis anno 2013.

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Duterme, Sophie; Vanhoof, Raymond; Vanderpas, Jean; Pierard, Denis; Huygen, Kris

2016-04-01

Report on the pitfalls of serodiagnosis of pertussis in Belgium for 2013 by the NRC Bordetella. Determine cases of acute infection using an anti-pertussis toxin (PT) IgG antibody ELISA. A total of 2471 serum samples were received. Clinical information on the duration of cough (at moment of blood sampling) is essential for a reliable interpretation of the results. In order to avoid false negative results, 213 samples for which this information was lacking were not tested. For a total of 2179 patients tested, 520 (23.9%) had antibody levels indicative of an acute infection, 261 (12%) samples were diagnosed as positive (indicative of a pertussis infection or vaccination during the last year), 143 (6.7%) samples were classified as doubtful and 752 (34,5%) (35.5%) were diagnosed as negative. The serodiagnosis of pertussis has limited value for the early diagnosis of the disease and PCR analysis on nasopharyngeal swabs is the method of choice during the first 2 weeks and always for young children pertussis, serum samples should preferentially be collected 3 weeks after onset of symptoms.

Bordetella bronchiseptica antigen enhances the production of Mycoplasma hyopneumoniae antigen-specific immunoglobulin G in mice.

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Yim, Seol-Hwa; Hahn, Tae-Wook; Joo, Hong-Gu

2017-09-30

We previously demonstrated that Bordetella ( B .) bronchiseptica antigen (Ag) showed high immunostimulatory effects on mouse bone marrow cells (BMs) while Mycoplasma ( M .) hyopneumoniae Ag showed low effects. The focus of this study was to determine if B. bronchiseptica Ag can enhance the M. hyopneumoniae Ag-specific immune response and whether the host's immune system can recognize both Ags. MTT assay results revealed that each or both Ags did not significantly change BM metabolic activity. Flow cytometry analysis using carboxyfluorescein succinimidyl ester showed that B. bronchiseptica Ag can promote the division of BMs. In cytokine and nitric oxide (NO) assays, B. bronchiseptica Ag boosted production of tumor necrosis factor-alpha in M. hyopneumoniae Ag-treated BMs, and combined treatment with both Ags elevated the level of NO in BMs compared to that from treatment of M. hyopneumoniae Ag alone. Immunoglobulin (Ig)G enzyme-linked immunosorbent assay using the sera of Ag-injected mice clearly indicated that B. bronchiseptica Ag can increase the production of M. hyopneumoniae Ag-specific IgG. This study provided information valuable in the development of M. hyopneumoniae vaccines and showed that B. bronchiseptica Ag can be used both as a vaccine adjuvant and as a vaccine Ag.

Cloning and sequencing of a gene encoding a 21-kilodalton outer membrane protein from Bordetella avium and expression of the gene in Salmonella typhimurium.

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Gentry-Weeks, C R; Hultsch, A L; Kelly, S M; Keith, J M; Curtiss, R

1992-01-01

Three gene libraries of Bordetella avium 197 DNA were prepared in Escherichia coli LE392 by using the cosmid vectors pCP13 and pYA2329, a derivative of pCP13 specifying spectinomycin resistance. The cosmid libraries were screened with convalescent-phase anti-B. avium turkey sera and polyclonal rabbit antisera against B. avium 197 outer membrane proteins. One E. coli recombinant clone produced a 56-kDa protein which reacted with convalescent-phase serum from a turkey infected with B. avium 197. In addition, five E. coli recombinant clones were identified which produced B. avium outer membrane proteins with molecular masses of 21, 38, 40, 43, and 48 kDa. At least one of these E. coli clones, which encoded the 21-kDa protein, reacted with both convalescent-phase turkey sera and antibody against B. avium 197 outer membrane proteins. The gene for the 21-kDa outer membrane protein was localized by Tn5seq1 mutagenesis, and the nucleotide sequence was determined by dideoxy sequencing. DNA sequence analysis of the 21-kDa protein revealed an open reading frame of 582 bases that resulted in a predicted protein of 194 amino acids. Comparison of the predicted amino acid sequence of the gene encoding the 21-kDa outer membrane protein with protein sequences in the National Biomedical Research Foundation protein sequence data base indicated significant homology to the OmpA proteins of Shigella dysenteriae, Enterobacter aerogenes, E. coli, and Salmonella typhimurium and to Neisseria gonorrhoeae outer membrane protein III, Haemophilus influenzae protein P6, and Pseudomonas aeruginosa porin protein F. The gene (ompA) encoding the B. avium 21-kDa protein hybridized with 4.1-kb DNA fragments from EcoRI-digested, chromosomal DNA of Bordetella pertussis and Bordetella bronchiseptica and with 6.0- and 3.2-kb DNA fragments from EcoRI-digested, chromosomal DNA of B. avium and B. avium-like DNA, respectively. A 6.75-kb DNA fragment encoding the B. avium 21-kDa protein was subcloned into the

Molecular epidemiology of Bordetella pertussis in Cambodia determined by direct genotyping of clinical specimens.

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Moriuchi, Takumi; Vichit, Ork; Vutthikol, Yong; Hossain, Md Shafiqul; Samnang, Chham; Toda, Kohei; Grabovac, Varja; Hiramatsu, Yukihiro; Otsuka, Nao; Shibayama, Keigo; Kamachi, Kazunari

2017-09-01

This study sought to determine the genotypes of circulating Bordetella pertussis, the causative agent of pertussis, in Cambodia by direct molecular typing of clinical specimens. DNA extracts from nasopharyngeal swabs obtained from 82 pertussis patients in 2008-2016 were analyzed by multilocus variable-number tandem repeat analysis (MLVA). B. pertussis virulence-associated allelic genes (ptxA, prn, and fim3) and the pertussis toxin promoter ptxP were also investigated by DNA sequence-based typing. Forty-four DNA extracts (54%) yielded a complete MLVA profile, and these were sorted into 8 MLVA types (MT18, MT26, MT27, MT29, MT43, MT72, MT95, and MT200). MT27 and MT29, which are common in developed countries, were the predominant strain types (total 73%). The predominant profile of virulence-associated allelic genes was the combination of ptxP3/ptxA1/prn2/fim3A (48%). MT27 strains were detected during the entire study period, whereas MT29 strains were only found in 2014-2016. The B. pertussis population in Cambodia, where a whole-cell pertussis vaccine (WCV) has been continuously used, resembled those observed previously in developed countries where acellular pertussis vaccines are used. Circulating B. pertussis strains in Cambodia were distinct from those in other countries using WCVs. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

Modulation of pulmonary defense mechanisms by acute exposures to nitrogen dioxide. [Staphylococcus aureus; Proteus mirabilis; Pasteurella pneumotropica

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Jakab, G.J.

1987-02-01

The effect of acute exposures to NO/sub 2/ on the antibacterial defenses of the murine lung was assessed following inhalation challenges with Staphylococcus aureus, Proteus mirabilis, and Pasteurella pneumotropica. With S. aureus pulmonary antibacterial defenses were suppressed at NO/sub 2/ levels of 4.0 ppm and greater. Exposure to 10.0 ppm enhanced the intrapulmonary killing of P. mirabilis which correlated with an increase in the phagocytic cell populations lavaged from the lungs; at 20.0 ppm bactericidal activity against P. mirabilis was impaired. Pulmonary antibacterial defenses against P. pneumotropica were impaired at 10.0 ppm which correlated with a decrease in the retrieved phagocytic lung cell population. Reversing the order of treatment (ie., NO/sub 2/ exposure prior to bacterial challenge) raised the threshold concentration for NO/sub 2/-induced impairment of intrapulmonary bacterial killing. With S. aureus the effect was not observed at 5.0 ppm but at 10.0 ppm and with P. mirabilis not at 20.0 ppm but at 30.0 ppm intrapulmonary killing was enhanced. Exposures up to 20.0 ppm of NO/sub 2/ did not effect the physical translocation mechanisms of the lung as quantitated by declines in pulmonary radiotracer activity following aerogenic challenge with /sup 32/P-labeled staphylococci.

[Accumulation of the bvg- Bordetella pertussis a virulent mutants in the process of experimental whooping cough in mice].

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Medkova, A Iu; Siniashina, L N; Rumiantseva, Iu P; Voronina, O L; Kunda, M S; Karataev, G I

2013-01-01

The duration of the persistence and dynamics of accumulation of insertion bvg- Bordetella pertussis mutants were studied in lungs of laboratory mice after intranasal and intravenous challenge by virulent bacteria of the causative agent of whooping cough. The capability of the virulent B. pertussis bacteria to long-term persistence in the body of mice was tested. Using the real-time PCR approximately hundred genome equivalents of the B. pertussis DNA were detected in lungs of mice in two months after infection regardless of the way of challenge. Using the bacterial test bacteria were identified during only four weeks after challenge. Bvg- B. pertussis avirulent mutants were accumulated for the infection time. The percentage of the avirulent bacteria in the B. pertussis population reached 50% in 7-9 weeks after challenge. The obtained results show that the laboratory mice can be used for study of the B. pertussis insertion mutant formation dynamics in vivo and confirm the hypothesis about insertional bvg- B. pertussis virulent mutants accumulation during development of pertussis infection in human.

SNP-based typing: a useful tool to study Bordetella pertussis populations.

Directory of Open Access Journals (Sweden)

Marjolein van Gent

Full Text Available To monitor changes in Bordetella pertussis populations, mainly two typing methods are used; Pulsed-Field Gel Electrophoresis (PFGE and Multiple-Locus Variable-Number Tandem Repeat Analysis (MLVA. In this study, a single nucleotide polymorphism (SNP typing method, based on 87 SNPs, was developed and compared with PFGE and MLVA. The discriminatory indices of SNP typing, PFGE and MLVA were found to be 0.85, 0.95 and 0.83, respectively. Phylogenetic analysis, using SNP typing as Gold Standard, revealed false homoplasies in the PFGE and MLVA trees. Further, in contrast to the SNP-based tree, the PFGE- and MLVA-based trees did not reveal a positive correlation between root-to-tip distance and the isolation year of strains. Thus PFGE and MLVA do not allow an estimation of the relative age of the selected strains. In conclusion, SNP typing was found to be phylogenetically more informative than PFGE and more discriminative than MLVA. Further, in contrast to PFGE, it is readily standardized allowing interlaboratory comparisons. We applied SNP typing to study strains with a novel allele for the pertussis toxin promoter, ptxP3, which have a worldwide distribution and which have replaced the resident ptxP1 strains in the last 20 years. Previously, we showed that ptxP3 strains showed increased pertussis toxin expression and that their emergence was associated with increased notification in The Netherlands. SNP typing showed that the ptxP3 strains isolated in the Americas, Asia, Australia and Europe formed a monophyletic branch which recently diverged from ptxP1 strains. Two predominant ptxP3 SNP types were identified which spread worldwide. The widespread use of SNP typing will enhance our understanding of the evolution and global epidemiology of B. pertussis.

SNP-Based Typing: A Useful Tool to Study Bordetella pertussis Populations

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van der Heide, Han G. J.; Heuvelman, Kees J.; Kallonen, Teemu; He, Qiushui; Mertsola, Jussi; Advani, Abdolreza; Hallander, Hans O.; Janssens, Koen; Hermans, Peter W.; Mooi, Frits R.

2011-01-01

To monitor changes in Bordetella pertussis populations, mainly two typing methods are used; Pulsed-Field Gel Electrophoresis (PFGE) and Multiple-Locus Variable-Number Tandem Repeat Analysis (MLVA). In this study, a single nucleotide polymorphism (SNP) typing method, based on 87 SNPs, was developed and compared with PFGE and MLVA. The discriminatory indices of SNP typing, PFGE and MLVA were found to be 0.85, 0.95 and 0.83, respectively. Phylogenetic analysis, using SNP typing as Gold Standard, revealed false homoplasies in the PFGE and MLVA trees. Further, in contrast to the SNP-based tree, the PFGE- and MLVA-based trees did not reveal a positive correlation between root-to-tip distance and the isolation year of strains. Thus PFGE and MLVA do not allow an estimation of the relative age of the selected strains. In conclusion, SNP typing was found to be phylogenetically more informative than PFGE and more discriminative than MLVA. Further, in contrast to PFGE, it is readily standardized allowing interlaboratory comparisons. We applied SNP typing to study strains with a novel allele for the pertussis toxin promoter, ptxP3, which have a worldwide distribution and which have replaced the resident ptxP1 strains in the last 20 years. Previously, we showed that ptxP3 strains showed increased pertussis toxin expression and that their emergence was associated with increased notification in the Netherlands. SNP typing showed that the ptxP3 strains isolated in the Americas, Asia, Australia and Europe formed a monophyletic branch which recently diverged from ptxP1 strains. Two predominant ptxP3 SNP types were identified which spread worldwide. The widespread use of SNP typing will enhance our understanding of the evolution and global epidemiology of B. pertussis. PMID:21647370

Effect of different forms of adenylate cyclase toxin of Bordetella pertussis on protection afforded by an acellular pertussis vaccine in a murine model.

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Cheung, Gordon Y C; Xing, Dorothy; Prior, Sandra; Corbel, Michael J; Parton, Roger; Coote, John G

2006-12-01

Four recombinant forms of the cell-invasive adenylate cyclase toxin (CyaA) of Bordetella pertussis were compared for the ability to enhance protection against B. pertussis in mice when coadministered with an acellular pertussis vaccine (ACV). The four forms were as follows: fully functional CyaA, a CyaA form lacking adenylate cyclase enzymatic activity (CyaA*), and the nonacylated forms of these toxins, i.e., proCyaA and proCyaA*, respectively. None of these forms alone conferred significant (P > 0.05) protection against B. pertussis in a murine intranasal challenge model. Mice immunized with ACV alone showed significant (P protection was only significant (P protection provided by CyaA* was due to an augmentation of both Th1 and Th2 immune responses to B. pertussis antigens.

Crystallization and preliminary X-ray diffraction analysis of two extracytoplasmic solute receptors of the DctP family from Bordetella pertussis

International Nuclear Information System (INIS)

Rucktooa, Prakash; Huvent, Isabelle; Antoine, Rudy; Lecher, Sophie; Jacob-Dubuisson, Françoise; Villeret, Vincent; Bompard, Coralie

2006-01-01

Sample preparation, crystallization and preliminary X-ray analysis are reported for two B. pertussis extracytoplasmic solute receptors. DctP6 and DctP7 are two Bordetella pertussis proteins which belong to the extracytoplasmic solute receptors (ESR) superfamily. ESRs are involved in the transport of substrates from the periplasm to the cytosol of Gram-negative bacteria. DctP6 and DctP7 have been crystallized and diffraction data were collected using a synchrotron-radiation source. DctP6 crystallized in space group P4 1 2 1 2, with unit-cell parameters a = 108.39, b = 108.39, c = 63.09 Å, while selenomethionyl-derivatized DctP7 crystallized in space group P2 1 2 1 2 1 , with unit-cell parameters a = 64.87, b = 149.83, c = 170.65 Å. The three-dimensional structure of DctP7 will be determined by single-wavelength anomalous diffraction, while the DctP6 structure will be solved by molecular-replacement methods

Global Population Structure and Evolution of Bordetella pertussis and Their Relationship with Vaccination

Science.gov (United States)

Bart, Marieke J.; Harris, Simon R.; Advani, Abdolreza; Arakawa, Yoshichika; Bottero, Daniela; Bouchez, Valérie; Cassiday, Pamela K.; Chiang, Chuen-Sheue; Dalby, Tine; Fry, Norman K.; Gaillard, María Emilia; van Gent, Marjolein; Guiso, Nicole; Hallander, Hans O.; Harvill, Eric T.; He, Qiushui; van der Heide, Han G. J.; Heuvelman, Kees; Hozbor, Daniela F.; Kamachi, Kazunari; Karataev, Gennady I.; Lan, Ruiting; Lutyńska, Anna; Maharjan, Ram P.; Mertsola, Jussi; Miyamura, Tatsuo; Octavia, Sophie; Preston, Andrew; Quail, Michael A.; Sintchenko, Vitali; Stefanelli, Paola; Tondella, M. Lucia; Tsang, Raymond S. W.; Xu, Yinghua; Yao, Shu-Man; Zhang, Shumin; Mooi, Frits R.

2014-01-01

ABSTRACT Bordetella pertussis causes pertussis, a respiratory disease that is most severe for infants. Vaccination was introduced in the 1950s, and in recent years, a resurgence of disease was observed worldwide, with significant mortality in infants. Possible causes for this include the switch from whole-cell vaccines (WCVs) to less effective acellular vaccines (ACVs), waning immunity, and pathogen adaptation. Pathogen adaptation is suggested by antigenic divergence between vaccine strains and circulating strains and by the emergence of strains with increased pertussis toxin production. We applied comparative genomics to a worldwide collection of 343 B. pertussis strains isolated between 1920 and 2010. The global phylogeny showed two deep branches; the largest of these contained 98% of all strains, and its expansion correlated temporally with the first descriptions of pertussis outbreaks in Europe in the 16th century. We found little evidence of recent geographical clustering of the strains within this lineage, suggesting rapid strain flow between countries. We observed that changes in genes encoding proteins implicated in protective immunity that are included in ACVs occurred after the introduction of WCVs but before the switch to ACVs. Furthermore, our analyses consistently suggested that virulence-associated genes and genes coding for surface-exposed proteins were involved in adaptation. However, many of the putative adaptive loci identified have a physiological role, and further studies of these loci may reveal less obvious ways in which B. pertussis and the host interact. This work provides insight into ways in which pathogens may adapt to vaccination and suggests ways to improve pertussis vaccines. PMID:24757216

Multiplex PCR-based assay for detection of Bordetella pertussis in nasopharyngeal swab specimens.

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Wadowsky, R M; Michaels, R H; Libert, T; Kingsley, L A; Ehrlich, G D

1996-11-01

A multiplex PCR-based assay was developed for the detection of Bordetella pertussis in nasopharyngeal swab specimens. The assay simultaneously amplified two separate DNA targets (153 and 203 bp) within a B. pertussis repetitive element and a 438-bp target within the beta-actin gene of human DNA (PCR amplification control). PCR products were detected by a sensitive and specific liquid hybridization gel retardation assay. A total of 496 paired nasopharyngeal swab specimens were tested by both the PCR-based assay and culture. Although 30 (6%) of the specimens inhibited the amplification of the beta-actin target, in all 29 specimens studied, the inhibition disappeared on repeat testing or was easily overcome with a 1:8 dilution or less of specimen digest. Of the 495 specimen pairs yielding a final evaluable result by the PCR-based assay, 19.0% were positive by the PCR-based assay, whereas 13.9% were positive by culture (P < 0.0001). After resolving the PCR-positive, culture-negative results by testing an additional aliquot from these specimens by the multiplex PCR-based assay, the PCR-based assay had a sensitivity and specificity of 98.9 and 99.7%, respectively, compared with values of 73.4 and 100%, respectively, for culture. In comparison with patients with culture-confirmed pertussis, those with PCR-positive, culture-negative results were older and more likely to have had prolonged cough, immunization with pertussis vaccine, or treatment with erythromycin. This multiplex PCR-based assay is substantially more sensitive than culture and identifies specimens that contain inhibitors of PCR.

The relationship between mucosal immunity, nasopharyngeal carriage, asymptomatic transmission and the resurgence of Bordetella pertussis

Science.gov (United States)

Gill, Christopher; Rohani, Pejman; Thea, Donald M

2017-01-01

The incidence of whooping cough in the US has been rising slowly since the 1970s, but the pace of this has accelerated sharply since acellular pertussis vaccines replaced the earlier whole cell vaccines in the late 1990s. A similar trend occurred in many other countries, including the UK, Canada, Australia, Ireland, and Spain, following the switch to acellular vaccines. The key question is why. Two leading theories (short duration of protective immunologic persistence and evolutionary shifts in the pathogen to evade the vaccine) explain some but not all of these shifts, suggesting that other factors may also be important. In this synthesis, we argue that sterilizing mucosal immunity that blocks or abbreviates the duration of nasopharyngeal carriage of Bordetella pertussis and impedes person-to-person transmission (including between asymptomatically infected individuals) is a critical factor in this dynamic. Moreover, we argue that the ability to induce such mucosal immunity is fundamentally what distinguishes whole cell and acellular pertussis vaccines and may be pivotal to understanding much of the resurgence of this disease in many countries that adopted acellular vaccines. Additionally, we offer the hypothesis that observed herd effects generated by acellular vaccines may reflect a modification of disease presentation leading to reduced potential for transmission by those already infected, as opposed to inducing resistance to infection among those who have been exposed. PMID:28928960

The role of lipooligosaccharide phosphorylcholine in colonization and pathogenesis of Histophilus somni in cattle

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Elswaifi Shaadi F

2012-06-01

Full Text Available Abstract Histophilus somni is a Gram-negative bacterium and member of the Pasteurellaceae that is responsible for respiratory disease and other systemic infections in cattle. One of the bacterium’s virulence factors is antigenic phase variation of its lipooligosaccharide (LOS. LOS antigenic variation may occur through variation in composition or structure of glycoses or their substitutions, such as phosphorylcholine (ChoP. However, the role of ChoP in the pathogenesis of H. somni disease has not been established. In Haemophilus influenzae ChoP on the LOS binds to platelet activating factor on epithelial cells, promoting bacterial colonization of the host upper respiratory tract. However, ChoP is not expressed in the blood as it also binds C-reactive protein, resulting in complement activation and killing of the bacteria. In order to simulate the susceptibility of calves with suppressed immunity due to stress or previous infection, calves were challenged with bovine herpes virus-1 or dexamethazone 3 days prior to challenge with H. somni. Following challenge, expression of ChoP on the LOS of 2 different H. somni strains was associated with colonization of the upper respiratory tract. In contrast, lack of ChoP expression was associated with bacteria recovered from systemic sites. Histopathology of cardiac tissue from myocarditis revealed lesions containing bacterial clusters that appeared similar to a biofilm. Furthermore, some respiratory cultures contained substantial numbers of Pasteurella multocida, which were not present on preculture screens. Subsequent biofilm experiments have shown that H. somni and P. multocida grow equally well together in a biofilm, suggesting a commensal relationship may exist between the two species. Our results also showed that ChoP contributed to, but was not required for, adhesion to respiratory epithelial cells. In conclusion, expression of ChoP on H. somni LOS contributed to colonization of the bacteria to the

Potential probiotic attributes of a new strain of Bacillus coagulans CGMCC 9951 isolated from healthy piglet feces.

Science.gov (United States)

Gu, Shao-Bin; Zhao, Li-Na; Wu, Ying; Li, Shi-Chang; Sun, Jian-Rui; Huang, Jing-Fang; Li, Dan-Dan

2015-06-01

A new strain of Bacillus coagulans CGMCC 9551, which has a broad range of antibacterial activities against six main pathogenic bacteria including Escherichia coli O8, Staphylococcus aureus, Salmonella enterica subsp. enterica serovar enteritidis, Streptococcus suis, Listeria monocytogenes and Pasteurella multocida, was isolated from healthy piglet feces. In adhesion assay, the isolate exhibited a stronger adhesion to pig intestinal mucus than that of B. subtilis JT143 and L. acidophilus LY24 respectively isolated from BioPlus(®)2B and FloraFIT(®) Probiotics (P coagulans CGMCC 9551 was reduced by only 20% at 4 h exposure under 0.9% w/v bile salt. The strain was fully resistant to pH 2 for 2 h with 90.1 ± 3.5% survival and susceptible to 15 antibiotics commonly used in veterinary medicine. Additionally, the bacteria showed amylase, protease and cellulase activities. The safety assessment demonstrated the lack of toxicity potential in B. coagulans CGMCC 9551 by ligated rabbit ileal loop assay, acute and subchronic toxicity test. These results implied that that the new strain of B. coagulans CGMCC 9951 isolated from healthy piglet feces has promising probiotic characteristics and offers desirable opportunities for its successful commercialization as one excellent candidate probiotic.

Occurrence and severity of lung lesions in slaughter pigs vaccinated against Mycoplasma hyopneumoniae with different strategies.

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Hillen, Sonja; von Berg, Stephan; Köhler, Kernt; Reinacher, Manfred; Willems, Hermann; Reiner, Gerald

2014-03-01

Different vaccination strategies against Mycoplasma hyopneumoniae have been adopted worldwide. Reports from the field indicate varying levels of protection among currently available vaccines. The goal of the present study was to compare the efficacies of three widespread commercial vaccination strategies against M. hyopneumoniae under field conditions. 20 farms were included. 14 farms used different single dose vaccines (vaccine 1 [V1], 8 herds; vaccine 2 [V2], 6 herds); another 6 farms (V3) used a two dose vaccination strategy. Gross lesions of 854 lungs and histopathology from 140 lungs were quantified, and a quantitative PCR was applied to detect M. hyopneumoniae and porcine circovirus 2 (PCV2) DNA in lung tissue (n=140). In addition, porcine reproductive and respiratory disease virus (PRRSV), swine influenza virus (SIV), Actinobacillus pleuropneumoniae, Haemophilus parasuis and Pasteurella multocida were tested by qualitative PCR. 53% of lungs were positive for M. hyopneumoniae. 55.9% of lungs showed macroscopic enzootic pneumonia (EP)-like lesions. Lung lesion scores (Phyopneumoniae-loads (Phyopneumoniae indicating that the applied diagnostic tools are valuable in confirming the prevalence and severity of M. hyopneumoniae infections. Comparing different vaccination strategies against M. hyopneumoniae indicates varying levels of protection. M. hyopneumoniae is still a major problem despite the widely applied vaccination. Copyright © 2014 Elsevier B.V. All rights reserved.

Crystallization and preliminary X-ray diffraction analysis of two extracytoplasmic solute receptors of the DctP family from Bordetella pertussis

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Rucktooa, Prakash; Huvent, Isabelle [UMR8161 CNRS Institut de Biologie de Lille, Laboratoire de Cristallographie Macromoléculaire, 1 Rue du Professeur Calmette, BP 447, 59021 Lille CEDEX (France); IFR 142, Institut Pasteur de Lille, 1 Rue du Professeur Calmette, BP 245, 59021 Lille CEDEX (France); Antoine, Rudy; Lecher, Sophie; Jacob-Dubuisson, Françoise, E-mail: francoise.jacob@ibl.fr [IFR 142, Institut Pasteur de Lille, 1 Rue du Professeur Calmette, BP 245, 59021 Lille CEDEX (France); INSERM-U629, Lille (France); Institut Pasteur de Lille, 1 Rue du Professeur Calmette, BP 245, 59021 Lille CEDEX (France); Villeret, Vincent, E-mail: francoise.jacob@ibl.fr; Bompard, Coralie [UMR8161 CNRS Institut de Biologie de Lille, Laboratoire de Cristallographie Macromoléculaire, 1 Rue du Professeur Calmette, BP 447, 59021 Lille CEDEX (France); IFR 142, Institut Pasteur de Lille, 1 Rue du Professeur Calmette, BP 245, 59021 Lille CEDEX (France)

2006-10-01

Sample preparation, crystallization and preliminary X-ray analysis are reported for two B. pertussis extracytoplasmic solute receptors. DctP6 and DctP7 are two Bordetella pertussis proteins which belong to the extracytoplasmic solute receptors (ESR) superfamily. ESRs are involved in the transport of substrates from the periplasm to the cytosol of Gram-negative bacteria. DctP6 and DctP7 have been crystallized and diffraction data were collected using a synchrotron-radiation source. DctP6 crystallized in space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = 108.39, b = 108.39, c = 63.09 Å, while selenomethionyl-derivatized DctP7 crystallized in space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 64.87, b = 149.83, c = 170.65 Å. The three-dimensional structure of DctP7 will be determined by single-wavelength anomalous diffraction, while the DctP6 structure will be solved by molecular-replacement methods.

Pasteurella pneumotropica evades the human complement system by acquisition of the complement regulators factor H and C4BP.

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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.

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

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

2014-01-01

/Kmvalue for the enzyme using 3-sialyllactose as the donor for 6-sialyllactose synthesis at pH 5.4 and 40◦Cwas determined to be 23.22 ± 0.7 M−1s−1. Moreover, the enzyme was capable of catalyzing the synthesisof both 3- and 6-sialylated galactooligosaccharides, when galactooligosaccharides served as acceptors....

Susceptibility of Rocky Mountain bighorn sheep and domestic sheep to pneumonia induced by bighorn and domestic livestock strains of Pasteurella haemolytica.

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Onderka, D K; Rawluk, S A; Wishart, W D

1988-10-01

Bighorn sheep were inoculated intratracheally with suspensions of nonhemolytic Pasteurella haemolytica biotype T (10(12) organisms) unique to wild bighorns, with beta-hemolytic P. haemolytica biotype T (10(12) organisms) isolated from clinically normal domestic sheep or intradermally with half a dose of a cattle vaccine containing P. haemolytica biotype A (10(5) organisms). The bighorn strain caused lobar necrotizing bronchopneumonia whereas both domestic livestock strains precipitated fatal septicemia and fibrinous bronchopneumonia. The serotypes given were T3, T4, T15 and A1 and these were recovered from lung lesions and other organs. In three trials, domestic sheep were inoculated intratracheally with suspensions of bighorn sheep pneumonic lungs, and two concentrations of the P. haemolytica bighorn strain (10(4) and 10(12) organisms). One of these sheep was inoculated intrabronchially. The domestic sheep experienced a transient fever and elevated white blood cell counts. After six days, none of the sheep had lung lesions and inoculated organisms could not be recovered. It is suggested that bighorn sheep are very susceptible to P. haemolytica from domestic livestock and should not be allowed in contact with sheep or cattle.

Susceptibility of Dall sheep (Ovis dalli dalli) to pneumonia caused by Pasteurella haemolytica.

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Foreyt, W J; Silflow, R M; Lagerquist, J E

1996-10-01

We evaluated susceptibility of Dall sheep (Ovis dalli dalli) to bacterial pneumonia induced by two strains of Pasteurella haemolytica of domestic sheep origin by evaluating the sensitivity of blood neutrophils of eight Dall sheep to lysis by cytotoxins of P. haemolytica, and by intratracheal inoculation of three Dall sheep, two bighorn sheep (Ovis canadensis), and two domestic sheep with 3.7 x 10(6) or 2.5 x 10(7) colony forming units of P. haemolytica. Neutrophils from the Dall sheep were more sensitive to lysis by cytotoxins from supernatants of a P. haemolytica, biotype A, serotype 2 (A2), of domestic sheep origin, than were neutrophils from six bighorn sheep. This cytotoxic bacterium was the same isolate that was used for intratracheal inoculation of two Dall sheep and two domestic sheep. Inoculation of this cytotoxic P. haemolytica A2 resulted in fatal fibrinopurulent pleuropneumonia in the first Dall sheep within 24 hr of inoculation, and pneumonic lesions in the second Dall sheep before it was euthanized 52 hr after inoculation. This strain of P. haemolytica A2 did not cause respiratory disease when inoculated into two domestic sheep. A noncytotoxic strain of P. haemolytica; biotype T, serotype 3,4,10 of domestic sheep origin did not result in pneumonia in the third Dall sheep or two bighorn sheep. Prior to inoculation, P. haemolytica, biotype T isolates were obtained from all three Dall sheep, but none of these isolates was cytotoxic. At necropsy, cytotoxic P. haemolytica A2 was isolated from lungs and other tissues of the two pneumonic Dall sheep. Based on these results, we conclude that Dall sheep appear to be at least as sensitive as bighorn sheep to pneumonia caused by P. haemolytica A2 of domestic sheep origin. Because in vitro and in vivo results appear closely correlated in this and other studies, we believe with additional evaluation and standardization, neutrophil cytotoxicity tests may serve as a substitute for live animal challenges in future

A new topology of the HK97-like fold revealed in Bordetella bacteriophage by cryoEM at 3.5 Å resolution

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Zhang, Xing; Guo, Huatao; Jin, Lei; Czornyj, Elizabeth; Hodes, Asher; Hui, Wong H; Nieh, Angela W; Miller, Jeff F; Zhou, Z Hong

2013-01-01

Bacteriophage BPP-1 infects and kills Bordetella species that cause whooping cough. Its diversity-generating retroelement (DGR) provides a naturally occurring phage-display system, but engineering efforts are hampered without atomic structures. Here, we report a cryo electron microscopy structure of the BPP-1 head at 3.5 Å resolution. Our atomic model shows two of the three protein folds representing major viral lineages: jellyroll for its cement protein (CP) and HK97-like (‘Johnson’) for its major capsid protein (MCP). Strikingly, the fold topology of MCP is permuted non-circularly from the Johnson fold topology previously seen in viral and cellular proteins. We illustrate that the new topology is likely the only feasible alternative of the old topology. β-sheet augmentation and electrostatic interactions contribute to the formation of non-covalent chainmail in BPP-1, unlike covalent inter-protein linkages of the HK97 chainmail. Despite these complex interactions, the termini of both CP and MCP are ideally positioned for DGR-based phage-display engineering. DOI: http://dx.doi.org/10.7554/eLife.01299.001 PMID:24347545

Phenotypical and genotypical characterization of Bordetella pertussis strains isolated in São Paulo, Brazil, 1988-2002 Caracterização fenotípica e genética de cepas de Bordetella pertussis isoladas em São Paulo, Brasil, 1988-2002

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Célia R. Gonçalves

2007-04-01

Full Text Available Whooping cough or pertussis was a major cause of childhood morbidity and mortality in the world until the introduction of a whole-cell vaccine in the 1940's. However, since the early 1980's whooping cough cases have increased in many countries, becoming an important problem of public health. This increase may be due to accuracy of laboratory diagnosis and reporting of the disease, a decline in immunity over time, demographic changes, and adaptation of the bacterial population to vaccine-induced immunity. The purpose of this study was to analyze phenotypically and genotypically a collection of 67 Bordetella pertussis isolates recovered during the period 1988-2002 in São Paulo State, Brazil to determine their characteristics and relatedness. All isolates were submitted to susceptibility testing to erythromycin, serotyping, and 56 isolates were analyzed by Pulsed Field Gel Electrophoresis (PFGE. All isolates were susceptible to erythromycin and the majority of them belonged to serotype 1,3. The 56 isolates were classified into 11 PFGE profiles according to the differences in banding patterns. Although more than 60% of the isolates were recovered from patients aged less than three months, almost 15% of them were isolated from adolescents/adults evidencing the increase in the incidence of pertussis among this group of age.A coqueluche ou pertussis foi a maior causa de morbidade e mortalidade infantil em todo o mundo até a introdução de uma vacina na década de 1940. Entretanto, desde a década de 1980, a coqueluche tornou-se, em muitos países , um importante problema de saúde pública. Este acontecimento pode ser atribuído à melhoria do diagnóstico laboratorial e da notificação da doença, declínio da imunidade no decorrer do tempo, mudanças demográficas ou adaptação da população bacteriana à imunidade induzida pela vacina. O objetivo deste estudo foi analisar as características fenotípicas e genotípicas de uma coleção de 67

Identifying the age cohort responsible for transmission in a natural outbreak of Bordetella bronchiseptica.

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Gráinne H Long

2010-12-01

Full Text Available Identifying the major routes of disease transmission and reservoirs of infection are needed to increase our understanding of disease dynamics and improve disease control. Despite this, transmission events are rarely observed directly. Here we had the unique opportunity to study natural transmission of Bordetella bronchiseptica--a directly transmitted respiratory pathogen with a wide mammalian host range, including sporadic infection of humans--within a commercial rabbitry to evaluate the relative effects of sex and age on the transmission dynamics therein. We did this by developing an a priori set of hypotheses outlining how natural B. bronchiseptica infections may be transmitted between rabbits. We discriminated between these hypotheses by using force-of-infection estimates coupled with random effects binomial regression analysis of B. bronchiseptica age-prevalence data from within our rabbit population. Force-of-infection analysis allowed us to quantify the apparent prevalence of B. bronchiseptica while correcting for age structure. To determine whether transmission is largely within social groups (in this case litter, or from an external group, we used random-effect binomial regression to evaluate the importance of social mixing in disease spread. Between these two approaches our results support young weanlings--as opposed to, for example, breeder or maternal cohorts--as the age cohort primarily responsible for B. bronchiseptica transmission. Thus age-prevalence data, which is relatively easy to gather in clinical or agricultural settings, can be used to evaluate contact patterns and infer the likely age-cohort responsible for transmission of directly transmitted infections. These insights shed light on the dynamics of disease spread and allow an assessment to be made of the best methods for effective long-term disease control.

Bordetella Pertussis virulence factors in the continuing evolution of whooping cough vaccines for improved performance.

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Dorji, Dorji; Mooi, Frits; Yantorno, Osvaldo; Deora, Rajendar; Graham, Ross M; Mukkur, Trilochan K

2018-02-01

Despite high vaccine coverage, whooping cough caused by Bordetella pertussis remains one of the most common vaccine-preventable diseases worldwide. Introduction of whole-cell pertussis (wP) vaccines in the 1940s and acellular pertussis (aP) vaccines in 1990s reduced the mortality due to pertussis. Despite induction of both antibody and cell-mediated immune (CMI) responses by aP and wP vaccines, there has been resurgence of pertussis in many countries in recent years. Possible reasons hypothesised for resurgence have ranged from incompliance with the recommended vaccination programmes with the currently used aP vaccine to infection with a resurged clinical isolates characterised by mutations in the virulence factors, resulting in antigenic divergence with vaccine strain, and increased production of pertussis toxin, resulting in dampening of immune responses. While use of these vaccines provide varying degrees of protection against whooping cough, protection against infection and transmission appears to be less effective, warranting continuation of efforts in the development of an improved pertussis vaccine formulations capable of achieving this objective. Major approaches currently under evaluation for the development of an improved pertussis vaccine include identification of novel biofilm-associated antigens for incorporation in current aP vaccine formulations, development of live attenuated vaccines and discovery of novel non-toxic adjuvants capable of inducing both antibody and CMI. In this review, the potential roles of different accredited virulence factors, including novel biofilm-associated antigens, of B. pertussis in the evolution, formulation and delivery of improved pertussis vaccines, with potential to block the transmission of whooping cough in the community, are discussed.

The importance of subcutaneous abscess infection by Pasteurella spp. and Staphylococcus aureus as a cause of meat condemnation in slaughtered commercial rabbits

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A. Ferreira

2014-12-01

Full Text Available Subcutaneous abscesses are lesions frequently reported in commercial rabbits. Both at farm and slaughterhouse level, these lesions are responsible for economic losses and a potential decrease in meat quality. The present study was devised to identify the main causes of meat condemnation in slaughtered commercial rabbits and assess the importance of abscess lesions in this domain. For these purposes, 281423 rabbits were evaluated during meat inspection at the slaughterhouse. The results achieved showed that subcutaneous abscesses were the major cause of condemnation, being responsible for the rejection of 1355 (0.48% rabbit carcasses. The main affected area was the hind limbs (31.37%, followed by the cervical area (23.10%. Microbiological analyses of 27 abscess samples indicated Pasteurella spp. as the bacteria mostly isolated (59.3%, followed by Staphylococcus aureus (25.9%. These results enable us to advise the industry on the significance of abscesses as an important cause of economic losses, due to meat condemnation during post mortem inspection, and highlight the importance of implementing monitoring plans as a way to control this pathological problem.

Genotypic characterization by multi locus variable number of tandem repeats analysis international Bordetella pertussis vaccine strains

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M. Fatah Moghadam

2017-10-01

Full Text Available Background: In 1930's first whole cell pertussis vaccines became available to the public heralding a dramatic success in overcoming the global burden of the disease. To date only a handful of B. pertussis strains have been used by international/local pertussis vaccine manufacturers. Inevitable well-documented genetic changes in the world population of this pathogen have prompted serious questions on suitability of traditional vaccine strains protect human against currently circulating wild isolates of Bordetella pertussis. Objective: Analyzing the genetic diversity within the most frequently-used vaccine strains of B. pertussis in the world Methods: A recently developed multi locus variable number of tandem repeats analysis (MLVA genotyping system along with a bioinforamtic piece of analysis was conducted on 11 strain / substrains of B137, B203 (10536, C393, Cs, E476, Tohama I, J445 (134, B202 and J446 (509 plus 2 sub-strains of 134 and 509 that are used at Razi institute for preparation of pertussis vaccine. In this study have used 6 individual loci of VNTR1, VNTR3a, VNTR3b, VNTR4, VNTR5 and VNTR6. Findings: Six distinct genotypes were recognized among the examined strains by comparing our data with the Dutch MLVA databank. These were all new and not reported before in the database. Conclusion: This observation reiterates on necessity for detection of predominant native strains to include in vaccine preparations suitable for different countries.

ORF Alignment: NC_002663 [GENIUS II[Archive

Lifescience Database Archive (English)

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ORF Alignment: NC_002663 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available multocida ... subsp. multocida str. Pm70] ... Length = 159 ... Query: 2 ... TTNRQEVLQNRLGPEIQELKAQ...CKTVMLATVGEDGNPNVSYAPFAINNGEYQVFISTIAR 61 ... TTNRQEVLQNRLGPEIQELKAQCKTVML...ATVGEDGNPNVSYAPFAINNGEYQVFISTIAR Sbjct: 1 ... TTNRQEVLQNRLGPEIQELKAQCKTVMLATVGEDGNPNVSYAPFAINNGEYQVFISTIAR 60

[Immunogenicity and protective efficacy of pertactin recombinants against Bordetella bronchiseptica challenge].

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Zhao, Zhanqin; Wang, Chen; Xue, Yun; Ding, Ke; Zhang, Chunjie; Cheng, Xiangchao; Li, Yinju; Liu, Yichen; Wu, Tingcai

2010-09-01

In this study we showed the immunogenicity and protective efficacy of five pertactin recombinants against Bordetella bronchiseptica (Bb) challenge. The complete coding sequence (2040 bp) of the prn gene (PRN) and its fragments,5'-terminal 1173 bp fragment (PN),3'-terminal 867 bp fragment (PC), two copies of region I (654 bp; PR I) in PN, and 2 copies of region II (678 bp; PR II) in PC, were separately cloned into the prokaryotic expression vector pGEX-KG, and expressed in the Eschierichia coli BL21 (DE3) using induction by isopropyl-beta-D-thiogalactopyranoside. The recombinant proteins were named GST-PRN, GST-PN, GST-PC, GST-2PR I and GST-2PR II. All five recombinant proteins showed immunological reactivity in the Western-blot analysis. Mice, immunized subcutaneously with two doses of the purified proteins mixed with an equal volume of Freund's adjuvant,produced robust PRN-specific IgG antibody levels. When challenged, 6 of 9 mice in GST-2PR I group and all 9 mice in the other groups survived intranasal challenge with three times the 50% lethal dose (LD50) of virulent Bb HH0809. After challenge with 10 LD50 7/9,3/9,6/9,1/10 and 6/10 of the mice survived. Furthermore, complete protection against intraperitoneal (i.p.) challenge with 10 LD50 of HH0809 was observed in mice that were injected i.p. with 0.5 ml rabbit anti-GST-PRN, GST-PN,GST-PC or GST-2PR II serum. Only 1 of 10 mice survived in the group of mice that received anti-GST-2PR I, and no survivors were noted in the group of mice that received PRN-absorbed rabbit antiserum (0/5). In this study,we showed that all of five pertactin recombinants had differential immunogenicity and protective efficacy against Bb challenge. Mice immunized with GST-PC had better survival against fatal Bb challenge than did those immunized with GST-PN. In addition, GST-2PR II and GST-2PR I provided the similar results These data may have implications for the development of safe and efficacious subunit vaccines for the prevention of

ORF Alignment: NC_002663 [GENIUS II[Archive

Lifescience Database Archive (English)

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Substantial gaps in knowledge of Bordetella pertussis antibody and T cell epitopes relevant for natural immunity and vaccine efficacy

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Vaughan, Kerrie; Seymour, Emily; Peters, Bjoern; Sette, Alessandro

2016-01-01

The recent increase in whooping cough in vaccinated populations has been attributed to waning immunity associated with the acellular vaccine. The Immune Epitope Database (IEDB) is a repository of immune epitope data from the published literature and includes T cell and antibody epitopes for human pathogens. The IEDB conducted a review of the epitope literature, which revealed 300 Bordetella pertussis-related epitopes from 39 references. Epitope data are currently available for six virulence factors of B. pertussis: pertussis toxin, pertactin, fimbrial 2, fimbrial 3, adenylate cyclase and filamentous hemagglutinin. The majority of epitopes were defined for antibody reactivity; fewer T cell determinants were reported. Analysis of available protective correlates data revealed a number of candidate epitopes; however few are defined in humans and few have been shown to be protective. Moreover, there are a limited number of studies defining epitopes from natural infection versus whole cell or acellular/subunit vaccines. The relationship between epitope location and structural features, as well as antigenic drift (SNP analysis) was also investigated. We conclude that the cumulative data is yet insufficient to address many fundamental questions related to vaccine failure and this underscores the need for further investigation of B. pertussis immunity at the molecular level. PMID:24530743

Chronic intestinal pseudo-obstruction associated with enteric ganglionitis in a Persian cat

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Jeremy Mortier

2016-06-01

Full Text Available Case summary A 7-year-old neutered male Persian cat was presented for acute vomiting and inappetence. Physical examination revealed severe abdominal distension. Radiographs demonstrated pneumoperitoneum, megaoesophagus and generalised gaseous distension of the digestive tract. Exploratory coeliotomy was performed, revealing markedly distended and thickened small and large intestines with no observable peristalsis. No intestinal perforation was present. Bacteriological and cytological analysis of abdominal fluid revealed a septic peritonitis involving Pasteurella multocida . Full-thickness intestinal biopsies demonstrated lymphocytic ganglioneuritis localised to the enteric nervous system, in association with glandular atrophy and muscular layer hypertrophy. Amoxicillin-clavulanate and analgesics were given. The cat’s general condition gradually improved after the addition of pyridostigmine bromide (0.5 mg/kg q12h PO, initiated 3 days postsurgery. Vomiting resolved and did not recur. Follow-up radiographs at 15 days, and 1 and 6 months showed persistent intestinal ileus, milder than on the pretreatment radiographs. Thirty months after presentation the cat is still alive, without clinical signs, and receives 1 mg/kg q12h pyridostigmine. Relevance and novel information To our knowledge, this is the first case of ganglioneuritis of the myenteric plexus described in cats, as well as the first one successfully treated with pyridostigmine. Chronic intestinal pseudo-obstruction is a very rare condition in cats but should be included in the differential diagnosis of generalised gastrointestinal ileus.

Isolation and Antibiogram of Aerobic Nasal Bacterial Flora of Apparently Healthy West African Dwarf Goats

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B. O. Emikpe

2009-01-01

Full Text Available Goats are important in the livestock economy by their adaptability to adverse environmental conditions as they are good sources of protein and income for the rural poor. Studies conducted on the bacterial flora of the respiratory tract in goats focused on the pneumonic lungs, with fewer studies on the apparently normal nasal passage and antibiogram of isolated organisms. This study was carried out on 60 apparently healthy West African Dwarf goats. The nasal swab from each goat was analyzed using standard methods. The disc diffusion technique was used for the antibiotic sensitivity test. Three hundred and twenty-eight isolates were obtained. The most frequently isolated species was Streptococcus spp., while Escherichia coli and Staphylococcus aureus were the second dominant bacteria. Other species were isolated at relatively lower rates. The isolation of Mannheimia haemolytica and Pasteurella multocida from the nasal cavity of apparently healthy goats in this study reflects their possible role in most common respiratory diseases encountered in small ruminants. Most of the bacteria were found to be susceptible to streptomycin, quinolones (perfloxacin, ciprofloxacin and ofloxacin and gentamicin, while they were resistant to tetracycline, augmentin and erythromycin. This study shows the relationship between misuse or unrestricted use of antibiotics and drug resistance. Therefore, there is a need for practitioners and researchers to be informed of the appropriate antibiotics to be used in respiratory infections and during control programs.

Chronic intestinal pseudo-obstruction associated with enteric ganglionitis in a Persian cat.

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Mortier, Jeremy; Elissalt, Estelle; Palierne, Sophie; Semin, Marie Odile; Delverdier, Maxence; Diquélou, Armelle

2016-01-01

Case summary A 7-year-old neutered male Persian cat was presented for acute vomiting and inappetence. Physical examination revealed severe abdominal distension. Radiographs demonstrated pneumoperitoneum, megaoesophagus and generalised gaseous distension of the digestive tract. Exploratory coeliotomy was performed, revealing markedly distended and thickened small and large intestines with no observable peristalsis. No intestinal perforation was present. Bacteriological and cytological analysis of abdominal fluid revealed a septic peritonitis involving Pasteurella multocida . Full-thickness intestinal biopsies demonstrated lymphocytic ganglioneuritis localised to the enteric nervous system, in association with glandular atrophy and muscular layer hypertrophy. Amoxicillin-clavulanate and analgesics were given. The cat's general condition gradually improved after the addition of pyridostigmine bromide (0.5 mg/kg q12h PO), initiated 3 days postsurgery. Vomiting resolved and did not recur. Follow-up radiographs at 15 days, and 1 and 6 months showed persistent intestinal ileus, milder than on the pretreatment radiographs. Thirty months after presentation the cat is still alive, without clinical signs, and receives 1 mg/kg q12h pyridostigmine. Relevance and novel information To our knowledge, this is the first case of ganglioneuritis of the myenteric plexus described in cats, as well as the first one successfully treated with pyridostigmine. Chronic intestinal pseudo-obstruction is a very rare condition in cats but should be included in the differential diagnosis of generalised gastrointestinal ileus.

Quantification and characterization of enzymatically produced hyaluronan with fluorophore-assisted carbohydrate electrophoresis.

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Kooy, Floor K; Ma, Muyuan; Beeftink, Hendrik H; Eggink, Gerrit; Tramper, Johannes; Boeriu, Carmen G

2009-01-15

Hyaluronan (HA) is a polysaccharide with high-potential medical applications, depending on the chain length and the chain length distribution. Special interest goes to homogeneous HA oligosaccharides, which can be enzymatically produced using Pasteurella multocida hyaluronan synthase (PmHAS). We have developed a sensitive, simple, and fast method, based on fluorophore-assisted carbohydrate electrophoresis (FACE), for characterization and quantification of polymerization products. A chromatographic pure fluorescent template was synthesized from HA tetrasaccharide (HA4) and 2-aminobenzoic acid. HA4-fluor and HA4 were used as template for PmHAS-mediated polymerization of nucleotide sugars. All products, fluorescent and nonfluorescent, were analyzed with gel electrophoresis and quantified using lane densitometry. Comparison of HA4- and HA4-fluor-derived polymers showed that the fluorophore did not negatively influence the PmHAS-mediated polymerization. Only even-numbered oligosaccharide products were observed using HA4-fluor or HA4 as template. The fluorophore intensity was linearly related to its concentration, and the limit of detection was determined to be 7.4pmol per product band. With this assay, we can now differentiate oligosaccharides of size range DP2 (degree of polymerization 2) to approximately DP400, monitor the progress of polymerization reactions, and measure subtle differences in polymerization rate. Quantifying polymerization products enables us to study the influence of experimental conditions on HA synthesis.

Serologic survey for selected arboviruses and other potential pathogens in wildlife from Mexico.

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Aguirre, A A; McLean, R G; Cook, R S; Quan, T J

1992-07-01

During 1988 and 1989, a serologic survey of wildlife was conducted in northeastern Mexico to determine the presence, prevalence, and distribution of arboviruses and other selected disease agents. Eighty mammal specimens were tested. Antibodies to vesicular stomatitis-Indiana, Venezuelan equine encephalitis-Mena II, Rio Grande virus, and vesicular stomatitis-New Jersey were detected predominantly in small mammals. Deer and mouflon (Ovis musimon) had antibodies to bluetongue and epizootic hemorrhagic disease. Two species had serologic evidence of recent exposure to Francisella tularensis. A white-tailed deer (Odocoileus virginianus) had antibodies to Anaplasma marginale. All specimens tested for antibodies against Yersinia pestis and Brucella abortus were negative. Sera from 315 birds were tested for antibody against five equine encephalitis viruses and six avian pathogens. During 1988, antibodies to Venezuelan equine encephalitis-Mena II, Venezuelan equine encephalitis-TC83, St. Louis encephalitis, eastern equine encephalitis, and western equine encephalitis were detected in birds of several species. Antibodies to Pasteurella multocida and Newcastle disease virus were also detected. Birds from five species presented antibodies to Mycoplasma meleagridis. Specimens tested for M. gallisepticum, M. synoviae, and Chlamydia psittaci were negative. To the best of our knowledge, this survey represents the first serologic evidence of bluetongue, Cache Valley virus, epizootic hemorrhagic disease, Jamestown Canyon virus, vesicular stomatitis-Indiana, vesicular stomatitis-New Jersey, Rio Grande virus, and tularemia reported among wildlife in Mexico.

Genomic content of Bordetella pertussis clinical isolates circulating in areas of intensive children vaccination.

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Valérie Bouchez

Full Text Available BACKGROUND: The objective of the study was to analyse the evolution of Bordetella pertussis population and the influence of herd immunity in different areas of the world where newborns and infants are highly vaccinated. METHODOLOGY: The analysis was performed using DNA microarray on 15 isolates, PCR on 111 isolates as well as GS-FLX sequencing technology on 3 isolates and the B. pertussis reference strain, Tohama I. PRINCIPAL FINDINGS: Our analyses demonstrate that the current circulating isolates are continuing to lose genetic material as compared to isolates circulating during the pre-vaccine era whatever the area of the world considered. The lost genetic material does not seem to be important for virulence. Our study confirms that the use of whole cell vaccines has led to the control of isolates that were similar to vaccine strains. GS-FLX sequencing technology shows that current isolates did not acquire any additional material when compared with vaccine strains or with isolates of the pre-vaccine era and that the sequenced strain Tohama I is not representative of the isolates. Furthermore, this technology allowed us to observe that the number of Insertion Sequence elements contained in the genome of the isolates is temporally increasing or varying between isolates. CONCLUSIONS: B. pertussis adaptation to humans is still in progress by losing genetic material via Insertion Sequence elements. Furthermore, recent isolates did not acquire any additional material when compared with vaccine strains or with isolates of the pre-vaccine era. Herd immunity, following intensive vaccination of infants and children with whole cell vaccines, has controlled isolates similar to the vaccine strains without modifying significantly the virulence of the isolates. With the replacement of whole cell vaccines by subunit vaccines, containing only few bacterial antigens targeting the virulence of the bacterium, one could hypothesize the circulation of isolates

The importance of Bordetella pertussis strains which do not produce virulence factors in the epidemiology of pertussis

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Maciej Polak

2017-05-01

Full Text Available Bordetella pertussis strains, which have lost the ability to produce antigens, such as pertactin - Prn, pertussis toxin - Ptx, filamentous haemagglutinin - FHA, fimbriae type 2 and 3 - Fim 2 and 3, tracheal colonization factor - TcfA, have recently been isolated in countries with a high vaccination coverage. The emergence of such isolates might have resulted from B. pertussis natural evolution course or adaptive mechanisms, allowing increased circulation of the pathogen in vaccinated populations. So far, the majority of described mutants were deficient in the Prn production. Prn deficient isolates were found in countries which use acellular pertussis vaccines (aP in routine immunization programmes. The increase of frequency of Prn¯ strains isolation was correlated with the period of routine vaccination with aP vaccines. In most countries, the spread of these isolates has resulted from independent mutations rather than from the expansion of a single clone. Prn¯ isolates were collected from patients showing typical clinical symptoms of pertussis found for Prn+ strains. Results of in vitro and in vivo studies have shown that Prn¯, Ptx¯ and FHA¯ isolates retain cytotoxic properties, and besides Ptx¯ isolates, were lethal in intranasally infected mice. Further explanation of the impact of antigen deficiencies on virulence and transmission of B. pertussis in the context of the continuous increase of pertussis incidence is necessary to develop a new, optimized strategy of pertussis prevention.

Cloning, expression, purification and preliminary crystallographic analysis of the short-chain dehydrogenase enzymes WbmF, WbmG and WbmH from Bordetella bronchiseptica

International Nuclear Information System (INIS)

Harmer, Nicholas J.; King, Jerry D.; Palmer, Colin M.; Preston, Andrew; Maskell, Duncan J.; Blundell, Tom L.

2007-01-01

The expression, purification, and crystallisation of the short-chain dehydrogenases WbmF, WbmG and WbmH from B. bronchiseptica are described. Native diffraction data to 1.5, 2.0, and 2.2 Å were obtained for the three proteins, together with complexes with nucleotides. The short-chain dehydrogenase enzymes WbmF, WbmG and WbmH from Bordetella bronchiseptica were cloned into Escherichia coli expression vectors, overexpressed and purified to homogeneity. Crystals of all three wild-type enzymes were obtained using vapour-diffusion crystallization with high-molecular-weight PEGs as a primary precipitant at alkaline pH. Some of the crystallization conditions permitted the soaking of crystals with cofactors and nucleotides or nucleotide sugars, which are possible substrate compounds, and further conditions provided co-complexes of two of the proteins with these compounds. The crystals diffracted to resolutions of between 1.50 and 2.40 Å at synchrotron X-ray sources. The synchrotron data obtained were sufficient to determine eight structures of the three enzymes in complex with a variety of cofactors and substrate molecules

Pericardite em suínos ao abate no Rio Grande Sul: avaliação de agentes bacterianos e lesões associadas

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Carolini F. Coelho

2014-07-01

Full Text Available O objetivo do presente estudo foi identificar a frequência de lesões macroscópicas e microscópicas e dos agentes bacterianos envolvidos em pericardites em suínos no abate no Estado do Rio Grande do Sul. As amostras foram coletadas em frigoríficos de suínos com Serviço de Inspeção Federal (SIF entre fevereiro a outubro de 2010 e a condenação por pericardite dos animais acompanhados foi de 3,9% (299/7.571. No total foram investigados 91 casos de pericardites, 89% deles foram classificados como crônicos por histopatologia e pleurite crônica foi observada em 47% dos pulmões correspondentes, todavia não houve associação significativa entre as duas lesões. Os agentes bacterianos isolados a partir dos corações foram Streptococcus spp., Pasteurella multocida, Haemophilus parasuis e Streptococcus suis. DNA bacterianos mais detectados pela PCR foram de Mycoplasma hyopneumoniae e Actinobacillus pleuropneumoniae. Houve associação significativa entre isolamento de P. multocida e Streptococcus sp. nos corações e pulmões correspondentes. Esses resultados sugerem que a infecção no pulmão possa ter servido de porta de entrada para a colonização do pericárdio adjacente. Apesar de M. hyopneumoniae ter sido o agente detectado com maior frequência pela PCR em corações e pulmões correspondentes, não houve associação significativa da detecção dos agentes nos órgãos. Isto sugere que as infecções foram eventos independentes. Os demais agentes investigados não apresentaram associação significativa entre isolamento ou detecção de DNA em coração e pulmão correspondente. Outro achado importante foi a presença de coinfecções bacterianas em 2% dos corações e por PCR foi detectado DNA bacteriano de dois ou mais agentes em 16,5% dos corações. Esses resultados sugerem que as coinfecções em pericardites precisam ser melhor estudadas.

Both CD4+ and CD8+ Lymphocytes Participate in the IFN-γ Response to Filamentous Hemagglutinin from Bordetella pertussis in Infants, Children, and Adults

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Violette Dirix

2012-01-01

Full Text Available Infant CD4+ T-cell responses to bacterial infections or vaccines have been extensively studied, whereas studies on CD8+ T-cell responses focused mainly on viral and intracellular parasite infections. Here we investigated CD8+ T-cell responses upon Bordetella pertussis infection in infants, children, and adults and pertussis vaccination in infants. Filamentous hemagglutinin-specific IFN-γ secretion by circulating lymphocytes was blocked by anti-MHC-I or -MHC-II antibodies, suggesting that CD4+ and CD8+ T lymphocytes are involved in IFN-γ production. Flow cytometry analyses confirmed that both cell types synthesized antigen-specific IFN-γ, although CD4+ lymphocytes were the major source of this cytokine. IFN-γ synthesis by CD8+ cells was CD4+ T cell dependent, as evidenced by selective depletion experiments. Furthermore, IFN-γ synthesis by CD4+ cells was sometimes inhibited by CD8+ lymphocytes, suggesting the presence of CD8+ regulatory T cells. The role of this dual IFN-γ secretion by CD4+ and CD8+ T lymphocytes in pertussis remains to be investigated.

Conservación por congelación de Bordetella pertussis y Corynebacterium diphtheriae, empleados en la producción de vacunas para uso humano

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Yilian Plasencia,

2000-11-01

Full Text Available En el presente estudio se evaluó el método de congelación a –70ºC para la preservación de Bordetella pertussis y Corynebacterium diphtheriae. Para verificar el sustento de los cultivos se realizó un adecuado control de calidad, que incluyó comprobación de pureza, viabilidad y estabilidad de las propiedades de interés. En este trabajo se probaron diferentes formulaciones. Se seleccionó la que arrojó los mejores resultados y se realizó un estudio de mantenimiento de las características evaluadas durante el tiempo. Para medir determinados parámetros se realizaron procesos a escala industrial, empleándose para esto un biorreactor Chemap de 35 L. Se tomaron como referencia los valores obtenidos por las cepas conservadas por liofilización. De esta forma se buscaron alternativas y soluciones a problemas presentados en su conservación. Los resultados obtenidos sugieren la posible inclusión en el Programa de Mantenimiento establecido.

Purification and assay of cell-invasive form of calmodulin-sensitive adenylyl cyclase from Bordetella pertussis

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Masure, H.R.; Donovan, M.G.; Storm, D.R.

1991-01-01

An invasive form of the CaM-sensitive adenylyl cyclase from Bordetella pertussis can be isolated from bacterial culture supernatants. This isolation is achieved through the use of QAE-Sephadex anion-exchange chromatography. It has been demonstrated that the addition of exogenous Ca 2+ to the anion-exchange gradient buffers will affect elution from the column and will thereby affect the isolation of invasive adenylyl cyclase. This is probably due to a Ca2(+)-dependent interaction of the catalytic subunit with another component in the culture supernatant. Two peaks of adenylyl cyclase activity are obtained. The Pk1 adenylyl cyclase preparation is able to cause significant increases in intracellular cAMP levels in animal cells. This increase occurs rapidly and in a dose-dependent manner in both N1E-115 mouse neuroblastoma cells and human erythrocytes. The Pk2 adenylyl cyclase has catalytic activity but is not cell invasive. This material can serve, therefore, as a control to ensure that the cAMP which is measured is, indeed, intracellular. A second control is to add exogenous CaM to the Pk1 adenylyl cyclase preparation. The 45-kDa catalytic subunit-CaM complex is not cell invasive. Although the mechanism for membrane translocation of the adenylyl cyclase is unknown, there is evidence that the adenylyl cyclase enters animal cells by a mechanism distinct from receptor-mediated endocytosis. Calmodulin-sensitive adenylyl cyclase activity can be removed from preparations of the adenylyl cyclase that have been subjected to SDS-polyacrylamide gel electrophoresis. This property of the enzyme has enabled purification of the catalytic subunit to apparent homogeneity. The purified catalytic subunit from culture supernatants has a predicted molecular weight of 45,000. This polypeptide interacts directly with Ca 2+ and this interaction may be important for its invasion into animal cells

Purification and assay of cell-invasive form of calmodulin-sensitive adenylyl cyclase from Bordetella pertussis

Energy Technology Data Exchange (ETDEWEB)

Masure, H.R.; Donovan, M.G.; Storm, D.R.

1991-01-01

An invasive form of the CaM-sensitive adenylyl cyclase from Bordetella pertussis can be isolated from bacterial culture supernatants. This isolation is achieved through the use of QAE-Sephadex anion-exchange chromatography. It has been demonstrated that the addition of exogenous Ca{sup 2}{sup +} to the anion-exchange gradient buffers will affect elution from the column and will thereby affect the isolation of invasive adenylyl cyclase. This is probably due to a Ca2(+)-dependent interaction of the catalytic subunit with another component in the culture supernatant. Two peaks of adenylyl cyclase activity are obtained. The Pk1 adenylyl cyclase preparation is able to cause significant increases in intracellular cAMP levels in animal cells. This increase occurs rapidly and in a dose-dependent manner in both N1E-115 mouse neuroblastoma cells and human erythrocytes. The Pk2 adenylyl cyclase has catalytic activity but is not cell invasive. This material can serve, therefore, as a control to ensure that the cAMP which is measured is, indeed, intracellular. A second control is to add exogenous CaM to the Pk1 adenylyl cyclase preparation. The 45-kDa catalytic subunit-CaM complex is not cell invasive. Although the mechanism for membrane translocation of the adenylyl cyclase is unknown, there is evidence that the adenylyl cyclase enters animal cells by a mechanism distinct from receptor-mediated endocytosis. Calmodulin-sensitive adenylyl cyclase activity can be removed from preparations of the adenylyl cyclase that have been subjected to SDS-polyacrylamide gel electrophoresis. This property of the enzyme has enabled purification of the catalytic subunit to apparent homogeneity. The purified catalytic subunit from culture supernatants has a predicted molecular weight of 45,000. This polypeptide interacts directly with Ca{sup 2}{sup +} and this interaction may be important for its invasion into animal cells.

Differences in Bordetella pertussis DNA load according to clinical and epidemiological characteristics of patients with whooping cough.

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Brotons, Pedro; de Paz, Hector D; Toledo, Diana; Villanova, Marta; Plans, Pedro; Jordan, Iolanda; Dominguez, Angela; Jane, Mireia; Godoy, Pere; Muñoz-Almagro, Carmen

2016-04-01

To identify associations between nasopharyngeal Bordetella pertussis DNA load and clinical and epidemiological characteristics and evaluate DNA load prognostic value in pertussis severity. Prospective observational multi-centre study including nasopharyngeal samples positive to pertussis DNA by real-time PCR collected from children and adult patients in more than 200 health centres of Catalonia (Spain) during 2012-2013. B. pertussis load was inversely correlated with age (rho = -0.32, p < 0.001), time to diagnosis (rho = -0.33, p < 0.001) and number of symptoms (rho = 0.13, p = 0.002). Median bacterial load was significantly higher in inpatients versus outpatients (4.91 vs. 2.55 log10 CFU/mL, p < 0.001), patients with complications versus those without (6.05 vs. 2.82 log10 CFU/mL, p < 0.001), disease incidence in summer and autumn versus spring and winter (3.50 vs. 2.21 log10 CFU/mL, p = 0.002), and unvaccinated-partially vaccinated patients versus vaccinated (4.20 vs. 2.76 log10 CFU/mL, p = 0.004). A logistic regression model including bacterial load and other candidate prognostic factors showed good prediction for hospital care (AUC = 0.94) although only age and unvaccinated status were found to be prognostic factors. We observed strong positive associations of nasopharyngeal bacterial load with severity outcomes of hospitalisation and occurrence of complications. Bacterial load and other independent variables contributed to an accurate prognostic model for hospitalisation. Copyright © 2016 The British Infection Association. Published by Elsevier Ltd. All rights reserved.

[PCR testing for Bordetella pertussis in household contacts as a diagnostic tool for atypical whooping cough in unvaccinated young infants].

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Cosnes-Lambe, Cecile; Raymond, Josette; Vallet, Christelle; Armengaud, Jean-Baptiste; Bosdure, Emmanuelle; Catalano-Pons, Charlotte; Chalumeau, Martin; El Hajje, Marie-Joelle; Moulin, Florence; de Suremain, Nathalie; Reglier-Poupet, Hélène; Poyart, Claire; Gendrel, Dominique

2008-10-01

False-negative findings of polymerase chain reaction (PCR) for genuine pertussis as well as the numerous atypical forms of whooping cough make it difficult to diagnose this disease in young babies. For two years, real-time PCR was performed to test for Bordetella pertussis in 86 infants younger than 6 months hospitalized for apnea or paroxysmal and/or vomiting cough and in 205 of their household contacts, whether or not they coughed. Group 1 included 30 infants for whom PCR detected B. pertussis (25 of whom were also RSV+). PCR was also positive for at least one household contact in 25/30 families. This group included 16 babies with apnea and 12 who developed a whooping cough during follow-up. Group 2 comprised 12 infants whose PCR was negative while at least one household contact had positive results. Five of these infants had severe apnea and 6 developed a whooping cough. Group 3 included 44 infants (28 RSV +) for whom PCR was negative in the index case and in the household contacts: none developed a whooping cough during follow-up. Only 3 of the 54 positive household contacts had a paroxysmal cough or a typical whooping cough and 12 had no cough at all. Positive PCR in a household contact, symptomatic or not, is helpful for the diagnosis of atypical whooping cough in young infants.

Pulsed-Field Gel Electrophoresis Analysis of Bordetella pertussis Isolates Circulating in Europe from 1998 to 2009

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Advani, Abdolreza; Hallander, Hans O.; Dalby, Tine; Krogfelt, Karen Angeliki; Guiso, Nicole; Njamkepo, Elisabeth; von Könnig, Carl Heinz Wirsing; Riffelmann, Marion; Mooi, Frits R.; Sandven, Per; Lutyńska, Anna; Fry, Norman K.; Mertsola, Jussi

2013-01-01

Between 1998 and 2009, Bordetella pertussis clinical isolates were collected during three periods, i.e., 1998 to 2001 (n = 102), 2004 to 2005 (n = 154), and 2007 to 2009 (n = 140), from nine countries with distinct vaccination programs, i.e., Denmark, Finland, France, Germany, The Netherlands, Norway, Poland, Sweden, and the United Kingdom. Pulsed-field gel electrophoresis (PFGE) analysis was performed according to standardized recommendations for epidemiological typing of B. pertussis. There were 81 different PFGE profiles, five of which (BpSR3, BpSR5, BpSR10, BpSR11, and BpSR12) were observed in 61% of the 396 isolates and shown to be predominant in almost all countries. The major profile, BpSR11, showed a decreasing trend from 25% to 30% in 1998 to 2005 to 13% in 2007 to 2009, and there were increases in BpSR3 and BpSR10 from 0% and 8% to 21% and 22%, respectively. One difference between these profiles is that BpSR11 contains isolates harboring the fim3-2 allele and BpSR3 and BpSR10 contain isolates harboring the fim3-1 allele. The total proportion of the five predominant profiles increased from 44% in 1998 to 2001 to 63% in 2004 to 2005 to 70% in 2007 to 2009. In conclusion, common PFGE profiles were identified in B. pertussis populations circulating in European countries with different vaccination programs and different vaccine coverages. These prevalent isolates contain the novel pertussis toxin promoter ptxP3 allele. However, there is evidence for diversifying selection between ptxP3 strains characterized by distinct PFGE profiles. This work shows that, even within a relatively short time span of 10 years, successful isolates which spread through Europe and cause large shifts in B. pertussis populations may emerge. PMID:23175253

Tiamulin activity against fastidious and nonfastidious veterinary and human bacterial isolates: initial development of in vitro susceptibility test methods.

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Jones, Ronald N; Pfaller, Michael A; Rhomberg, Paul R; Walter, Donald H

2002-02-01

Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine. This report summarizes studies to establish standardized susceptibility testing methods, interpretive criteria, and reagent details for use in veterinary methods recently developed by the National Committee for Clinical Laboratory Standards (NCCLS) (standards M31-A and M37-A, NCCLS, Wayne, Pa., 1999). A total of 636 fastidious and nonfastidious animal and human pathogens were processed by using media and procedures described by the NCCLS. Tiamulin disk diffusion tests used a 30-microg disk concentration, and the proposed MIC breakpoints corresponding to levels achievable in animal target tissues (lung) were or =32 microg/ml for resistance. Correlate zone diameters for specific nonfastidious species were as follows: for Pasteurella multocida and staphylococci tested on Mueller-Hinton agar, susceptibility at > or =19 mm and resistance at or =16 mm and resistance at or =9 mm) was suggested for veterinary fastidious medium broth and enriched chocolate Mueller-Hinton agar. Absolute categorical agreement between NCCLS dilution and disk diffusion test results with these criteria ranged from 90.5 to 96.2%. Tiamulin susceptibility testing methods appear to be accurate in their categorical classification for indicated species, and their availability will allow immediate testing of animal isolates to guide therapy via appropriate levels of dosing and to monitor the development of resistance for agents in this unique class.

Isolation and polymerase chain reaction-based identification of Riemerella anatipestifer from ducks in Kerala, India

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Manju Soman

2014-10-01

Full Text Available Aim: The aim was to isolate and characterize Riemerella anatipestifer organisms from disease outbreaks in ducks in Kerala. Materials and Methods: Ducklings, suspected of Riemerella infection, were sacrificed and subjected to post-mortem examination. Heart blood smears and impression smears from liver and spleen were examined for the presence of pathogenic organisms. Heart blood, lung, liver, and spleen collected aseptically from the birds were subjected to isolation trials in brain heart infusion agar and 10% bovine blood agar. The isolates were characterized based on morphology, cultural characteristics and biochemical tests, and their identity were confirmed by polymerase chain reaction (PCR and the PCR amplified DNA was sequenced. The antibiotic sensitivity testing of the isolates were carried out using six antibiotics viz ciprofloxacin, chloramphenicol, enrofloxacin, amoxycillin, cotrimoxazole, and gentamicin. Results: Colonies suggestive of Riemerella organisms could be isolated on blood agar. Biochemical characterization and PCR confirmed the identity of isolates as R. anatipestifer. The nucleotide sequence of the PCR product showed 99% homology to the R. anatipestifer sequences in the NCBI. The antibiogram revealed that the organisms were sensitive to ciprofloxacin, enrofloxacin, and gentamicin. Conclusion: The present study suggests that the PCR assay can facilitate fast and proper identification of R. anatipestifer infection in ducks. The assay can also differentiate between R. anatipestifer and Pasteurella multocida and can replace the traditional methods of differentiation which are cumbersome and time-consuming.

Pharmacokinetic/pharmacodynamic evaluation of marbofloxacin as a single injection for Pasteurellaceae respiratory infections in cattle using population pharmacokinetics and Monte Carlo simulations.

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Paulin, A; Schneider, M; Dron, F; Woehrle, F

2018-02-01

Population pharmacokinetic of marbofloxacin was investigated with 52 plasma concentration-time profiles obtained after intramuscular administration of Forcyl® in cattle. Animal's status, pre-ruminant, ruminant, or dairy cow, was retained as a relevant covariate for clearance. Monte Carlo simulations were performed using a stratification by status, and 1000 virtual disposition curves were generated in each bovine subpopulation for the recommended dosage regimen of 10 mg/kg as a single injection. The probability of target attainment (PTA) of pharmacokinetic/pharmacodynamic (PK/PD) ratios associated with clinical efficacy and prevention of resistance was determined in each simulated subpopulation. The cumulative fraction of response (CFR) of animals achieving a PK/PD ratio predictive of positive clinical outcome was then calculated for the simulated dosage regimen, taking into account the minimum inhibitory concentration (MIC) distribution of Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni. When considering a ratio of AUC 0-24 hr /MIC (area under the curve/minimum inhibitory concentration) greater than 125 hr, CFRs ranging from 85% to 100% against the three Pasteurellaceae in each bovine subpopulation were achieved. The PTA of the PK/PD threshold reflecting the prevention of resistances was greater than 90% up to MPC (mutant prevention concentration) values of 1 μg/ml in pre-ruminants and ruminants and 0.5 μg/ml in dairy cows. © 2017 The Authors. Journal of Veterinary Pharmacology and Therapeutics Published by John Wiley & Sons Ltd.

A bibliography of references to avian cholera

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Wilson, Sonoma S.

1979-01-01

Mrs. Wilson has made a genuine effort to include in this bibliography every significant reference to avian cholera since Louis Pasteur's articles appeared in 1880, although she recognizes the likelihood that a few have been overlooked. New listings have been added throughout 1978, but comprehensive coverage of the literature cannot be claimed beyond June of that year.Textbook accounts, because they are generally summaries of work published elsewhere, are excluded. Papers dealing primarily with the biology of Pasteurella multocida, as opposed to the disease it induces in birds, are also excluded, unless they report information of diagnostic usefulness. Short abstracts are not included unless the journals in which they are published are more widely available than those in which the complete articles appear or they are English summaries of foreign language articles.In compiling this bibliography, Mrs. Wilson has made extensive use of Biological Abstracts, the Pesticide Documentation Bulletin, and printouts generated by Bibliographic Retrieval Services, Inc. The "Literature Cited" sections of textbooks and journal articles pertinent to the subject were sources of many additional references. Regardless of the origin of the citation, its accuracy was confirmed by comparison with the original publication, except in those few instances (marked with an asterisk) when the journal was not on the shelves of the libraries accessible to us.The author will be grateful to users of the bibliography who point out errors or omissions.Wayne I. JensenMicrobiologist In Charge

Disclosing respiratory co-infections: a broad-range panel assay for avian respiratory pathogens on a nanofluidic PCR platform.

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Croville, Guillaume; Foret, Charlotte; Heuillard, Pauline; Senet, Alexis; Delpont, Mattias; Mouahid, Mohammed; Ducatez, Mariette F; Kichou, Faouzi; Guerin, Jean-Luc

2018-06-01

Respiratory syndromes (RS) are among the most significant pathological conditions in edible birds and are caused by complex coactions of pathogens and environmental factors. In poultry, low pathogenic avian influenza A viruses, metapneumoviruses, infectious bronchitis virus, infectious laryngotracheitis virus, Mycoplasma spp. Escherichia coli and/or Ornithobacterium rhinotracheale in turkeys are considered as key co-infectious agents of RS. Aspergillus sp., Pasteurella multocida, Avibacterium paragallinarum or Chlamydia psittaci may also be involved in respiratory outbreaks. An innovative quantitative PCR method, based on a nanofluidic technology, has the ability to screen up to 96 samples with 96 pathogen-specific PCR primers, at the same time, in one run of real-time quantitative PCR. This platform was used for the screening of avian respiratory pathogens: 15 respiratory agents, including viruses, bacteria and fungi potentially associated with respiratory infections of poultry, were targeted. Primers were designed and validated for SYBR green real-time quantitative PCR and subsequently validated on the Biomark high throughput PCR nanofluidic platform (Fluidigm©, San Francisco, CA, USA). As a clinical assessment, tracheal swabs were sampled from turkeys showing RS and submitted to this panel assay. Beside systematic detection of E. coli, avian metapneumovirus, Mycoplasma gallisepticum and Mycoplasma synoviae were frequently detected, with distinctive co-infection patterns between French and Moroccan flocks. This proof-of-concept study illustrates the potential of such panel assays for unveiling respiratory co-infection profiles in poultry.

A bighorn sheep die-off in southern Colorado involving a Pasteurellaceae strain that may have originated from syntopic cattle.

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Wolfe, Lisa L; Diamond, Brandon; Spraker, Terry R; Sirochman, Michael A; Walsh, Daniel P; Machin, Chandra M; Bade, Donald J; Miller, Michael W

2010-10-01

We investigated a pasteurellosis epizootic in free-ranging bighorn sheep (Ovis canadensis) wherein a Pasteurellaceae strain carried by syntopic cattle (Bos taurus) under severe winter conditions appeared to contribute to pneumonia in affected bighorns. Twenty-one moribund or dead bighorn sheep were found on the "Fossil Ridge" herd's winter range, Colorado, USA, between 13 December 2007 and 29 February 2008. Eight carcasses examined showed gross or microscopic evidence of acute to subacute fibrinous bronchopneumonia. All eight carcasses yielded at least one β-hemolytic Mannheimia haemolytica biogroup 1(±(G)) strain, and seven also yielded a β-hemolytic Bibersteinia trehalosi biogroup 4 (CDS) strain; evidence of Pasteurella multocida, Mycoplasma ovipneumoniae, and parainfluenza 3 and bovine respiratory syncytial viruses was also detected. Isolates of β-hemolytic Manneimia haemolytica biogroup 1(G) from a bighorn carcass and a syntopic cow showed 99.5% similarity in genetic fingerprints; B. trehalosi biogroup 4(CDS) isolates were ≥94.9% similar to an isolate from a nearby bighorn herd. Field and laboratory observations suggested that pneumonia in affected bighorns may have been caused by a combination of pathogens including two pathogenic Pasteurellaceae strains--one likely of cattle origin and one likely of bighorn origin--with infections in some cases perhaps exacerbated by other respiratory pathogens and severe weather conditions. Our and others' findings suggest that intimate interactions between wild sheep and cattle should be discouraged as part of a comprehensive approach to health management and conservation of North American wild sheep species.

Fusobacterium necrophorum in North American Bighorn Sheep ( Ovis canadensis ) Pneumonia.

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Shanthalingam, Sudarvili; Narayanan, Sanjeevkumar; Batra, Sai Arun; Jegarubee, Bavananthasivam; Srikumaran, Subramaniam

2016-07-01

Fusobacterium necrophorum has been detected in pneumonic bighorn sheep (BHS; Ovis canadensis ) lungs, in addition to the aerobic respiratory pathogens Mannheimia haemolytica , Bibersteinia trehalosi , Pasteurella multocida , and Mycoplasma ovipneumoniae . Similar to M. haemolytica , F. necrophorum produces a leukotoxin. Leukotoxin-induced lysis and degranulation of polymorphonuclear leukocytes (PMNs) and macrophages are responsible for acute inflammation and lung tissue damage characteristic of M. haemolytica -caused pneumonia. As one approach in elucidating the role of F. necrophorum in BHS pneumonia, we determined the frequency of the presence of F. necrophorum in archived pneumonic BHS lung tissues, and susceptibility of BHS leukocytes to F. necrophorum leukotoxin. A species-specific PCR assay detected F. necrophorum in 37% of pneumonic BHS lung tissues (total tested n=70). Sequences of PCR amplicons were similar to the less virulent F. necrophorum subsp. funduliforme. Fusobacterium necrophorum leukotoxin exhibited cytotoxicity to BHS PMNs and peripheral blood mononuclear cells. As with the M. haemolytica leukotoxin, F. necrophorum leukotoxin was more toxic to BHS PMNs than domestic sheep PMNs. It is likely that F. necrophorum enters the lungs after M. haemolytica and other aerobic respiratory pathogens enter the lungs and initiate tissue damage, thereby creating a microenvironment that is conducive for anaerobic bacterial growth. In summary, Fusobacterium leukotoxin is highly toxic for BHS leukocytes; however, based on the PCR findings, it is unlikely to play a direct role in the development of BHS pneumonia.

Evaluation of Phenolic Compounds and Antioxidant and Antimicrobial Activities of Some Common Herbs

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Muhammad Abdul Qadir

2017-01-01

Full Text Available The study was designed to evaluate the phenolic, flavonoid contents and antioxidant and antimicrobial activities of onion (Allium cepa, garlic (Allium sativum, mint (Mentha spicata, thyme (Thymus vulgaris, oak (Quercus, aloe vera (Aloe barbadensis Miller, and ginger (Zingiber officinale. All extracts showed a wide range of total phenolic contents, that is, 4.96 to 98.37 mg/100 g gallic acid equivalents, and total flavonoid contents, that is, 0.41 to 17.64 mg/100 g catechin equivalents. Antioxidant activity (AA was determined by measuring reducing power, inhibition of peroxidation using linoleic acid system, and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH scavenging activity. Different extracts inhibited oxidation of linoleic acid by 16.6–84.2% while DPPH radical scavenging activity (IC50 values ranged from 17.8% to 79.1 μg/mL. Reducing power at 10 mg/mL extract concentration ranged from 0.11 to 0.84 nm. Furthermore the extracts of these medicinal herbs in 80% methanol, 80% ethanol, 80% acetone, and 100% water were screened for antimicrobial activity by disc diffusion method against selected bacterial strains, Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Pasteurella multocida, and fungal strains, Aspergillus niger, Aspergillus flavus, Rhizopus solani, and Alternaria alternata. The extracts show better antimicrobial activity against bacterial strains as compared to fungal strains. Results of various assays were analyzed statistically by applying appropriate statistical methods.

Adenovirus 2, Bordetella bronchiseptica, and Parainfluenza Molecular Diagnostic Assay Results in Puppies After vaccination with Modified Live Vaccines.

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Ruch-Gallie, R; Moroff, S; Lappin, M R

2016-01-01

Canine adenovirus 2, parainfluenza, and Bordetella bronchiseptica cause respiratory disease in dogs, and each has a modified live intranasal vaccine available. Molecular diagnostic assays to amplify specific nucleic acids are available for each of these agents. If positive molecular diagnostic assay results are common after vaccination, the positive predictive value of the diagnostic assays for disease would be decreased. To determine the impact of administration of commercially available modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza vaccine has on the results of a commercially available PCR panel. Eight puppies from a research breeding facility negative for these pathogens. Blinded prospective pilot study. Puppies were vaccinated with a single dose of modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza and parenteral dose of adenovirus 2, canine distemper virus, and parvovirus. Nasal and pharyngeal swabs were collected on multiple days and submitted for PCR assay. Nucleic acids of all 3 organisms contained in the topical vaccine were detected from both samples multiple times through 28 days after vaccination with higher numbers of positive samples detected between days 3 and 10 after vaccination. Vaccine status should be considered when interpreting respiratory agent PCR results if modified live vaccines have been used. Development of quantitative PCR and wild-type sequencing are necessary to improve positive predictive value of these assays by distinguishing vaccinate from natural infection. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

A versatile, non genetically modified organism (GMO)-based strategy for controlling low-producer mutants in Bordetella pertussis cultures using antigenic modulation.

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Goffin, Philippe; Slock, Thomas; Smessaert, Vincent; De Rop, Philippe; Dehottay, Philippe

2015-08-01

The uncontrolled presence of non-producer mutants negatively affects bioprocesses. In Bordetella pertussis cultures, avirulent mutants emerge spontaneously and accumulate. We characterized the dynamics of accumulation using high-throughput growth assays and competition experiments between virulent and avirulent (bvg(-) ) isolates. A fitness advantage of bvg(-) cells was identified as the main driver for bvg(-) accumulation under conditions of high virulence factor production. Conversely, under conditions that reduce their expression (antigenic modulation), bvg(-) takeover could be avoided. A control strategy was derived, which consists in applying modulating conditions whenever virulence factor production is not required. It has a wide range of applications, from routine laboratory operations to vaccine manufacturing, where pertussis toxin yields were increased 1.4-fold by performing early pre-culture steps in modulating conditions. Because it only requires subtle modifications of the culture medium and does not involve genetic modifications, this strategy is applicable to any B. pertussis isolate, and should facilitate regulatory acceptance of process changes for vaccine production. Strategies based on the same concept, could be derived for other industrially relevant micro-organisms. This study illustrates how a sound scientific understanding of physiological principles can be turned into a practical application for the bioprocess industry, in alignment with Quality by Design principles. Copyright © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Pesquisa de antigenos aglutinantes "major" 1, 2 e 3 em cepas de Bordetella pertussis, isoladas de crianças com coqueluche atendidas no Hospital de Isolamento Emílio Ribas de São Paulo, Brasil Determination of 1, 2 and 3 major antigens in Bordetella pertussis strains isolated from Brazilian children with whooping-cough

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Sebastião Timo Iaria

1977-09-01

Full Text Available Em 30 cepas de Bordetella pertussis isoladas de crianças com coqueluche, atendidas no Hospital de Isolamento Emílio Ribas de São Paulo, foram pesquisados os antígenos aglutinantes ''major" 1, 2 e 3. Levando-se em conta a presença combinada dos três antígenos, as provas de soro-aglutinação rápida em lamina revelaram que 25 (83,3% cepas possuiam os fatores 1, 2 e 3, enquanto que 3 (10,0% e 2 (6,7% foram positivas, somente, para 1, 2 e 1, 3, respectivamente. Os resultados foram discutidos, considerando-se a importância deste antígeno no preparo de vacinas.The presence of major antigens, 1, 2 and 3 were determined in 30 strains of B. pertussis isolated from children with whooping-cough hospitalized at the Hospital Emílio Ribas, São Paulo Brazil. The method used was the slide-agglutination test. Tests showed that 25(83.3% of strains were positives for factors 1, 2 and 3. Factores 1 and 3 alone were present in 3 (10% of strains and 1 and 2 alone in 2 (6.7%.

Detection of Bordetella pertussis using a PCR test in infants younger than one year old hospitalized with whooping cough in five Peruvian hospitals.

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Castillo, María Esther; Bada, Carlos; Del Aguila, Olguita; Petrozzi-Helasvuo, Verónica; Casabona-Ore, Verónica; Reyes, Isabel; Del Valle-Mendoza, Juana

2015-12-01

To report the incidence, epidemiology, and clinical features of Bordetella pertussis in Peruvian infants under 1 year old. A prospective cross-sectional study was conducted in five hospitals in Peru from January 2010 to July 2012. A total of 392 infants under 1 year old were admitted with a clinical diagnosis of whooping cough and tested for B. pertussis by PCR. The pertussis toxin and IS481 genes were detected in 39.54% (155/392) of the cases. Infants aged less than 3 months were the most affected, with a prevalence of 73.55% (114/155). The most common household contact was the mother, identified in 20% (31/155) of cases. Paroxysm of coughing (89.03%, 138/155), cyanosis (68.39%, 106/155), respiratory distress (67.09%, 104/155), and breastfeeding difficulties (39.35%, 61/155) were the most frequent symptoms reported. An increase in pertussis cases has been reported in recent years in Peru, despite national immunization efforts. Surveillance with PCR for B. pertussis is essential, especially in infants less than 1 year old, in whom a higher rate of disease-related complications and higher mortality have been reported. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Exhaust Air Dust Monitoring is Superior to Soiled Bedding Sentinels for the Detection of Pasteurella pneumotropica in Individually Ventilated Cage Systems.

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Miller, Manuel; Ritter, Brbel; Zorn, Julia; Brielmeier, Markus

2016-11-01

Reliable detection of unwanted organisms is essential for meaningful health monitoring in experimental animal facilities. Currently, most rodents are housed in IVC systems, which prevent the aerogenic transmission of pathogens between cages. Typically soiled-bedding sentinels (SBS) exposed to soiled bedding collected from a population of animals within an IVC rack are tested as representatives, but infectious agents often go undetected due to inefficient transmission. Pasteurellaceae are among the most prevalent bacterial pathogens isolated from experimental mice, and the failure of SBS to detect these bacteria is well established. In this study, we investigated whether analysis of exhaust air dust (EAD) samples by using a sensitive and specific real-time PCR assay is superior to conventional SBS monitoring for the detection of Pasteurella pneumotropica (Pp) infections. In a rack with a known prevalence of Pp-positive mice, weekly EAD sampling was compared with the classic SBS method over 3 mo. In 6 rounds of testing, with a prevalence of 5 infected mice in each of 7 cages in a rack of 63 cages, EAD PCR detected Pp at every weekly time point; SBS failed to detect Pp in all cases. The minimal prevalence of Pp-infected mice required to obtain a reliable positive result by EAD PCR testing was determined to be 1 in 63 cages. Reliable detection of Pp was achieved after only 1 wk of exposure. Analysis of EAD samples by real-time PCR assay provides a sensitive, simple, and reliable approach for Pp identification in laboratory mice.

Bordetella pertussis Isolates from Argentinean Whooping Cough Patients Display Enhanced Biofilm Formation Capacity Compared to Tohama I Reference Strain.

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Arnal, Laura; Grunert, Tom; Cattelan, Natalia; de Gouw, Daan; Villalba, María I; Serra, Diego O; Mooi, Frits R; Ehling-Schulz, Monika; Yantorno, Osvaldo M

2015-01-01

Pertussis is a highly contagious disease mainly caused by Bordetella pertussis. Despite the massive use of vaccines, since the 1950s the disease has become re-emergent in 2000 with a shift in incidence from infants to adolescents and adults. Clearly, the efficacy of current cellular or acellular vaccines, formulated from bacteria grown in stirred bioreactors is limited, presenting a challenge for future vaccine development. For gaining insights into the role of B. pertussis biofilm development for host colonization and persistence within the host, we examined the biofilm forming capacity of eight argentinean clinical isolates recovered from 2001 to 2007. All clinical isolates showed an enhanced potential for biofilm formation compared to the reference strain Tohama I. We further selected the clinical isolate B. pertussis 2723, exhibiting the highest biofilm biomass production, for quantitative proteomic profiling by means of two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) coupled with mass spectrometry, which was accompanied by targeted transcriptional analysis. Results revealed an elevated expression of several virulence factors, including adhesins involved in biofilm development. In addition, we observed a higher expression of energy metabolism enzymes in the clinical isolate compared to the Tohama I strain. Furthermore, all clinical isolates carried a polymorphism in the bvgS gene. This mutation was associated to an increased sensitivity to modulation and a faster rate of adhesion to abiotic surfaces. Thus, the phenotypic biofilm characteristics shown by the clinical isolates might represent an important, hitherto underestimated, adaptive strategy for host colonization and long time persistence within the host.

Identifying long-term memory B-cells in vaccinated children despite waning antibody levels specific for Bordetella pertussis proteins.

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Hendrikx, Lotte H; Oztürk, Kemal; de Rond, Lia G H; Veenhoven, Reinier H; Sanders, Elisabeth A M; Berbers, Guy A M; Buisman, Anne-Marie

2011-02-04

Whooping cough is a respiratory disease caused by Bordetella pertussis. Since the 1950s in developed countries pertussis vaccinations are included in the national immunization program. However, antibody levels rapidly wane after both whole cell and acellular pertussis vaccination. Therefore protection against pertussis may depend largely on long-term B- and T-cell immunities. We investigated long-term pertussis-specific memory B-cell responses in children who were primed at infant age with the Dutch wP-vaccine (ISRCTN65428640). Purified B-cells were characterized by FACS-analysis and after polyclonal stimulation memory B-cells were detected by ELISPOT-assays specific for pertussis toxin, filamentous haemagglutinin, pertactin and tetanus. In addition, plasma IgG levels directed to the same antigens were measured by a fluorescent bead-based multiplex immunoassay. Two and 3 years after wP priming as well as 2 and 5 years after the aP booster at the age of 4, low plasma IgG levels to the pertussis proteins were found. At the same time, however pertussis protein-specific memory B-cells could be detected and their number increased with age. The number of tetanus-specific memory B-cells was similar in all age groups, whereas IgG-tetanus levels were high 2 years after tetanus booster compared to pre- and 5 years post-booster levels. This study shows the presence of long-term pertussis protein-specific memory B-cells in children despite waning antibody levels after vaccination, which suggests that memory B-cells in addition to antibodies may contribute to protection against pertussis. Copyright © 2010 Elsevier Ltd. All rights reserved.

Mucosal and systemic immune responses elicited by recombinant Lactococcus lactis expressing a fusion protein composed of pertussis toxin and filamentous hemagglutinin from Bordetella pertussis.

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Torkashvand, Ali; Bahrami, Fariborz; Adib, Minoo; Ajdary, Soheila

2018-05-05

We constructed a food-grade expression system harboring a F1S1 fusion protein of Bordetella pertussis to be produced in Lactococcus lactis NZ3900 as a new oral vaccine model against whooping cough, caused by B. pertussis. F1S1 was composed of N-terminally truncated S1 subunit of pertussis toxin and type I immunodominant domain of filamentous hemagglutinin which are both known as protective immunogens against pertussis. The recombinant L. lactis was administered via oral or intranasal routes to BALB/c mice and the related specific systemic and mucosal immune responses were then evaluated. The results indicated significantly higher levels of specific IgA in the lung extracts and IgG in sera of mucosally-immunized mice, compared to their controls. It was revealed that higher levels of IgG2a, compared to IgG1, were produced in all mucosally-immunized mice. Moreover, immunized mice developed Th1 responses with high levels of IFN-γ production by the spleen cells. These findings provide evidence for L. lactis to be used as a suitable vehicle for expression and delivery of F1S1 fusion protein to mucosa and induction of appropriate systemic and mucosal immune responses against pertussis. Copyright © 2018 Elsevier Ltd. All rights reserved.

Characterization of a bordetella pertussis diaminopimelate (DAP) biosynthesis locus identifies dapC, a novel gene coding for an N-succinyl-L,L-DAP aminotransferase.

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Fuchs, T M; Schneider, B; Krumbach, K; Eggeling, L; Gross, R

2000-07-01

The functional complementation of two Escherichia coli strains defective in the succinylase pathway of meso-diaminopimelate (meso-DAP) biosynthesis with a Bordetella pertussis gene library resulted in the isolation of a putative dap operon containing three open reading frames (ORFs). In line with the successful complementation of the E. coli dapD and dapE mutants, the deduced amino acid sequences of two ORFs revealed significant sequence similarities with the DapD and DapE proteins of E. coli and many other bacteria which exhibit tetrahydrodipicolinate succinylase and N-succinyl-L,L-DAP desuccinylase activity, respectively. The first ORF within the operon showed significant sequence similarities with transaminases and contains the characteristic pyridoxal-5'-phosphate binding motif. Enzymatic studies revealed that this ORF encodes a protein with N-succinyl-L,L-DAP aminotransferase activity converting N-succinyl-2-amino-6-ketopimelate, the product of the succinylase DapD, to N-succinyl-L,L-DAP, the substrate of the desuccinylase DapE. Therefore, this gene appears to encode the DapC protein of B. pertussis. Apart from the pyridoxal-5'-phosphate binding motif, the DapC protein does not show further amino acid sequence similarities with the only other known enzyme with N-succinyl-L,L-DAP aminotransferase activity, ArgD of E. coli.

Swamp Buffalo in South Kalimantan : Problem, Disease and Control

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Lily Natalia

2006-12-01

Full Text Available In recent years, several studies have been carried out to evaluate and investigate the important diseases of swamp buffaloes (Bubalus carabanensis in Kalimantan . More attention has been focused on the case of acute infectious diseases and sudden death in the buffaloes . Fasciolosis black disease, acute enteritis, especially fatal enterotoxaemia haemorrhagic septicaemia . and trypanosomiasis (Surra, are some of the important diseases found in these animals . Black disease caused by toxigenic Clostridium novyi occurs in the presence of the organism in the liver and the degree of liver fluke Fasciola gigantica infestation . In regions where black disease is enzootic, Cl. novvi can be isolated from livers of normal healthy animals . In Hulu Sungai Utara district, South Kalimantan, the prevalence of fasciolosis caused by Fasciola gigantica in swamp buffalo was 77% in 1991 . A gross sudden change in diet due to seasonal changes could induce rumen and intestinal stasis, which provide a favourable environment for the rapid proliferation of commensal toxigenic Clostridium perfringens in the small intestine . Subsequent absorption of the toxin produced through the gut wall and its generalized dissemination culminated in a fatal enterotoxaemia . Haemorrhagic septicaemia (HS is an acute, fatal disease affecting swamp buffalo, and caused by Pasteurella multocida B : 2 . The swamp buffalo is particularly susceptible for HS, and the reported greatest losses of swamp buffalo in Kalimantan due to HS is recorded in 1980s. The clinical signs of Surra in swamp buffalo were also found in certain areas in Danau Panggang area . Hulu Sungai Utara district . Vaccination is the accepted method for controlling Black disease, enterotoxaemia and HS. Multi component vaccine, alum adjuvant containing at least 5 types of clostridial toxoids and P. multocida B2 bacterin have been used and provide good protection to the animals . Control and treatment of liver fluke infestation

Immunological and protective effects of Bordetella bronchiseptica subunit vaccines based on the recombinant N-terminal domain of dermonecrotic toxin.

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Wang, Chuanwen; Liu, Liping; Zhang, Zhen; Yan, Zhengui; Yu, Cuilian; Shao, Mingxu; Jiang, Xiaodong; Chi, Shanshan; Wei, Kai; Zhu, Ruiliang

2015-10-01

Dermonecrotic toxin (DNT) produced by Bordetella bronchiseptica (B. bronchiseptica) can cause clinical turbinate atrophy in swine and induce dermonecrotic lesions in model mice. We know that the N-terminal of DNT molecule contains the receptor-binding domain, which facilitates binding to the target cells. However, we do not know whether this domain has sufficient immunogenicity to resist B. bronchiseptica damage and thereby to develop a subunit vaccine for the swine industry. In this study, we prokaryotically expressed the recombinant N-terminal of DNT from B. bronchiseptica (named DNT-N) and prepared it for the subunit vaccine to evaluate its immunogenicity. Taishan Pinus massoniana pollen polysaccharide (TPPPS), a known immunomodulator, was used as the adjuvant to examine its immune-conditioning effects. At 49 d after inoculation, 10 mice from each group were challenged with B. bronchiseptica, and another 10 mice were intradermally challenged with native DNT, to examine the protection imparted by the vaccines. The immune parameters (T-lymphocyte counts, cytokine secretions, serum antibody titers, and survival rates) and skin lesions were determined. The results showed that pure DNT-N vaccine significantly induced immune responses and had limited ability to resist the B. bronchiseptica and DNT challenge, whereas the mice administered with TPPPS or Freund's incomplete adjuvant vaccine could induce higher levels of the above immune parameters. Remarkably, the DNT-N vaccine combined with TPPPS adjuvant protected the mice effectively to prevent B. bronchiseptica infection. Our findings indicated that DNT-N has potential for development as an effective subunit vaccine to counteract the damage of B. bronchiseptica infection, especially when used conjointly with TPPPS. Copyright © 2015 Elsevier B.V. All rights reserved.

[Serological evaluation of Bordetella pertussis infection in adults with prolonged cough].

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Sönmez, Cemile; Çöplü, Nilay; Gözalan, Ayşegül; Yılmaz, Ülkü; Bilekli, Selen; Demirci, Nilgün Yılmaz; Biber, Çiğdem; Erdoğan, Yurdanur; Esen, Berrin; Çöplü, Lütfi

2016-07-01

Pertussis is a vaccine-preventable disease that is transmitted from infected to susceptible individuals by respiratory route. Bordetella pertussis infection may occur at any age as neither vaccine nor natural infection induced immunity lasts life-long. This study was planned to demonstrate the serological evidence of infection among adults, to raise awareness among clinicians and to provide data for the development of strategies to protect vulnerable infants. A total of 538 patients (345 female, 193 male) ages between 18-87 years who had a complain of prolonged cough for more than two weeks were included in the study. Anti-pertussis toxin (PT) IgG and anti-filamentous hemagglutinin (FH) IgG levels from single serum samples were measured by an in-house ELISA test which was standardized and shown to be efficient previously. Anti-PT IgG antibody levels of ≥ 100 EU/ml were considered as acute/recent infection with B.pertussis. In our study, 9.7% (52/538) of the patients had high levels of anti-PT IgG (≥ 100 EU/ml) and among those patients 43 (43/52; 82.7%) also had high (≥ 100 EU/ml) anti-FHA IgG levels. There were no statistically significant differences in terms of age, gender, education level, DPT (diphtheria-pertussis-tetanus) vaccination history, smoking history or average daily cigarette consumption (p> 0.05) between the cases with high antibody levels (n= 52). When the symptoms and the presence of cases with high antibody levels were evaluated, it was detected that no one parameter was significantly different from others, except that 24.1% of the cases with inspiratory whooping had high anti-PT levels. There was also no statistically significant difference between high anti-PT levels ≥ 100 EU/ml and the patients with risk factors [smoking (21/200; 10.5%), presence of disease that cause chronic cough and/or drug usage (19/171; %11.1), and whole factors which cause chronic cough (32/306; %10.5)] and without risk factors (p= 0.581; p= 0.357; p= 0

Antioxidant, antimicrobial, antitumor, and cytotoxic activities of an important medicinal plant (Euphorbia royleana from Pakistan

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Aisha Ashraf

2015-03-01

Full Text Available The aim of present study was to evaluate antioxidant, antimicrobial, and antitumor activities of methanol, hexane, and aqueous extracts of fresh Euphorbia royleana. Total phenolic and flavonoid contents were estimated as gallic acid and querectin equivalents, respectively. Antioxidant activity was assessed by scavenging of free 2,2′- diphenyl-1-picrylhydrazyl radicals and reduction of ferric ions, and it was observed that inhibition values increase linearly with increase in concentration of extract. The results of ferric reducing antioxidant power assay showed that hexane extract has maximum ferric reducing power (12.70 ± 0.49 mg gallic acid equivalents/g of plant extract. Maximum phenolic (47.47 ± 0.71 μg gallic acid equivalents/mg of plant extract and flavonoid (63.68 ± 0.43 μg querectin equivalents/mg of plant extract contents were also found in the hexane extract. Furthermore, we examined antimicrobial activity of the three extracts (methanol, hexane, aqueous against a panel of microorganisms (Escherichia coli, Bacillus subtillis, Pasteurella multocida, Aspergillus niger, and Fusarium solani by disc-diffusion assay, and found the hexane extract to be the best antimicrobial agent. Hexane extract was also observed as to be most effective in a potato disc assay. As hexane extract showed potent activity in all the investigated assays, it was targeted for cytotoxic assessment. Maximum cytotoxicity (61.66% by hexane extract was found at 800 μg/mL. It is concluded that investigated extracts have potential for isolation of antioxidant and antimicrobial compounds for the pharmaceutical industry.

Avaliação de bacterina e Lactobacillus plantarum frente à infecção experimental por Vibrio harveyi em pós-larvas de Litopenaeus vannamei

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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.

Plasma concentrations resulting from florfenicol preparations given to pigs in their drinking water.

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Gutiérrez, L; Vargas, D; Ocampo, L; Sumano, H; Martinez, R; Tapia, G

2011-09-01

Florfenicol administered through the drinking water has been recommended as a metaphylactic antibacterial drug to control outbreaks of respiratory diseases in pigs caused by strains of Actinobacillus pleuropneumoniae and Pasteurella multocida, yet it is difficult to pinpoint in practice when the drug is given metaphylactically or therapeutically. Further, pigs are likely to reject florfenicol-medicated water, and plasma concentrations of the drug are likely to be marginal for diseases caused by Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. The reported minimal inhibitory concentration (MIC) values for these organisms show a breakpoint of 2 to 3 μg/mL. An experiment was conducted during September and October 2009. One hundred twenty healthy crossbred pigs (Landrace-Yorkshire), weighing 23 ± 6.2 kg, were used in this trial. They were randomly assigned to 5 groups, with 3 replicates of 8 animals/group. Two commercial preparations of florfenicol were administered through the drinking water at 2 concentrations (0.01 and 0.015%). Water intake was measured before and after medication, and plasma concentrations of florfenicol were determined by HPLC. Considerable rejection of florfenicol-medicated water was observed. However, plasma florfenicol concentrations were of a range sufficient for a methaphylaxis approach to preventing disease by bacteria, with MIC breakpoints of ≤ 0.25 μg/mL. Decreased efficacy as a metaphylactic medication should be expected for bacteria with MIC >0.25 μg/mL, considering the reported existence of bacteria resistant to florfenicol and the natural resistance of Streptococcus suis or E. coli to this drug.

Optimizing polymerase chain reaction testing for the diagnosis of pertussis: current perspectives

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Arbefeville S

2015-09-01

Full Text Available Sophie Arbefeville, Patricia Ferrieri Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN, USA Abstract: Nucleic acid testing has revolutionized the diagnosis of pertussis in the clinical microbiology laboratory and has become the main avenue of testing for pertussis infection. Real-time polymerase chain reaction (RT-PCR is an important tool for timely diagnosis of pertussis and is more sensitive than culture. The most commonly amplified targets are the insertion-sequence (IS genes, which are found in multiple copies in the genome of Bordetella species. Some strains of Bordetella pertussis have more than 200 copies of IS481 in their genome. This high number of repeats allows RT-PCR assays to be very sensitive and makes nucleic acid testing two to three times more sensitive than culture. Despite these advantages, RT-PCR can give inaccurate results due to contamination or lack of specificity. Contamination can easily happen during specimen collection, DNA extraction, or nucleic acid amplification steps. To avoid contamination, laboratories need to have quality controls and good workflows in place. The poor specificity of the nucleic acid assays amplifying the IS genes is because they are found in various Bordetella species and, thus, not unique to a specific species. Bordetella holmesii, a more recently described Bordetella species found to be responsible for respiratory symptoms similar to pertussis in adolescents and adults, can be misidentified as B. pertussis in RT-PCR assays that amplify only the IS481 target. Use of multiple targets may improve specificity of RT-PCR assays for pertussis. In the past few years, the US Food and Drug Administration has cleared three commercial assays for the detection of B. pertussis in respiratory specimens. Several commercial assays and analyte-specific reagents, which are not US Food and Drug Administration cleared, are available for the detection of one

Biofilm formation and cellulose expression by Bordetella avium 197N, the causative agent of bordetellosis in birds and an opportunistic respiratory pathogen in humans.

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McLaughlin, Kimberley; Folorunso, Ayorinde O; Deeni, Yusuf Y; Foster, Dona; Gorbatiuk, Oksana; Hapca, Simona M; Immoor, Corinna; Koza, Anna; Mohammed, Ibrahim U; Moshynets, Olena; Rogalsky, Sergii; Zawadzki, Kamil; Spiers, Andrew J

2017-06-01

Although bacterial cellulose synthase (bcs) operons are widespread within the Proteobacteria phylum, subunits required for the partial-acetylation of the polymer appear to be restricted to a few γ-group soil, plant-associated and phytopathogenic pseudomonads, including Pseudomonas fluorescens SBW25 and several Pseudomonas syringae pathovars. However, a bcs operon with acetylation subunits has also been annotated in the unrelated β-group respiratory pathogen, Bordetella avium 197N. Our comparison of subunit protein sequences and GC content analyses confirms the close similarity between the B. avium 197N and pseudomonad operons and suggests that, in both cases, the cellulose synthase and acetylation subunits were acquired as a single unit. Using static liquid microcosms, we can confirm that B. avium 197N expresses low levels of cellulose in air-liquid interface biofilms and that biofilm strength and attachment levels could be increased by elevating c-di-GMP levels like the pseudomonads, but cellulose was not required for biofilm formation itself. The finding that B. avium 197N is capable of producing cellulose from a highly-conserved, but relatively uncommon bcs operon raises the question of what functional role this modified polymer plays during the infection of the upper respiratory tract or survival between hosts, and what environmental signals control its production. Copyright © 2017 Institut Pasteur. All rights reserved.

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002927 gi|33603734 >1kmoA 3 661 60 754 2e-50 ... ref|NP_891294.1| exogenous ferric... siderophore receptor [Bordetella bronchiseptica ... RB50] gb|AAB51774.1| exogenous ferric siderophor...e ... receptor emb|CAE35124.1| exogenous ferric siderophore ... receptor [Bordetella bronchise

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002929 gi|33592330 >1uynX 1 299 352 647 5e-40 ... ref|NP_879974.1| tracheal colonization... factor precursor [Bordetella pertussis Tohama ... I] emb|CAA08832.2| tracheal colonization fac...tor ... [Bordetella pertussis] emb|CAE41497.1| tracheal ... colonization factor precursor [Bor

Epidemiology of pertussis in Casablanca (Morocco): contribution of conventional and molecular diagnosis tools.

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Katfy, Khalid; Guiso, Nicole; Diawara, Idrissa; Zerouali, Khalid; Slaoui, Bouchra; Jouhadi, Zineb; Zineddine, Abdelhadi; Belabbes, Houria; Elmdaghri, Naima

2017-05-16

Pertussis, a vaccine preventable disease, is still responsible of significant morbidity and mortality around the world, mostly in newborns. The aim of the present study was (1) to introduce pertussis surveillance in the major pediatric hospital of Casablanca (2) to analyze the prevalence of pertussis among children under 14 years of age and their entourage in Casablanca, Morocco. This is a prospective and non-case controlled study, including children suspected of Pertussis admitted at the Abderrahim Harouchi Pediatric Hospital in Casablanca, from January 2013 to June 2015. Nasopharyngeal samples were obtained for Bordetella spp. culture and Real time PCR detection (RT-PCR) with specific primers of Bordetella spp., B. pertussis, B. parapertussis and B. holmesii. The detection of Bordetella spp. was also performed in some household contacts of the children suspected of pertussis. During the 2.5-years period, a total of 282 samples were collected from hospitalized children (156) and in some of their contacts (126). Among 156 samples from the children (from whom 57% were under 2 month of age), Bordetella DNA was detected in 61% (96/156) by RT-PCR. Among these positive samples, 91.7% (88/96) corresponded to B. pertussis DNA. Furthermore, in 39.5% (38/96) of the Bordetella positive samples, B. holmesii DNA was also detected. B. parapertussis DNA was detected in only one sample (1/156). Out of the 156 samples collected from the hospitalized children, only 48 were tested by culture, and 4 B. pertussis were isolated (8.3%). Among the 126 samples from the contacts of the children, mostly mothers (115 cases), Bordetella DNA was detected in 47% (59/126), 90% (53/59) being B. pertussis DNA. Moreover, B. holmesii DNA was also detected in 18.6% (11/59) of the Bordetella positive samples, and coexistence of B. pertussis and B. holmesii DNA in 36.5% (35/96). Two B. pertussis were isolated by culture performed on 43 samples of the contacts of the children (4.6%). This study

Disease: H01077 [KEGG MEDICUS

Lifescience Database Archive (English)

Full Text Available o the genus Bordetella that is isolated from poultry with respiratory disease. B. hinzii may cause disease i...sters K, Blackall PJ ... TITLE ... Bordetella hinzii sp. nov., isolated from poultry and humans. ... JOURNAL ... Int J Syst Bacteriol 45:37-45 (1995) DOI:10.1099/00207713-45-1-37

In vitro activity of five tetracyclines and some other antimicrobial agents against four porcine respiratory tract pathogens.

Science.gov (United States)

Pijpers, A; Van Klingeren, B; Schoevers, E J; Verheijden, J H; Van Miert, A S

1989-09-01

The minimal inhibitory concentrations (MIC) of five tetracyclines and ten other antimicrobial agents were determined for four porcine bacterial respiratory tract pathogens by the agar dilution method. For the following oxytetracycline-susceptible strains, the MIC50 ranges of the tetracyclines were: P. multocida (n = 17) 0.25-0.5 micrograms/ml; B. bronchiseptica (n = 20) 0.25-1.0 micrograms/ml; H. pleuropneumoniae (n = 20) 0.25-0.5 micrograms/ml; S. suis Type 2 (n = 20) 0.06-0.25 micrograms/ml. For 19 oxytetracycline-resistant P. multocida strains the MIC50 of the tetracyclines varied from 64 micrograms/ml for oxytetracycline to 0.5 micrograms/ml for minocycline. Strikingly, minocycline showed no cross-resistance with oxytetracycline, tetracycline, chlortetracycline and doxycycline in P. multocida and in H. pleuropneumoniae. Moreover, in susceptible strains minocycline showed the highest in vitro activity followed by doxycycline. Low MIC50 values were observed for chloramphenicol, ampicillin, flumequine, ofloxacin and ciprofloxacin against P. multocida and H. pleuropneumoniae. B. bronchiseptica was moderately susceptible or resistant to these compounds. As expected tiamulin, lincomycin, tylosin and spiramycin were not active against H. pleuropneumoniae. Except for flumequine, the MIC50 values of nine antimicrobial agents were low for S. suis Type 2. Six strains of this species showed resistance to the macrolides and lincomycin.

Occurrence of antimicrobial resistance in bacteria from diagnostic samples from dogs.

Science.gov (United States)

Pedersen, Karl; Pedersen, Kristina; Jensen, Helene; Finster, Kai; Jensen, Vibeke F; Heuer, Ole E

2007-10-01

To study the occurrence of antimicrobial resistance among common bacterial pathogens from dogs and relate resistance patterns to data on consumption of antimicrobials. The antimicrobial susceptibility patterns of 201 Staphylococcus intermedius, 37 Streptococcus canis, 39 Pseudomonas aeruginosa, 25 Pasteurella multocida, 29 Proteus spp. and 449 Escherichia coli isolates from clinical submissions from dogs were determined by a broth-dilution method for determination of minimal inhibitory concentration. Data for consumption of antimicrobials were retrieved from VetStat, a national database for reporting antimicrobial prescriptions. The majority of the antimicrobials prescribed for dogs were broad-spectrum compounds, and extended-spectrum penicillins, cephalosporins and sulphonamides + trimethoprim together accounted for 81% of the total amount used for companion animals. Resistance to cephalosporins and amoxicillin with clavulanic acid was very low for all bacterial species examined, except for P. aeruginosa, and resistance to sulphonamides and trimethoprim was low for most species. Among the S. intermedius isolates, 60.2% were resistant to penicillin, 30.2% to fusidic acid and 27.9% to macrolides. Among E. coli isolates, the highest level of resistance was recorded for ampicillin, sulphonamides, trimethoprim, tetracyclines and streptomycin. Certain differences in resistance patterns between isolates from different sites or organs were noticed for E. coli, S. intermedius and Proteus isolates. This investigation provided data on occurrence of antimicrobial resistance in important pathogenic bacteria from dogs, which may be useful for the small animal practitioner. Resistance was low to the compounds that were most often used, but unfortunately, these compounds were broad-spectrum. Data on resistance and usage may form a background for the establishment of a set of recommendations for prudent use of antimicrobials for companion animals.

Aquatic bird disease and mortality as an indicator of changing ecosystem health

Science.gov (United States)

Newman, Scott H.; Chmura, Aleksei; Converse, Kathy; Kilpatrick, A. Marm; Patel, Nikkita; Lammers, Emily; Daszak, Peter

2007-01-01

We analyzed data from pathologic investigations in the United States, collected by the USGS National Wildlife Health Center between 1971 and 2005, into aquatic bird mortality events. A total of 3619 mortality events was documented for aquatic birds, involving at least 633 708 dead birds from 158 species belonging to 23 families. Environmental causes accounted for the largest proportion of mortality events (1737 or 48%) and dead birds (437 258 or 69%); these numbers increased between 1971 and 2000, with biotoxin mortalities due to botulinum intoxication (Types C and E) being the leading cause of death. Infectious diseases were the second leading cause of mortality events (20%) and dead birds (20%), with both viral diseases, including duck plague (Herpes virus), paramyxovirus of cormorants (Paramyxovirus PMV1) and West Nile virus (Flavivirus), and bacterial diseases, including avian cholera (Pasteurella multocida), chlamydiosis (Chalmydia psittici), and salmonellosis (Salmonella sp.), contributing. Pelagic, coastal marine birds and species that use marine and freshwater habitats were impacted most frequently by environmental causes of death, with biotoxin exposure, primarily botulinum toxin, resulting in mortalities of both coastal and freshwater species. Pelagic birds were impacted most severely by emaciation and starvation, which may reflect increased anthropogenic pressure on the marine habitat from over-fishing, pollution, and other factors. Our study provides important information on broad trends in aquatic bird mortality and highlights how long-term wildlife disease studies can be used to identify anthropogenic threats to wildlife conservation and ecosystem health. In particular, mortality data for the past 30 yr suggest that biotoxins, viral, and bacterial diseases could have impacted >5 million aquatic birds.

Genome Structural Diversity among 31 Bordetella pertussis Isolates from Two Recent U.S. Whooping Cough Statewide Epidemics.

Science.gov (United States)

Bowden, Katherine E; Weigand, Michael R; Peng, Yanhui; Cassiday, Pamela K; Sammons, Scott; Knipe, Kristen; Rowe, Lori A; Loparev, Vladimir; Sheth, Mili; Weening, Keeley; Tondella, M Lucia; Williams, Margaret M

2016-01-01

During 2010 and 2012, California and Vermont, respectively, experienced statewide epidemics of pertussis with differences seen in the demographic affected, case clinical presentation, and molecular epidemiology of the circulating strains. To overcome limitations of the current molecular typing methods for pertussis, we utilized whole-genome sequencing to gain a broader understanding of how current circulating strains are causing large epidemics. Through the use of combined next-generation sequencing technologies, this study compared de novo, single-contig genome assemblies from 31 out of 33 Bordetella pertussis isolates collected during two separate pertussis statewide epidemics and 2 resequenced vaccine strains. Final genome architecture assemblies were verified with whole-genome optical mapping. Sixteen distinct genome rearrangement profiles were observed in epidemic isolate genomes, all of which were distinct from the genome structures of the two resequenced vaccine strains. These rearrangements appear to be mediated by repetitive sequence elements, such as high-copy-number mobile genetic elements and rRNA operons. Additionally, novel and previously identified single nucleotide polymorphisms were detected in 10 virulence-related genes in the epidemic isolates. Whole-genome variation analysis identified state-specific variants, and coding regions bearing nonsynonymous mutations were classified into functional annotated orthologous groups. Comprehensive studies on whole genomes are needed to understand the resurgence of pertussis and develop novel tools to better characterize the molecular epidemiology of evolving B. pertussis populations. IMPORTANCE Pertussis, or whooping cough, is the most poorly controlled vaccine-preventable bacterial disease in the United States, which has experienced a resurgence for more than a decade. Once viewed as a monomorphic pathogen, B. pertussis strains circulating during epidemics exhibit diversity visible on a genome structural

Acquisition of C1 inhibitor by Bordetella pertussis virulence associated gene 8 results in C2 and C4 consumption away from the bacterial surface.

Science.gov (United States)

Hovingh, Elise S; van den Broek, Bryan; Kuipers, Betsy; Pinelli, Elena; Rooijakkers, Suzan H M; Jongerius, Ilse

2017-07-01

Whooping cough, or pertussis, is a contagious disease of the respiratory tract that is re-emerging worldwide despite high vaccination coverage. The causative agent of this disease is the Gram-negative Bordetella pertussis. Knowledge on complement evasion strategies of this pathogen is limited. However, this is of great importance for future vaccine development as it has become apparent that a novel pertussis vaccine is needed. Here, we unravel the effect of Virulence associated gene 8 (Vag8) of B. pertussis on the human complement system at the molecular level. We show that both recombinant and endogenously secreted Vag8 inhibit complement deposition on the bacterial surface at the level of C4b. We reveal that Vag8 binding to human C1-inhibitor (C1-inh) interferes with the binding of C1-inh to C1s, C1r and MASP-2, resulting in the release of active proteases that subsequently cleave C2 and C4 away from the bacterial surface. We demonstrate that the depletion of these complement components in the bacterial surrounding and subsequent decreased deposition on B. pertussis leads to less complement-mediated bacterial killing. Vag8 is the first protein described that specifically prevents C1s, C1r and MASP-2 binding to C1-inh and thereby mediates complement consumption away from the bacterial surface. Unravelling the mechanism of this unique complement evasion strategy of B. pertussis is one of the first steps towards understanding the interactions between the first line of defense complement and B. pertussis.

Compatibility of a bivalent modified-live vaccine against Bordetella bronchiseptica and CPiV, and a trivalent modified-live vaccine against CPV, CDV and CAV-2.

Science.gov (United States)

Jacobs, A A C; Bergman, J G H E; Theelen, R P H; Jaspers, R; Helps, J M; Horspool, L J I; Paul, G

2007-01-13

Eight puppies (group 1) were vaccinated once with a bivalent modified-live vaccine against infectious tracheobronchitis by the intranasal route and at the same time with an injectable trivalent vaccine against canine parvovirus, canine distemper virus and canine adenovirus; a second group of eight puppies (group 2) was vaccinated only with the intranasal bivalent vaccine, and a further eight puppies (group 3) were vaccinated only with the injectable trivalent vaccine. Three weeks later they were all challenged with wildtype Bordetella bronchiseptica and canine parainfluenza virus by the aerosol route, and their antibody responses to the five vaccine organisms were determined. Oronasal swabs were taken regularly before and after the challenge for the isolation of bacteria and viruses, and the puppies were observed for clinical signs for three weeks after the challenge. There were no significant differences in the puppies' titres against canine parvovirus, canine distemper virus and canine adenovirus type 2 between the groups vaccinated with or without the bivalent intranasal vaccine. After the challenge the mean clinical scores of the two groups vaccinated with the intranasal vaccine were nearly 90 per cent lower (P=0.001) than the mean score of the group vaccinated with only the trivalent injectable vaccine, and the puppies in this group all became culture-positive for B bronchiseptica and canine parainfluenza virus. There were only small differences between the rates of isolation of B bronchiseptica from groups 1, 2 and 3, but significantly lower yields of canine parainfluenza virus were isolated from groups 1 and 2 than from group 3.

Acquisition of C1 inhibitor by Bordetella pertussis virulence associated gene 8 results in C2 and C4 consumption away from the bacterial surface

Science.gov (United States)

Hovingh, Elise S.; Kuipers, Betsy; Pinelli, Elena; Rooijakkers, Suzan H. M.

2017-01-01

Whooping cough, or pertussis, is a contagious disease of the respiratory tract that is re-emerging worldwide despite high vaccination coverage. The causative agent of this disease is the Gram-negative Bordetella pertussis. Knowledge on complement evasion strategies of this pathogen is limited. However, this is of great importance for future vaccine development as it has become apparent that a novel pertussis vaccine is needed. Here, we unravel the effect of Virulence associated gene 8 (Vag8) of B. pertussis on the human complement system at the molecular level. We show that both recombinant and endogenously secreted Vag8 inhibit complement deposition on the bacterial surface at the level of C4b. We reveal that Vag8 binding to human C1-inhibitor (C1-inh) interferes with the binding of C1-inh to C1s, C1r and MASP-2, resulting in the release of active proteases that subsequently cleave C2 and C4 away from the bacterial surface. We demonstrate that the depletion of these complement components in the bacterial surrounding and subsequent decreased deposition on B. pertussis leads to less complement-mediated bacterial killing. Vag8 is the first protein described that specifically prevents C1s, C1r and MASP-2 binding to C1-inh and thereby mediates complement consumption away from the bacterial surface. Unravelling the mechanism of this unique complement evasion strategy of B. pertussis is one of the first steps towards understanding the interactions between the first line of defense complement and B. pertussis. PMID:28742139

Exposure to Bordetella pertussis adenylate cyclase toxin affects integrin-mediated adhesion and mechanics in alveolar epithelial cells.

Science.gov (United States)

Angely, Christelle; Nguyen, Ngoc-Minh; Andre Dias, Sofia; Planus, Emmanuelle; Pelle, Gabriel; Louis, Bruno; Filoche, Marcel; Chenal, Alexandre; Ladant, Daniel; Isabey, Daniel

2017-08-01

The adenylate cyclase (CyaA) toxin is a major virulent factor of Bordetella pertussis, the causative agent of whooping cough. CyaA toxin is able to invade eukaryotic cells where it produces high levels of cyclic adenosine monophosphate (cAMP) affecting cellular physiology. Whether CyaA toxin can modulate cell matrix adhesion and mechanics of infected cells remains largely unknown. In this study, we use a recently proposed multiple bond force spectroscopy (MFS) with an atomic force microscope to assess the early phase of cell adhesion (maximal detachment and local rupture forces) and cell rigidity (Young's modulus) in alveolar epithelial cells (A549) for toxin exposure 95%) at CyaA concentration of 0.5 nM, but a significant effect (≈81%) at 10 nM. MFS performed on A549 for three different concentrations (0.5, 5 and 10 nM) demonstrates that CyaA toxin significantly affects both cell adhesion (detachment forces are decreased) and cell mechanics (Young's modulus is increased). CyaA toxin (at 0.5 nM) assessed at three indentation/retraction speeds (2, 5 and 10 μm/s) significantly affects global detachment forces, local rupture events and Young modulus compared with control conditions, while an enzymatically inactive variant CyaAE5 has no effect. These results reveal the loading rate dependence of the multiple bonds newly formed between the cell and integrin-specific coated probe as well as the individual bond kinetics which are only slightly affected by the patho-physiological dose of CyaA toxin. Finally, theory of multiple bond force rupture enables us to deduce the bond number N which is reduced by a factor of 2 upon CyaA exposure (N ≈ 6 versus N ≈ 12 in control conditions). MFS measurements demonstrate that adhesion and mechanical properties of A549 are deeply affected by exposure to the CyaA toxin but not to an enzymatically inactive variant. This indicates that the alteration of cell mechanics triggered by CyaA is a consequence of the increase in

Avian Cholera emergence in Arctic-nesting northern Common Eiders: using community-based, participatory surveillance to delineate disease outbreak patterns and predict transmission risk

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Samuel A. Iverson

2016-12-01

Full Text Available Emerging infectious diseases are a growing concern in wildlife conservation. Documenting outbreak patterns and determining the ecological drivers of transmission risk are fundamental to predicting disease spread and assessing potential impacts on population viability. However, evaluating disease in wildlife populations requires expansive surveillance networks that often do not exist in remote and developing areas. Here, we describe the results of a community-based research initiative conducted in collaboration with indigenous harvesters, the Inuit, in response to a new series of Avian Cholera outbreaks affecting Common Eiders (Somateria mollissima and other comingling species in the Canadian Arctic. Avian Cholera is a virulent disease of birds caused by the bacterium Pasteurella multocida. Common Eiders are a valuable subsistence resource for Inuit, who hunt the birds for meat and visit breeding colonies during the summer to collect eggs and feather down for use in clothing and blankets. We compiled the observations of harvesters about the growing epidemic and with their assistance undertook field investigation of 131 colonies distributed over >1200 km of coastline in the affected region. Thirteen locations were identified where Avian Cholera outbreaks have occurred since 2004. Mortality rates ranged from 1% to 43% of the local breeding population at these locations. Using a species-habitat model (Maxent, we determined that the distribution of outbreak events has not been random within the study area and that colony size, vegetation cover, and a measure of host crowding in shared wetlands were significantly correlated to outbreak risk. In addition, outbreak locations have been spatially structured with respect to hypothesized introduction foci and clustered along the migration corridor linking Arctic breeding areas with wintering areas in Atlantic Canada. At present, Avian Cholera remains a localized threat to Common Eider populations in the

Cryostored autologous skull bone for cranioplasty? A study on cranial bone flaps' viability and microbial contamination after deep-frozen storage at -80°C.

Science.gov (United States)

Chan, David Yuen Chung; Mok, Yi Tan; Lam, Ping Kuen; Tong, Cindy See Wai; Ng, Stephanie Chi Ping; Sun, Tin Fung David; Poon, Wai Sang

2017-08-01

Craniectomy is a life-saving procedure. Subsequent cranioplasty with autologous skull bone has a bone resorption rate from 4% to 22.8% and an infection rate from 3.3% to 26%. There are concerns with their viability and the potential microbial contamination as they were explanted for a long period of time. Eighteen cranial bone flaps stored at Prince of Wales Hospital Skull Bone Bank during the period from June 2011 to March 2016 were identified. Ethics approval was obtained. Bone chips and deep bone swabs were collected for osteoblast culture and microbial culture. Skull Bone Bank was kept at -80°C under strict aseptic technique during the study period. The storage period ranged from 4months to 55months. For the osteoblast culture, all eighteen bone flaps had no viable osteoblast growth. For the bacterial culture, five had positive bacteria growth (27.8%). Three were Pasteurella multocida and two were Methicillin-resistant Staphylococcus aureus. The mean duration of storage of the infected bone flap was 32.9months (±15.1months) versus 19.9months (±17.9months) of those bone flaps with no bacterial growth (p=0.1716). The mean size of the infected versus non-infected bone flaps was 117.7cm 2 (±44.96cm 2 ) versus 76.8cm 2 (±50.24cm 2 ) respectively (p=0.1318). Although in this study statistical significance was not reached, it was postulated that infected bone flaps tended to be larger in size and had a longer duration of storage. In conclusion, cryostored skull bone flaps beyond four months showed no viable osteoblasts. Bacterial contamination rate of bone flaps was 27.8% in this study. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Primera incursión en la obtención de curieles libre de patógenos específicos en Cuba

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Bárbara O González Navarro

2014-01-01

Full Text Available Título corto: Primera incursión en la obtención de curieles SPF en Cuba Título en ingles: First foray into the production of specific pathogen-free guinea pigs in Cuba Resumen: Resultados confiables y económicos solo son obtenidos cuando los animales de experimentación son aislados de factores ambientales y biológicos, implantándose en el biomodelo una microbiota normal, lejos de la presencia de microorganismos patógenos. El objetivo de la investigación fue obtener curieles libre de patógenos específicos por cesárea aséptica, mantenidos en aisladores y alimentados con dietas estériles. Se realizaron 26 histerectomías. Los animales fueron alimentados con una fórmula modificada (L-477 en forma de papilla hasta los 21 días y permanentemente después la C-484 sólida y granulada, esterilizadas a 121oC/20 minutos o a 1,5Mrad. Además fueron suplementados con vitamina C y B1. El forraje o heno fue consumido a partir de la primera generación. La microbiota gastrointestinal se administró por vía oral en 0,5 ml de una dilución de 10-6/g de contenido de la porción final del íleon, ciego y principio del colon de curieles, a las 24 y 48 horas del nacimiento. Se utilizó para el monitoreo microbiológico caldo Tioglicolato, caldo Triptona Soya y caldo Saboraud incubados aeróbicamente a temperatura de 55, 37y 25oC respectivamente. Se obtuvieron 51 neonatos. La mortalidad más alta se registró entre los primeros 10 días de edad (58,8%. Se lograron 12 animales (3 machos y 9 hembras, 6 de las hembras se reprodujeron aproximadamente a los 9 meses de edad, lográndose 11 crías por parto normal. La metodología aplicada permitió obtener curieles libres de Salmonella sp, Pasteurella sp, Streptococcus del tipo A y C, Bordetella bronchiseptica, Toxoplasma gondii, Virus Sendai y parásitos internos y externos. Palabras clave: curieles, libre de patógenos específicos, aisladores, histerectomía y dietas estériles. Abstract

Evaluation of antibacterial properties of some medicinal plants used in Iran.

Science.gov (United States)

Bonjar, Shahidi

2004-10-01

Forty-five species of 29 plant families used in the traditional medicine by Iranian people, showed antibacterial activities against one or more of the bacterial species: Bacillus cereus, Bacillus pumilus, Bordetella bronchiseptica, Escherichia coli, Klebsiella pneumoniae, Micrococcus luteus, Pseudomonas aeruginosa, Pseudomonas fluorescens, Serratia marcescens, Staphylococcus aureus and Staphylococcus epidermidis. No plant showed activity against Serratia marcescens; Bordetella bronchiseptica being the most susceptible species. All extracts showed the same activity 18 months later.

Characterization of two Achromobacter xylosoxidans isolates from patients with pertussis-like symptoms

Institute of Scientific and Technical Information of China (English)

Fiorella Orellana-Peralta; Michelle Jacinto; Maria J Pons; Cludia Gomes; Carlos Bada; Isabel Reyes; Juana del Valle Mendoza; Joaquim Ruiz

2015-01-01

Objective:To characterize two Achromobacter xylosoxidans recovered from 2 patients diagnosed with pertussis during a Bordetella pertussis surveillance program. Methods:Nasopharyngeal swabs from 2 children under 1 year of age with clinical suspicion of pertussis were analyzed by culture and PCR. Results:Two Achromobacter xylosoxidans A8, closely related to Bordetella spp. were recovered from 2 patients diagnosed of pertussis, both carrying the ptxA gene and IS418 the pertussis toxin encoding gene. Subsequently, antibiotic susceptibility was evaluated by disk-diffusion method and by PCR. Conclusions:Although more detailed studies are needed, the present data highlight the possibility that Achromobacter xylosoxidans, closely related Bordetella pertussis microorganisms and not covered under the vaccine umbrella, might also result in cases of whooping cough. Thereby further surveillance is necessary to determine the extension and relevance of their pathogenic role in order to discriminate their real public health implication.

Antimicrobial effect of probiotics on bacterial species from dental plaque.

Science.gov (United States)

Zambori, Csilla; Morvay, Attila Alexandru; Sala, Claudia; Licker, Monica; Gurban, Camelia; Tanasie, Gabriela; Tirziu, Emil

2016-03-31

The antimicrobial role of probiotic Lactobacillus casei subspecies casei DG (L. casei DG) and of the mix culture of probiotic Lactobacillus acidophilus LA-5 and Bifidobacterium BB-12 was tested on species of Staphylococcus, Streptococcus, Pasteurella, and Neisseria genera from supragingival sites from dogs with dental disease of different breed, age, sex, weight, and diet. The research was conducted on these four genera because of their importance in zoonotic infections after dog bites. Species from Staphylococcus, Streptococcus, Pasteurella, and Neisseria genera were isolated and identified. To test the antimicrobial efficacy of L. casei DG and the mixed culture of probiotic L. acidophilus LA-5 and Bifidobacterium bifidum BB-12 on the pathogenic species, the agar overlay method was used. L. casei DG had a bactericidal effect on all analyzed species isolated from Staphylococcus, Streptococcus, Pasteurella, and Neisseria genera after 24 hours of incubation. The mixed probiotic culture made up of L. acidophilus LA-5 and Bifidobacterium BB-12 species had no bactericidal effect on the species of Staphylococcus and Streptococcus genera, which were resistant. However, it had a bacteriostatic effect on several species of Pasteurella and Neisseria genera. This work highlights the antimicrobial potential of probiotics in vitro, demonstrating that the probiotic L. casei DG has a bactericidal effect on all analyzed species isolated from dental plaque and that the mix culture of probiotic L. acidophilus LA-5 and Bifidobacterium BB-12 has only a bacteriostatic effect.

Biochemical and Structural Characterization of WlbA from Bordetella pertussis and Chromobacterium violaceum: Enzymes Required for the Biosynthesis of 2,3-Diacetamido-2,3-dideoxy-d-mannuronic Acid

Energy Technology Data Exchange (ETDEWEB)

Thoden, James B.; Holden, Hazel M. (UW)

2011-12-22

The unusual sugar 2,3-diacetamido-2,3-dideoxy-d-mannuronic acid, or ManNAc3NAcA, has been observed in the lipopolysaccharides of both pathogenic and nonpathogenic Gram-negative bacteria. It is added to the lipopolysaccharides of these organisms by glycosyltransferases that use as substrates UDP-ManNAc3NAcA. Five enzymes are ultimately required for the biosynthesis of UDP-ManNAc3NAcA starting from UDP-N-acetylglucosamine. The second enzyme in the pathway, encoded by the wlba gene and referred to as WlbA, catalyzes the NAD-dependent oxidation of the C-3' hydroxyl group of the UDP-linked sugar. Here we describe a combined structural and functional investigation of the WlbA enzymes from Bordetella pertussis and Chromobacterium violaceum. For this investigation, ternary structures were determined in the presence of NAD(H) and substrate to 2.13 and 1.5 {angstrom} resolution, respectively. Both of the enzymes display octameric quaternary structures with their active sites positioned far apart. The octamers can be envisioned as tetramers of dimers. Kinetic studies demonstrate that the reaction mechanisms for these enzymes are sequential and that they do not require {alpha}-ketoglutarate for activity. These results are in sharp contrast to those recently reported for the WlbA enzymes from Pseudomonas aeruginosa and Thermus thermophilus, which function via ping-pong mechanisms that involve {alpha}-ketoglutarate. Taken together, the results reported here demonstrate that there are two distinct families of WlbA enzymes, which differ with respect to amino acid sequences, quaternary structures, active site architectures, and kinetic mechanisms.

Development of a PCR test for identification of Haemophilus somnus in pure and mixed cultures

DEFF Research Database (Denmark)

Angen, Øystein; Ahrens, Peter; Tegtmeier, Conny

1998-01-01

a single colony of H. somnus in the presence of 10(9) CFU of P. multocida even after 2 days of incubation. In conclusion, the present PCR test has been shown to represent a specific test for identification of H. somnus both in pure and mixed cultures. It represents a quick, sensitive and reliable method...... rise to an amplicon in the PCR test. The performance of the test on mixed cultures was evaluated by adding P. multocida to serial dilutions of H. somnus and incubating the agarplates for 1 and 2 days. This showed that the PCR test applied to the harvest from an agarplate can be expected to detect...

Ca2+ Influx and Tyrosine Kinases Trigger Bordetella Adenylate Cyclase Toxin (ACT) Endocytosis. Cell Physiology and Expression of the CD11b/CD18 Integrin Major Determinants of the Entry Route

Science.gov (United States)

Etxebarria, Aitor; González-Bullón, David; Gómez-Bilbao, Geraxane; Ostolaza, Helena

2013-01-01

Humans infected with Bordetella pertussis, the whooping cough bacterium, show evidences of impaired host defenses. This pathogenic bacterium produces a unique adenylate cyclase toxin (ACT) which enters human phagocytes and catalyzes the unregulated formation of cAMP, hampering important bactericidal functions of these immune cells that eventually cause cell death by apoptosis and/or necrosis. Additionally, ACT permeabilizes cells through pore formation in the target cell membrane. Recently, we demonstrated that ACT is internalised into macrophages together with other membrane components, such as the integrin CD11b/CD18 (CR3), its receptor in these immune cells, and GM1. The goal of this study was to determine whether ACT uptake is restricted to receptor-bearing macrophages or on the contrary may also take place into cells devoid of receptor and gain more insights on the signalling involved. Here, we show that ACT is rapidly eliminated from the cell membrane of either CR3-positive as negative cells, though through different entry routes, which depends in part, on the target cell physiology and characteristics. ACT-induced Ca2+ influx and activation of non-receptor Tyr kinases into the target cell appear to be common master denominators in the different endocytic strategies activated by this toxin. Very importantly, we show that, upon incubation with ACT, target cells are capable of repairing the cell membrane, which suggests the mounting of an anti-toxin cell repair-response, very likely involving the toxin elimination from the cell surface. PMID:24058533

Ca2+ influx and tyrosine kinases trigger Bordetella adenylate cyclase toxin (ACT endocytosis. Cell physiology and expression of the CD11b/CD18 integrin major determinants of the entry route.

Directory of Open Access Journals (Sweden)

Kepa B Uribe

Full Text Available Humans infected with Bordetella pertussis, the whooping cough bacterium, show evidences of impaired host defenses. This pathogenic bacterium produces a unique adenylate cyclase toxin (ACT which enters human phagocytes and catalyzes the unregulated formation of cAMP, hampering important bactericidal functions of these immune cells that eventually cause cell death by apoptosis and/or necrosis. Additionally, ACT permeabilizes cells through pore formation in the target cell membrane. Recently, we demonstrated that ACT is internalised into macrophages together with other membrane components, such as the integrin CD11b/CD18 (CR3, its receptor in these immune cells, and GM1. The goal of this study was to determine whether ACT uptake is restricted to receptor-bearing macrophages or on the contrary may also take place into cells devoid of receptor and gain more insights on the signalling involved. Here, we show that ACT is rapidly eliminated from the cell membrane of either CR3-positive as negative cells, though through different entry routes, which depends in part, on the target cell physiology and characteristics. ACT-induced Ca(2+ influx and activation of non-receptor Tyr kinases into the target cell appear to be common master denominators in the different endocytic strategies activated by this toxin. Very importantly, we show that, upon incubation with ACT, target cells are capable of repairing the cell membrane, which suggests the mounting of an anti-toxin cell repair-response, very likely involving the toxin elimination from the cell surface.

Relative Contribution of Th1 and Th17 Cells in Adaptive Immunity to Bordetella pertussis: Towards the Rational Design of an Improved Acellular Pertussis Vaccine

Science.gov (United States)

Ross, Pádraig J.; Allen, Aideen C.; Walsh, Kevin; Misiak, Alicja; Lavelle, Ed C.; McLoughlin, Rachel M.; Mills, Kingston H. G.

2013-01-01

Whooping cough caused by Bordetella pertussis is a re-emerging infectious disease despite the introduction of safer acellular pertussis vaccines (Pa). One explanation for this is that Pa are less protective than the more reactogenic whole cell pertussis vaccines (Pw) that they replaced. Although Pa induce potent antibody responses, and protection has been found to be associated with high concentrations of circulating IgG against vaccine antigens, it has not been firmly established that host protection induced with this vaccine is mediated solely by humoral immunity. The aim of this study was to examine the relative contribution of Th1 and Th17 cells in host immunity to infection with B. pertussis and in immunity induced by immunization with Pw and Pa and to use this information to help rationally design a more effective Pa. Our findings demonstrate that Th1 and Th17 both function in protective immunity induced by infection with B. pertussis or immunization with Pw. In contrast, a current licensed Pa, administered with alum as the adjuvant, induced Th2 and Th17 cells, but weak Th1 responses. We found that IL-1 signalling played a central role in protective immunity induced with alum-adsorbed Pa and this was associated with the induction of Th17 cells. Pa generated strong antibody and Th2 responses, but was fully protective in IL-4-defective mice, suggesting that Th2 cells were dispensable. In contrast, Pa failed to confer protective immunity in IL-17A-defective mice. Bacterial clearance mediated by Pa-induced Th17 cells was associated with cell recruitment to the lungs after challenge. Finally, protective immunity induced by an experimental Pa could be enhanced by substituting alum with a TLR agonist that induces Th1 cells. Our findings demonstrate that alum promotes protective immunity through IL-1β-induced IL-17A production, but also reveal that optimum protection against B. pertussis requires induction of Th1, but not Th2 cells. PMID:23592988

[Serum immunoglobulin IgG subclass distribution of antibody responses to pertussis toxin and filamentous hemagglutinin of Bordetella pertussis in patients with whooping cough].

Science.gov (United States)

Rastawicki, Waldemar; Smietańska, Karolina; Rokosz-Chudziak, Natalia; Jagielski, Marek

2013-01-01

The present study was aimed at determining the IgG subclass distribution against pertussis toxin (PT) and filamentous hemagglutinin (FHA) of Bordetella pertussis in patients with whooping cough. The total number of 222 serum samples obtained from patients suspected in clinical investigation for pertussis were tested separately by in-house ELISA for the presence of IgG antibodies to pertussis toxin and filamentous hemagglutinin. The percentage distribution of specific anti-PT and anti-FHA IgG subclass response was calculated only on the basis of group of sera confirmed in the present study as positive for total IgG antibodies (183 sera to PT antigen and 129 to FHA antigen). Paired serum specimens were obtained from 36 patients. Based on the results of determining the level of antibodies in the sera of 40 blood donors, the cut-off limit of serum antibodies for each subclass was set at arithmetic mean plus two standard deviations. Antibodies of IgG1 to pertussis toxin and filamentous hemagglutinin were diagnosed in 151 (82.5%) and 99 (76.7%), IgG2 in 72 (39.0%) and 50 (38.8%), IgG3 in 17 (9.3%) and 43 (33.3%), IgG4 in 55 (30.1%) and 53 (41.1%) serum samples, respectively. There were no significant differences in percentage of sera with IgG1, IgG2 and IgG3 in relation to age of the patients. However, the frequency of occurrence of IgG4 antibodies was highest in the group of the youngest children to the age of 6 years old (61.8% for PT and 68.0% for FHA), and decrease with age, reaching the minimum in the group of patients above 40 years old (13.2% and 4.2% for PT and FHA, respectively). We also found significantly higher frequency of IgG4 to PT and FHA antigens in men than in women. Statistically significant, essential changes in the pattern of IgG subclass during the course of infection were not found. In conclusion, this study showed that all four subclasses of IgG antibodies to pertussis toxin and filamentous hemagglutinin are produced during whooping cough.

Masked rat: an x-ray-induced mutant with chronic blepharitis, alopecia, and pasteurellosis

International Nuclear Information System (INIS)

Kent, R.L.; Lutzner, M.A.; Hansen, C.T.

1976-01-01

An autosomal recessive mutation had been previously x-ray-induced in the rat and named the masked rat (genotype mk/mk). This study describes the mutant's appearance, histology, and microflora. The rat's eyelids were swollen, often to the point of closure, and its face was partially covered by a brownish crust, giving the mutant a mask-like appearance. The chronic blepharitis was also accompanied by alopecia that appeared as bare patches across the mutant's back. Pasteurella pneumotropica was found in eyelids and on skin from all masked rats. The normal rat demonstrated a resistance to Pasteurella pneumotropica infection, or, conversely, the masked rat appeared to be genetically predisposed to pasteurellosis

Pets and Pasteurella Infections

Science.gov (United States)

... present in some children, including an infection of the joints ( arthritis ), bones (osteomyelitis), and tendons (tenosynovitis). Less frequently, youngsters may have pneumonia , urinary tract ...

Detection of Staphylococcus Aureus Enterotoxin A and B Genes with PCR-EIA and a Hand-Held Electrochemical Sensor

National Research Council Canada - National Science Library

Aitichou, Mohamed; Henkins, Robert; Sultana, Afroz M; Ulrich, Robert G; Ibrahim, M. S

2004-01-01

... S. aureus DNA, and genomic DNA from Alcaligens, Bacillus, Bacteroides, Bordetella, Burkholderia, Clostridium, Comanonas, Enterobacter, Enterococcus, Escherichia, Francisella, Haemophilus, Klebsiella...

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella pertussis Tohama I] ... Length = 106 ... Query: 21 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQEQVR...QAFDNLNAILAAAGCSFEDVVDVTVF 80 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQEQVRQAFDNL...NAILAAAGCSFEDVVDVTVF Sbjct: 1 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQEQVRQAFDNLNAILAAAGCSFEDVVDVTVF 60 ...

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella bronchiseptica RB50] ... Length = 106 ... Query: 21 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQ...VRQAFDNLNAILAAAGCGXXXXXXXXXX 80 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQVRQAFD...NLNAILAAAGCG ... Sbjct: 1 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQVRQAFDNLNAILAAAGCGFDDVVDVTVF 60 ...

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella bronchiseptica RB50] ... Length = 106 ... Query: 21 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQ...VRQAFDNLNAILAAAGCGXXXXXXXXXX 80 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQVRQAFD...NLNAILAAAGCG ... Sbjct: 1 ... YSPAIRSNGFLFVSGQVGSQEDGSPKQGLQDQVRQAFDNLNAILAAAGCGFDDVVDVTVF 60 ...

21 CFR 520.2261b - Sulfamethazine powder.

Science.gov (United States)

2010-04-01

... and bovine respiratory disease complex (shipping fever complex) (Pasteurella spp.), colibacillosis... (F. necrophorum), acute mastitis (Streptococcus spp.), and acute metritis (Streptococcus spp.) (iii...

Microbial flora of odontogenic abscesses in pet guinea pigs.

Science.gov (United States)

Minarikova, A; Hauptman, K; Knotek, Z; Jekl, V

2016-10-01

Abscesses of odontogenic origin in guinea pigs pose a serious health problem and need to be treated with a combination of surgical and medical therapy. The aim of this prospective study was to describe the microbial flora of odontogenic abscesses associated with osteomyelitis in 24 pet guinea pigs, to perform antibiotic sensitivity testing, and to make recommendations for practitioners on the antibiotics of first choice. Inclusion criteria for the study included the animal being diagnosed with an odontogenic abscess which underwent surgery and was not pre-treated with an antibiotic. Inclusion criteria matched for 24 guinea pigs. Samples (pus, capsule and affected tooth/bone) for bacteriological examination were collected under sterile conditions during the surgical procedure. The most commonly isolated bacteria from abscesses of odontogenic origin were Bacteroides fragilis in 12.8 per cent (6/47) of cases, Pasteurella multocida in 10.6 per cent (5/47) and Peptostreptococcus anaerobius in 8.5 per cent (4/47). Aerobic bacterial species only were isolated in 29.2 per cent (7/24) of cases, anaerobic bacteria only were isolated in 33.3 per cent (8/24), and mixed infection with anaerobic and aerobic bacterial species was seen in 37.5 per cent (9/24). Aerobes (n=20) were sensitive to enrofloxacin and marbofloxacin in 100 per cent of samples, benzylpenicillin potassium (penicillin G, PNCG) in 90 per cent, cephalotin in 85 per cent, amoxicillin-clavulanate in 75 per cent, doxycycline in 70 per cent, gentamicin in 65 per cent and trimethoprim-sulfamethoxazole in 55 per cent. Anaerobes (n=27) were sensitive to amoxicillin-clavulanate in 100 per cent of cases, clindamycin in 96.3 per cent, metronidazole in 92.6 per cent, PNCG in 92.6 per cent and cephalotin in 74.1 per cent. As guinea pigs are strictly herbivorous animals, based on the results of this study the recommended antibiotic treatment for odontogenic abscesses is a combination of fluoroquinolones and metronidazole

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available licase ... [Bordetella pertussis Tohama I] ... Length = 447 ... Query: 17 ... LPILHRLMPLANTSASPARHPV...RALILTPTRELADQVYESVKRYSLHTPLRSAVVFGGVD 76 ... LPILHRLMPLANTSASPARHPVRALILT...PTRELADQVYESVKRYSLHTPLRSAVVFGGVD Sbjct: 56 ... LPILHRLMPLANTSASPARHPVRALILTPTRELADQVYESVKRYSLHTPLRSAVVFGGVD 115

Neonatal extracorporeal membrane oxygenation: Initial experience of Hospital de São João

Directory of Open Access Journals (Sweden)

G. Rocha

2014-11-01

Full Text Available The purpose of this series is to report the initial ECMO experience of the Neonatal Intensive Care Unit of Hospital de São João. The first three clinical cases are reported. Case report 1: a 39 weeks gestational age girl with severe lung hypoplasia secondary to a bilateral congenital diaphragmatic hernia. Case report 2: a 39 weeks gestational age girl with a right congenital diaphragmatic hernia and a tracheal stenosis. Case report 3: a 34 weeks gestational age boy, with 61 days of life, with a Bordetella pertussis pneumonia, severe pulmonary hypertension, shock, hyperleukocytosis and seizures. Resumo: O objetivo desta série é apresentar a experiência inicial da Unidade de Cuidados Intensivos Neonatais do Hospital de São João com ECMO no recém-nascido. São apresentados os 3 primeiros casos. Caso 1: recém-nascido de 39 semanas de idade gestacional, com hipoplasia pulmonar severa secundária a hérnia diafragmática congénita bilateral. Caso 2: recém-nascido de 39 semanas de idade gestacional, com hérnia diafragmática congénita direita e estenose traqueal. Caso 3: pré-termo de 34 semanas de idade gestacional, sexo masculino, com 61 dias de vida, com pneumonia por Bordetella pertussis, hipertensão pulmonar severa, choque, hiperleucocitose e convulsões. Keywords: Extracorporeal membrane oxygenation, Newborn, Congenital diaphragmatic hernia, Tracheal stenosis, Bordetella pertussis infection, Palavras-chave: Oxigenação por membrana extracorporal, Recém-nascido, Hérnia diafragmática congénita, Estenose traqueal, Infeção por Bordetella pertussis

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available a factor ... [Bordetella pertussis Tohama I] ... Length = 146 ... Query: 6 ... EALIVACIPSLRRYARGLTGERD...RADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 65 ... EALIVACIPSLRRYARGLTGERDRAD...DLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ Sbjct: 1 ... EALIVACIPSLRRYARGLTGERDRADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 6

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available a factor ... [Bordetella pertussis Tohama I] ... Length = 146 ... Query: 6 ... EALIVACIPSLRRYARGLTGERD...RADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 65 ... EALIVACIPSLRRYARGLTGERDRAD...DLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ Sbjct: 1 ... EALIVACIPSLRRYARGLTGERDRADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 6

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available a factor ... [Bordetella pertussis Tohama I] ... Length = 146 ... Query: 6 ... EALIVACIPSLRRYARGLTGERD...RADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 65 ... EALIVACIPSLRRYARGLTGERDRAD...DLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ Sbjct: 1 ... EALIVACIPSLRRYARGLTGERDRADDLVQDTLERAWSRFSRWQRRGELRAWMFGIMHNQ 6

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella bronchiseptica RB50] ... Length = 157 ... Query: 653 RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMR...ELTPRMVARYTQVDYHRELALVAATQVP 712 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPR...MVARYTQVDYHRELALVAATQVP Sbjct: 1 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPRMVARYTQVDYHRELALVAATQVP 60 ... Query: 7

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella bronchiseptica RB50] ... Length = 157 ... Query: 653 RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMR...ELTPRMVARYTQVDYHRELALVAATQVP 712 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPR...MVARYTQVDYHRELALVAATQVP Sbjct: 1 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPRMVARYTQVDYHRELALVAATQVP 60 ... Query: 7

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella bronchiseptica RB50] ... Length = 157 ... Query: 653 RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMR...ELTPRMVARYTQVDYHRELALVAATQVP 712 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPR...MVARYTQVDYHRELALVAATQVP Sbjct: 1 ... RPIRPEDGEPLQEFIRGLSERSRYMRFVSMMRELTPRMVARYTQVDYHRELALVAATQVP 60 ... Query: 7

Serological study of Bordetella Pertussis, Mycoplasma Pneumonia and Chlamydia Pneumonia in Iranian hajj pilgrims with prolonged cough illnesses: A follow-up study.

Science.gov (United States)

Rahimian, Masoud; HosseiniB, Mahdi

2017-11-01

Hajj pilgrimage is the biggest and longest mass gathering in the Muslim world. Annually, about 50% of more than 2.5 million pilgrims participating in this ritual get involved in severe devastating coughs. Most coughs continue, so the pilgrims turn back home and transmit them to family members and other people. Despite the high prevalence of coughs for many years, what causes them remains unknown. Considering the pertussis-like clinical picture of the so-called "hajj coughs", the researchers conducted a study to measure antibodies against three known common atypical bacteria, namely Bordetella Pertussis, Chlamydia Pneumonia and Mycoplasma Pneumonia. The study was done on three out of eleven groups of pilgrims from Yazd province, central Iran. The sample was selected randomly and consisted of 202 pilgrims who completed an informed consent. Their blood samples were taken, and the plasma was separated and then stored at -70 °C. After turning back from the journey, the pilgrims had their second blood samples taken. As many as 52 pilgrims failed to come for the second sampling, and two samples were broken during transportation. The final analysis was performed on the remaining 148 pairs of samples. Antibodies were already elevated in many pilgrims before the journey probably due to their old age (causing more exposure to pathogens) or unplanned pertussis vaccination. After their return, antibody elevation was only mild, again probably due to the old age of the participants (i.e. due to their weaker immune systems). Some antibodies even fell down without any known reason. In this study, previous hajj journey was assumed as a prophylactic factor, due to acquisition of immunity. Coughs with a prolonged pertussis-like picture were also presumed to be more related than other types of coughs to atypical pathogens. Statistical tests showed that the history of previous journeys had no prophylactic effect. Also, no correlation was found between the clinical pictures of coughs

Untitled

African Journals Online (AJOL)

cholera result in significant economic losses to susceptibility to ... standardized single disk method and minimal multocida .... using a panel of 16 reference antibodies against rhamnose .... horizontal transmission within the bacterial on our in ...

21 CFR 520.2170 - Sulfabromomethazine sodium boluses.

Science.gov (United States)

2010-04-01

... pneumonia and bovine respiratory disease complex (shipping fever complex) associated with Pasteurella spp.; acute metritis and acute mastitis caused by Streptococcus spp. (3) Limitations. Administer orally...

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ly ... RNA polymerase sigma factor [Bordetella pertussis Tohama ... I] ... Length = 185 ... Query: 31 ... SAFETI...MRRHNRLLFRTARSILHSDAEAEDALQEAYLRAWRALDSFRADARLSTWLVRIV 90 ... SAFETI...MRRHNRLLFRTARSILHSDAEAEDALQEAYLRAWRALDSFRADARLSTWLVRIV Sbjct: 18 ... SAFETIMRRHNRLLFRTARSILHSDAEAE

Calcium-dependent disorder-to-order transitions are central to the secretion and folding of the CyaA toxin of Bordetella pertussis, the causative agent of whooping cough.

Science.gov (United States)

O'Brien, Darragh P; Perez, Ana Cristina Sotomayor; Karst, Johanna; Cannella, Sara E; Enguéné, Véronique Yvette Ntsogo; Hessel, Audrey; Raoux-Barbot, Dorothée; Voegele, Alexis; Subrini, Orso; Davi, Marilyne; Guijarro, J Inaki; Raynal, Bertrand; Baron, Bruno; England, Patrick; Hernandez, Belen; Ghomi, Mahmoud; Hourdel, Véronique; Malosse, Christian; Chamot-Rooke, Julia; Vachette, Patrice; Durand, Dominique; Brier, Sébastien; Ladant, Daniel; Chenal, Alexandre

2018-01-12

The adenylate cyclase toxin (CyaA) plays an essential role in the early stages of respiratory tract colonization by Bordetella pertussis, the causative agent of whooping cough. Once secreted, CyaA invades eukaryotic cells, leading to cell death. The cell intoxication process involves a unique mechanism of translocation of the CyaA catalytic domain directly across the plasma membrane of the target cell. Herein, we review our recent results describing how calcium is involved in several steps of this intoxication process. In conditions mimicking the low calcium environment of the crowded bacterial cytosol, we show that the C-terminal, calcium-binding Repeat-in-ToXin (RTX) domain of CyaA, RD, is an extended, intrinsically disordered polypeptide chain with a significant level of local, secondary structure elements, appropriately sized for transport through the narrow channel of the secretion system. Upon secretion, the high calcium concentration in the extracellular milieu induces the refolding of RD, which likely acts as a scaffold to favor the refolding of the upstream domains of the full-length protein. Due to the presence of hydrophobic regions, CyaA is prone to aggregate into multimeric forms in vitro, in the absence of a chaotropic agent. We have recently defined the experimental conditions required for CyaA folding, comprising both calcium binding and molecular confinement. These parameters are critical for CyaA folding into a stable, monomeric and functional form. The monomeric, calcium-loaded (holo) toxin exhibits efficient liposome permeabilization and hemolytic activities in vitro, even in a fully calcium-free environment. By contrast, the toxin requires sub-millimolar calcium concentrations in solution to translocate its catalytic domain across the plasma membrane, indicating that free calcium in solution is actively involved in the CyaA toxin translocation process. Overall, this data demonstrates the remarkable adaptation of bacterial RTX toxins to the

A Comparative Study of the Oral Microbiome Compositions of ...

African Journals Online (AJOL)

2017-12-05

Dec 5, 2017 ... external factors such as oral hygiene practices,[6] types. Original Article ..... exclusive bacterial species; Porphyromonas gingivalis,. Pasteurella pneumotropica, Tannerella .... oral health and disease. Virulence 2014;5:424-32.

Usage of antimicrobials and occurrence of antimicrobial resistance among bacteria from mink

DEFF Research Database (Denmark)

Pedersen, Karl; Hammer, Anne Sofie; Sørensen, Charlotte Mark

2009-01-01

, whereas resistance to other antimicrobials was rare. All P aeruginosa were sensitive to gentamicin and colistin and sensitive or intermediate to enrofloxacin. whereas most isolates were resistant to all other antimicrobials. All P. multocida and haemolytic streptococci were sensitive to penicillin...

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella pertussis Tohama I] ... Length = 153 ... Query: 41 ... LVAKDGVVLARGVNRMLADHDPTAHXXXXXXXXXX...XXXXXXXXDGCVVYASGQPCPMCLA 100 ... LVAKDGVVLARGVNRMLADHDPTAH ... ... ... DGCVVYASGQPCPMCLA Sbjct: 27 ... LVAKDGVVLARGVNRMLADHDPTAHTELLALREAGRALRSARLDGCVVYASGQPCPMCLA 86 ... Query: 161 EWQDGPQ 167 ... EWQDGPQ Sbjct: 147 EWQDGPQ 153

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella pertussis Tohama I] ... Length = 153 ... Query: 41 ... LVAKDGVVLARGVNRMLADHDPTAHTEXXXXXXXX...XXXXXXXXDGCVVYASGQPCPMCLA 100 ... LVAKDGVVLARGVNRMLADHDPTAHTE ... ... ... DGCVVYASGQPCPMCLA Sbjct: 27 ... LVAKDGVVLARGVNRMLADHDPTAHTELLALREAGRALRSARLDGCVVYASGQPCPMCLA 86 ... Query: 161 EWQDGPQ 167 ... EWQDGPQ Sbjct: 147 EWQDGPQ 153

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available ... [Bordetella pertussis Tohama I] ... Length = 153 ... Query: 41 ... LVAKDGVVLARGVNRMLADHDPTAHXXXXXXXXXX...XXXXXXXXDGCVVYASGQPCPMCLA 100 ... LVAKDGVVLARGVNRMLADHDPTAH ... ... ... DGCVVYASGQPCPMCLA Sbjct: 27 ... LVAKDGVVLARGVNRMLADHDPTAHTELLALREAGRALRSARLDGCVVYASGQPCPMCLA 86 ... Query: 161 EWQDGPQ 167 ... EWQDGPQ Sbjct: 147 EWQDGPQ 153

The diagnostic activity on wild animals through the description of a ...

African Journals Online (AJOL)

The diagnostic activity on wild animals through the description of a model case report (caseous lymphadenitis by Corynebacterium pseudotuberculosis associated with Pasteurella spp and parasites infection in an alpine ibex – Capra ibex )

Disease: H01066 [KEGG MEDICUS

Lifescience Database Archive (English)

Full Text Available Bordetella, B. petrii is an environmental species but sometimes can infect immunocompetent humans. Infectiou... petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pat

Pertussis Tests

Science.gov (United States)

... of Conditions Not Listed? Not Listed? Acidosis and Alkalosis Adrenal Insufficiency and Addison Disease Alcoholism Allergies Alzheimer ... tested? Pertussis, commonly called whooping cough, is a respiratory infection caused by the bacteria Bordetella pertussis . These ...

21 CFR 520.90b - Ampicillin trihydrate tablets.

Science.gov (United States)

2010-04-01

..., tonsillitis, and bronchitis due to Streptococcus spp., Staphylococcus spp., Escherichia coli, Proteus mirabilis, and Pasteurella spp., urinary tract infections (cystitis) due to Streptococcus spp., Staphylococcus spp., E., coli, P. mirabilis, and Enterococcus spp.; gastrointestinal infections due to...

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002928 gi|33596933 >1kmoA 7 661 65 702 8e-60 ... ref|NP_884576.1| putative ferrisi...derophore receptor [Bordetella parapertussis 12822] ... emb|CAE37631.1| putative ferrisiderophore rec

ORF Alignment: NC_002929 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002929 gi|33592999 >1kmoA 7 661 65 702 1e-60 ... ref|NP_880643.1| putative ferrisi...derophore receptor [Bordetella pertussis Tohama I] ... emb|CAE42244.1| putative ferrisiderophore rece

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002927 gi|33600770 >1kmoA 7 661 65 702 2e-60 ... ref|NP_888330.1| putative ferrisi...derophore receptor [Bordetella bronchiseptica RB50] ... emb|CAE32282.1| putative ferrisiderophore rec

A rapid minor groove binder PCR method for distinguishing the vaccine strain Brucella abortus 104M.

Science.gov (United States)

Nan, Wenlong; Qin, Lide; Wang, Yong; Zhang, Yueyong; Tan, Pengfei; Chen, Yuqi; Mao, Kairong; Chen, Yiping

2018-01-24

Brucellosis is a widespread zoonotic disease caused by Gram-negative Brucella bacteria. Immunisation with attenuated vaccine is an effective method of prevention, but it can interfere with diagnosis. Live, attenuated Brucella abortus strain 104M has been used for the prevention of human brucellosis in China since 1965. However, at present, no fast and reliable method exists that can distinguish this strain from field strains. Single nucleotide polymorphism (SNP)-based assays offer a new approach for such discrimination. SNP-based minor groove binder (MGB) and Cycleave assays have been used for rapid identification of four Brucella vaccine strains (B. abortus strains S19, A19 and RB51, and B. melitensis Rev1). The main objective of this study was to develop a PCR assay for rapid and specific detection of strain 104M. We developed a SNP-based MGB PCR assay that could successfully distinguish strain 104M from 18 representative strains of Brucella (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae, and B. ovis), four Brucella vaccine strains (A19, S19, S2, M5), and 55 Brucella clinical field strains. The assay gave a negative reaction with four non-Brucella species (Escherichia coli, Pasteurella multocida, Streptococcus suis and Pseudomonas aeruginosa). The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 220 fg for the 104M strain and 76 fg for the single non-104M Brucella strain tested (B. abortus A19). The assay was also reproducible (intra- and inter-assay coefficients of variation = 0.006-0.022 and 0.012-0.044, respectively). A SNP-based MGB PCR assay was developed that could straightforwardly and unambiguously distinguish B. abortus vaccine strain 104M from non-104M Brucella strains. Compared to the classical isolation and identification approaches of bacteriology, this real-time PCR assay has substantial advantages in terms of

Antioxidant, antibacterial activity, and phytochemical characterization of Melaleuca cajuputi extract.

Science.gov (United States)

Al-Abd, Nazeh M; Mohamed Nor, Zurainee; Mansor, Marzida; Azhar, Fadzly; Hasan, M S; Kassim, Mustafa

2015-10-24

The threat posed by drug-resistant pathogens has resulted in the increasing momentum in research and development for effective alternative medications. The antioxidant and antibacterial properties of phytochemical extracts makes them attractive alternative complementary medicines. Therefore, this study evaluated the phytochemical constituents of Melaleuca cajuputi flower and leaf (GF and GL, respectively) extracts and their antioxidant and antibacterial activities. Radical scavenging capacity of the extracts was estimated using 2,2-diphenyl-2-picrylhydrazyl and Fe(2+)-chelating activity. Total antioxidant activity was determined using ferric reducing antioxidant power assay. Well diffusion, minimum inhibitory concentration, and minimum bactericidal concentration assays were used to determine antibacterial activity against eight pathogens, namely Staphylococcus aureus, Escherichia coli, Bacillus cereus, Staphylococcus epidermidis, Salmonella typhimurium, Klebsiella pneumonia, Streptococcus pneumoniae, and Pasteurella multocida. We identified and quantified the phytochemical constituents in methanol extracts using liquid chromatography/mass spectrometry (LC/MS) and gas chromatography (GC)/MS. This study reports the antioxidant and radical scavenging activity of M. cajuputi methanolic extracts. The GF extract showed better efficacy than that of the GL extract. The total phenolic contents were higher in the flower extract than they were in the leaf extract (0.55 ± 0.05 and 0.37 ± 0.05 gallic acid equivalent per mg extract dry weight, respectively). As expected, the percentage radical inhibition by GF was higher than that by the GL extract (81 and 75 %, respectively). A similar trend was observed in Fe(2+)-chelating activity and β-carotene bleaching tests. The antibacterial assay of the extracts revealed no inhibition zones with the Gram-negative bacteria tested. However, the extracts demonstrated activity against B. cereus, S. aureus, and S. epidermidis. In

Pharmacokinetics (PK), pharmacodynamics (PD), and PK-PD integration of ceftiofur after a single intravenous, subcutaneous and subcutaneous-LA administration in lactating goats.

Science.gov (United States)

Fernández-Varón, Emilio; Cárceles-García, Carlos; Serrano-Rodríguez, Juan Manuel; Cárceles-Rodríguez, Carlos M

2016-10-13

Bacterial pneumonia in goats is usually caused by Mannheimia haemolytica and Pasteurella multocida. Another important infection disease in lactating goats is intramammary infection producing mastitis, usually associated with coagulase-negative Staphylococcus spp. However, treatment of bacterial pneumonia in goats not affected by mastitis problems should be restricted to antimicrobials with scant penetration to milk in order to avoid long withdrawal times. Ceftiofur is a third-generation cephalosporin antimicrobial with activity against various gram-positive and gram-negative, aerobic and anaerobic bacteria encountered by domestic animals. The objectives of the present study were to establish the serum concentration-time profile for ceftiofur in lactating goats after intravenous, subcutaneous and a SC-long-acting ceftiofur formulation; to determine ceftiofur penetration into milk; to determine in vitro and ex vivo activity of ceftiofur establishing MIC, MBC, MPC and time-kill profiles against field strains of M. haemolytica and finally to calculate the main surrogate markers of efficacy. The pharmacokinetics studies revealed an optimal PK properties for the SC-LA formulation tested. Ceftiofur was well absorbed following SC and SC-LA administration, with absolute bioavailabilities (F) of 85.16 and 84.43 %, respectively. After ceftiofur analysis from milk samples, no concentrations were found at any sampling time. The MIC, MBC and MPC data of ceftiofur against five M. haemolytica strains isolated from goats affected by pneumonia were tested showing excelent sensitivity of ceftiofur against this pathogen. For PK-PD analysis, ratios were calculated suggesting a high level of bacterial kill against the five strains of M. haemolytica tested. The systemic ceftiofur exposure achieved in lactating goats following IV, SC and especially with the SC-LA administration is consistent with the predicted PK-PD ratios needed for a positive therapeutic outcome for M. haemolytica

Nigerian Veterinary Journal - Vol 35, No 2 (2014)

African Journals Online (AJOL)

A Case Report of Respiratory Mannheimiosis in Sheep and Goat Complicated by Bordetella parapertussis · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. PS Ekong, BO Akanbi, MO Odugbo ...

ORF Alignment: NC_002928 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002928 gi|33596750 >1v7zA 3 253 1 251 8e-40 ... ref|NP_884393.1| putative creatini...ne amidohydrolase [Bordetella parapertussis 12822] ... emb|CAE37436.1| putative creatinine amidohydro

Characterisation of a novel Mannheimia sp from Australian feedlot cattle

DEFF Research Database (Denmark)

Blackall, P.J.; Angen, Øystein; Fegan, N.

2001-01-01

glucosida, M granulomatis, M ruminalis and M varigena - using a range of phenotypic and genotypic methods. Results Phenotypic characterisation indicated that the isolates belonged to the trehalose-negative [Pasteurella] haemolytica complex. This complex has recently been reorganised into five species within...

ORF Alignment: NC_002927 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available NC_002927 gi|33600518 >1v7zA 3 253 1 251 1e-39 ... ref|NP_888078.1| putative creatini...ne amidohydrolase [Bordetella bronchiseptica RB50] ... emb|CAE32030.1| putative creatinine amidohydro

Myelin/oligodendrocyte glycoprotein-induced autoimmune encephalomyelitis in common marmosets : the encephalitogenic T cell epitope pMOG24-36 is presented by a monomorphic MHC class II molecule

NARCIS (Netherlands)

Brok, H.P.M.; Uccelli, A.; Kerlero De Rosbo, N.; Bontrop, R.E.; Roccatagliata, L.; Groot, de N.G.; Capello, E.; Laman, J.D.; Nicolay, K.; Mancardi, G.L.; Ben-Nun, A.; Hart, 't L.A.

2000-01-01

Immunization of common marmosets (Callithrix jacchus) with a single dose of human myelin in CFA, without administration of Bordetella pertussis, induces a form of autoimmune encephalomyelitis (EAE) resembling in its clinical and pathological expression multiple sclerosis in humans. The EAE incidence

An evidence synthesis approach to estimating the incidence of symptomatic pertussis infection in the Netherlands, 2005-2011

NARCIS (Netherlands)

McDonald, Scott A; Teunis, Peter; van der Maas, Nicoline; de Greeff, Sabine; de Melker, Hester; Kretzschmar, MEE

2015-01-01

BACKGROUND: Despite high vaccination coverage, infection with Bordetella pertussis is a current public health concern in the Netherlands and other European Union member states. Because surveillance data are subject to extensive under-ascertainment and under-reporting, incidence is difficult to

Läkaköha - aktuaalne uurimisteema / Marje Oona

Index Scriptorium Estoniae

Oona, Marje, 1963-

2012-01-01

TÜ peremeditsiini õppetooli töötajate poolt algatatud uurimistööst, mille eesmärgiks on uurida Bordetella spp. infektsioonide epidemioloogiat, molekulaargeneetikat ja kliinilisi eripärasid ning selgitada läkaköha sagedasema diagnsimise põhjusi Eestis

Cellular and humoral immunity after infection with B. pertussis : the role of age, antigen and vaccination history

NARCIS (Netherlands)

van Twillert, I

2017-01-01

Pertussis (whooping cough), is a bacterial disease of the respiratory tract, caused by the human pathogen Bordetella pertussis. Vaccination against pertussis has dramatically lowered pertussis incidence and mortality rates; however pertussis still occurs. The duration of immunity to B. pertussis

TMFunction data: 2097 [TMFunction[Archive

Lifescience Database Archive (English)

Full Text Available H, Clantin B, Locht C, Molle G, Jacob-Dubuisson F, Saint N J Biol Chem. 2006 Jan 6;281(1):158-66 Electropho... d(221-228) ... transport ... TpsB Transporter FhaC Bordetella pertussis ... M?li AC, Hodak

TMFunction data: 2114 [TMFunction[Archive

Lifescience Database Archive (English)

Full Text Available H, Clantin B, Locht C, Molle G, Jacob-Dubuisson F, Saint N J Biol Chem. 2006 Jan 6;281(1):158-66 Electropho... d(221-228) ... transport ... TpsB Transporter FhaC Bordetella pertussis ... M?li AC, Hodak

鸭多杀性巴氏杆菌外膜蛋白A基因表达及抗原性鉴定%Cloning and Expression of the omp A Gene of pasteurella multocida

Institute of Scientific and Technical Information of China (English)

孙龚; 郭东春; 刘家森; 姜骞; 司昌德; 林欢; 韩凌霞; 崔玉东; 曲连东

2010-01-01

根据已发表的Pm 70株的序列(GenBank登录号:AE004439)设计了两对引物,用PCR方法扩增了鸭多杀性巴氏杆菌标准株C48-102的外膜蛋白基因A(omp A),扩增的片段为1 062 bp.将测序结果与GenBank中多杀性巴氏杆菌P52、PM 70、T931317、194289的omp A序列比对结果表明,核苷酸水平上同源性为89.0%～98.9%;在氨基酸水平上同源性为90.7%～99.2%.全omp A序列在大肠杆菌中干扰蛋白的正常表达,因此截断omp A蛋白的信号肽序列,将去信号肽的omp A基因插入到pPRO-EX-Htb载体上,构建了原核表达载体Htb-omp A,转化BL21,并诱导表达,SDS-PAGE结果表明,表达蛋白约为35 kDa,与预期的分子量大小相符.Western-blot结果表明,表达的蛋白具有良好的抗原活性.本研究重组蛋白omp A的获得,为敲除omp A基因多杀性巴氏杆菌突变株的血清学检测奠定基础.

Alexandria Journal of Medicine - Vol 52, No 2 (2016)

African Journals Online (AJOL)

Purification of heat labile toxin from Bordetella pertussis vaccine strain 134 employed indigenous technology · EMAIL FREE FULL TEXT EMAIL FREE FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT. KC Shivanandappa, S Jagannathan, S Pandiyarajan, P Tamilvanan, T Umadevi, Jeeva Kalaiselvan, ...

Bacillus anthracis Edema Toxin Inhibits Staphylococcus aureus Enterotoxin B Effects in Vitro: A Potential Protein Therapeutic?

Science.gov (United States)

2005-10-01

5). Inherent characteristics of edema toxin and other procaryotic adenylate cyclases from Bordetella pertussis, Pseudomonas aeruginosa, and Yersinia...by mouse peritoneal macrophages: the role of cellular cyclic AMP. Immunology 64:719–724. 12. Krakauer, T. 1999. Induction of CC chemokines in human

TMFunction data: 2122 [TMFunction[Archive

Lifescience Database Archive (English)

Full Text Available d(3-206) ... transport ... TpsB Transporter FhaC Bordetella pertussis ... M?li AC, Hodak H, Clantin B, Locht C, Mol...le G, Jacob-Dubuisson F, Saint N J Biol Chem. 2006 Jan 6;281(1):158-66 Electrophore