-
Journal of Virology Oct 1997Cleavage of influenza A virus hemagglutinin (HA) is required for expression of fusion activity and virus entry into cells. Extracellular proteases are responsible for...
Cleavage of influenza A virus hemagglutinin (HA) is required for expression of fusion activity and virus entry into cells. Extracellular proteases are responsible for the proteolytic cleavage activation of avirulent avian and mammalian influenza viruses and contribute to pathogenicity and tissue tropism. The relative contributions of host and microbial proteases to cleavage activation in natural infection remain to be established. We examined 23 respiratory bacterial pathogens and 150 aerobic bacterial isolates cultured from the nasal cavities of pigs for proteolytic activity. No evidence of secreted proteases was found for the bacterial pathogens, including Haemophilus parasuis, Pasteurella multocida, Actinobacillus pleuropneumoniae, Bordetella bronchiseptica, and Streptococcus suis. Proteolytic bacteria were isolated from 7 of 11 swine nasal samples and included Staphylococcus chromogenes, Staphylococcus hyicus, Aeromonas caviae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Enterococcus sp. Only P. aeruginosa secreted a protease, elastase, that cleaved influenza virus HA. However, compared to trypsin, the site of cleavage by elastase was shifted one amino acid in the carboxy-terminal direction and resulted in inactivation of the virus. Under the conditions of this study, we identified several bacterial isolates from the respiratory tracts of pigs that secrete proteases in vitro. However, none of these proteolytic isolates demonstrated direct cleavage activation of influenza virus HA.
Topics: Animals; Bacteria; Bacterial Infections; Endopeptidases; Hemagglutinin Glycoproteins, Influenza Virus; Influenza A virus; Nasal Mucosa; Pancreatic Elastase; Pseudomonas aeruginosa; Respiratory Tract Infections; Swine; Swine Diseases
PubMed: 9311838
DOI: 10.1128/JVI.71.10.7579-7585.1997 -
Infection and Immunity Apr 2009Host-pathogen interactions are of great importance in understanding the pathogenesis of infectious microorganisms. We developed in vitro models to study the...
Host-pathogen interactions are of great importance in understanding the pathogenesis of infectious microorganisms. We developed in vitro models to study the host-pathogen interactions of porcine respiratory tract pathogens using two immortalized epithelial cell lines, namely, the newborn pig trachea (NPTr) and St. Jude porcine lung (SJPL) cell lines. We first studied the interactions of Actinobacillus pleuropneumoniae, an important swine pathogen, using these models. Under conditions where cytotoxicity was absent or low, we showed that A. pleuropneumoniae adheres to both cell lines, stimulating the induction of NF-kappaB. The NPTr cells consequently secrete interleukin 8, while the SJPL cells do not, since they are deprived of the NF-kappaB p65 subunit. Cell death ultimately occurs by necrosis, not apoptosis. The transcriptomic profile of A. pleuropneumoniae was determined after contact with the porcine lung epithelial cells by using DNA microarrays. Genes such as tadB and rcpA, members of a putative adhesin locus, and a gene whose product has high homology to the Hsf autotransporter adhesin of Haemophilus influenzae were upregulated, as were the genes pgaBC, involved in biofilm biosynthesis, while capsular polysaccharide-associated genes were downregulated. The in vitro models also proved to be efficient with other swine pathogens, such as Actinobacillus suis, Haemophilus parasuis, and Pasteurella multocida. Our results demonstrate that interactions of A. pleuropneumoniae with host epithelial cells seem to involve complex cross talk which results in regulation of various bacterial genes, including some coding for putative adhesins. Furthermore, our data demonstrate the potential of these in vitro models in studying the host-pathogen interactions of other porcine respiratory tract pathogens.
Topics: Actinobacillus pleuropneumoniae; Animals; Apoptosis; Bacterial Adhesion; Bacterial Proteins; Cell Line; Cells, Cultured; Epithelial Cells; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Host-Pathogen Interactions; Lung; Molecular Sequence Data; Oligonucleotide Array Sequence Analysis; Swine; Trachea
PubMed: 19139196
DOI: 10.1128/IAI.00297-08 -
Journal of Clinical Microbiology Sep 1995The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various...
The MICs of ceftiofur and other antimicrobial agents, tested for comparison, for 515 bacterial isolates of pigs from the United States, Canada, and Denmark with various diseases were compared. The organisms tested included Actinobacillus pleuropneumoniae, Escherichia coli, Pasteurella multocida, Salmonella choleraesuis, Salmonella typhimurium, Streptococcus suis, Streptococcus dysgalactiae subsp. equisimilis, Streptococcus equi subsp. equi, and Streptococcus equi subsp. zooepidemicus. In addition to ceftiofur, the following antimicrobial agents or combinations were tested: enrofloxacin, ampicillin, sulfamethazine, trimethoprim-sulfadiazine (1:19), erythromycin, lincomycin, spectinomycin, lincomycin-spectinomycin (1:8), tilmicosin, and tetracycline. Tilmicosin was only tested against the U.S. isolates. Overall, ceftiofur and enrofloxacin were the most active antimicrobial agents tested against all isolates, with MICs inhibiting 90% of isolates tested (MIC90s) of < or = 2.0 and < or = 1.0 microgram/ml, respectively. Erythromycin, sulfamethazine, spectinomycin, and lincomycin demonstrated limited activity against all of the organisms tested, with MIC90s of > or = 8.0, > or = 256.0, > or = 32.0, and > or = 16.0 micrograms/ml, respectively. Trimethoprim-sulfadiazine was active against isolates of A. pleuropneumoniae, S. choleraesuis, S. typhimurium, P. multocida, S. equi, and S. suis (MIC90s, < or = 0.5 microgram/ml) but was less active against the E. coli strains tested (MIC90, > 16.0 micrograms/ml). Ampicillin was active against the P. multocida, S. suis, and S. equi isolates tested (MIC90s, 0.5, 0.06, and 0.06 micrograms/ml, respectively) and was moderately active against S. typhimurium (MIC90s, 2.0 micrograms/ml). However, this antimicrobial agent was much less active when it was tested against A. pleuropneumoniae, S. cholerae-suis, and E. coli (MIC90s, 16.0, > 32.0, and 32.0 micrograms/ml, respectively). Against the U.S. isolates of A. pleuropneumoniae and P. multocida, tilmicosin was moderately active (MIC90s, 4.0 and 8.0 micrograms/ml, respectively). However, this compound was not active against the remaining U.S. isolates (MIC90s, > 64.0 micrograms/ml). Differences in the MICs from one country to another were not detected with enrofloxacin, ceftiofur, or lincomycin for the strains tested, but variations in the MICs of the remaining antimicrobial agents were observed.
Topics: Animals; Bacteria; Canada; Cephalosporins; Denmark; Microbial Sensitivity Tests; Swine; United States
PubMed: 7494042
DOI: 10.1128/jcm.33.9.2435-2444.1995 -
The Canadian Veterinary Journal = La... Oct 1976
Topics: Actinobacillus Infections; Alberta; Animals; Swine; Swine Diseases
PubMed: 974978
DOI: No ID Found -
The Canadian Veterinary Journal = La... Jun 1990Actinobacillus suis was isolated from tissues of 39 pigs, 2 porcine lungs, and 1 uterine swab submitted for diagnostic evaluation from 24 farms in southwestern Ontario...
Actinobacillus suis was isolated from tissues of 39 pigs, 2 porcine lungs, and 1 uterine swab submitted for diagnostic evaluation from 24 farms in southwestern Ontario between 1985 and 1988. These isolates represented a gradually increasing incidence of herd outbreaks caused by A. suis in southwestern Ontario. The outbreaks were typified by sudden death in suckling or recently weaned pigs; 87% of the affected pigs examined at the laboratory were between two and 28 days old. Petechial to ecchymotic hemorrhages in the thoracic and abdominal organs accompanied by serofibrinous exudates in both cavities were the most common gross lesions. The lesions were characterized histologically by bacterial thromboembolism and necrosis randomly scattered in thoracic and abdominal organs. Occasionally, bacterial thromboemboli were surrounded by centrifugally radiating, eosinophilic, club-like colonies. Diffuse necrohemorrhagic myocarditis that was more severe in the atria, and diffuse subacute meningoencephalitis, were less frequent but distinctive lesions. Multiple litters were affected in most herd outbreaks, and mortality often approached 50% in affected litters. Although the A. suis organism was susceptible to nearly every antibiotic against which it was tested, the suddenness of herd outbreaks precluded attempts at treatment.
PubMed: 17423607
DOI: No ID Found -
Canadian Journal of Veterinary Research... Oct 1990Capsular polysaccharide antigens of serotypes 1, 2, 5 and 7 of Actinobacillus pleuropneumoniae were used in enzyme-linked immunosorbent assays (ELISAs) to test sera from...
Capsular polysaccharide antigens of serotypes 1, 2, 5 and 7 of Actinobacillus pleuropneumoniae were used in enzyme-linked immunosorbent assays (ELISAs) to test sera from experimentally infected and field pigs. Specific reactions were found in sera of experimental pigs with antigens of serotypes 1, 5 and 7 whereas the serotype 2 antigen was cross-reactive. A 1:200 serum dilution was used for testing of 300 sera from 21 swine herds in southern Ontario. Cases of pleuropneumonia had occurred in 11 of these herds, but not in the others. The negative cut-off value was the mean optical density at 405 nm (OD405) + three standard deviations (SD) for 16 negative reference sera. Sera from four pigs naturally infected with Actinobacillus suis were tested and found to react to varying degrees with each of the antigens. Therefore a second cut-off value was determined as the mean OD405 + 2 SD for the A. suis sera. Sera which, in the ELISA produced OD readings above the latter cut-off were considered positive for antibodies to A. pleuropneumoniae; those which were lower than the former cut-off were considered negative. Readings between the two cut-off values may have been due to low positive titers or cross-reactivity, possibly with A. suis, and could not be used to predict pleuropneumonia. Of the pleuropneumonia-free herds, none had positive reactors to serotypes 5 or 7, whereas one and two herds had positive reactors to serotypes 1 and 2, respectively. Of the pleuropneumonia positive herds, six had positive reactors to serotype 1, one to serotype 2, four to serotype 5, and eight to serotype 7.
Topics: Actinobacillus; Actinobacillus Infections; Animals; Antibodies, Bacterial; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Pleuropneumonia; Polysaccharides, Bacterial; Serotyping; Swine; Swine Diseases
PubMed: 2249177
DOI: No ID Found -
Cell Death & Disease Sep 2019Actinobacillus pleuropneumoniae (A.pp) causes severe pneumonia associated with enormous economic loss in pigs. Peracute diseased pigs die in <24 h with pneumonia....
Actinobacillus pleuropneumoniae (A.pp) causes severe pneumonia associated with enormous economic loss in pigs. Peracute diseased pigs die in <24 h with pneumonia. Neutrophils are the prominent innate immune cell in this infection that massively infiltrate the infected lung. Here we show that neutrophils release neutrophil extracellular traps (NETs) as response to A.pp infection. Numerous NET-markers were identified in bronchoalveolar lavage fluid (BALF) of A.pp-infected piglets in vivo, however, most NET fibers are degraded. Importantly, A.pp is able to enhance its growth rate in the presence of NETs that have been degraded by nucleases efficiently. A.pp itself releases no nuclease, but we identified host nucleases as sources that degrade NETs after A.pp infection. Furthermore, the nucleases of co-infecting pathogens like Streptococcus suis increase growth of A.pp in presence of porcine NETs. Thus, A.pp is not only evading the antimicrobial activity of NETs, A.pp is rather additionally using parts of NETs as growth factor thereby taking advantage of host nucleases as DNase1 or nucleases of co-infecting bacteria, which degrade NETs. This effect can be diminished by inhibiting the bacterial adenosine synthase indicating that degraded NETs serve as a source for NAD, which is required by A.pp for its growth. A similar phenotype was found for the human pathogen Haemophilus (H.) influenzae and its growth in the presence of human neutrophils. H. influenzae benefits from host nucleases in the presence of neutrophils. These data shed light on the detrimental effects of NETs during host immune response against certain bacterial species that require and/or efficiently take advantage of degraded DNA material, which has been provided by host nuclease or nucleases of other co-infecting bacteria, as growth source.
Topics: Actinobacillus Infections; Actinobacillus pleuropneumoniae; Animals; Extracellular Traps; Lung; Pneumonia, Bacterial; Swine; Swine Diseases
PubMed: 31506432
DOI: 10.1038/s41419-019-1895-4 -
The Journal of Veterinary Medical... May 2015The genetic organization of the gene involved in the capsular polysaccharide (CPS) biosynthesis of Actinobacillus pleuropneumoniae serotype 14 has been determined. The...
The genetic organization of the gene involved in the capsular polysaccharide (CPS) biosynthesis of Actinobacillus pleuropneumoniae serotype 14 has been determined. The DNA region for the CPS biosynthesis of serotype 14 (cps14) comprised 9 open reading frames, designated as cps14AB1B2B3CDEFG genes, encoding Cps14A to Cps14G protein, respectively. Cps14A was similar to CpsA of A. pleuropneumoniae serotypes 1, 4 and 12; the Cps14B1 and Cps14B2 were similar to CpsB of A. pleuropneumoniae serotypes 1, 4 and 12, suggesting that CPS structure of A. pleuropneumoniae serotype 14 would belong to Group I including A. pleuropneumoniae serotypes 1, 4, 12 and 15. Surprisingly, the overall nucleotide sequence, deduced amino acid sequence, and the genetic organization of the cps14 were nearly identical to those of Actinobacillus suis. This study will provide the molecular basic knowledge for development of diagnostics and vaccine of A. pleuropneumoniae serotype 14.
Topics: Actinobacillus pleuropneumoniae; Bacterial Capsules; DNA, Bacterial; Gene Expression Regulation, Bacterial; Polysaccharides, Bacterial
PubMed: 25648373
DOI: 10.1292/jvms.14-0174 -
Canadian Journal of Veterinary Research... Jan 1990The minimum inhibitory concentrations (MIC) of ciprofloxacin, enrofloxacin, and norfloxacin were tested for approximately ten clinical isolates of each of Actinobacillus...
The minimum inhibitory concentrations (MIC) of ciprofloxacin, enrofloxacin, and norfloxacin were tested for approximately ten clinical isolates of each of Actinobacillus pleuropneumoniae, Actinobacillus suis, Actinomyces pyogenes, Corynebacterium pseudotuberculosis, Erysipelothrix rhusiopathiae, Haemophilus parasuis, Haemophilus somnus, Pasteurella haemolytica, Pasteurella multocida, Rhodococcus equi, Streptococcus equi, Streptococcus suis and Streptococcus zooepidemicus. Ciprofloxacin and enrofloxacin had similar activity and were more active than norfloxacin. All isolates had an MIC of 1.0 microgram/mL or less for ciprofloxacin and enrofloxacin, and these drugs had particularly marked activity against the gram-negative bacteria tested.
Topics: Actinobacillus; Actinomyces; Anti-Infective Agents; Ciprofloxacin; Corynebacterium; Enrofloxacin; Erysipelothrix; Fluoroquinolones; Haemophilus; In Vitro Techniques; Microbial Sensitivity Tests; Norfloxacin; Pasteurella; Quinolones; Rhodococcus; Streptococcus
PubMed: 2306672
DOI: No ID Found -
Animals : An Open Access Journal From... Aug 2020Data on the scope of bacterial pathogens present and the frequency of antimicrobial resistance (AMR) in New Zealand's pigs are limited. This study describes bacterial...
Data on the scope of bacterial pathogens present and the frequency of antimicrobial resistance (AMR) in New Zealand's pigs are limited. This study describes bacterial isolates, antimicrobial susceptibility data, and multidrug resistance (MDR; resistance to ≥3 antimicrobial classes) from New Zealand pig submissions. Porcine test data from June 2003 to February 2016 were obtained from commercial veterinary pathology laboratory records. In total, 470/477 unique submissions resulted in bacterial growth, yielding 779 isolates. Sample type was recorded for 360/477 (75.5%); lung (79/360; 21.9%), faecal (61/360; 16.9%) and intestinal (45/360; 12.5%) were most common. The most common isolates were (186/779, 23.9%), (43/779; 5.5%), (43/779; 5.5%), unidentified spp. (38/779; 4.9%), alpha haemolytic (32/779; 4.1%), coagulase negative S spp. (26/779; 3.3%), and (25/779; 3.2%). Susceptibility results were available for 141/779 (18.1%) isolates from 62/470 (13.2%) submissions. Most were susceptible to trimethoprim-sulphonamide (75/81; 92.6%), but fewer were susceptible to penicillin (37/77; 48.1%), tilmicosin (18/43; 41.9%), or tetracyclines (41/114; 36.0%). No susceptibility data were available for spp., spp., or spp. isolates. MDR was present in 60/141 (42.6%) isolates. More data on sample submission drivers, antimicrobial drug use, and susceptibilities of important porcine bacterial isolates are required to inform guidelines for prudent antimicrobial use, to reduce their prevalence, human transmission, and to minimise AMR and MDR.
PubMed: 32824043
DOI: 10.3390/ani10081427