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Emerging Infectious Diseases Mar 2004An outbreak of bovine necrotic vulvovaginitis associated with Porphyromonas levii, an emerging animal and human pathogen, affected 32 cows on a dairy farm in the...
An outbreak of bovine necrotic vulvovaginitis associated with Porphyromonas levii, an emerging animal and human pathogen, affected 32 cows on a dairy farm in the northeast of Israel. Five animals had to be culled. This report appears to be the first that associates P. levii with bovine necrotic vulvovagnitis.
Topics: Animals; Bacteroidaceae Infections; Cattle; Cattle Diseases; Disease Outbreaks; Female; Israel; Porphyromonas; Vulvovaginitis
PubMed: 15109423
DOI: 10.3201/eid1003.020592 -
American Journal of Veterinary Research May 2002To examine the host response toward Porphyromonas levii, by evaluating chemotaxis, phagocytosis, and oxidative burst of bovine macrophages in vitro.
OBJECTIVE
To examine the host response toward Porphyromonas levii, by evaluating chemotaxis, phagocytosis, and oxidative burst of bovine macrophages in vitro.
SAMPLE POPULATION
Cultured bovine macrophages obtained from monocytes harvested from blood samples of 15 Holstein steers. Porphyromonas levii was isolated from the foot rot lesion of an acutely affected feedlot steer.
PROCEDURE
Monocytes were cultured for macrophage differentiation over 7 days. Porphyromonas levii was cultured in strict anaerobic conditions for experimentation. Chemotaxis was evaluated by quantifying macrophage migration toward P. levii in Boyden chambers. Phagocytosis was assessed by quantification of macrophages engulfing P. levii following incubation with or without anti-P. levii serum or purified IgG. Oxidative burst was measured by use of the nitroblue tetrazolium reduction assay.
RESULTS
Chemotaxis toward P. levii was not significantly different from control values at any of the tested bacterial concentrations. Phagocytosis of P. levii was approximately 10% at a 10:1 bacterium to macrophage ratio and did not change significantly over time. When higher proportions of P. levii were tested for phagocytosis, the 1,000:1 bacterium to macrophage ratio had a significant increase, compared with the 10:1 test group. Opsonization of P. levii with high-titeranti-P. levii serum or anti-P. levii IgG produced a significant increase in macrophage phagocytosis. Oxidative production significantly increased compared with control in the 1,000:1 test group only.
CONCLUSIONS AND CLINICAL RELEVANCE
Porphyromonas levii may evade host detection by decreased chemotaxis, phagocytosis, and oxidative burst by macrophages. Acquired immunity may be beneficial for clearance of P. levii in foot rot lesions in cattle.
Topics: Animals; Antibodies, Bacterial; Cattle; Cattle Diseases; Chemotaxis; Foot Rot; Indicators and Reagents; Macrophages; Male; Microscopy, Electron; N-Formylmethionine Leucyl-Phenylalanine; Nitroblue Tetrazolium; Phagocytosis; Pseudomonas; Pseudomonas Infections; Respiratory Burst
PubMed: 12013480
DOI: 10.2460/ajvr.2002.63.757 -
Canadian Journal of Veterinary Research... Apr 2002The objective was to evaluate the pro-inflammatory response of bovine macrophages towards Porphyromonas levii, an etiologic agent of acute interdigital phlegmon, by...
The objective was to evaluate the pro-inflammatory response of bovine macrophages towards Porphyromonas levii, an etiologic agent of acute interdigital phlegmon, by evaluating the mRNA expression of tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), and interleukin 8 (IL-8). Bovine macrophages detect the presence of bacteria, such as P. levii, and respond by upregulating transcription of the genes for pro-inflammatory cytokines TNF-alpha and IL-1beta in addition to the neutrophil chemoattractant IL-8. Monocytes were isolated from blood obtained from Holstein steers. These cells were cultured and differentiated into macrophages over 7 d, followed by exposure to P. levii, Escherichia coli lipopolysaccharide (LPS), or tissue culture medium for 1, 1.5, 2, 4, 6, 8,12, or 24 h. Total RNA was extracted, and reverse transcriptase polymerase chain reaction was conducted to examine the presence of TNF-alpha, IL-1beta, or IL-8 mRNA. Products were visualized on agarose gels to determine the presence or absence of cytokine mRNA amplified DNA. Bovine macrophages, when exposed to P. levii or E. coli LPS, produced mRNA for TNF-alpha, IL-1beta, and IL-8. Expression of all 3 cytokines was higher in the P. levii and LPS-exposed macrophages at all time points examined, compared with tissue culture medium-treated cells. Expression of these cytokines by macrophages is likely directly involved in orchestration of the immune response, and particularly in neutrophil recruitment to affected tissues in acute interdigital phlegmon.
Topics: Animals; Bacteroidaceae Infections; Cattle; Cattle Diseases; Cells, Cultured; Cytokines; Interleukin-1; Interleukin-8; Lipopolysaccharides; Macrophage Activation; Macrophages; Male; Porphyromonas; RNA, Messenger; Reverse Transcriptase Polymerase Chain Reaction; Time Factors; Tumor Necrosis Factor-alpha
PubMed: 11989740
DOI: No ID Found -
The Journal of Antimicrobial... May 2000Gram-negative anaerobes belonging to the genera Fusobacterium, Prevotella and Porphyromonas were investigated for the presence of tetQ and ermF, which have been shown to...
Gram-negative anaerobes belonging to the genera Fusobacterium, Prevotella and Porphyromonas were investigated for the presence of tetQ and ermF, which have been shown to be spread by conjugal elements. One hundred isolates from either sites of infection or various body sites in healthy subjects were studied. PCR was used to detect tetQ, and DNA-DNA hybridization studies on EcoRI chromosomal digests were undertaken to detect the presence of tetQ and ermF. Antibiotic sensitivity assays were performed on selected isolates to detect tetracycline, erythromycin and penicillin resistance. Twenty Fusobacterium isolates lacked tetQ, and were tetracycline sensitive. Twenty per cent of Prevotella spp. isolates both from clinical specimens and from healthy subjects were found to possess tetQ. Of 20 Porphyromonas isolates tested, one (Porphyromonas levii) from a case of bacterial vaginosis was shown to possess tetQ in the chromosome. The presence of tetQ was always associated with tetracycline resistance. Four isolates of Prevotella melaninogenica and one isolate of Prevotella were ermF-positive, although expression of erythromycin resistance was not consistently associated with detection of this gene. Antibiotic resistance phenotypes of Prevotella isolates were shown to be related to specific chromosomal restriction patterns by hybridization studies: tetracycline resistance and tetracycline/erythromycin resistance are conferred by Bacteroides tetracycline-resistant ERL elements, whereas the tetracycline/penicillin resistance phenotype could be due to spread of elements identified in Prevotella only. Tetracycline/erythromycin-resistant and tetracycline/erythromycin/penicillin-resistant P. melaninogenica isolates were found in this study. It appeared that the presence of tetQ and ermF in Bacteroides and Prevotella contributed to the persistence of antibiotic resistance isolates within the host and to potential spread to other organisms through conjugal elements.
Topics: Anti-Bacterial Agents; Bacteroidaceae Infections; Drug Resistance, Microbial; Erythromycin; Fusobacterium; Fusobacterium Infections; Genes, Bacterial; Humans; Microbial Sensitivity Tests; Nucleic Acid Hybridization; Penicillins; Polymerase Chain Reaction; Porphyromonas; Prevotella; Tetracycline; Tetracycline Resistance
PubMed: 10797077
DOI: 10.1093/jac/45.5.577 -
Canadian Journal of Veterinary Research... Apr 1999Acute interdigital phlegmon (AIP) is a commonly occurring anaerobic bacterial infection in cattle. This study examined in vitro the interaction of bovine...
Acute interdigital phlegmon (AIP) is a commonly occurring anaerobic bacterial infection in cattle. This study examined in vitro the interaction of bovine polymorphonuclear granulocytic neutrophils (PMN) from blood with bacterial species involved in AIP. Polymorphonuclear neutrophils were purified from whole bovine blood, exposed to one of the three putative etiologic agents of AIP and comparatively assessed for phagocytosis using light microscopy. Fusobacterium necrophorum and Prevotella intermedia were effectively phagocytosed by PMN, but Porphyromonas levii was phagocytosed significantly less effectively by PMN. The effect of high titre anti-P. levii bovine serum on antibody-mediated phagocytosis by PMN was also evaluated. High titre serum increased the efficiency of phagocytosis of P. levii by bovine PMN. This was independent of heat labile complement factors. Antibodies specific for P. levii were assessed for protease activity capable of cleaving bovine immunoglobulins (IgG, IgG1, IgG2, and IgM). Partially purified supernatant from broth cultures of P. levii were incubated with biotinylated immunoglobulins (Igs). Samples were taken from times 0 to 72 h and examined using SDS-PAGE followed by Western blot analysis. Streptavidin-alkaline phosphatase and NBT-BCIP were used to visualize the Igs for heavy and light chains as well as lower molecular weight fragments of these glycoproteins. Porphyromonas levii produced an immunoglobulin protease which readily cleaved bovine IgG into fragments, but did not act against IgM. Specifically, the enzyme may be a significant virulence factor as it may act to neutralize the antibodies demonstrated necessary for effective PMN-mediated phagocytosis.
Topics: Animals; Bacterial Proteins; Biotinylation; Cattle; Fusobacterium; Immunoglobulin G; Immunoglobulin M; Male; Metalloendopeptidases; Neutrophils; Phagocytosis; Porphyromonas; Prevotella; Substrate Specificity
PubMed: 10369568
DOI: No ID Found -
Archives of Oral Biology Dec 1997The ability of three Porphyromonas spp., seven Prevotella spp., seven Fusobacterium spp. and two related Bacteroides spp. (B. levii and B. macacae) to degrade an...
The ability of three Porphyromonas spp., seven Prevotella spp., seven Fusobacterium spp. and two related Bacteroides spp. (B. levii and B. macacae) to degrade an extensive range of synthetic endo-, amino- and diamino peptidase substrates linked to the fluorescent leaving group 7-amido-4-methylcoumarin (NHMec) was investigated. Many more species than was previously recognized exhibited peptidase activities, albeit at lower levels than those already described for Porphyromonas gingivalis. Detection of chymotrypsin-like activity was dependent on which of three NHMec-linked substrates was used, but all species exhibited degradative activity with at least one of these substrates. Elastase-like activity was detected in all species though not all species reacted with each of the elastase substrates. Glycylprolyl peptidase activity was detected in all of the species tested with the exception of F. mortiferum, F. gonidiaformans, F. naviforme and F. necrophorum. While the detection of peptidase activities does not appear to be useful for the differentiation of species within the genera Bacteroides and Prevotella, its ability to differentiate species of the genus Porphyromonas or Fusobacterium warrants further investigation.
Topics: Alanine; Aminopeptidases; Bacteroides; Chymotrypsin; Coumarins; Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; Endopeptidases; Fluorescent Dyes; Fusobacterium; Fusobacterium necrophorum; Humans; Indicators and Reagents; Lysine; Mouth; Oligopeptides; Pancreatic Elastase; Phenylalanine; Porphyromonas; Porphyromonas gingivalis; Prevotella; Proline; Tyrosine; Virulence
PubMed: 9460536
DOI: 10.1016/s0003-9969(97)00078-2 -
Journal of the American Veterinary... Jan 1998To determine whether ceftiofur sodium would be useful for treatment of acute interdigital phlegmon (foot rot) in cattle. (Clinical Trial)
Clinical Trial Comparative Study Randomized Controlled Trial
OBJECTIVE
To determine whether ceftiofur sodium would be useful for treatment of acute interdigital phlegmon (foot rot) in cattle.
DESIGN
Randomized controlled trial.
ANIMALS
308 cross-bred yearling steers with clinical signs of acute interdigital phlegmon (i.e., lameness with interdigital swelling, interdigital lesions, or both).
PROCEDURE
Steers were randomly assigned to 1 of 3 treatment groups: ceftiofur at a dosage of 0.1 mg/kg (0.045 mg/lb) of body weight, IM, every 24 hours; ceftiofur at a dosage of 1.0 mg/kg (0.45 mg/lb), IM, every 24 hours, and oxytetracycline at a dosage of 6.6 mg/kg (3 mg/lb), IM, every 24 hours. All animals were treated for 3 days; treatment was considered successful if animals were no longer lame on day 4. Biopsy specimens were collected prior to treatment from 5 animals in each group and submitted for anaerobic bacterial culture and histologic examination.
RESULTS
Success rates for the high-dosage ceftiofur (94/129; 73%) and oxytetracycline (87/128; 68%) groups were significantly higher than that for the low-dosage ceftiofur group (5/50; 10%), but there were no significant differences between the high-dosage ceftiofur and oxytetracycline groups. Anaerobic bacteria most frequently isolated from biopsy specimens were Porphyromonas levii and Provetella intermedia.
CLINICAL IMPLICATIONS
Use of ceftiofur at a dosage of 1.0 mg/kg for treatment of cattle with acute interdigital phlegmon was as effective as use of oxytetracycline at a dosage of 6.6 mg/kg. However, ceftiofur has a negligible withdrawal time and, therefore, may be a better choice for treatment of near-market weight animals.
Topics: Acute Disease; Animals; Anti-Bacterial Agents; Bacteroidaceae Infections; Biopsy; Cattle; Cattle Diseases; Cephalosporins; Dose-Response Relationship, Drug; Foot; Foot Rot; Lameness, Animal; Oxytetracycline; Porphyromonas; Time Factors
PubMed: 9448831
DOI: No ID Found -
Clinical Infectious Diseases : An... Sep 1997There is still inadequate information on the role of certain newly described or reclassified anaerobes in disease processes, on their normal sites of carriage, and on... (Review)
Review
There is still inadequate information on the role of certain newly described or reclassified anaerobes in disease processes, on their normal sites of carriage, and on their antimicrobial susceptibilities. Herein, we summarize this information (most of the literature reviewed is from the past 5 years, but a few of the articles are approximately 10 years old). Porphyromonas species had seemed to be relatively nonpathogenic, but recent work indicates that this belief is incorrect. P. gingivalis, P levii-like organisms, and P. endodontalis-like organisms have been recovered from a variety of oral and extraoral infections. P. macacae has been recovered from infected cat bite wounds. Sutterella wadsworthensis, recently differentiated from Campylobacter gracilis, has been found in a variety of infections. Bilophila wadsworthia has also been recovered from a wide variety of infections. Newly described anaerobic cocci, gram-positive nonsporeforming rods, and clostridia have also been isolated from various infections.
Topics: Bacteria, Anaerobic; Gram-Negative Anaerobic Bacteria; Gram-Positive Bacteria; Humans
PubMed: 9310641
DOI: 10.1086/516237 -
Clinical Infectious Diseases : An... Sep 1997A new method of identifying bacteria, phylogenetic 16S rRNA sequencing, has led to major reorganizations among most genera of anaerobic bacteria. The pigmented... (Review)
Review
A new method of identifying bacteria, phylogenetic 16S rRNA sequencing, has led to major reorganizations among most genera of anaerobic bacteria. The pigmented Prevotella species now comprise seven species including P. nigrescens and P. tannerae; P. intermedia/P. nigrescens-like organisms await inclusion. The former Mitsuokella dentalis and Hallella seregens were transferred to Prevotella as one species, P. dentalis. P. enoeca is a new nonpigmenting Prevotella. The genus Porphyromonas currently includes 11 pigmented species and one nonpigmented species, P. catoniae; P. levii-like and P. endodontalis-like organisms are candidates for the genus. Fusobacterium nucleatum currently has five subspecies, and F. varium includes the former F. pseudonecrophorum. Former Wolinella recta and Wolinella curva now are Campylobacter rectus and Campylobacter curvus; Campylobacter showae is a new species. Isolates included in the bile-sensitive former Bacteroides gracilis now are Campylobacter gracilis; the bile-resistant B. gracilis isolates were transferred to a new genus, Sutterella, as S. wadsworthensis. The new Actinomyces species include two subspecies of the A. neuii and the A. radingae-A. turicensis complex. The genus Eubacterium sensu stricto is represented by E. limosum, and the former E. alactolyticum was reclassified in a new genus, Pseudoramibacter, as P. alactolyticus. Recent entries include E. saphenum, E. minutum, E. exiguum, E. infirmum, and E. tardum. A new genus, Atopobium houses some former lactobacilli and streptococci. The genus Peptostreptococcus also have four new species; P. hydrogenalis, P. lacrimalis, P. lactolyticus, and P. vaginalis.
Topics: Bacteria, Anaerobic; Gram-Negative Anaerobic Bacteria; Gram-Positive Bacteria; Humans
PubMed: 9310640
DOI: 10.1086/516227 -
Endodontics & Dental Traumatology Feb 1997Lipopolysaccharides (LPSs) of 11 bacterial strains from the type species of the genera Bacteroides (B. fragilis), Prevotella (Pr. melaninogenica), Porphyromonas (Po.... (Comparative Study)
Comparative Study
Lipopolysaccharides (LPSs) of 11 bacterial strains from the type species of the genera Bacteroides (B. fragilis), Prevotella (Pr. melaninogenica), Porphyromonas (Po. gingivalis), Campylobacter (C. fetus subsp. fetus), and Wolinella (W. succinogenes), and from the type strains of B. distasonis, B. forsythus, B. ureolyticus, Po. levii, Po. macacae, and C. gracilis, were extracted with hot water-phenol (Westphal method). S-form LPSs, obtained from all organisms, were well resolved with tricine-sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and visualized by silver staining. Lipid A was not stained. Also profiles from LPS of Escherichia coli, serotypes 0111:B4 and 055:B5, could be distinguished. While W. succinogenes showed a relatively short S-form LPS on electrophoregrams, the other bacteria, including B. fragilis, exhibited long-ladder LPSs. Po. gingivalis displayed the largest number of bands and the longest O-chain. The long O-chain of this bacterium may be important for its virulence.
Topics: Antigens, Bacterial; Bacteroides; Campylobacter; Electrophoresis, Polyacrylamide Gel; Glycine; Gram-Negative Anaerobic Bacteria; Lipopolysaccharides; Molecular Weight; Porphyromonas; Prevotella melaninogenica; Virulence; Wolinella
PubMed: 9206383
DOI: 10.1111/j.1600-9657.1997.tb00003.x