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An epitope shared by enterobacterial and neisserial porin proteins.

A murine monoclonal antibody (MAb F9-16) raised against a porin protein epitope called Po I of an E. coli 055 strain showed broad cross-reactivity with bacteria within the Enterobacteriaceae, and also recognized neisseriae and moraxellae. In an immunodot assay, the antibody was bound by 32/33 strains of neisseriae and moraxellae after SDS treatment of the bacteria. Testing intact bacteria, 11/33 isolates showed definite MAb binding, including serogroup A and B meningococci. In Western blotting, the anti-Po I MAb targeted the gonococcal porin proteins PIA and PIB, and class 1, class 2, and class 3 porins of meningococci. The MAb showed no reactivity against decapeptides which corresponded to the whole length of a meningococcal class 1 porin protein of the subtype P1, 7, 16. These findings accord with the inference that enterobacterial, neisserial and moraxellae porin proteins share an epitope (Po I) which is determined by the three-dimensional rather than by the primary structure of the proteins and that this epitope is shielded in most isolates but surface-exposed in some isolates, including some strains of meningococci. Since Po I is broadly distributed among commensal and pathogenic bacteria and has demonstrated immunogenicity in humans, this epitope may play a role in elicitation of "normal" antibodies with immunoprotective activity.

Antibodies, Bacterial↗

Characterization of radiation-resistant vegetative bacteria in beef.

Ground beef contains numerous microorganisms of various types. The commonly recognized bacteria are associated with current problems of spoilage. Irradiation, however, contributes a new factor through selective destruction of the microflora. The residual microorganisms surviving a nonsterilizing dose are predominantly gram-negative coccobacilli. Various classifications have been given, e.g., Moraxella, Acinetobacter, Achromobacter, etc. For a more detailed study of these radiation-resistant bacteria occurring in ground beef, an enrichment procedure was used for isolation. By means of morphological and biochemical tests, most of the isolates were found to be Moraxella, based on current classifications. The range of growth temperatures was from 2 to 50 C. These bacteria were relatively heat sensitive, e.g., D10 of 5.4 min at 70 C or less. The radiation resistance ranged from D10 values of 273 to 2,039 krad. Thus, some were more resistant than any presently recognized spores. A reference culture of Moraxella osloensis was irradiated under conditions comparable to the enrichment procedure used with the ground beef. The only apparent changes were in morphology and penicillin sensitivity. However, after a few subcultures these bacteria reverted to the characteristics of the parent strain. Thus, it is apparent that these isolates are a part of the normal flora of ground beef and not aberrant forms arising from the irradiation procedure. The significance, if any, of these bacteria is not presently recognized.

Dose-Response Relationship, Radiation↗

Preliminary study on relationships among strains forming a bacterial community selected on naphthalene from a marine sediment.

Two bacterial strains were isolated from a bacterial community formed of nine strains, selected from a marine sediment on a seawater medium with naphthalene as sole carbon source. The two strains studied in the present work were the only strains of this community able to grow in pure culture on naphthalene; therefore, they were called "primary" strains. The seven other strains were maintained in the community by using metabolic intermediates of the two primary strains; they were called "auxiliary" strains. Regulation of naphthalene metabolism was studied for the two primary strains. They oxidized naphthalene into catechol, which was degraded only by the meta pathway. For Pseudomonas Lav. 4, naphthalene oxygenase and salicylate hydroxylase were inducible; catechol 2,3-dioxygenase was constitutive. For Moraxella Lav. 7, naphthalene oxygenase was constitutive; salicylate hydroxylase and catechol 2,3-oxygenase were inducible. The Moraxella strain carries two cryptic plasmids, about 63- and 85-kb in molecular size. In the bacterial community culture medium, Moraxella Lav. 7 prevented accumulation of 2-hydroxymuconate semialdehyde formed by Pseudomonas Lav. 4. The auxiliary strains take up formic, acetic, pyruvic, propionic, and succinic acids released by the two primary strains.

Biodegradation, Environmental↗

Bacteriological investigation of infectious keratoconjunctivitis in Norwegian sheep.

Contagious keratoconjunctivitis is a rather common disease in Norwegian sheep. Since the knowledge of its aetiology is limited, the present study was performed to determine the microorganisms involved. Local veterinarians throughout the country collected conjunctival swabs from both sick (n = 43) and healthy (n = 42) sheep on 15 farms with outbreaks of ovine keratoconjunctivitis, and further from healthy sheep (n = 50) on 17 farms not showing any signs of conjunctival disease. All samples were cultivated for bacteria and mycoplasma. Listeria monocytogenes was isolated from 3 cases (1%) in one single herd. Staphylococcus aureus (5%), Corynebacterium spp. (2%) and Escherichia coli (4%) were isolated only in herds with keratoconjunctivitis, but from both sick and healthy animals. Moraxella (Branhamella) ovis was isolated from 28% of sampled animals in affected herds and from 10% of sampled animals in healthy herds. The corresponding numbers for Moraxella spp. were 9%/12%, for Pseudomonas spp. 7%/8%, for Staphylococcus spp. 22//22%, for Bacillus spp. 12%/14%, for Micrococcus spp. 6%/2% and for Streptococcus/Enterococcus spp. 2%/2%. Mycoplasma conjunctivae was isolated from 16 animals with keratoconjunctivitis (37%) and from 3 animals without clinical signs (7%) in farms with keratoconjunctivitis. In farms without clinical signs of keratoconjunctivitis, M. conjunctivae was isolated in 4 animals (8%). To our knowledge, this is the first time M. conjunctivae has been isolated in Norway. Other predisposing agents found were Moraxella (Branhamella) ovis and Listeria monocytogenes. The etiological importance of different microorganisms in ovine keratoconjunctivitis seems to vary; some are probably only present as secondary invaders. Other possible causes of ovine keratoconjunctivitis in Norway, such as Chlamydia psittaci, remain to be investigated.

Animals↗

[A clinicobacteriologic study on clavulanic acid/amoxicillin in pediatric acute otitis media].

We carried out clinical and bacteriological studies on clavulanic acid/amoxicillin and amoxicillin in pediatric acute otitis media at 14 general practice settings. The results are summarized as follows. 1. The major isolated organisms from content of middle ear effusion were Streptococcus pneumoniae 31.8%, Haemophilus influenzae 35.8% and Moraxella subgenus Branhamella catarrhalis 1.5%. Similar results were observed for the major isolates organisms from content of nasopharynx Streptococcus pneumoniae 31.1%, Haemophilus influenzae 33.9% and Moraxella subgenus Branhamella catarrhalis 19.2%. 2. 42.2% of S. pneumoniae isolated from middle ear effusion were drug resistant S. pneumoniae (PISP, PRSP) and they were increasing year by year. 3. 46.7% of S. pneumoniae isolated from nasopharyngeal swab were drug resistant S. pneumoniae (PISP, PRSP) and they were increasing year by year. The incidence of drug resistant S. pneumoniae isolated from all cases and organisms were 26.3% and 14.5%, respectively. 4. On MIC90, antimicrobial activity of CVA/AMPC against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella subgenus Branhamella catarrhalis was superior to SBTPC. 5. In the evaluation of clinical efficacy, bacteriological efficacy and utility, CVA/AMPC-treated group was significantly superior to AMPC-treated group. 6. Adverse reactions were observed in 22% of CVA/AMPC-treated group, involving diarrhea and loose stool.

Acute Disease↗

Microbial pathogens of adult community-acquired pneumonia in Southern Estonia.

Prospective analysis of the etiology of community-acquired pneumonia in 209 inpatients (mean age 56.2+/-11.2 years) was performed at the Lung Hospital of Tartu University, Estonia. Majority of the patients (87.6%) had at least one significant modifying or risk factor and 30.6% had severe community-acquired pneumonia. Streptococcus pneumoniae, beta-hemolytic streptococci, Klebsiella pneumoniae, and Moraxella catarrhalis were the most frequent pathogens (22.0, 12.2, 11.4, and 10.2%, respectively). Combined etiology was detected in 17.2%. Gram-negative pathogens prevailed over gram-positives and were significantly more frequent in patients aged > or =60 years (p=0.002) as in those with underlying diseases (p=0.01). An age of > or =60 years and previous antibacterial therapy were significant risk factors for Klebsiella pneumoniae as a pathogen (p=0.005 and p=0.002, respectively). Susceptibility of Streptococcus pneumoniae to penicillin, erythromycin, and doxycycline was 79.5, 90.2, and 100%, respectively, whereas all penicillin-resistant isolates were cefuroxime- and ceftriaxone-susceptible. Klebsiella pneumoniae was 100, 88.5, 84.2, and 81.8% susceptible to ceftazidime, gentamicin, ampicillin-sulbactam, and ciprofloxacin, and Moraxella catarrhalis was 100, 100, 100, 93.8, and 93.3% susceptible to ampicillin-sulbactam, cephalexin, ceftriaxone, ciprofloxacin, and erythromycin, respectively. In conclusion, Streptococcus pneumoniae, Moraxella catarrhalis, and Klebsiella pneumoniae were the major pathogens of community-acquired pneumonia in Southern Estonia. Although gram-negatives were more prevalent, the major pathogens were favorably susceptible without a noticeable cross-resistance. Nevertheless, improved surveillance studies are needed because of a pressure for the rise in antibiotic resistance in the society.

Adolescent↗

Taxonomy of pasteurella anatipestifer. 2. Cellular fatty-acid profile by gas chromatography.

Methylated cellular fatty acids of representative strains of Pasteurella spp., Moraxella spp., and P. anatipestifer were subjected to gas chromatography in an attempt to further support the independence of P. anatipestifer from both Pasteurella and Moraxella. All Pasteurella spp. and Moraxella spp. revealed group characteristics specific for each genus that could be easily differentiated from the unique profile of P. anatipestifer. All P. anatipestifer strains tested showed similar fatty-acid profiles in gas chromatography, regardless of host of origin.

Animals↗

The role of face flies in an episode of infectious bovine keratoconjunctivitis.

Cattle partially protected from face flies (Musca autumnalis) had much less infectious bovine keratoconjunctivitis and yielded fewer isolated of hemolytic Moraxella bovis than did similar cattle that received no face fly protection. The number of new isolated of M bovis from the unprotected cattle was highly correlated with the number of face files observed on the cattle. Close physical proximity to cattle infected with M bovis was important in the spread of the bacteria to previously uninfected cattle; however, it did not require physical contact among the cattle to spread from herd to herd. Moraxella bovis first began to spread from herd to herd after face fly populations exceeded 10/animal for 1 month. Keratoconjunctivitis was not observed in 53% of the cattle from which M bovis was isolated. Moraxella bovis was isolated from 4 of 45 pools of face files collected from animals in the untreated herds.

Animals↗

Studies of Pasteurella anatipestifer: an approach to its classification.

Moraxella spp., Pasteurella spp., and strains of P. anatipestifer were tested for biochemical reactions, growth temperature, viability, antibacterial sensitivity, and DNA base composition. P. anatipestifer was viable for shorter periods at 37 C, showed high resistance to polymyxin B and kanamycin, and had lower base composition than reported for Moraxella and Pasteurella spp. Because of these conditions, P. anatipestifer should be excluded from the genera Pasteurella and Moraxella.

Animals↗

Characterization of some fish and shrimp spoiling bacteria.

The classification of some important groups of bacteria involved in fish and shrimp spoilage was studied. Trimethylamine is produced by Pseudomonas putrefaciens, a "non-defined" group resembling Ps. putrefaciens, Photobacterium spp. and some Moraxella-like bacteria. Hypoxanthine is produced by the same groups of bacteria except the last named and also by the "typical shrimp spoilers" (presumptive Alteromonas). Strong off-odours are produced on fresh fish by Ps. putrefaciens, dextrose-oxidative Pseudomonas spp. (Groups I and II according to Shewan, Hobbs and Hodgkiss, 1960), the above mentioned "non-defined" group and by only some of the "typical shrimp spoilers", whereas Moraxella-like bacteria and Photobacterium spp. failed to produce strong odours. Strong off-odours are produced on boiled shrimp by the "typical shrimp spoilers" (presumptive Altermonas). Ps. putrefaciens, the dextrose-oxidative Pseudomonas spp. and the "non-defined" group; Moraxella-like bacteria produced less offensive odours or none, nor did Photobacterium.

Alcaligenes↗

Isolation and characterization by conventional methods and genetic transformation of Psychrobacter and Acinetobacter from fresh and spoiled meat, milk and cheese.

Of 126 samples of fresh and spoiled meat and dairy products, 40% were positive for the presence of Moraxella-like bacteria and 64% of Acinetobacter; 279 and 466 strains, respectively, were isolated and a part of these were tested by biochemical methods and DNA transformation assays. In some cases, the Moraxellaceae in the samples examined reached considerable quantitative levels, but their percentage in the microflora was generally low. Moraxella-like bacteria were predominant in fresh meat, Acinetobacter in spoiled meat and milk. Most acinetobacters belonged to biotype lwoffii (sensu lato) and all 90 strains tested were positive for DNA transformation with an auxotrophic Acinetobacter. Moraxella-like bacteria were identified as Psychrobacter immobilis in 96% of 103 transformation assays. Moraxellaceae show lipolytic activity but they are considered of low incidence in food spoilage. Only 3.7% of acinetobacters from dairy sources was able to produce ropy milk. Unlike strains from clinical isolates, psychrobacters and acinetobacters isolated from food often do not grow at 37 degrees C.

Acinetobacter↗

Riemerella anatipestifer gen. nov., comb. nov., the causative agent of septicemia anserum exsudativa, and its phylogenetic affiliation within the Flavobacterium-Cytophaga rRNA homology group.

The phylogenetic position of the causative agent of septicemia anserum exsudativa, now most often referred to as [Moraxella] anatipestifer (brackets indicate a generically misnamed taxon) or "[Pasteurella] anatipestifer," was established by performing rRNA cistron similarity studies. [Moraxella] anatipestifer belongs to rRNA superfamily V, together with the genera Flavobacterium, Cytophaga, Flexibacter, Weeksella, Capnocytophaga, and Sphingobacterium. The detailed structure of rRNA superfamily V, which now contains five major rRNA homology groups, is described. An analysis of various phenotypic parameters, including new data (cellular proteins and fatty acids) and previously published data (respiratory quinones, enzyme activities, and classical phenotypic features), revealed that [Moraxella] anatipestifer differs in many aspects from its closest relatives, Flavobacterium indologenes, Flavobacterium gleum, Flavobacterium indoltheticum, Flavobacterium balustinum, Flavobacterium meningosepticum, and Weeksella zoohelcum. The combined genotypic and phenotypic data indicate that this organism should be placed in a separate genus; the name Riemerella anatipestifer gen. nov., comb. nov. is proposed for this bacterium. The specific epithet anatipestifer is kept in order to avoid nomenclatural confusion. However, it should be emphasized that the illness caused by this organism is a septicemic disease which is not restricted to ducks.

Animals↗

Microbiological characteristics of Pacific shrimp (Pandalus jordani).

Microorganisms associated with Pacific shrimp (Pandalus jordani) were isolated and identified. Those on the iced raw shrimp, which yielded an average count of 1.6 x 10(6), were predominantly Moraxella, Pseudomonas, Acinetobacter, Arthrobacter, and Flavobacterium-Cytophaga spp. The blanching and peeling reduced the microbial level to 3.3 x 10(4) and also selectively eliminated Moraxella spp. The microbial flora changed after each processing sequence, and the heat sensitivity and growth characteristics of the representative microbial groups suggested that the presence of Arthrobacter and Acinetobacter spp. in peeled shrimp may indicate inadequate cleaning of raw shrimp or a shorter blanching time. The presence of Moraxella and Flavobacterium-Cytophaga spp. would indicate the degree of secondary contamination, and the presence of Pseudomonas spp. would indicate the shelf-age of the processed shrimp.

Animals↗

Numerical taxonomy of gram-negative, nonmotile, nonfermentative bacteria isolated during chilled storage of lamb carcasses.

A numerical taxonomic study using 75 characters was performed with 132 strains of gram-negative, nonmotile, nonfermentative bacteria selected on the basis of lack of motility and Gram reaction among 1,200 cultures isolated during aerobic storage of lamb carcasses. At the 80% similarity level (SSM), eight clusters were formed. Strains in clusters 1 to 6 could be identified as members of the family Moraxellaceae and, more specifically, as members of the Psychrobacter-[Moraxella] phenylpyruvica subgroup. Of these strains, clusters 1 and 2 (88 strains) were identified as [Moraxella] phenylpyruvica and cluster 3 (15 strains) was identified as Psychrobacter immobilis. Clusters 4, 5, and 6 were not identifiable with any species. Clusters 7 and 8 consisted of 14 strains considered nonmotile variants of Pseudomonas fragi. The highest separation indices corresponded to acid production from certain carbohydrates (melibiose, L-arabinose, and cellobiose). Although strains of Psychrobacter-Moraxella clusters were relatively frequently identified at the completion of slaughter, very few cultures were detected on spoiled carcasses. It appears, therefore, that this group of organisms has only low spoilage potential.

Animals↗

Role of neutral metabolites in microbial conversion of 3beta-acetoxy-19-hydroxycholest-5-ene into estrone.

Biotransformation of 3beta-acetoxy-19-hydroxycholest-5-ene (19-HCA, 6 g) by Moraxella sp. was studied. Estrone (712 mg) was the major metabolite formed. Minor metabolites identified were 5alpha-androst-1-en-19-ol-3,17-dione (33 mg), androst-4-en-19-ol-3,17-dione (58 mg), androst-4-en-9alpha,19-diol-3,17-dione (12 mg), and androstan-19-ol-3,17-dione (1 mg). Acidic metabolites were not formed. Time course experiments on the fermentation of 19-HCA indicated that androst-4-en-19-ol-3,17-dione was the major metabolite formed during the early stages of incubation. However, with continuing fermentation its level dropped, with a concomitant increase in estrone. Fermentation of 19-HCA in the presence of specific inhibitors or performing the fermentation for a shorter period (48 h) did not result in the formation of acidic metabolites. Resting-cell experiments carried out with 19-HCA (200 mg) in the presence of alpha,alpha'-bipyridyl led to the isolation of three additional metabolites, viz., cholestan-19-ol-3-one (2 mg), cholest-4-en-19-ol-3-one (10 mg), and cholest-5-en-3beta,19-diol (12 mg). Similar results were also obtained when n-propanol was used instead of alpha,alpha'-bipyridyl. Resting cells grown on 19-HCA readily converted both 5alpha-androst-1-en-19-ol-3,17-dione and androst-4-en-19-ol-3,17-dione into estrone. Partially purified 1,2-dehydrogenase from steroid-induced Moraxella cells transformed androst-4-en-19-ol-3,17-dione into estrone and formaldehyde in the presence of phenazine methosulfate, an artificial electron acceptor. These results suggest that the degradation of the hydrocarbon side chain of 19-HCA does not proceed via C(22) phenolic acid intermediates and complete removal of the C(17) side chain takes place prior to the aromatization of the A ring in estrone. The mode of degradation of the sterol side chain appears to be through the fission of the C(17)-C(20) bond. On the basis of these observations, a new pathway for the formation of estrone from 19-HCA in Moraxella sp. has been proposed.

Journal Article↗

On the in-vitro antimicrobial activity of oleuropein and hydroxytyrosol.

Secoiridoides (oleuropein and derivatives), one of the major classes of polyphenol contained in olives and olive oil, have recently been shown to inhibit or delay the rate of growth of a range of bacteria and microfungi but there are no data in the literature concerning the possible employment of these secoiridoides as antimicrobial agents against pathogenic bacteria in man. In this study five ATCC standard bacterial strains (Haemophilus influenzae ATCC 9006, Moraxella catarrhalis ATCC 8176, Salmonella typhi ATCC 6539, Vibrio parahaemolyticus ATCC 17802 and Staphylococcus aureus ATCC 25923) and 44 fresh clinical isolates (Haemophilus influenzae, eight strains, Moraxella catarrhalis, six strains, Salmonella species, 15 strains, Vibrio cholerae, one strain, Vibrio alginolyticus, two strains, Vibrio parahaemolyticus, one strain, Staphylococcus aureus, five penicillin-susceptible strains and six penicillin-resistant strains), causal agents of intestinal or respiratory tract infections in man, were tested for in-vitro susceptibility to two olive (Olea europaea) secoiridoides, oleuropein (the bitter principle of olives) and hydroxytyrosol (derived from oleuropein by enzymatic hydrolysis and responsible for the high stability of olive oil). The minimum inhibitory concentrations (MICs) calculated in our study are evidence of the broad antimicrobial activity of hydroxytyrosol against these bacterial strains (MIC values between 0.24 and 7.85 microg mL(-1) for ATCC strains and between 0.97 and 31.25 microg mL(-1) for clinically isolated strains). Furthermore oleuropein also inhibited (although to a much lesser extent) the growth of several bacterial strains (MIC values between 62.5 and 500 microg mL(-1) for ATCC strains and between 31.25 and 250 microg mL(-1) for clinical isolates); oleuropein was ineffective against Haemophilus influenzae and Moraxella catarrhalis. These data indicate that in addition to the potential employment of its active principles as food additives or in integrated pest-management programs, Olea europaea can be considered a potential source of promising antimicrobial agents for treatment of intestinal or respiratory tract infections in man.

Anti-Bacterial Agents↗

Biochemical analysis of lactoferrin receptors in the Neisseriaceae: identification of a second bacterial lactoferrin receptor protein.

Bacterial transferrin receptors that have been described in the families Pasteurellaceae and Neisseriaceae are composed of two receptor proteins, transferrin binding proteins 1 and 2 (Tbp1 and Tbp2). In contrast, bacterial lactoferrin receptors have only been described for human pathogens in the family Neisseriaceae, and were believed to consist of a single protein, Lbp1, which is highly homologous to Tbp1. We describe a modified affinity isolation procedure that facilities isolation of a second lactoferrin receptor protein Lbp2 (a presumptive Tbp2 homologue) from Neisseria meningitidis, Moraxella catarrhalis and Moraxella bovis using immobilized lactoferrin. Antiserum specific for either the M. catarrhalis Tbp1+2 molecules, the M. catarrhalis Lbp1 molecule, or for a commercial preparation of human lactoferrin did not react on western blots with the same organisms' affinity purified Lbp2. In addition, the M. catarrhalis Lbp2 could be isolated in a functional form without contaminating Lbp1 or Tbp1+2. We also demonstrate that the bovine pathogen, M. bovis, produces functional transferrin and lactoferrin receptors specific for the bovine forms of these glycoproteins. A putative lbpB gene, recently speculated to reside immediately upstream of the N. meningitidis Lbp1 structural gene, lbpA, likely encodes the newly isolated Lbp2 protein from this bacterial species.

Animals↗

Susceptibility of European respiratory tract isolates to trovafloxacin, ciprofloxacin, clarithromycin, azithromycin and ampicillin.

As part of the Artemis project, 11500 isolates (3000 from patients with respiratory tract infections) were collected throughout six European countries between 1994 and 1996. Twenty-seven hospitals or laboratories participated in this first phase of the study. The activities of three classes of antimicrobial agents (fluoroquinolones, beta-lactam agents, macrolides) are presented for the six most frequently isolated pathogens (Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Pseudomonas aeruginosa, Klebsiella pneumoniae). Overall, trovafloxacin and ciprofloxacin activities were similar for Haemophilus influenzae, Moraxella catarrhalis and Klebsiella pneumoniae isolates. Of the Streptococcus pneumoniae isolates, 6% were resistant to penicillin. Trovafloxacin had the highest activity against the Streptococcus pneumoniae isolates, with a minimum inhibitory concentration of 0.25 mg/l for 90% of isolates (MIC90); all strains tested were susceptible to trovafloxacin. The MIC90 of ciprofloxacin for Streptococcus pneumoniae was 3 mg/l, and overall 52% of the strains were susceptible; 9% were resistant. Azithromycin and clarithromycin exhibited similar activity against all collected pathogens, except Haemophilus influenzae. All strains of Haemophilus influenzae were susceptible to azithromycin compared with 79% for clarithromycin, with respective MIC90s of 2 and 16 mg/l. The data presented demonstrate differences in the susceptibility patterns of six major respiratory tract pathogens in Europe.

Ampicillin↗