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Influence of lipopolysaccharides and lipids A from some marine bacteria on spontaneous and Escherichia coli LPS-induced TNF-alpha release from peripheral human blood cells.

Some endotoxic properties of lipopolysaccharides (LPS) and lipids A (LA) from the marine bacteria Marinomonas communis ATCC 27118(T), Marinomonas mediterranea ATCC 700492(T), and Chryseobacterium indoltheticum CIP 103168(T) were studied. The preparations tested were shown to have high 50% lethal doses (4 microg per mouse for LPS from M. mediterranea and more than 12 microg per mouse for two other LPS and LA from C. indoltheticum) and were moderate (371 +/- 37 pg/ml at 10 microg/ml of C. indoltheticum LPS), weak (148 +/- 5 pg/ml at 1 microg/ml of M. mediterranea LPS), and zero (LA and LPS from M. communis and LA from C. indoltheticum) inducers of tumor necrosis factor alpha (TNF-alpha) release from peripheral human blood cells. The capacity of the LA and LPS samples from marine bacteria to inhibit TNF-alpha release induced by LPS from Escherichia coli O55 : B5 (10 ng/ml) was also studied.

Animals↗

In vitro antimicrobial activity of gatifloxacin compared with other quinolones against clinical isolates from cancer patients.

Owing to the predominance of gram-positive pathogens in neutropenic cancer patients, newer generation quinolones with an expanded gram-positive spectrum and enhanced potency, may have a role to play for prophylaxis and/or empiric therapy in such patients. The in vitro activity of gatifloxacin was compared with that of ciprofloxacin, levofloxacin and trovafloxacin against 848 recent clinical isolates from cancer patients. Against gram-positive organisms, gatifloxacin was the most active agent tested inhibiting all Aerococcus, Listeria monocytogens, Micrococcus, Stomatococcus mucilaginous, Bacillus, and Rhodococcus equi strains at < or =2 mg/l, its designated susceptibility breakpoint. It was also very active against methicillin-susceptible staphylococci and Streptococcus spp. (including penicillin nonsusceptible Streptococcus pneumoniae and viridans streptococci). It had moderate activity against methicillin-resistant staphylococci and Enterococcus faecalis, inhibiting 68-80% of these strains at < or =2 mg/l. Gatifloxacin also had good activity against the Enterobacteriaceae (although ciprofloxacin was more potent) inhibiting >95% of isolates at < or =1 mg/l. Nonfermentative gram-negative organisms were less susceptible to all 4 agents. Gatifloxacin was very active against Acinetobacter lwoffi (MIC100 0.12 mg/l) and had moderate activity against Acinetobacter baumanii, Chryseobacterium spp., Stenotrophomonas maltophilia, Pseudomonas aeruginosa and other Pseudomonas species. Alcaligenes xylosoxidans strains were relatively resistant to all 4 agents.

Anti-Bacterial Agents↗

Culture-independent analysis of midgut microbiota in the arbovirus vector Culicoides sonorensis (Diptera: Ceratopogonidae).

Differences in midgut microbial communities inhabiting Culicoides spp., insect vectors of virus pathogens, may affect the variation observed in the ability of these biting midges to propagate arthropod-borne viruses. As a first step toward addressing this hypothesis, midgut bacterial communities were compared between Culicoides species expected to be efficient and inefficient vectors of virus pathogens. We used 16S rDNA sequence and restriction fragment information to provisionally identify 36 bacterial genera from guts of wild adult female biting midges, Culicoides sonorensis Wirth and Jones and Culicoides variipennis (Coquillet), from two geographical locations. Bacterial identification was made by sequence analysis of 16S rDNA fragments and by terminal restriction fragment length polymorphism analysis of polymerase chain reaction-amplified 16S rDNA fragments from adult guts. Of 36 bacterial genera identified, 12 had been previously identified in other insects: Comomonas, Enterobacter, Klebsiella, Acinetobacter, Pseudomonas, Stenotrophomonas, Staphylococcus, Chryseobacterium, Moraxella, Acholeplasma, Flavobacterium, and Rickettsia, Significant differences in bacterial community composition were found between all three groups of wild adult females analyzed: live-trapped C. sonorensis, laboratory-emerged C. sonorensis, and laboratory-emerged C. variipennis.

Animals↗

Cutaneous and pulmonary mycosis in green anacondas (Euncectes murinus).

Two dead, captive green anacondas (Eunectes murinus), including one male and one female, submitted for necropsy were in poor body condition, having multiple, scattered, dark red foci on the scales and mottled lungs. Both snakes had severe mycotic dermatitis. In addition, the male snake had mycotic stomatitis, and the female snake had mycotic pneumonia. Trichophyton sp., Verticillium sp., and Alternaria sp. were isolated from the dermal lesions. The pulmonary lesions were morphologically consistent with Aspergillus sp. Bacterial organisms isolated from skin and internal organs included Chryseobacterium meningosepticum, Stenotrophomonas maltophilia, Aeromonas hydrophila, and Providencia rettgeri. Mycotic diseases can be devastating to reptiles, and suboptimal husbandry and captivity were likely the predisposing factors that led to opportunistic invasion in these snakes.

Alternaria↗

Amoebae-resisting bacteria isolated from human nasal swabs by amoebal coculture.

Amoebae feed on bacteria, and few bacteria can resist their microbicidal ability. Amoebal coculture could therefore be used to selectively grow these amoebae-resisting bacteria (ARB), which may be human pathogens. To isolate new ARB, we performed amoebal coculture from 444 nasal samples. We recovered 7 (1.6%) ARB from 444 nasal swabs, including 4 new species provisionally named Candidatus Roseomonas massiliae, C. Rhizobium massiliae, C. Chryseobacterium massiliae, and C. Amoebinatus massiliae. The remaining isolates were closely related to Methylobacterium extorquens, Bosea vestrii, and Achromobacter xylosoxidans. Thus, amoebal coculture allows the recovery of new bacterial species from heavily contaminated samples and might be a valuable approach for the recovery of as-yet unrecognized emerging pathogens from clinical specimens.

Adult↗

Psychrobacters and related bacteria in freshwater fish.

Three phenotypic identification systems were employed to identify 106 strains of gram-negative, nonmotile, aerobic bacteria obtained during iced storage of wild (Salmo trutta and Esox lucius) and farmed (Oncorhynchus mykiss) freshwater fish. Using diagnostic tables and computer-assisted identification, the isolates were Psychrobacter (64 strains), Acinetobacter (24 strains), Moraxella (6 strains), Chryseobacterium (5 strains), Myroides odoratus (2 strains), Flavobacterium (1 strain), Empedobacter (1 strain), and unidentified (3 strains). Overall similarities of all strains were determined for 108 characters by numerical analysis (simple matching coefficient of similarity [S] and clustering by unweighted pair group average linkage [UPGMA]). At the 77% similarity level, 92 strains formed nine major clusters (3 or more strains) and four small clusters (2 strains). Cluster 1 (25 isolates divided into two main subclusters) could be assigned to Psychrobacter phenylpyruvicus, clusters 2 and 3 (26 isolates) were designated as Psychrobacter immobilis, and clusters 4 (3 isolates) and 7 (4 isolates) were identified as Psychrobacter urativorans and Psychrobacter spp., respectively. Clusters 5 (five isolates), 6 (three isolates), and 9 (five isolates) were labeled as Acinetobacter spp., Acinetobacter johnsonii, and Acinetobacter lwoffii, respectively. Cluster 8 (12 isolates), with a high resemblance to Thornley's phenon 4 (a heterogeneous group of bacteria isolated from poultry and related to Acinetobacter), remained unnamed. The restriction pattern was identical for strains grouped into clusters 2 and 3 (P. immobilis) but was different for the remaining Psychrobacter isolates. A large proportion of isolates belonging to the family Moraxellaceae were closely related. Psychrobacters and A. johnsonii were present in freshly caught fish and river water. In the latter stages of storage, P. phenylpyruvicus and acinetobacters tended to decrease, whereas P. immobilis increased.

Animals↗

Sulfobacins A and B, novel von Willebrand factor receptor antagonists. I. Production, isolation, characterization and biological activities.

Sulfobacins A and B, novel von Willebrand factor (vWF) receptor antagonists, have been isolated from the culture broth of Chryseobacterium sp. (Flavobacterium sp.) NR 2993 by ethyl acetate extraction, and by Sephadex LH-20 and silica gel column chromatographies. The physico-chemical properties of the sulfobacins indicate that their structures are completely different from that of aurintricarboxylic acid, the one known vWF receptor antagonist. Sulfobacins A and B inhibit the binding of vWF to its receptor with IC50S of 0.47 and 2.2 microM, respectively. Sulfobacin A also inhibits ristocetin-induced agglutination in human platelets fixed with paraformaldehyde with an IC50 of 0.58 microM.

Alkanesulfonic Acids↗

Sulfobacins A and B, novel von Willebrand factor receptor antagonists. II. Structural elucidation.

Sulfobacins A and B are novel von Willebrand factor (vWF) receptor antagonists produced by Chryseobacterium sp. NR 2993. The structures of sulfobacins A and B have been determined to be (2R,3R)-3-hydroxy-2-[(R)-3-hydroxy-15-methylhexadecanamido]-15- methylhexadecanesulfonic acid and (2R,3R)-3-hydroxy-15-methyl-2-[13-methyltetradecanamido]- hexadecanesulfonic acid, respectively, by various 2D NMR experiments and by methanolysis. The absolute configurations of the sulfobacins were determined by a modified MOSHER's method. The structures are related to sulfonolipids, major components of the cell envelope of gliding bacteria of the genus Cytophaga.

Alkanesulfonic Acids↗

Characteristics of neonatal bacterial meningitis in a teaching hospital in Taiwan from 1984-1997.

During the period from 1984 to 1997, 85 bacterial meningitis neonates with positive cerebrospinal fluid cultures were treated. The ages of these patients ranged from 1 to 28 days. The male to female ratio was 1.7 to 1. The most common causative agent was group B beta-hemolytic streptococci (GBS, 31.8%), followed by Escherichia coli (20%), Proteus mirabilis (7.1%), Enterobacter cloacae (5.9%), other streptococci excluding Streptococcus pneumoniae (5.9%), Chryseobacterium meningosepticum (5.9%), enterococci (4.7%), and Klebsiella pneumoniae (3.5%). Among the 85 patients treated, 51 (60%) were younger than 7 days old. Among them, dyspnea was the most common clinical manifestation. In contrast, fever and diarrhea were seen more frequently in neonates with late onset of disease (after seven days of age). Ampicillin and cefotaxime were the most commonly used antibiotics. The most frequently encountered complications were hydrocephalus and seizures. Since 1991, GBS has overtaken E. coli as the leading cause of neonatal bacterial meningitis. This was accompanied by a fall in the mortality rate, but a sustained high incidence of complications and sequelae. The results of this study highlight the importance of developing strategies to prevent group B streptococcal infection.

Female↗

[Non-fermenting Gram-negative bacilli: distribution in clinical specimens and antimicrobial susceptibility].

In the present work was studied the prevalence, distribution in clinical specimens, and antimicrobial susceptibility of non-fermentative Gram-negative bacilli (NFGNB) from patients attended at Hospital "Angela I. de Llano" (Corrientes, Argentina). A total of 125 strains of NFGNB were recovered from various clinical specimens from July, 1997 to December, 1998. Isolates were identified by classical biochemical tests. Drug sensitivity was performed by standard methods with cefotaxime (CTX), ceftazidime (CAZ), piperacillin (PIP), ampicillin-sulbactam (AMS), piperacillin/tazobactam (TAZ), imipenem (IMP), amikacin (AKN), gentamicin (GEN) and ciprofloxacin (CIP). The most common isolates were Pseudomonas aeruginosa (48.8%); Acinetobacter baumannii (16.8%), Acinetobacter spp. (6.4%), Chryseobacterium spp. (5.6%), Stenotrophomonas maltophilia (4%), and others (18.4%). Most of them were recovered from respiratory secretions (36.0%), and urine (26.4%). IMP was the most effective antimicrobial. Many species of NFGNB showed resistance to several antibiotics tested (CTX, GEN, AMS, and CIP). Due to multiresistance found by more prevalent NFGNB, constant survey of antibacterial sensibility are essential for a correct control and management of nosocomial infections, and ambulatory patients with some risk factors.

Argentina↗

Polyamine distribution profiles in newly validated genera and species within the Flavobacterium-Flexibacter-Cytophaga-Sphingobacterium complex.

Cellular polyamines of 58 strains belonging to the Flavobacterium-Flexibacter-Cytophaga-Sphingobacterium complex were analysed by HPLC. Homospermidine was found in all species of Flavobacterium, Chryseobacterium, Empedobacter, Myroides, Cellulophaga, Salegentibacter, Psychroserpens and Gelidibacter of the family Flavobacteriaceae. Flavobacterium ferrugineum located outside of this family also contained homospermidine. Cytophaga fermentans and C. xylanolytica belonging to the family Bacteroidaceae contained spermidine. Cytophaga marinoflava and C. latercula belonging to Flavobacteriaceae contained homospermidine. The Cytophaga hutchinsonii/C. aurantiaca group contained homospermidine which was the major polyamine in Flexibacter maritimus/ F. ovolyticus of the family Flavobacteriaceae. The Flexibacter sancti/F filiformis/ Cytophaga arvensicola group, F. elegans, F. ruber, F. canadensis, F. flexilis and F. tractuosus, were located separately in different six clusters, and contained homospermidine. The Flexibacter litoralis/F. polymorphus/F. aggregans group contained spermidine, which was detected in Flexibacter roseolus belonging to a divergent cluster. Sphingobacterium and Pedobacter species of the family Sphingobacteriaceae contained homospermidine. Polyamine profiles serve, as a phenotypic chemotaxonomic marker, for the classification of this complex.

Bacteria↗

Characteristics of nosocomial bacterial meningitis in children.

Nosocomial meningitis is uncommon in children. We reviewed the medical records of all children who developed bacterial meningitis at least 72 hours after admission to Mackay Memorial Hospital for the period July 1992 through June 2000. Clinical manifestations, predisposing factors, pathogens, and outcomes were analyzed. Twenty-two cases of nosocomial meningitis were identified, comprising 9.2% (22/239) of all pediatric cases of bacterial meningitis during the study period. The male-to-female ratio was 14:8. All patients were younger than 6 months of age except for one, who was 7 years old. The mean duration between admission and onset of meningitis was 15.3 days (range, 3 to 58 days). Twenty-two organisms were isolated, including 13 Gram-negative bacteria (59%) and 9 Gram-positive bacteria (41%). The most common pathogen was Escherichia coli (5 cases), followed by Enterobacter cloacae (3), Staphylococcus aureus (3), and Chryseobacterium meningosepticum (3). Seventeen patients (77%) had concomitant bacteremia. Predisposing factors for acquisition of nosocomial meningitis included previous treatment with broad-spectrum antibiotics (68%), prematurity with very low birth weight (41%), and total parenteral nutrition (32%). Two patients (9%) had previous neurosurgical intervention. Four patients (18%) died, 3 of whom were low birth weight premature infants. Nine patients (41%) had sequelae, including developmental delay, hydrocephalus, hearing impairment, and epilepsy. Neurosurgery was not a significant risk factor for the development of nosocomial meningitis, while very low birth weight played an important role. Previous intraventricular hemorrhage or hydrocephalus, prematurity with very low birth weight, infection with Gram-negative bacteria, and prior broad-spectrum antibiotic administration were associated with poor outcome.

Bacterial Infections↗

Identification of carbohydrate degrading bacteria in sub-tropical regions.

Several bacteria from the islands of Puerto Rico, Vieques and Trinidad were isolated for their carbohydrate degrading activities. These terrestrial and marine bacterium were collected from pineapple agricultural lands, tropical rain forests, coastlands and mangrove swamps. Organisms were screened for activity using chromogenic substrates (AZCL Megazyme International Ltd., Ireland). The media composition for the effective culturing of some of the marine organisms has also been standardized. Gram-negative organisms were identified by sequence analysis of the PCR- amplified partial small subunit rRNA gene. Results indicate that the majority of the marine organisms tested belong to the genera Vibrio and Pseudoalteromonas and in the terrestrial environments Chryseobacterium predominated. These experiments reveal that sub-tropical environments are potentially good sources of microorganisms with novel carbohydrase activities.

Carbohydrate Metabolism↗

Gram-negative bacteria from the camel tick Hyalomma dromedarii (Ixodidae) and the chicken tick Argas persicus (Argasidae) and their antibiotic sensitivities.

A total of nine species of gram-negative bacteria were isolated from organs and haemolymph of the hard tick Hyalomma (Hyalomma) dromedarii and the soft tick Argas (Persicargas) persicus. Four species namely Serratia liquefaciens, Stenotrophomonas maltophilia, Klebsiella ornithinolytica and Aeromonas hydrophila were isolated from H. dromedarii and five species namely Rahnella aquatilis, Pseudomonas fluorescens, Enterobacter cloacae, Chryseomonas luteola and Chryseobacterium meningosepticum were isolated from A. persicus. Isolated bacteria were identified using the analytical profile index 20E. Disk diffusion test was carried out on all isolated bacteria to determine antibiotic sensitivity of chloramphenicol, amoxillin/clavulanic acid, neomycin, streptomycin, triplesulphur tetracycline and nitrofurantion. The results were discussed.

Animals↗

['In vitro' activity of different antimicrobial agents on Gram-negative nonfermentative bacilli, excluding Pseudomonas aeruginosa and Acinetobacter spp].

Gram-negative nonfermentative bacilli (NFB) are widely spread in the environment. Besides of difficulties for identification, they often have a marked multiresistance to antimicrobial agents, including those active against Pseudomonas aeruginosa. The objective of this study was to evaluate the 'in vitro' activity of different antimicrobial agents on 177 gram-negative nonfermentative bacilli isolates (excluding Pseudomonas aeruginosa and Acinetobacter spp.) isolated from clinical specimens. Minimum inhibitory concentrations (MIC) were determined according to the Mueller Hinton agar dilution method against the following antibacterial agents: ampicillin, piperacillin, piperacillin-tazobactam, sulbactam, cefoperazone, cefoperazone-sulbactam, ceftazidime, cefepime, aztreonam, imipenem, meropenem, colistin, gentamicin, amikacin, trimethoprim-sulfamethoxazole, chloramphenicol, erythromycin, rifampin, norfloxacin, ciprofloxacin and minocycline. Seven isolates: Sphingobacterium multivorum (2), Sphingobacteriumspiritivorum (1), Empedobacterbrevis (1), Weeksella virosa (1), Bergeyella zoohelcum (1) and Oligella urethralis (1), were tested for amoxicillin-clavulanic acid and ampicillin-sulbactam susceptibility, and susceptibility to cefoperazone or sulbactam was not determined. Multiresistance was generally found in Stenotrophomonas maltophilia, Burkholderia cepacia, Chryseobacterium spp., Myroides spp., Achromobacter xylosoxidans, and Ochrobactrum anthropi isolates. On the other hand, Pseudomonas stutzeri, Shewanella putrefaciens-algae, Sphingomonas paucimobilis, and Pseudomonas oryzihabitans, Bergeyella zoohelcum, Weeksella virosa and Oligella urethralis were widely susceptible to the antibacterial agents tested. As a result of the wide variation in antimicrobial susceptibility shown by different species, a test on susceptibility to different antibacterial agents is essential in order to select an adequate therapy. The marked multiresistance evidenced by some species, prompts the need to develop new antimicrobial agents active against this group of bacteria and to search for synergistic combinations.

Anti-Bacterial Agents↗

[Rapid identification of Riemerella anatipestifer on the basis of specific PCR amplifying 16S rDNA].

Riemerella anatipestifer (RA) infection is the main disease causing severe losses in duck production. Because RA is characterized more by the absence than by the presence of specific phenotypic properties and different scholar had the different results of biochemical detection, it can't always be identified quickly and correctly only by the phenotypic properties or biochemical characteristics. The research object was to develop a species specific PCR method for RA detection. Because of the conserved structure of rRNA and appropriate size of 16S rRNA, a multiple alignment of 16S rDNA (gene coding 16S rRNA) was processed among RA, Escherichia coli, Pasteurella multocida, Salmonella enteritidis and Salmonella gallinarum, which are the main bacteria causing duck diseases. A pair of species specific primers named 190f and 843r were selected from the variable regions of 16S rDNA depending on the result of multiple alignment. Using BLAST on NCBI website for a sequence similarity search, the results showed that this pair of primers had very high specificity, except for having a lower sequence similarity with some species of Flavobacterium and Chryseobacterium. A PCR assay was performed and the template was extracted using the bacteria genomic DNA extraction kit and boiling method respectively. A chelate resin named Chelex 100 was used in the boiling method at the same time. Under the annealing temperature of 60 degrees C, all the 26 RA strains, including 19 representative strains of serotypes 1 to approximately 19 and 7 domestic isolated strains, showed the same 654bp fragment after PCR, while there was no amplification with isolates of other bacterial species. Also a series of sensitivity experiments were performed and proved that the detection limit of this method was 50pg genomic DNA, 1.5 x 10(6) CFU/mL and 15 CFU/mL, when the template was prepared with genomic DNA extraction kit, only boiling method and boiling method with Chelex 100 respectively. 12 clinical cases which were probably infected with RA were chosen to identify the accuracy and sensitivity of this PCR method. Several other conventional detecting methods including bacteria isolating, differentiating culture, biochemical experiments and serotyping were used at the same time. The templates of PCR were extracted from brains or livers by boiling method with Chelex 100. Finally, 3 cases were identified as RA infection by the conventional methods and 4 by the PCR method, which proved the good accuracy and sensitivity of the PCR method. Thus, this PCR assay provides a rapid and accurate method for identification of Riemerella anatipestifer. It will help to make the final decision in clinical diagnose or species identification, especially when a new serotype or sub-serotype of RA comes up.

Animals↗

Immunization with bacterial antigens: Flavobacterium and Flexibacter infections.

Seven bacterial species belonging to the Flavobacterium-Cytophaga group are currently considered to be pathogenic for fish. Because they were only recently described and/or because the disease they provoke has little economic significance, no study has been performed concerning immunization against Chryseobacterium scophthalmum, Flavobacterium johnsoniae and Flexibacter ovolyticus. Immunization with Flexibacter maritimus has not been investigated, and fish surviving a natural infection appear to remain equally susceptible to the disease during subsequent outbreaks. On the other hand, interesting data are available concerning immunization against Flavobacterium branchiophilum, Flavobacterium columnare and Flavobacterium psychrophilum because these bacterial species have been known for many years and are responsible for heavy losses in many countries. Surviving fish are usually protected against further infection by the same pathogens. Extensive serological studies have been performed and virulence mechanisms have also been investigated. The review of immunization trials against these three bacterial species shows important variations depending on the species of fish and the route of administration, but in several cases vaccinated fish were successfully protected by high titre of specific antibodies. However, no vaccine is commercially available.

Animals↗

Fermentation, purification, formulation, and pharmacological evaluation of a prolyl endopeptidase from Myxococcus xanthus: implications for Celiac Sprue therapy.

Celiac Sprue is a multi-factorial disease characterized by an inflammatory response to ingested wheat gluten and similar proteins in rye and barley. Proline-rich gluten peptides from wheat, rye, and barley are relatively resistant to gastrointestinal digestion, and therefore persist in the intestinal lumen to elicit immunopathology in genetically susceptible individuals. In this study, we characterize the in vitro gluten detoxifying properties of a therapeutically promising prolyl endopeptidase from Myxococcus xanthus (MX PEP), and describe the development of a prototypical enteric-coated capsule containing a pharmacologically useful dose of this enzyme. A high-cell density fed-batch fermentation process was developed for overproduction of recombinant MX PEP in E. coli, yielding 0.25-0.4 g/L purified protein. A simple, scalable purification and lyophilization procedure was established that yields >95% pure, highly active and stable enzyme as a dry powder. The dry powder was blended with excipients and encapsulated in a hard gelatin capsule. The resulting capsule was enteric coated using Eudragit L30-D55 polymer coat, which provided sufficient resistance to gastric conditions (> 1 h in 0.01 M HCl, pH 2 with pepsin) and rapid release under duodenal conditions (15-30 min release in pH 6.0 in the presence of trypsin and chymotrypsin). In conjunction with pancreatic enzymes, MX PEP breaks down whole gluten into a product mixture that is virtually indistinguishable from that generated by the Flavobacterium meningosepticum (FM) PEP as judged by chromatographic assays. Competitive studies involving selected immunogenic peptides mixed with whole gluten reveal that both PEPs have a wide range of substrate specificity. Our results support further in vitro and in vivo evaluation of the MX PEP capsule as an oral therapeutic agent for Celiac Sprue patients.

Celiac Disease↗