Flavobacterium

Acta Pathol Microbiol Immunol Scand {B}, 1987 Feb, 95(1), 41 - 7
Phenotypic characterization of Flavobacterium meningosepticum strains identified by DNA-DNA hybridization; Bruun B et al.; Fifty-two strains found to belong to Flavobacterium meningosepticum on the basis of DNA-DNA hybridization analyses were characterized and found to be phenotypically homogeneous . All strains were oxidase, catalase, indole, gelatinase and beta-galactosidase positive, and produced acid from glucose, mannose, fructose, maltose and mannitol; nitrate was not reduced . F . meningosepticum could be differentiated from Flavobacterium group IIb by its ability to produce beta-galactosidase, and by the latter taxon's ability to produce a bright yellow pigment in contrast to the weak or non-existing pigmentation of F . meningosepticum . Phenotypic characteristics that could differentiate between the two main DNA relatedness groups of F . meningosepticum were not discovered, wherefore a subdivision of the present species into two species cannot be recommended . Strains from the DNA relatedness groups comprising 19 of 20 CSF isolates were found to be able to grow at 40 degrees C and to produce a weak yellow pigment; in contrast, strains from the three other DNA relatedness groups were unable to grow at 40 degrees C and produced no pigment.

Acta Pathol Microbiol Immunol Scand {B}, 1987 Feb, 95(1), 33 - 9
Genetic heterogeneity of Flavobacterium meningosepticum demonstrated by DNA-DNA hybridization; Ursing J et al.; DNA-DNA reassociation studies on 52 strains of Flavobacterium meningosepticum showed two main hybridization groups about 40-55% interrelated and comprising 4 and 48 strains respectively . The larger group could be further divided into four subgroups if differences in thermal stability of the reassociated duplexes were taken into consideration . Of a total of 20 strains isolated from cerebrospinal fluid, 18 were found in one subgroup of 28 strains, which also contained seven blood isolates . The results indicate that genetically defined subgroups within the species might differ with regard to pathogenic significance.

Med Microbiol Immunol (Berl), 1987, 176(2), 103 - 11
Flavobacterium group IIb bacteremia: report of a case and review of Flavobacterium infections; Siegman-Igra Y et al.; A case of nonfatal Flavobacterium CDC group IIb bacteremia in a hospitalized woman is presented . The portal of entry of the organism was either an intravenous line or an open, eroded wound associated with bilateral breast carcinoma . Flavobacterium infections in the post-neonatal age group are reviewed.

J Appl Bacteriol, 1987 Jan, 62(1), 29 - 41
Identification and distribution of Flavobacterium meningosepticum in clinical material; Holmes B; During the 19 year period ending December 1984, 82 (1.7%) of 4840 strains of Gram-negative non-fermentative bacteria submitted to the National Collection of Type Cultures (NCTC) for computer-assisted identification were Flavobacterium meningosepticum . These figures suggest either that F . meningosepticum occurs only rarely in clinical material in the UK, or that when it does occur it is not always recognized . The sources from which the NCTC strains were isolated are reported and also the characteristics by which the species may be identified . The clinical significance of F . meningosepticum is reviewed, particularly its role in neonatal meningitis, of which there have been but two reported cases in the UK, both imported . The susceptibility of the species to antimicrobial agents is discussed and also antimicrobial therapy of neonatal meningitis due to this species.

Scand J Infect Dis Suppl, 1987, 52, 26 - 31
Intramuscular imipenem/cilastatin for treatment of mild and moderately severe bacterial infections; Keller CA et al.; The efficacy and safety of intramuscularly administered imipenem/cilastatin was studied in 70 patients with mild or moderately severe bacterial infections (skin and soft tissue infections, respiratory tract infections, urinary tract infections and pelvic infections) . Doses of imipenem/cilastatin ranged from 0.5 to 0.75 g twice daily . Fifty-five patients were evaluable for bacteriological efficacy; in the remaining 15 patients no pathogens were isolated or susceptibility data were lacking . MIC50 and MIC90 of imipenem were 0.12 mg/l and 0.5 mg/l, respectively, for Gram-negative pathogens isolated and 0.25 mg/l and 0.5 mg/l, respectively, for Gram-positive pathogens . Only one strain (a Flavobacterium odoratum) was resistant to imipenem . Clinical cure and bacteriological elimination was achieved in 94% of evaluable patients while 3% showed marked clinical improvement . Two patients were considered therapeutic failures . No clinical adverse effects were noted . Abnormal liver transaminases were recorded in 23% of the patients and 11% developed eosinophilia . In no patient was imipenem/cilastatin discontinued due to adverse effects . It is concluded that intramuscular imipenem/cilastatin in these patients was well tolerated and efficacious.

Biochem Biophys Res Commun, 1986 Dec 15, 141(2), 825 - 30
Catabolism of pentachlorophenol by a Flavobacterium sp; Steiert JG et al.; The pathway employed for pentachlorophenol (PCP) degradation by an aerobic, chlorophenol-utilizing Flavobacterium sp . was initiated by conversion of PCP to tetrachloro-p-hydroquinone (TCH) . 18O labelling experiments demonstrated that the first dechlorination, where a hydroxyl replaced the chlorine at PCP ring position number 4, involved a hydrolytic reaction . Then two reductive dechlorinations of TCH followed to yield firstly trichlorohydroquinone (TrCH) and then 2,6-dichlorohydroquinone (DCH) . Thus, the initial steps in catabolism of PCP by the Flavobacterium were: PCP----TCH----TrCH.

FEBS Lett, 1986 Dec 15, 209(2), 235 - 7
Specificity of prolyl endopeptidase; Nomura K; A series of tetrapeptides, Cbz(Bz)-Gly-X-Leu-Gly, were synthesized and the kinetic parameters, kcat and kcat/Km, determined for their hydrolyses by prolyl endopeptidase from Flavobacterium . The peptides with X = N-Me-Ala, Sar and Ala as well as the standard substrate (X = Pro) were found to be good substrates, while those with X = alpha-aminobutyryl, Hyp, Ser and Gly were poor substrates, and those with X = pipecolyl, alpha-aminoisobutyryl, N-Me-Val, N-Me-Leu, Hyp(O-Bzl) and Ser(O-Bzl) were not cleaved at all . These results suggest that the specificity-determining site or S1 subsite of the enzyme is designed to fit exactly the proline residue of the substrate with allowance for the residues carrying substituents at the N and/or C alpha which must not exceed the size of the pyrrolidine ring of proline.

Thromb Res, 1986 Dec 1, 44(5), 599 - 610
Removal of the anticoagulant activities of the low molecular weight heparin fractions and fragments with flavobacterial heparinase; Yang VC et al.; Recently, the development of low molecular weight heparin fractions and fragments (LMHF) as potential antithrombotic agents has gained increased attention . However, the lack of antagonists to neutralize the anticoagulant effects of these drugs may seriously exclude them from possible uses in extracorporeal therapy . This is mainly because of the concern that the high dosage of the drugs employed in extracorporeal therapy could lead to serious bleeding risks . Our earlier work has demonstrated that immobilized heparinase can remove polydisperse heparin both in vitro and in vivo . To examine whether such a system may be used as a novel approach to neutralize the anticoagulant effects of LMHF, different LMHF were tested using heparinase . In vitro data showed that both the APTT and anti-FXa activities of the LMHF including Kabi 2165, PK 10169, Cy 216 and CY 222 were nearly completely eliminated by heparinase in less than 20 min . This study suggests that an immobilized heparinase system may be an useful element for the acceptance of the LMHF for their use in extracorporeal therapy.

J Appl Bacteriol, 1986 Nov, 61(5), 421 - 6
A note on starch hydrolysis and beta-glucuronidase activity among flavobacteria; Petzel JP et al.; Most flavobacteria tested with the fluorogenic substrate 4-methylumbelliferyl-beta-D-glucuronide possessed beta-glucuronidase (GUD), but when some of the same strains were tested with the API ZYM gallery, all were negative for GUD . Conflicting reports also appear in the literature about starch hydrolysis among flavobacteria . We observed that the results obtained can depend on the medium used and the length of incubation . Our results indicate that GUD activity and starch hydrolysis are more widely distributed in the genus Flavobacterium than previously reported.

Atherosclerosis, 1986 Nov, 62(2), 151 - 8
Effect of very low molecular weight heparin-derived oligosaccharides on lipoprotein lipase release in rabbits; Merchant ZM et al.; Oligosaccharide fragments of heparin were prepared using flavobacterial heparinase . Following sizing, these oligosaccharide fractions were administered (i.v.) to rabbits and were examined for their ability to release lipoprotein lipase . The decasaccharides (dp = 10, Mr avg = 2,800) were the smallest oligosaccharides which resulted in substantial lipase release . The plasma lipase levels obtained with decasaccharides were comparable to low molecular weight heparin and one-third those obtained when heparin was administered at an equivalent dose . The peak plasma lipase concentration was observed 10 min following heparinization and fell off rapidly over the 60-min time course . The lipase release activity paralleled the in vivo pharmacokinetics of the heparin and decasaccharide sample as determined by monitoring their anti-Factor Xa activity . No activation of purified bovine milk lipoprotein lipase or plasma lipase was detectable at the concentrations studied, indicating that the increase in circulating lipolytic activity was due entirely to release . Lipoprotein lipase accounted for a major portion of the released activity with hepatic triglyceride lipase representing the remainder of the lipolytic activity . The sized decasaccharide sample was characterized with regards to its structure and anticoagulant activity . The decasaccharides exhibited reduced anticoagulant activity possibly making it a better drug candidate in the treatment of atherosclerosis.

Arch Microbiol, 1986 Nov, 146(2), 125 - 9
Biodegradation of polyethylene glycol by symbiotic mixed culture (obligate mutualism); Kawai F et al.; Neither Flavobacterium sp . nor Pseudomonas sp . grew on a polyethylene glycol (PEG) 6000 medium containing the culture filtrate of their mixed culture on PEG 6000 . The two bacteria did not grow with a dialysis culture on a PEG 6000 medium . Flavobacterium sp . grew well on a dialysis culture containing a tetraethylene glycol medium supplemented with a small amount of PEG 6000 as an inducer, while poor growth of Pseudomonas sp . was observed . Three enzymes involved in the metabolism of PEG, PEG dehydrogenase, PEG-aldehyde dehydrogenase and PEG-carboxylate dehydrogenase (ether-cleaving) were present in the cells of Flavobacterium sp . The first two enzymes were not found in the cells of Pseudomonas sp . PEG 6000 was degraded neither by intact cells of Flavobacterium sp . nor by those of Pseudomonas sp., but it was degraded by their mixture . Glyoxylate, a metabolite liberated by the ether-cleaving enzyme, inhibited the growth of the mixed culture . The ether-cleaving enzyme was remarkably inhibited by glyoxylate . Glyoxylate was metabolized faster by Pseudomonas sp . than by Flavobacterium sp., and seemed to be a key material for the symbiosis.

J Biochem (Tokyo), 1986 Oct, 100(4), 1049 - 55
Purification and properties of glucoside 3-dehydrogenase from Flavobacterium saccharophilum; Takeuchi M et al.; A membrane-bound glucoside 3-dehydrogenase {EC 1.1.99.13}, which oxidizes validoxylamine A to the 3-keto derivative, was solubilized from the membrane fraction of Flavobacterium saccharophilum by Triton X-100 and purified about 280-fold with an overall yield of 30% from the membrane fraction by column chromatography on DEAE- and CM-Sepharose CL-6B and gel filtration on Sephacryl S-300 . The purified enzyme exhibited a single protein band on disc gel electrophoresis, and FAD was shown to be the prosthetic group . The enzyme had a molecular weight of 270,000 as determined by gel filtration on Sephacryl S-300 and consisted of 4 identical subunits each with a molecular weight of 66,000 . The enzyme reacted with various artificial electron acceptors such as 2,6-dichlorophenolindophenol (DCIP), phenazine methosulfate, and ferricyanide . The optimum pH for DCIP reductase activity was 6.0 . The enzyme was inhibited by Hg2+ and p-chloromercuribenzoate . D-Glucose and methyl-alpha- and beta-D-glucoside showed the highest susceptibility to the enzyme, and were converted to the corresponding 3-keto sugars.

J Gen Microbiol, 1986 Oct, 132 ( Pt 10), 2723 - 32
Characterization of Flavobacterium species by analysis of volatile fatty acid production; Rasoamananjara D et al.; Seventy-four Flavobacterium strains were characterized by gas-liquid chromatographic analysis of volatile fatty acids produced in the culture medium . Principal components analysis permitted the graphic representation of the relative positions of the different strains, and aggregation according to the variance enabled a hierarchical classification to be established . The study revealed three subgroups each for F . meningosepticum and F . odoratum . Our F . breve, Flavobacterium sp . group IIb and F . multivorum strains appeared to be homogeneous . These results tallied with those of previous studies on DNA base composition and reassociation, electrophoretic protein profiles and cellular fatty acid composition.

J Biol Chem, 1986 Sep 15, 261(26), 12000 - 5
Transfer of glycerol by Endo-beta-N-acetylglucosaminidase F to oligosaccharides during chitobiose core cleavage; Trimble RB et al.; N-Linked oligosaccharides, when hydrolyzed by glycerol-containing preparations of endo-beta-N-acetylglucosaminidase (Endo) F from Flavobacterium meningosepticum were found to have glycerol attached to their reducing ends . The absence of a reducing end was confirmed by high-field 1H NMR spectroscopy, and the incorporated glycerol was verified through mass spectrometry and collisionally activated decomposition fast atom bombardment/mass spectrometry/mass spectrometry techniques . Periodate oxidation of {1(3)-14C}glycerol-labeled oligosaccharides indicated glycerol was glycosidically linked via its 1(3) carbon to the C1 of the reducing end N-acetylglucosamine . In a second, less favored reaction, the glycerol glycoside was hydrolyzed by Endo F using water as the terminal nucleophile, thus regenerating the N-acetylglucosamine reducing end . Glycerol could be removed from Endo F preparations without affecting enzyme stability, and chitobiosyl core hydrolysis in its absence provided intact oligosaccharides with normal N-acetylglucosamine reducing ends . The incorporation of labeled glycerol may provide a useful method for monitoring of Endo F release of oligosaccharides.

Quad Sclavo Diagn, 1986 Sep, 22(3), 318 - 29
{Isolation of Flavobacterium odoratum from human matter}; Andreoni S; Described the most important taxonomical, bacteriological, clinical and epidemiological aspects of microorganisms of Flavobacterium genus, the author refer about 8 strains of Flavobacterium odoratum isolated from urine (6), sputum (1), surgical exudate (1) . In some cases, etiopathogenetic possible correlation between isolates and clinical disease discussed, the author emphasizes the large antibiotic resistance kind of this species of microorganism non fermenter.

Mikrobiologiia, 1986 Sep-Oct, 55(5), 872 - 4
{Coagulation of bacteria by the action of montmorrillonite}; Nikovskaia GN et al.; Conditions were studied for the coagulation of aggregation-stable Bacillus subtilis, Flavobacterium rigens and Escherichia coli cell dispersions . Their aggregative stability decreased when montmorillonite, a clay mineral, was added to a weakly acid medium or in the presence of Al3+ ions . Bacterial heterocoagulation under the action of montmorillonite can be used as a universal strategy for biomass isolation from aqueous dispersion media.

J Biochem (Tokyo), 1986 Sep, 100(3), 773 - 80
Substrate specificity of endo-beta-galactosidases from Flavobacterium keratolyticus and Escherichia freundii is different from that of Pseudomonas sp; Ito M et al.; The substrate specificity of endo-beta-galactosidase of Pseudomonas sp . was found to differ from that of Flavobacterium keratolyticus or Escherichia freundii, based on the following experimental results . The endo-beta-galactosidases from these three bacteria released 6-O-sulfo-GlcNAc beta 1-3Gal as one of the major products from keratan sulfates from different sources . In addition to the sulfated disaccharide, Flavobacterium and Escherichia enzymes produced GlcNAc beta 1-3Gal, which is also an integral repeating unit of keratan sulfate, whereas the Pseudomonas enzyme did not release any non-sulfated disaccharide . Tetrasaccharides were prepared from the teleost skin keratan sulfate by digestion with Pseudomonas enzyme followed by gel filtration on Sephadex G-50 chromatography . A part of the tetrasaccharide fraction was hydrolyzed by Flavobacterium enzyme to produce 6-O-sulfo-GlcNAc beta 1-3Gal and GlcNAc beta 1-3Gal, whereas the fraction was completely resistant to retreatment with the Pseudomonas enzyme . Endo-beta-galactosidases from F . keratolyticus and E . freundii hydrolyzed the internal beta-1,4-galactosyl linkage of various neolacto-type glycosphingolipids to produce glucosylceramides . However, these glycosphingolipids were completely resistant to the Pseudomonas enzyme . These findings clearly show that the sulfation on the N-acetylglucosamine adjacent to galactose in the lactosaminoglycans is essential for expression of the Pseudomonas enzyme, but not for that of the Flavobacterium or Escherichia enzyme.

Biochimie, 1986 Sep, 68(9), 1087 - 96
Isolation and some properties of an arylamidase from Flavobacterium IIb; Milliere JB et al.; A constitutive L-leucylarylamidase (EC 3.4.11) hydrolase able to cleave L-aminoacyl-beta naphthylamide and L-aminoacyl-4 nitroanilide substrates, was isolated from sonicated cells of Flavobacterium IIb and partially purified with a 0.9% yield and a 159-fold recovery . Its molecular weight was estimated to be about 170,000 +/- 10% . This arylamidase exhibited optimum activity at pH 7.0 and 28 degrees C for the hydrolysis of L-leucine-4NA and is inhibited strongly by metal chelating agents, and to a weaker extent, by some sulfhydryl and reducing agents . Heavy metal ions: Cd2+, Zn2+, Cu2+, Hg2+ and Co2+, markedly inhibit it, and Zn2+ is a competitive inhibitor . This metalloenzyme, free of carboxypeptidase, proteinase and L-leucine aminopeptidase (L-leucylglycine substrate) activities, hydrolyzes aminoacyl-beta NA, aminoacyl-4NA and some dipeptides with unsubstituted amino groups of the L-configuration . The lowest Km values are associated with substrates having neutral or basic residues, with large side chains.

Appl Environ Microbiol, 1986 Jul, 52(1), 92 - 7
Pentachlorophenol degradation: a pure bacterial culture and an epilithic microbial consortium; Brown EJ et al.; The steady-state growth of a Flavobacterium strain known to utilize pentachlorophenol (PCP) was examined when cellobiose and PCP simultaneously limited its growth rate in continuous culture . A concentration of 600 mg of PCP per liter in influent medium could be continuously degraded without affecting steady-state growth . We measured specific rates of PCP carbon degradation as high as 0.15 +/- 0.01 g (dry weight) of C per h at a growth rate of 0.045 h-1 . Comparable specific rates of PCP degradation were obtained and maintained by PCP-adapted, natural consortia of epilithic microorganisms . The consortium results suggest that a fixed-film bioreactor containing a PCP-adapted natural microbial population could be used to treat PCP-contaminated water.

J Biochem (Tokyo), 1986 Jun, 99(6), 1571 - 7
Chemical modification by diethylpyrocarbonate of an essential histidine residue in 3-ketovalidoxylamine A C-N lyase; Takeuchi M et al.; 3-Ketovalidoxylamine A C-N lyase of Flavobacterium saccharophilum is a monomeric protein with a molecular weight of 36,000 . Amino acid analysis revealed that the enzyme contains 5 histidine residues and no cysteine residue . The enzyme was inactivated by diethylpyrocarbonate (DEP) following pseudo-first order kinetics . Upon treatment of the inactivated enzyme with hydroxylamine, the enzyme activity was completely restored . The difference absorption spectrum of the modified versus native enzyme exhibited a prominent peak around 240 nm, but there was no absorbance change above 270 nm . The pH-dependence of inactivation suggested the involvement of an amino acid residue having a pKa of 6.8 . These results indicate that the inactivation is due to the modification of histidine residues . Substrates of the lyase, p-nitrophenyl-3-ketovalidamine, p-nitrophenyl-alpha-D-3-ketoglucoside, and methyl-alpha-D-3-ketoglucoside, protected the enzyme against the inactivation, suggesting that the modification occurred at or near the active site . Although several histidine residues were modified by DEP, a plot of log (reciprocal of the half-time of inactivation) versus log (concentration of DEP) suggested that one histidine residue has an essential role in catalysis.

Acta Pathol Microbiol Immunol Scand {B}, 1986 Jun, 94(3), 145 - 52
An investigation of a collection of yellow-pigmented Pseudomonas; Sogaard P et al.; Thirty-six strains of yellow-pigmented Pseudomonas from clinical as well as non-clinical material and 11 reference strains of Pseudomonas were investigated by means of conventional bacteriological methods (a total of 53 different tests) . Eighteen of the 36 yellow-pigmented strains could be classified as P . paucimobilis . Apart from the presence of lipid inclusions on beta-hydroxybutyrate, hydrolysis of DNA, and Tween 80 our results showed a high degree of accordance with other investigations . Eight strains showed characteristics compatible with inclusion in the CDC VE group; one orange-yellow strain showed the characteristics of P . vesicularis, and one was a pyoverdin negative, yellow P . putida . Eight strains remained unidentified . Strains of P . paucimobilis were most often resistant to antibiotics used for P . aeruginosa infections (viz . piperacillin, cefsulodin, ceftazidime) while the strains of the CDC VE group were often susceptible . Most strains were susceptible to the aminoglycosides . The difficulties in distinguishing yellow-pigmented strains of Pseudomonas from Flavobacterium spp . or Xanthomonas spp . are discussed.

Am J Physiol, 1986 May, 250(5 Pt 2), H879 - 88
Anticoagulantly active heparin-like molecules from mast cell-deficient mice; Marcum JA et al.; To assess the contribution of mast cells to the maintenance of blood fluidity, the hindlimb vasculature of mast cell-deficient mice (W/Wv) and littermates containing normal levels of mast cells (+/+), were perfused with purified human thrombin and antithrombin . Enzyme-inhibitor complex generation within the vasculature was enhanced to a comparable extent for W/Wv and +/+ mice over the uncatalyzed rate, that level of complex produced within a similar time interval in the absence of heparin . Perfusion of purified Flavobacterium heparinase prior to infusion of the hemostatic components, or perfusion of antithrombin modified at the heparin-binding domain, reduced W/Wv and +/+ hindlimb thrombin-antithrombin complex formation to the uncatalyzed rate . To further define the cellular source of the vascular-associated heparin-like molecules, endothelial cells isolated from epididymal fat pads of W/Wv and +/+ mice were grown in vitro . The acceleration of thrombin-antithrombin interactions in the presence of endothelial cell-derived glycosaminoglycans was similar for W/Wv and +/+ mice, was abolished with purified bacterial heparinase, and was expressed to only a minor extent when utilizing modified antithrombin . The biologically active mucopolysaccharides appear to be present on the cell surface.

Appl Environ Microbiol, 1986 May, 51(5), 926 - 30
Identification of a plasmid-borne parathion hydrolase gene from Flavobacterium sp . by southern hybridization with opd from Pseudomonas diminuta; Mulbry WW et al.; Parathion hydrolases have been previously described for an American isolate of Pseudomonas diminuta and a Philippine isolate of Flavobacterium sp . (ATCC 27551) . The gene which encodes the broad-spectrum organophosphate phosphotriesterase in P . diminuta has been shown by other investigators to be located on a 66-kilobase (kb) plasmid . The intact gene (opd, organophosphate-degrading gene) from this degradative plasmid was cloned into M13mp10 and found to express parathion hydrolase under control of the lac promoter in Escherichia coli . In Flavobacterium sp . strain ATCC 27551, a 43-kb plasmid was associated with the production of parathion hydrolase by curing experiments . The M13mp10-cloned fragment of the opd gene from P . diminuta was used to identify a homologous genetic region from Flavobacterium sp . strain ATCC 27551 . Southern hybridization experiments demonstrated that a genetic region from the 43-kb Flavobacterium sp . plasmid possessed significant homology to the opd sequence . Similar hybridization did not occur with three other native Flavobacterium sp . plasmids (approximately 23, 27, and 51 kb) present within this strain or with genomic DNA from cured strains . Restriction mapping of various recombinant DNA molecules containing subcloned fragments of both opd plasmids revealed that the restriction maps of the two opd regions were similar, if not identical, for all restriction endonucleases tested thus far . In contrast, the restriction maps of the cloned plasmid sequences outside the opd regions were not similar . Thus, it appears that the two discrete bacterial plasmids from parathion-hydrolyzing soil bacteria possess a common but limited region of sequence homology within potentially nonhomologous plasmid structures.

J Nutr Sci Vitaminol (Tokyo), 1986 Apr, 32(2), 137 - 45
Menaquinone (vitamin K2)-6 production by mutants of Flavobacterium meningosepticum; Tani Y et al.; Flavobacterium meningosepticum IFO 12535, a menaquinone (MK) producer, was mutagenized to improve productivity . A mutant, which was resistant to 1-hydroxy-2-naphthoate (HNA), was found to produce MK more abundantly: 34 mg/liter of culture broth and 5.5 mg/g of dry cells . The mutant was less sensitive to inhibition by HNA on MK biosynthesis than the wild-type strain . MK was isolated from cells of the mutant and identified as MK-6 based on its physicochemical characteristics . Mutants, which were given each KCN resistance, aromatic amino acid auxotrophy and no carotenoid productivity, did not show further increase of productivity.

South Med J, 1986 Apr, 79(4), 518 - 9
Sepsis from Flavobacterium meningosepticum, an uncommon pathogen with unusual susceptibility patterns; Burnakis TG et al.; Flavobacterium meningosepticum, because of its unusual susceptibility patterns and predilection for debilitated or immunocompromised hosts, can be a dangerous pathogen . Identification of the organism and proper susceptibility studies, which are essential in designing antibiotic therapy, helped in the selection of a regimen that proved curative in this case.

Infect Control, 1986 Apr, 7(4), 216 - 9
Microbial examination of kidney lithotripter tub water and epidural anesthesia catheters; Cooper GL et al.; Kidney lithotripsy patients frequently receive epidural anesthesia via indwelling epidural catheters . In our hospital, patients are immersed in a tub of warm, continuously-flowing tap water . The epidural catheter-entry site is covered by a transparent occlusive dressing . To determine the risk of microbial colonization of the epidural catheter during lithotripsy, we performed quantitative cultures of tub water and semiquantitative cultures of catheters in 63 lithotripsy procedures . Most of the tub water organisms were typical tap water and skin flora isolates . Total colony counts were generally low with no significant progression during the course of serial procedures . Forty-two epidural catheters were cultured; 34 (81%) were sterile, 8 (19%) were colonized with small numbers of flavobacteria or coagulase-negative staphylococci . Only four catheters had organisms present on catheter segments covered by the transparent occlusive dressing (in each case there was a single colony forming unit per semiquantitative plate) and these organisms were probable contaminants . We conclude that with our current lithotripsy procedures, the risk for the development of epidural catheter-associated infection seems to be low.

Appl Environ Microbiol, 1986 Mar, 51(3), 640 - 6
Sulfur regulation of heparinase and sulfatases in Flavobacterium heparinum; Cerbelaud EC et al.; Sulfur regulation of heparinase synthesis and sulfatase synthesis was studied in Flavobacterium heparinum . Heparinase synthesis was strongly repressed by sulfate and L-cysteine, while the activity of this enzyme showed little or no inhibition by these compounds . Heparinase was synthesized in the absence of heparin when L-methionine was used as the sole sulfur source . The sulfatases produced by F . heparinum, which include the sulfatases involved in heparin catabolism, were also studied . At least some of the sulfatase activity was regulated by sulfur compounds in a manner similar to heparinase regulation . L-Cysteic acid and taurine were not suitable sulfur sources to support the growth of F . heparinum.

Vet Microbiol, 1986 Mar, 11(3), 261 - 70
Phenotypic and genetic relationships of so-called Moraxella (Pasteurella) anatipestifer to the Flavobacterium/Cytophaga group; Piechulla K et al.; So-called Moraxella (or Pasteurella) anatipestifer and members of the Flavobacterium/Cytophaga group exhibit remarkable common features: lack of flagellation, low guanine + cytosine content of the chromosomal DNA, production of menaquinones and branched-chain fatty acids, absence of carbohydrate fermentation, and similar patterns of hydrolytic enzymes . Using the renaturation method of DNA:DNA hybridization two urease-negative European isolates and the urease-positive type strain (which was isolated in the United States) of M . P . anatipestifer were shown to have about 85% of their genome DNA base sequences in common; they may represent two subspecies . The type strain of this species was neither measurably related to the type species of the genus Moraxella nor to selected members of the family Pasteurellaceae (Pohl 1981) . On the other hand, low but significant degrees of DNA binding between selected strains of so-called M . anatipestifer, Cytophaga marinoflava, Flavobacterium meningosepticum, F . odoratum and F . pectinovorum were observed . On the basis of these findings the transfer of the so-called M . anatipestifer to the Flavobacterium/Cytophaga group (family Cytophagaceae) is proposed . More detailed investigations are required to establish its relationship at the genus level.

Carbohydr Res, 1986 Mar 1, 147(1), 87 - 100
Structural differences of dermatan sulfates from different origins; Poblacion CA et al.; The dermatan sulfates from hog, rat, rabbit, and beef liver, hog, rat, beef, and dog spleen, and hog skin were isolated and submitted to structural analysis . All of them migrated as single bands, close to the standard position for dermatan sulfate in agarose-gel electrophoresis . In polyacrylamide gel, however, each dermatan sulfate showed a characteristic electrophoretic migration-pattern: one, two, or three polydisperse bands, corresponding to different molecular weights, were obtained for the dermatan sulfates according to their origins . Chemical analysis showed that all of the dermatan sulfates here described are hybrid polymers composed of D-glucuronic and L-iduronic acid-containing disaccharide units . The relative position of these units in the polymer chains and the presence of 6-sulfated disaccharides were determined with the aid of chondroitinases B and AC from Flavobacterium heparinum . These studies show that each dermatan sulfate has a unique structure as regards the molecular weight, the presence of 6-sulfated disaccharide units, and also the relative amount and position of glucuronic and iduronic acid residues in the chains . These findings suggests a tissue- and species-specificity for the dermatan sulfates.

Proc Soc Exp Biol Med, 1986 Mar, 181(3), 432 - 7
Deglycosylation of gonadotropins with an endoglycosidase; Swedlow JR et al.; A commercially available endoglycosidase (N-glycanase, Genzyme, Boston, Mass.) purified from Flavobacterium meningosepticum with a specificity for cleaving asparagine-linked carbohydrate moieties in glycoproteins was tested on several pituitary and chorionic gonadotropins as substrates . All intact hormones tested were resistant to the action of the enzyme as were all beta subunits from the respective gonadotropins . All alpha subunits, however, were susceptible to the enzyme as evidenced by a decrease in molecular size when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) . Preparative experiments with ovine luteinizing hormone subunit (oLH alpha) indicated that only 35-40% of the carbohydrate was removed after N-glycanase treatment, suggesting that perhaps only one of the two carbohydrate moieties was cleavable under the conditions employed . The enzyme-modified subunit (DG-oLH alpha) was able to recombine with untreated oLH beta . An in vitro steroidogenic bioassay (rat Leydig cell) showed that the recombinant (DG-oLH alpha-oLH beta) was about 22% as potent as the native oLH, but in a testicular membrane binding assay for LH, it was equal in potency to the native hormone in competing with the radioligand.

J Clin Microbiol, 1986 Feb, 23(2), 267 - 73
Chemical characterization of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf; Dees SB et al.; The cellular fatty acid, sphingolipid, and isoprenoid quinone compositions of Flavobacterium odoratum, Flavobacterium breve, and Flavobacterium-like groups IIe, IIh, and IIf were determined, using thin-layer, gas-liquid, and reverse-phase high-performance liquid chromatography . The fatty acid data showed that groups IIe, IIh, and IIf were similar to recognized Flavobacterium species by the presence of relatively large amounts of iso-branched hydroxy and nonhydroxy acids . Groups IIe and IIh were essentially identical in fatty acid composition but were distinguished from group IIf, F . breve, and F . odoratum on the basis of minor qualitative and quantitative differences . All strains tested contained menaquinone 6 as the major isoprenoid quinone, and all lacked sphingolipids . Overall, the chemical data suggest that groups IIe, IIh, and IIf are additional Flavobacterium species and are different from sphingobacteria, which contain sphingolipid and menaquinone 7 as the major quinone.

Biochemistry, 1986 Jan 28, 25(2), 405 - 10
Effects of heparan sulfate removal on attachment and reattachment of fibroblasts and endothelial cells; Gill PJ et al.; Human skin fibroblasts and calf aorta endothelial cells were grown as tissue culture monolayers in the presence of {35S}sulfate in order to label the glycosaminoglycan portions of proteoglycans for investigation of their role in cell attachment . The {35S}glycosaminoglycans were then selectively removed from the cell monolayers by the addition of various glycosaminoglycan-degrading enzymes . As previously described, in contrast to trypsin treatment none of these enzymes removed any cells from the culture plates . Incubation with a preparation from Flavobacterium heparinum left only small stubs of {35S}glycosaminoglycans on the cell monolayers, indicating that all the cell-surface proteoheparan {35S}sulfate and proteochondroitin {35S}sulfate was accessible to this enzyme preparation . The treatment did not change the amount or time of incubation with trypsin necessary for release of the cells from the monolayers . Thus, cell attachment was not weakened by removal of heparan sulfate or chondroitin sulfate . In contrast, neither fibroblasts nor endothelial cells in suspension would reattach in the presence of the F . heparinum preparation while reattachment occurred readily in the presence of chondroitin ABC lyase . This provides evidence that heparan sulfate, but not chondroitin sulfate, is involved in the process of cell attachment even though neither is necessary for maintaining attachment.

J Biol Chem, 1986 Jan 5, 261(1), 172 - 7
Requirements of cleavage of high mannose oligosaccharides in glycoproteins by peptide N-glycosidase F; Chu FK; Peptide N-glycosidase from Flavobacterium meningosepticum cleaves complex as well as neutral glycoproteins (Plummer, T.H., Jr., Elder, J.H., Alexander, S., Phelan, A.W., and Tarentino, A.L . (1984) J . Biol . Chem . 259, 10700-10704) . Examples of neutral glycoprotein substrates include ribonuclease B (one high mannose oligosaccharide chain) and yeast external invertase (nine chains/invertase subunit) . The rate of deglycosylation by the glycosidase was greatly enhanced if the glycoprotein substrate was denatured prior to enzyme treatment, from a low of 11-fold for external invertase to a high of 844-fold for ribonuclease B . Peptide N-glycosidase F was unable to cleave the asparaginyl-N-acetylglucosamine bond in endo-beta-N-acetylglucosaminidase H-modified external invertase or ribonuclease B, although that in similarly modified glycopeptide substrate was cleaved . Ribonuclease B was digested sequentially with various exoglycosidases to produce an oligosaccharide chain of varied length . Using the resulting forms of ribonuclease B as substrates for peptide N-glycosidase F, the minimum oligosaccharide chain for cleavage was the di-N-acetyl-chitobiosyl core unit.

Ann Biol Clin (Paris), 1986, 44(1), 35 - 8
{Phenotype characteristics of 63 strains of Pseudomonas paucimobilis, mostly of hospital origin}; Richard C; Sixty-three strains of Pseudomonas paucimobilis, a Gram-negative, lemon yellow pigmented, glucose nonfermenting bacillus are phenotypically compared to the type strain of this new species (ATCC 29837) . The morphological, cultural, biochemical (ONPG-test, oxidase, aesculin positive) and nutritional (assimilation of many glucides) characteristics and susceptibility or resistance to antimicrobial agents (many antibiotics are active) are reported . The similarities and differences between P . paucimobilis and other yellow pigmented bacteria (Flavobacterium, especially group IIb, Xanthomonas, Pseudomonas maltophilia, stutzeri and cepacia) are presented . The occurrence of P . paucimobilis in diverse aquatic and aqueous sources and hospital environments and its significance in medical bacteriology (many isolates from blood cultures) are discussed.

Diagn Microbiol Infect Dis, 1986 Jan, 4(1), 65 - 9
Flavobacterium meningosepticum bacteremia in an adult with acute leukemia . Use of rifampin to clear persistent infection; Hirsh BE et al.; A case of Flavobacterium meningosepticum bacteremia complicating the course of a patient with leukemia is described . The patient was treated successfully when rifampin was added to the antibiotic therapy . Unusual organisms should be considered as agents of infection in immunocompromised hosts and susceptibility testing with drugs not commonly employed for gram-negative rods should be performed in complicated cases.

J Biochem (Tokyo), 1985 Dec, 98(6), 1631 - 8
Purification and properties of 3-ketovalidoxylamine A C-N lyase from Flavobacterium saccharophilum; Takeuchi M et al.; 3-Ketovalidoxylamine A C-N lyase was purified about 900-fold from the cell-free extract of Flavobacterium saccharophilum by ammonium sulfate fractionation, column chromatography on CM cellulose and gel filtration on Sephacryl S-200 . The purified enzyme was homogeneous as judged by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis . The molecular weight of the enzyme was estimated to be 36,000 by gel filtration on Sephacryl S-200 and by SDS polyacrylamide gel electrophoresis, indicating that the enzyme is a monomer . The optimum pH was found at 9.0 . The enzyme activity was inhibited by EDTA or ethyleneglycol bis(beta-aminoethylether)-N,N'-tetraacetic acid and the inhibition was reversed by Ca2+ ion . The enzyme was able to eliminate p-nitroaniline or p-nitrophenol from p-nitrophenyl-3-ketovalidamine (IV) or p-nitrophenyl-alpha-D-3-ketoglucoside (VI), but not from p-nitrophenyl-1-epi-3-ketovalidamine or p-nitrophenyl-beta-D-3-ketoglucoside . Apparent Km values for IV and VI were 0.24 mM and 0.5 mM, respectively.

Appl Environ Microbiol, 1985 Dec, 50(6), 1512 - 8
Isolation and characterization of Flavobacterium strains that degrade pentachlorophenol; Saber DL et al.; Bacteria able to mineralize 100 to 200 ppm of pentachlorophenol (PCP) were isolated by selective enrichment from PCP-contaminated soils from three geographic areas of Minnesota . Although differing somewhat in their responses to various biochemical and biophysical tests, all strains were assigned to the genus Flavobacterium . Five representative strains were examined in detail . All strains metabolized PCP as a sole source of carbon and energy; 73 to 83% of all carbon in the form of {U-14C}PCP was returned as 14CO2, with full liberation of chlorine as chloride . A comparison between strains in their ability to metabolize PCP showed some strains to be more efficient than others . Guanine-plus-cytosine contents of DNA ranged from 58.8 to 63.8%, and DNA/DNA hybridization studies with total DNA digests suggested substantial genetic homology between strains . All strains were shown to possess an 80- to 100-kilobase plasmid, and evidence suggested the presence of a larger plasmid (greater than 200 kilobases).

J Appl Bacteriol, 1985 Dec, 59(6), 561 - 6
A note on a selective agar medium for the enumeration of Flavobacterium species in water; Flint KP; A selective nutrient agar medium containing kanamycin at 50 micrograms/ml was developed for the isolation and enumeration of yellow-pigmented colonies from the River Sowe, Coventry . Such organisms were shown to be members of the heterogeneous genus Flavobacterium . Typically, yellow pigmented colonies constituted less than 10% of the colonies on nutrient agar alone but up to 70% on nutrient agar plus kanamycin . This medium is a useful addition to the range of media available for the isolation and further ecological study of particular species of this important group of micro-organisms.

J Biochem (Tokyo), 1985 Dec, 98(6), 1653 - 9
Occurrence of O-glycosidically peptide-linked oligosaccharides of poly-N-acetyllactosamine type (erythroglycan II) in the I-antigenically active Sendai virus receptor sialoglycoprotein GP-2; Suzuki Y et al.; Unique high molecular weight (M.W . 4,000-9,000) sugar chains termed erythroglycan II have been obtained from alkali/sodium borohydride digests of I-active asialoglycoprotein derived from sialoglycoprotein GP-2, which was isolated recently from bovine erythrocyte membranes as Sendai virus receptor (Suzuki, Y . et al . (1983) J . Biochem . 93, 1621-1633; (1984) ibid, 95, 1193-1200) . It was found that these sugar chains comprise about 40% of total alkali-labile oligosaccharides of asialo GP-2 and contain endo-beta-galactosidase (Flavobacterium keratolyticus)-resistant highly branched and heterogeneous oligosaccharides of poly-N-acetyllactosamine type which are linked O-glycosidically to the peptide backbone through N-acetylgalactosamine . Erythroglycan II also contains endo-beta-galactosidase-susceptible straight terminal polylactosaminyl side chains . A major oligosaccharide released by the enzyme cochromatographed with Gal beta 1-4GlcNAc beta 1-3Gal . Inhibitory activity of Sendai virus-mediated hemagglutination and the receptor activity for the virus were reduced significantly but not completely by the endo-beta-galactosidase . These results indicate that both linear and branched sialosylpolylactosamine sequences in erythroglycan II are important for the reception of the virus into the target cells.

Biochim Biophys Acta, 1985 Nov 22, 843(1-2), 1 - 7
Isolation and characterization of a heparin with high anticoagulant activity from Anomalocardia brasiliana; Dietrich CP et al.; The isolation, some structural features, physicochemical properties and pharmacological activities of a heparin from Anomalocardia brasiliana are reported . It is shown that the mollusc heparin is very similar to those present in mammalian tissues with regard to chemical composition, physicochemical properties, pharmacological activities and susceptibility to heparinase and heparitinase II from Flavobacterium heparinum, as well as to the types of products formed by the action of these enzymes . Three significant quantitative differences were observed for the mollusc heparin when compared with the ones from mammalian origin, namely, a higher degree of binding with antithrombin III (45%), higher molecular weight (27-43 kDa) and higher anticoagulant activity (320 I.U./mg) . The possible biological role of heparin is discussed in view of the present findings.

J Infect, 1985 Nov, 11(3), 233 - 8
Flavobacterium odoratum ventriculitis treated with intraventricular cefotaxime; Macfarlane DE et al.; A 6-week-old infant admitted to the University Hospital of the West Indies with hydrocephalus later developed ventriculitis . A heavy growth of Flavobacterium odoratum susceptible to gentamicin and cefotaxime was recovered from the ventricular fluid . Since intraventricular therapy was envisaged, a Pudenz reservoir was installed and ventricular fluid aspirated every 24 h to monitor treatment . Initial therapy consisted of intravenous cefotaxime, 50 mg/kg q.i.d . for 4 days . No significant reduction in the number of organisms in the ventricular fluid was achieved with this regimen . Intravenous therapy was therefore discontinued . On day 5 intraventricular therapy began with 5 mg cefotaxime 24 h for 6 days, followed by 1 mg/24 h for 4 days . Daily monitoring of intraventricular fluid indicated a high degree of antibacterial activity with rapid elimination of bacteria . Ventricular fluid remained sterile 10 days after therapy stopped . The Pudenz reservoir was removed, a ventriculoperitoneal shunt installed, and the patient discharged from hospital 4 days later without noticeable sequelae.

J Antibiot (Tokyo), 1985 Nov, 38(11), 1476 - 86
Studies on WB-3559 A, B, C and D, new potent fibrinolytic agents . II . Structure elucidation and synthesis; Uchida I et al.; The structures of WB-3559 A, B, C and D, new fibrinolytic agents isolated from Flavobacterium sp . No . 3559, have been elucidated to be as shown in 1, 2, 3 and 4, respectively, on the basis of chemical and spectroscopic evidence . Total synthesis of WB-3559 D (4) was achieved starting from the optically active aldehyde 14.

J Antibiot (Tokyo), 1985 Nov, 38(11), 1469 - 75
Studies on WB-3559 A, B, C and D, new potent fibrinolytic agents . I . Discovery, identification, isolation and characterization; Yoshida K et al.; WB-3559 A, B, C and D were produced by a bacterium which was classified as a member of the genus Flavobacterium . These compounds were purified by solvent extraction followed by chromatography on silica gel and then isolated by HPLC . The chemical structures were determined on the basis of chemical and spectroscopic evidence as in the succeeding papers . WB-3559 A, B, C and D stimulated rabbit plasma euglobulin clot lysis time . A chemically synthesized compound (WB-3559 D-syn) stimulated mouse plasma euglobulin clot lysis time when injected intravenously.

J Appl Bacteriol, 1985 Oct, 59(4), 311 - 23
A numerical taxonomic study of Flavobacterium-Cytophaga strains from dairy sources; Jooste PJ et al.; Phenotypic data on 203 Gram-negative non-fermentative bacteria of the Flavobacterium-Cytophaga group isolated from milk and butter were analyzed by numerical taxonomic techniques . Twenty reference strains including species of Flavobacterium, Cytophaga and strains of Pseudomonas paucimobilis were included in the study . Using the matching coefficient of Sokal & Michener with antibiotic susceptibility data included, 189 isolates were recovered in nine clusters . Six of these clusters were linked at or above the 85% S level while three were linked at or above the 79% S level . The largest cluster, representing 46.3% of the isolates, could be equated with Flavobacterium sp . Group IIb . Other clusters could be equated with Flavobacterium sp . L 16/1 (22.7% of isolates), F . balustinum (10.8% of isolates), F . breve (4.4%), F . multivorum (3.5%) and Cytophaga johnsonae (1.5%) . The cluster resembling Flavobacterium sp . L 16/1 and a smaller unclassified cluster, were exceptional in being susceptible to the antibiotics cephalothin and penicillin G.

Eur J Biochem, 1985 Oct 1, 152(1), 75 - 82
Flavobacterium heparinum 6-O-sulphatase for N-substituted glucosamine 6-O-sulphate; Bruce JS et al.; A specific glyco-6-O-sulphatase has been purified to homogeneity from Flavobacterium heparinum . The enzyme hydrolyses the 6-O-sulphates of 2-deoxy-2-sulphamido-6-O-sulpho-D-glucose (GlcNS-6S), 2-acetamido-2-deoxy-6-O-sulpho-D-glucose (GlcNAc-6S) and 2-amino-2-deoxy-6-O-sulphato-D-glucose (GlcN-6S) . The activity was purified 2100-fold by successive chromatography on CM-Sepharose CL-6B, Sepharose CL-4B, hydroxyapatite and blue-Sepharose CL-6B . Sodium dodecyl sulphate/polyacrylamide gel electrophoresis showed a protein of relative molecular mass 64000 . Four novel assays were developed using 35S-labelled and 14C-labelled monosaccharides . The purified enzyme was free of all other known heparin-degrading enzymes . In particular this was the first resolution of the 6-O-sulphatase from the sulphamidase . Optimal activity was at pH 7.5 . Enzyme activity was virtually unaffected by Na+ and K+ ions . Enhancements of activity of 12% and 30% were effected by Mg2+ and Ca2+ ions respectively . Inorganic phosphate and sulphate (both 0.005 mol dm-3) inhibited activity by 48% and 50% respectively . The Km value for the free amino substrate GlcN-6S was 1.35 mmol dm-3 . In contrast the Km values for the GlcNAc-6S and GlcNS-6S were 54 mumol dm-3 and 16 mumol dm-3 respectively.

J Biochem (Tokyo), 1985 Oct, 98(4), 975 - 9
Comparison of inhibitory effects of prolinal-containing peptide derivatives on prolyl endopeptidases from bovine brain and Flavobacterium; Yoshimoto T et al.; The inhibitory effects of proline-containing peptides and their derivatives on prolyl endopeptidases from Flavobacterium meningosepticum and bovine brain were compared . Replacement of the carboxyl terminal proline in N-blocked peptides with prolinal resulted in remarkable decreases in Ki values for both prolyl endopeptidases . Further reduction of the prolinal to prolinol led to a decrease in their inhibitory effects . Z-Pro-, Z-Val-, and Suc-Pro-prolinals were similarly inhibitory for both the enzymes with Ki values of nM order . However, the inhibitory effects of Z-Pyr-prolinal and Boc-Pro-prolinal on these enzymes were significantly distinguished: they strongly inhibited the mammalian prolyl endopeptidase with Ki values of nM order, while the Ki values of these compounds for the microbial enzyme were only of microM order . These results suggest that there are some structural differences in the S2 and S3 subsites between the two enzymes, though their substrate specificities are apparently indistinguishable.

Biochemistry, 1985 Sep 24, 24(20), 5587 - 92
Deglycosylation of alpha 1-proteinase inhibitor by endo-beta-N-acetylglucosaminidase F; Steube K et al.; The glycosidase endo-beta-N-acetylglucosaminidase F (endo F) from Flavobacterium meningosepticum was used for the deglycosylation of rat alpha 1-proteinase inhibitor (alpha 1 PI) . alpha 1 PI containing three oligosaccharide side chains of the complex type was isolated from rat serum or from the medium of rat hepatocyte primary cultures . High-mannose-type alpha 1 PI or hybrid-type alpha 1 PI was isolated from the media of hepatocytes treated with 1-deoxymannojirimycin or swainsonine, respectively . The susceptibility of complex-type alpha 1 PI to endo F was studied in the presence of various detergents . 3-{(3-Cholamidopropyl)dimethylammonio}-1-propanesulfonate and octyl glucopyranoside turned out to be most effective . In the absence of detergents, digestion of alpha 1 PI with high concentrations of endo F and/or long times of incubation led to the formation of alpha 1 PI with one and two oligosaccharide side chains . In the presence of 0.5% octyl glucopyranoside, the major cleavage products were unglycosylated alpha 1 PI and alpha 1 PI carrying one carbohydrate side chain . In contrast to the complex-type alpha 1 PI, the high-mannose type can be totally deglycosylated by endo F even in the absence of detergents . The susceptibility of the hybrid-type alpha 1 PI to endo F is between that of the complex and the high-mannose types.

Biochemistry, 1985 Aug 13, 24(17), 4665 - 71
Deglycosylation of asparagine-linked glycans by peptide:N-glycosidase F; Tarentino AL et al.; Endo-beta-N-acetylglucosaminidase F (Endo F) and peptide:N-glycosidase F (PNGase F) were purified from cultures of Flavobacterium meningosepticum by ammonium sulfate precipitation followed by gel filtration on TSK HW-55(S) . This system separated the two enzymes and provided PNGase F in a high state of purity, but the basis for the resolution appeared to be hydrophobic interaction and not molecular size . Studies using purified Endo F and PNGase F with defined glycopeptides demonstrated that Endo F was somewhat similar to Endo H in that it hydrolyzed many, but not all, high-mannose and hybrid oligosaccharides, as well as complex biantennary oligosaccharides . PNGase F, in contrast, hydrolyzed all classes of asparagine-linked glycans examined, provided both the alpha-amino and carboxyl groups of the asparagine residue were in peptide linkage . Deglycosylation studies with PNGase F revealed that many proteins in their native conformation were susceptible to this enzyme but that prior denaturation in sodium dodecyl sulfate greatly decreased the amount of enzyme required for complete carbohydrate removal.

Biochem J, 1985 Jul 15, 229(2), 369 - 77
Structure of heparin-derived tetrasaccharides; Merchant ZM et al.; The structure of heparin was examined by characterizing a disaccharide and five of the more than a dozen tetrasaccharide components obtained by its depolymerization with flavobacterial heparinase . Enzymic depolymerization of porcine mucosal heparin results in a mixture of di-, tetra-, hexa- and higher oligo-saccharides . The di- and tetra-saccharide components represent 75mol/100mol of these heparin fragments . Ion-exchange chromatography indicates the presence of only one disaccharide, deltaIdu2S(1----4)-alpha-D-GlcNS6S (where Idu is iduronic acid, deltaIdu is 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid, GlcN is glucosamine, GlcA is glucuronic acid and S is sulphate), but results in the isolation of five major and at least seven minor tetrasaccharide components . The structures of the disaccharide and five major tetrasaccharides were determined by chemical, enzymic, electrophoretic and spectroscopic methods, including 13C, 1H n.m.r . and fast atom bombardment-m.s . The structure of these five tetrasaccharides are: delta Idu2S(1----4)-alpha-D-GlcNS6S(1---4)-alpha-L-Idu2S(1-- --4)-alpha -D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS6S(1----4)-beta-D-GlcA(1--- -4)- alpha-D-GlcNS6S; delta Idu2S(1----4)-alpha-D-GlcNS(1----4)-beta-D-GlcA delta Idu2S(1----4)-alpha-D-GlcNAc(1----4)-beta-D-GlcA(1----4)- alpha-D-GlcNS6S; and delta Idu2S(1----4)-alpha- D-GlcNAc(1----4)-alpha-L-Idu(1----4)-alpha-D-GlcNS6S . Biological activity for the disaccharide and the five major tetrasaccharides was examined, and none of them were found to possess significant anticoagulant activity.

J Cell Physiol, 1985 Jul, 124(1), 113 - 9
Two distinct mechanisms for the interaction of cells with fibronectin substrata; Schwarz MA et al.; Fibronectin (Fn) was adsorbed onto neutral, sulfonated, imine-conjugated or gelatin coated polystyrene latex beads . In all cases, the Fn coated beads bound effectively to Chinese hamster ovary (CHO) cells in suspension . However, the binding of Fn coated neutral or positively charged imine conjugated bead was inhibited by low concentrations of heparin or heparan sulfate or by treatment of the cells with Flavobacterium heparanase . By contrast, binding of Fn coated sulfonated or gelatin beads was insensitive to inhibition by heparin and to heparanase treatment of cells . Adhesion of CHO cells to Fn coated tissue culture plastic was not sensitive to heparin, whereas adhesion of CHO cells to Fn-coated imine-conjugated plastic was sensitive to heparin . These observations imply that the functional status of Fn can be modulated by the nature of the surface to which the Fn is adsorbed . They further imply that, under some circumstances, the heparin/heparan sulfate binding domains of Fn can play a role in the attachment of Fn to the cell membrane via membrane proteoglycans . Under other circumstances, the interaction of Fn with the cell may primarily involve other receptors for Fn, presumably cell surface glycoproteins.

Ann Inst Pasteur Microbiol, 1985 May-Jun, 136A(3), 359 - 70
{Isolation and description of strains of Flavobacterium multivorum of telluric origin}; Pichinoty F et al.; Twenty encapsulated strains of Flavobacterium multivorum were isolated from soil by elective culture in minimal medium containing inulin as the sole source of carbon and energy . These strains were compared with the type strain and with 5 other strains (including 4 clinical strains) of F . multivorum . Of 168 substrates tested, 18 carbohydrates were used as the sole carbon and energy source by all 26 strains . A few amino acids were used by some strains . The yellow pigment produced was found to be a carotene and its production was photo-inducible . The presence of a cytochrome c oxidase of the aa3 type was suggested by absorption spectra . The major cell lipids were phosphatidylethanolamine and sphingolipids; the major fatty acids were 13-methyltetradecanoic and 2-hydroxy-13-methyltetradecanoic acids . Soil and clinical strains of F . multivorum showed roughly similar patterns of antibiotic multiresistance . The average G + C content of the DNA of 11 strains was 40.8 +/- 1.5 mol%.

Eur J Biochem, 1985 Apr 15, 148(2), 359 - 65
Flavobacterium heparinum 3-O-sulphatase for N-substituted glucosamine 3-O-sulphate; Bruce JS et al.; A novel bacterial sulphatase has been discovered in an extract of Flavobacterium heparinum . The enzyme hydrolyses the 3-O-sulphate from 2-deoxy-2-sulphamido-3-O-sulpho-D-glucose and 2-acetamido-2-deoxy-3-O-sulpho-D-glucose . The activity was purified 10 800-fold by chromatography successively on CM-Sepharose CL-6B, hydroxyapatite, taurine-Sepharose CL-4B and CM-Sepharose CL-6B . Sodium dodecylsulphate/polyacrylamide gel electrophoresis showed the enzyme to be homogeneous and of relative molecular mass 56 000 . Two novel assays were developed using 2-{14C}acetamido-2-deoxy-3-O-sulpho-D-glucose and 2-deoxy-2-sulphamido-3-O-sulpho-D-glucose as respective substrates . The purified 3-O-sulphatase was shown to be free of all other known heparin-degrading enzymes . Optimal activity was at pH 7.5 for the disulphated substrate and pH 8.0 for the N-acetylated substrate . Enzyme activity was virtually unaffected by Na+, K+ or Mg2+ ions . A 1.2-fold enhancement of activity was effected by 0.002 mol dm-3 Ca2+ . Inorganic phosphate and sulphate inhibited 3-O-sulphatase activity . The Km value of the N-acetylated substrate was determined to be 42 mumol dm-3 . No activity was detected with 2-amino-2-deoxy-3-O-sulpho-D-glucose.

J Biol Chem, 1985 Apr 10, 260(7), 3915 - 22
Purification and properties of creatinine iminohydrolase from Flavobacterium filamentosum; Esders TW et al.; Creatinine iminohydrolase (EC 3.5.4.21), which catalyzes hydrolysis of creatinine to N-methylhydantoin and ammonia, was purified from Flavobacterium filamentosum . The average molecular weight of the purified enzyme was 272,480, and the subunit molecular weight was 44,300 . Extensive specificity studies indicated that the enzyme utilized cytosine (Km, 0.62 mM; Vm, 20.1 units/mg) as well as creatinine (Km, 5.00 mM; Vm, 40.4 units/mg) as a substrate . Each was a competitive inhibitor toward hydrolysis of the other compound . Dialysis of creatinine iminohydrolase in the presence of 0.01 M Tris phosphate buffer, pH 7.5, containing 1,10-phenanthroline decreased activity by 98% . Reactivation was accomplished by incubating the apoenzyme in the presence of certain divalent metal chlorides, listed in decreasing order of effectiveness: iron(II), zinc, cobalt(II), cadmium, and nickel . The extent of reactivation depended on the substrate and on which metal ion was added to the apoenzyme . Creatinine to cytosine activity ratios varied from 1:3.75 (iron(II) chloride), to 1:0.9 (zinc chloride), to 1:0.06 (nickel chloride) . For different preparations of the holoenzyme that ratio ranged from 1:0.45 to 1:1.10 . Variable but significant quantities of zinc and iron were present in all preparations of the purified enzyme.

Appl Environ Microbiol, 1985 Apr, 49(4), 765 - 71
Determination of the concentration of maltose- and starch-like compounds in drinking water by growth measurements with a well-defined strain of a Flavobacterium species; van der Kooij D et al.; The growth kinetics of Flavobacterium sp . strain S12 specialized in the utilization of glycerol, and a number of oligo- and polysaccharides were determined in batch-culture experiments at 15 degrees C in pasteurized tap water supplied with very low amounts of substrates . Kss for the growth on maltotriose, maltotetraose, maltopentaose, and maltohexaose were 0.03 microM or less and below those for glucose (1.5 microM) and maltose (0.16 microM) . Kss for starch, amylose, and amylopectin were 8.4, 25.6, and 11.0 micrograms of C per liter, respectively . A yield of 2.3 X 10(7) CFU/micrograms of C on the oligo- and polysaccharides was calculated from the linear relationships observed between maximum colony counts in pasteurized tap water and the concentrations (usually below 25 micrograms of C per liter) of supplied compounds . The maximum colony counts of strain S12 grown in various types of raw water and tap water revealed that raw water contained only a few micrograms of maltose- and starch-like compounds per liter; in tap water the concentrations were all below 1 microgram of C and usually below 0.1 microgram of C per liter . The application of starch-based coagulant aids gave increased concentrations of maltose- and starch-like compounds in the water during treatment, but these concentrations were greatly reduced by coagulation and sedimentation, rapid sand filtration, and slow sand filtration.

Antimicrob Agents Chemother, 1985 Apr, 27(4), 612 - 4
Biochemical properties of beta-lactamase produced by Flavobacterium odoratum; Sato K et al.; A constitutively produced beta-lactamase was purified from Flavobacterium odoratum GN14053 . The purified enzyme gave a single protein band on polyacrylamide gel electrophoresis . The isoelectric point was 5.8, and the molecular weight was estimated to be about 26,000 . The enzyme activity was inhibited by EDTA, iodine, p-chloromercuribenzoate, HgCl2, and CuSO4 but not by clavulanic acid, sulbactam, imipenem, and cephamycin derivatives . The enzyme showed a broad substrate profile, hydrolyzing oxyiminocephalosporins, cephamycins, imipenem, and some penicillins.

J Clin Invest, 1985 Apr, 75(4), 1308 - 16
Evidence that cell surface heparan sulfate is involved in the high affinity thrombin binding to cultured porcine aortic endothelial cells; Shimada K et al.; It has been postulated that thrombin binds to endothelial cells through, at least in part, cell surface glycosaminoglycans such as heparan sulfate, which could serve as antithrombin cofactor on the endothelium . In the present study, we have directly evaluated the binding of 125I-labeled bovine thrombin to cultured porcine aortic endothelial cells . The thrombin binding to the cell surface was rapid, reversible, and displaced by enzymatically inactive diisopropylphosphoryl-thrombin . The concentration of thrombin at half-maximal binding was approximately 20 nM . Both specific and nonspecific binding of 125I-thrombin to the endothelial cell surface was partially inhibited in the presence of protamine sulfate, after the removal of cell surface heparan sulfate by the treatment of cells with crude Flavobacterium heparinum enzyme or purified heparitinase . The binding as a function of the concentration of thrombin revealed that the maximal amount of specific binding was reduced by approximately 50% with little alteration in binding affinity by these enzymatic treatments . The reversibility and active-site independence as well as the rate of the binding did not change after heparitinase treatment . Whereas removal of chondroitin sulfates by chondroitin ABC lyase treatment of cells did not affect the binding, identical enzymatic treatments of {35S}sulfate-labeled cells showed that either heparan sulfate or chondroitin sulfate was selectively and completely removed from the cell surface by heparitinase or chondroitin ABC lyase treatment, respectively . Furthermore, proteolysis of cell surface proteins by the purified glycosaminoglycan lyases was excluded by the identical enzymatic treatments of {3H}leucine-labeled or cell surface radioiodinated cells . Our results provide the first direct evidence that heparan sulfate on the cell surface is involved in the high-affinity, active site-independent thrombin binding by endothelial cells, and also suggest the presence of thrombin-binding sites that are not directly related to heparan sulfate.

J Biol Chem, 1985 Feb 10, 260(3), 1849 - 57
Purification and characterization of heparinase from Flavobacterium heparinum; Yang VC et al.; Heparinase (EC 4.2.2.7) isolated from Flavobacterium heparinum was purified to homogeneity by a combination of hydroxylapatite chromatography, repeated gel filtration chromatography, and chromatofocusing . Homogeneity was established by the presence of a single band on both sodium dodecyl sulfate and acid-urea gel electrophoretic systems . Amino acid analysis shows that the enzyme contains relatively high amounts of lysine residues (9%) consistent with its cationic nature (pI 8.5) but contains only 4 cysteine residues/polypeptide . The molecular weight of heparinase was estimated to be 42,900 +/- 1,000 daltons by gel filtration and 42,700 +/- 1,200 daltons by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The enzyme is very specific, acting only on heparin and heparan monosulfate out of 12 similar polysaccharide substrates tested . It has an activity maximum at pH 6.5 and 0.1 M NaCl and a stability maximum at pH 7.0 and 0.15 M NaCl . The Arrhenius activation energy was found to be 6.3 kcal/mol . However, the enzyme is very sensitive to thermal denaturation and loses activity very rapidly at temperatures over 40 degrees C . Kinetic studies of the heparinase reaction at 37 degrees C gave a Km of 8.04 X 10(-6) M and a Vm of 9.85 X 10(-5) M/min at a protein concentration of 0.5 microgram/ml . By adapting batch procedures of hydroxylapatite and QAE (quaternary aminoethyl)-Sephadex chromatography, gram quantities of heparinase that is nearly free of catalytic enzyme contaminants can be purified in 4-5 h.

Appl Environ Microbiol, 1985 Feb, 49(2), 382 - 7
Quantitative studies of heat-stable proteinase from Pseudomonas fluorescens P1 by the enzyme-linked immunosorbent assay; Birkeland SE et al.; Pseudomonas fluorescens P1 is a psychrotrophic bacterium isolated from milk . Proteinase P1, the main extracellular heat-stable proteinase fraction of P . fluorescens P1, has been purified to homogeneity . A procedure with a sandwich enzyme-linked immunosorbent assay, using microplates and alkaline phosphatase conjugate was shown to detect 0.25 ng of proteinase P1 in 1 ml of reconstituted skim milk or defatted cream . The method offers the combination of sensitivity and specificity for the detection of these enzymes in milk and dairy products . In reconstituted skim milk cultures proteinase P1 was detectable when the CFU approached 10(7)/ml . Cultures in milk diluted 1:10, 1:30, or 1:100 with water showed detectable proteinase at population densities close to 10(6) CFU/ml . Aeration stimulated proteinase production; thus, a skim milk culture with shaking at 5 degrees C had a proteinase level of 36,000 ng/ml after 7 days as compared to 80 ng/ml in a stationary culture . The rate of inactivation of proteinase P1 at 150 and 55 degrees C as expressed by residual antigenic activity determined by the enzyme-linked immunosorbent assay was somewhat different from the rate determined on the basis of residual proteolytic activity . The specificity of the enzyme-linked immunosorbent assay with proteinase P1 antibodies was identical for proteinase P1 and for enzymes from six other strains of P . fluorescens, one Chromobacterium strain, and one Flavobacterium strain . Some psychrotrophic strains produced immunologically unrelated proteinase(s) . These preliminary observations indicate that proteinase P1-related enzymes are common among psychrotrophs appearing as spoilage bacteria in milk.

Gastrointest Endosc, 1985 Feb, 31(1), 10 - 2
Bacteremia during esophageal variceal sclerotherapy: its cause and prevention; Brayko CM et al.; Eleven consecutive patients underwent a total of 34 esophageal variceal sclerotherapy (EVS) sessions for bleeding esophageal varices . Blood cultures were drawn pre-, intra-, and post-EVS . All pre- and post-EVS blood cultures were negative . Five of the initial nine patients studied were found to have positive blood cultures, drawn after a mean of six injections . Pseudomonas aeruginosa was cultured from the blood in four patients and Flavobacterium from one . The source of contamination was found to be contaminated water used during the sclerotherapy sessions . By instituting simple techniques to eliminate this contamination, patients undergoing the remaining 25 EVS sessions were culture negative.

Biochem Biophys Res Commun, 1985 Jan 16, 126(1), 365 - 72
Heparinlike molecules with anticoagulant activity are synthesized by cultured endothelial cells; Marcum JA et al.; Cultured microvascular endothelial cells isolated from rat epididymal fat pads produce glycosaminoglycans that accelerate thrombin-antithrombin complex formation . The heparinlike nature of these macromolecules was established by complete destruction of their anticoagulant activity employing purified Flavobacterium heparinase . Only 15% of the biologic activity of these complex carbohydrates was expressed when the heparin binding domain on the protease inhibitor was chemically modified at the Trp 49 residue . The anticoagulantly active species contains disaccharides which constitute the unique antithrombin binding region of the mucopolysaccharide . Removal of the biologically active heparinlike components from endothelial cells with 0.05% trypsin suggests that these molecular species are present on the cell surface.

Arzneimittelforschung, 1985, 35(8), 1215 - 9
Fractionation and structural features of two heparin families with high antithrombotic, antilipemic and anticoagulant activities; Bianchini P et al.; 15 heparin preparations from bovine intestine, pancreas and lung and hog intestine were fractionated in two main components by selective barium precipitation . The ones that precipitated at room temperature with barium (slow moving (SM)-heparins) had a high anticoagulant activity measured by the USP and APTT (activated partial thromboplastin time) assay and low antithrombotic activity by the Yin and Wessler method . The fractions precipitated at 5 degrees C with barium (fast moving (FM)-heparins) had a low anticoagulant action and high antithrombotic activity . The maximum anti-Xa activity (chromogenic method) was present in heparins with molecular weights around 12-15 X 10(3) daltons whereas high APTT and LPL releasing activities were present in SM-heparins with molecular weights of 30-40 X 10(3) and 15-25 X 10(3) daltons, respectively . FM-heparins had a higher anti-Xa activity and lower lipoprotein lipase (LPL)-releasing activity when compared with the SM-heparins with the same molecular weights . Significant structural differences were observed between SM- and FM-heparins by 13C-NMR spectra and enzymatic degradation with heparinase and heparitinase from Flavobacterium heparinum . Also, significant differences were observed for anti-Xa and anticoagulant activities for the two types of heparins depending on the pharmacological assay used.

J Infect, 1985 Jan, 10(1), 17 - 24
Septicaemia in a teaching hospital in Kuwait--I: Incidence and aetiology; Elhag KM et al.; During a period of 18 months of study, blood cultures were performed on 3845 patients in hospital with clinical signs of infection . Among these, 214 (5.6%) episodes of septicaemia were diagnosed which correspond to 10.9/1000 hospital admissions . About 80% of the episodes were due to Gram-negative organisms, most common of which were Escherichia coli (19.6%), Salmonella spp (16.5%) and Klebsiella spp (15.1%) . Gram-positive organisms implicated in 20% of episodes were mainly Staphylococcus aureus (9.3%) and enterococci (4.9%) . Of all the septicaemias 62.0% were community-acquired with Salmonella spp . being the organism most commonly implicated . Hospital-acquired infections were mainly due to Serratia spp, Pseudomonas spp and Flavobacterium spp . The antibiotic resistance pattern of the organisms showed that hospital-acquired organisms had relatively high resistance to most antibiotics as compared with community-acquired organisms.

J Clin Invest, 1985 Jan, 75(1), 272 - 9
Interaction of antithrombin III with bovine aortic segments . Role of heparin in binding and enhanced anticoagulant activity; Stern D et al.; Bovine antithrombin III (AT III) interaction with the luminal surface of bovine aortic segments with a continuous layer of endothelium was examined . Incubation of 125I-AT III with vessel segments, previously washed free of endogenous AT III, demonstrated specific, time-dependent binding to the protease inhibitor to the endothelium . Half-maximal binding was observed at an added AT III concentration of 14 nM . Binding of 125I-AT III to the vessel wall was reversible (50% dissociated in 4 min), and addition of either heparin or Factor Xa accelerated displacement of 125I-AT III from the vessel segment . Dissociation of 125I-AT III from the vessel segment in the presence of factor Xa coincided with the formation of a Factor Xa-125I-AT III complex . Inactivation of Factor IXa and Factor Xa by AT III was facilitated in the presence of vessel segments . Pretreatment of vessel segments with highly purified Flavobacterium heparinase precluded the vessel-dependent augmentation of AT III anticoagulant activity as well as specific binding of 125I-AT III to the vessel endothelium . In contrast, pretreatment of the vessel segments with chrondroitinases (ABC or AC) had no detectable effect on 125I-AT III binding or on AT III anticoagulant activity . AT III binding to vessel segments was competitively inhibited by increasing concentration of platelet factor 4 . Binding of the protease inhibitor to vessel segments was inhibited by chemical modification of AT III lysyl or tryptophan residues . These AT III derivatives retained progressive inhibitory activity . These data suggest that heparin-like molecules are present on the aortic vessel wall and mediate binding of AT III to the vessel surface, as well as enhancing the anticoagulant activity of AT III at these sites.

Int J Biochem, 1985, 17(11), 1179 - 83
Immuno-affinity purification of heparinase; Linhardt RJ et al.; Polyclonal IgG rabbit antibodies were prepared against a purified heparinase from Flavobacterium heparinum . Immuno-affinity purification of crude and partially purified heparinase is described . The resulting enzyme was of comparable purity to that prepared using the standard multistep purification scheme . The antibodies prepared were found to increase the activity of bound heparinase.

Dev Biol Stand, 1985, 61, 249 - 54
Characteristic cellular fatty acid composition and an ornithine-containing lipid as a new type of hemagglutinin in Bordetella pertussis; Kawai Y; The fatty acid composition of the total extractable cellular lipids of Bordetella pertussis was very characteristic and was mostly hexadecenoic and hexadecanoic acids (90%) in a ratio of about 1:1 . The fatty acid composition of Bordetella parapertussis and Bordetella bronchiseptica differed from that of B . pertussis . The two species were distinguished by the fatty acid composition of cell-bound lipids . The ornithine-containing lipid was characteristic of the genus Bordetella and its main structure was 3-hydroxyhexadecanoic acid amide-linked to ornithine and esterified to the second hexadecanoic acid . The lipid agglutinated human and some animal erythrocytes . The lipid is a new type of hemagglutinin and we proposed that hemagglutination occurred mainly by the hydrophobic interaction between the lipid moiety of the ornithine-containing lipid and phosphatidylcholine in the cell membrane of the erythrocytes . A relatively high content of ornithine-containing lipid was also found in opportunistic pathogens such as Flavobacterium meningosepticum which causes meningitis in babies and children . As the pathogenicity of the opportunistic pathogens is unclear, the ornithine-containing lipid may have an important role in pathogenicity.

Eur J Biochem, 1984 Dec 17, 145(3), 607 - 15
Flavobacterium heparinum 2-O-sulphatase for 2-O-sulphato-delta 4,5-glycuronate-terminated oligosaccharides from heparin; McLean MW et al.; The glycosulphatase which hydrolyses the 2-O-sulphate of the disaccharide, 4-deoxy-2-O-sulphato-alpha-L-threohex-4-enopyranosyl uronic acid-(1----4)-2-deoxy-2-sulphamido-6-O-sulphato-D-glucose (delta UA-2S----GlcNS-6S), has been isolated from the soluble fraction of disrupted Flavobacterium heparinum . The activity was purified 3300-fold by chromatography on CM-Sepharose CL-6B, hydroxyapatite, taurine-Sepharose CL-4B and blue-Sepharose CL-6B . From sodium dodecylsulphate/polyacrylamide gel electrophoresis, the enzyme was homogeneous and of 62000 Mr . A novel assay was devised using the de-N-sulphonated {1-3H}alditol, 4-deoxy-2-O-sulphato-alpha-L-threo-hex-4-enopyranosyl uronic acid-(1----4)-2-amino-2-deoxy-6-O-sulphato-D-{1-3H}glucitol (delta UA-2S----{1-3H}GlcNH2-ol-6S) . This alditol was shown by 13C-NMR to be desulphated in the analogous manner to the original reducing trisulphated disaccharide . The purified 2-O-sulphatase was completely free of heparinase I, heparinase II (heparitinase), chondroitinases AC, chondroitinase B, the delta 4,5-glycuronidase for heparin delta 4,5-disaccharides, the 6-O-sulphatase and the 2-sulphamidase . It was optimally active over the range pH 5.5-6.5 and was practically unaffected by Na, K, Ca or Mg ions . Inorganic phosphate inhibited the activity . The Km value for the alditol substrate was 1.22 mmol dm-3 . Using 13C-NMR, the 2-O-sulphatase was found to hydrolyse the analogous esters of higher delta 4,5-oligosaccharides from heparin . This contrasts with the findings of other authors {Dietrich, C . P., Silva, M . E., and Michelacci, Y . M . (1973) J . Biol . Chem . 248, 6408-6415}.

J Gen Microbiol, 1984 Dec, 130 ( Pt 12), 3335 - 8
Use of a synthetic oligonucleotide to identify a chromosomal gene for chloramphenicol acetyltransferase in a plasmid-bearing Flavobacterium; Beschle HG et al.; A micro-organism previously designated Flavobacterium sp . CB60 is resistant to chloramphenicol as a consequence of antibiotic acetylation by the enzyme chloramphenicol acetyltransferase and subsequent degradation of the acetylated product by co-metabolism . Although a 15.6 kb plasmid (pCB60) was demonstrated in this Flavobacterium strain, it did not appear to play a role in chloramphenicol acetylation . DNA hybridization was used to identify a fragment of DNA presumptively carrying the cat gene . The sequence of the synthetic probe was based upon known nucleotide sequences corresponding to the highly-conserved active site region of several chloramphenicol acetyltransferase variants . The structural gene for the enzyme in strain CB60 appeared to be chromosomal since the radioactive probe hybridized with a unique restriction fragment from chromosomal DNA but failed to do so with the DNA of plasmid pCB60.

J Trop Med Hyg, 1984 Dec, 87(6), 245 - 8
Meningitis in children of Riyadh; Babiker MA et al.; One hundred and forty cases of meningitis admitted to the Children's Hospital in Riyadh from June 1980 to the end of May 1982 were included in a prospective study . Sixty-seven percent were under 1 year of age and 90% were under 5 years old . One hundred and twenty-four (88.6%) had purulent CSF, 12 (8.6%) had viral and four (2.9%) had tuberculous meningitis . E . coli was the commonest causative organism in the neonates . After that age S . pneumoniae was the predominant isolate . The implication of increasing resistance of H . influenzae to ampicillin therapy were discussed and a further rare case of meningitis caused by Flavobacterium meningosepticum was described . Seventy-three (52%) were cured completely, 39 (28%) survived with immediate neurological deficits and 28 (20%) died . Mortality and neurological sequelae were found to be high in four categories of patients: neonates, patients who were stuporose or comatose on admission, those who were partially treated before admission and those who presented late after the onset of their symptoms.

Biochemistry, 1984 Nov 20, 23(24), 5801 - 12
Sequence variation in heparin octasaccharides with high affinity for antithrombin III; Atha DH et al.; We have isolated from nitrous acid cleavage products of heparin two major octasaccharide fragments which bind with high affinity to human antithrombin . Octasaccharide S, with the predominant structure iduronic acid----N-acetylglucosamine 6-O-sulfate----glucuronic acid-----N-sulfated glucosamine 3,6-di-O-sulfate----iduronic acid 2-O-sulfate----N-sulfated glucosamine 6-O-sulfate----iduronic acid 2-O-sulfate----anhydromannitol 6-O-sulfate, is sensitive to cleavage by Flavobacterium heparinase as well as platelet heparitinase and binds to antithrombin with a dissociation constant of (5-15) X 10(-8) M . Octasaccharide R, with the predominant structure iduronic acid 2-O-sulfate----N-sulfated glucosamine 6-O-sulfate----iduronic acid----N-acetylglucosamine 6-O-sulfate----glucuronic acid----N-sulfated glucosamine 3,6-di-O-sulfate----iduronic acid 2-O-sulfate----anhydromannitol 6-O-sulfate, is resistant to degradation by both enzymes and binds antithrombin with a dissociation constant of (4-18) X 10(-7) M . The occurrence of a 15-17% replacement of N-sulfated glucosamine 3,6-di-O-sulfate with N-sulfated glucosamine 3-O-sulfate and a 10-12% replacement of iduronic acid with glucuronic acid in both octasaccharides indicates that these substitutions have little or no effect on the binding of the oligosaccharides to the protease inhibitor . When bound to antithrombin, both octasaccharides produce a 40% enhancement in the intrinsic fluorescence of the protease inhibitor and a rate of human factor Xa inhibition of 5 X 10(5) M-1 s-1 as monitored by stopped-flow fluorometry . This suggests that the conformation of antithrombin in the region of the factor Xa binding site is similar when the protease inhibitor is complexed with either octasaccharide.

Appl Environ Microbiol, 1984 Nov, 48(5), 936 - 43
Isolation and characterization of a new Cytophaga species implicated in a work-related lung disease; Liebert CA et al.; A yellow-pigmented, gram-negative, gliding bacterium isolated from an industrial water spray air humidification system was implicated as a causative agent in several occurrences of lung disease with hypersensitivity pneumonitis-like symptoms . The bacterium, designated WF-164, lacked microcysts or fruiting bodies and had a DNA base composition of 34.8 mol% of guanine plus cytosine . Gliding, flexing, nonflagellated cells measuring 0.3 by 3.5 to 8.9 micron were observed by using light and electron microscopy . Tests to determine utilization of selected carbohydrates revealed an amylolitic, chitinoclastic, noncellulytic bacterium . A number of additional biochemical and physiological tests were performed . DNA homology studies detected a 77.8% similarity to Cytophaga aquatilis (ATCC 29551) . Comparisons of cellular fatty acid and carbohydrate contents of isolate WF-164 with a Flexibacter sp., several Cytophaga spp., and Flavobacterium reference strains revealed similar patterns to that of C . aquatilis . On the basis of these characteristics, isolate WF-164 was identified as a new Cytophaga sp.

J Gen Microbiol, 1984 Nov, 130 ( Pt 11), 2983 - 99
Differentiation between Xanthomonas campestris pv . oryzae, Xanthomonas campestris pv . oryzicola and the bacterial 'brown blotch' pathogen on rice by numerical analysis of phenotypic features and protein gel electrophoregrams; Vera Cruz CM et al.; Thirty-five Xanthomonas campestris pv . oryzae, fourteen X . campestris pv . oryzicola strains and six 'brown blotch' pathogens of rice, all of different geographical origin, were studied by numerical analysis of 133 phenotype features and gel electrophoregrams of soluble proteins, %G + C determinations and DNA:rRNA hybridizations . The following conclusions were drawn . (i) The Xanthomonas campestris pathovars oryzae and oryzicola display clearly distinct protein patterns on polyacrylamide gels and can be differentiated from each other by four phenotype tests . (ii) Both pathovars are indeed members of Xanthomonas which belongs to a separate rRNA branch of the second rRNA superfamily together with the rRNA branches of Pseudomonas fluorescens, Marinomonas, Azotobacter, Azomonas and Frateuria . (iii) 'Brown blotch' strains are considerably different from X . campestris pv . oryzae and oryzicola . They are not members of the genus Xanthomonas, but are more related to the generically misnamed . Flavobacterium capsulatum, Pseudomonas paucimobilis, Flavobacterium devorans and 'Pseudomonas azotocolligans' belonging in the fourth rRNA superfamily . (iv) No correlation was found between the virulence, pathogenic groups or geographical distribution of X . campestris pv . oryzae or oryzicola strains and any phenotypic or protein electrophoretic property or clustering.

J Nutr Sci Vitaminol (Tokyo), 1984 Oct, 30(5), 415 - 20
Vitamin B6 biosynthesis and glycolate reductase in Flavobacterium sp . 238-7; Tani Y et al.; An enzyme which reduces glycolate to glycolaldehyde as the first step of the biosynthesis of vitamin B6 was studied using the particulate fraction of Flavobacterium sp . 238-7, a vitamin B6 producer . The enzyme catalyzes the reduction of glycolate in the presence of NADPH, but not the oxidation of glycolaldehyde, and is designated as glycolate reductase . There is a positive correlation between the activity of the enzyme and vitamin B6 biosynthesis . The activity and the formation of the enzyme were not affected by vitamin B6 . This enzyme probably plays a role in the extraordinary accumulation of vitamin B6 by the bacterium.

Appl Environ Microbiol, 1984 Oct, 48(4), 690 - 3
Purification and partial characterization of Flavotoxin A; Hu WJ et al.; A heat-resistant, low-molecular-weight toxin was isolated from semisolid potato dextrose agar medium after inoculation with Flavobacterium farinofermentans sp . nov., which was isolated from fermented corn meal that caused some outbreaks of food poisoning in China . The toxin was purified by solvent partition, Sephadex LH-20 gel filtration, and C-18 reversed-phase column chromatography . Thin-layer chromatography and high-pressure liquid chromatographic methods were developed for the identification and analysis of the toxin . The purified toxin exhibited a single spot in thin-layer chromatography and a single peak in high-pressure liquid chromatography and had adsorption maxima at 232 and 267 nm . Mass spectral analysis indicated a molecular weight of 169 with an experimental formula of C9H13O3 . The 50% lethal dose of purified toxin in mice (oral) was less than 6.84 mg/kg, but greater than 0.68 mg/kg . Postmortem examination showed that the mice died of some type of neurological and cardiovascular system toxicity . The name Flavotoxin A is being assigned to the toxin.

J Biol Chem, 1984 Sep 10, 259(17), 10700 - 4
Demonstration of peptide:N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations; Plummer TH Jr et al.; Endo-beta-N-acetylglucosaminidase F preparations from Flavobacterium meningosepticum have been found to contain peptide:N-glycosidase activity . Only the second activity, designated as peptide:N-glycosidase F, readily cleaves the beta-aspartylglycosylamine linkage of a fetuin triantennary complex glycopeptide, as shown by the isolation of the corresponding carbohydrate-free peptide containing aspartic acid and of an intact oligosaccharide with a di-N-acetylchitobiosyl moiety at the reducing end . Both activities in the mixture will hydrolyze a high mannose octaglycopeptide from ovalbumin, with the type of product formed being influenced by pH . At pH 4.0, only the endo-beta-N-acetylglucosaminidase F activity is functional, releasing octapeptide-GlcNAc and oligosaccharide-GlcNAc . At pH 9.3, the predominant cleavage is by peptide:N-glycosidase F at the glycosylamine bond, releasing octapeptide and oligosaccharide-GlcNAc-GlcNAc . This latter oligosaccharide is then hydrolyzed by endo-beta-N-acetylglucosaminidase F to oligosaccharide-GlcNAc plus GlcNAc.

J Clin Invest, 1984 Aug, 74(2), 341 - 50
Acceleration of thrombin-antithrombin complex formation in rat hindquarters via heparinlike molecules bound to the endothelium; Marcum JA et al.; We have examined the role of heparinlike molecules in the regulation of coagulation by perfusing rat hindquarters with purified human thrombin and with its plasma inhibitor, antithrombin . Our data indicate that contact of the hemostatic components with the endothelium enhances the rate of thrombin-antithrombin complex formation by as much as 19-fold over the uncatalyzed rate of enzyme-inhibitor interaction . Heparinlike molecules are responsible for the antithrombin accelerating activity . The amount of thrombin-antithrombin complex generated within the hindlimb preparation after pretreatment of the vasculature with purified Flavobacterium heparinase or with addition of platelet Factor IV to the hemostatic components, was equal to the uncatalyzed levels . These heparinlike molecules appear to be tightly bound to the luminal surface of the endothelium, since they could not be detected within the physiologic buffer that was perfused through the animal . The above mucopolysaccharides function in a manner similar to commercial heparin, since modification of antithrombin at a site critical for heparin-dependent acceleration of the protease inhibitor resulted in a level of interaction product identical to the uncatalyzed amount . Finally, addition of diisofluorophosphate-thrombin to the enzyme perfusion stream reduced the amount of thrombin-antithrombin complex formed in the animal by 30-40%, which suggested that thrombin bound to the endothelium as well as enzyme free in solution are accessible to antithrombin that has interacted with heparinlike molecules present on the endothelium.

J Antibiot (Tokyo), 1984 Aug, 37(8), 859 - 67
Microbial degradation of validamycin A by Flavobacterium saccharophilum . Enzymatic cleavage of C-N linkage in validoxylamine A; Asano N et al.; The enzymatic cleavage of C-N linkage in the degradation of validamycin A by Flavobacterium saccharophilum was examined using N-p-nitrophenyl derivatives of validamine and valienamine as synthetic model substrates for validoxylamine A . Incubation of N-p-nitrophenylvalidamine with the membrane fraction from the organism led to formation of N-p-nitrophenyl-3-ketovalidamine, and succeeding cleavage of C-N linkage . As the products of the cleavage step, one was identified as p-nitroaniline and another keto compound could not be purified enough because of its instability . However, on the basis of its hydrogenation products, the structure of the keto compound could be established as 5D-(5/6)-5-C-(hydroxy-methyl)-2,6-dihydroxy-2-cyclohexen-1-one . The same experiment was carried out with N-p-nitrophenylvalienamine . In this case, N-p-nitrophenyl-3-ketovalienamine could be isolated as an intermediate but the desired keto compound from the cleavage step could not be isolated because of its instability . The participation of two enzymes, that is, a dehydrogenase and a C-N lyase on the cleavage of C-N linkage was assured, and moreover, the analysis of its products, together with those of the previous studies allow us to propose a degradation pathway of validamycin A by Flavobacterium saccharophilum.

J Antibiot (Tokyo), 1984 Jul, 37(7), 773 - 80
Bacterial production of 7-formamidocephalosporins . Isolation and structure determination; Singh PD et al.; Two new 7-formamidocephalosporins have been isolated as their acetyl derivatives (SQ 28,516 and SQ 28,517) from fermentations of a Flavobacterium sp . SC 12,154 . Structure 1 was deduced for SQ 28,516 from its spectroscopic properties while structure 2 was proposed for SQ 28,517 . SQ 28,516 exhibits weak antibacterial activity.

Arch Dis Child, 1984 Jun, 59(6), 582 - 4
Trimethoprim sulphamethoxazole in neonatal Flavobacterium meningosepticum infection; Linder N et al.; During an outbreak of Flavobacterium meningosepticum septicaemia in a neonatal intensive care unit 9 infants were treated with intravenous trimethoprim sulphamethoxazole . Bacteriological cure was achieved in 8 patients; one infant died of massive intraventricular haemorrhage on the first day of treatment . Apart from prolonged persistence of pre-existing thrombocytopenia there was no evidence of side effects . Trimethoprim sulphamethoxazole should be considered in the treatment of neonatal F meningosepticum sepsis in view of its activity against this organism, good penetration of the blood brain barrier, and the absence of serious side effects.

Proc Natl Acad Sci U S A, 1984 Jun, 81(12), 3781 - 5
Segmental homology and internal repetitiousness identified in putative nucleic acid polymerase and human hepatitis B surface antigen of human hepatitis B virus; Ohno S; In a previous paper, it was argued that only those coding sequences descended from oligomeric repeats (the number of bases in the oligomeric unit not being a multiple of 3) can retain sufficiently long alternative open reading frames, and that such alternative open reading frames serve as the reservoir for the sudden generation of new polypeptide chains with novel functions . It was suggested that plasmid-encoded 6-amino hexanoic acid linear oligomer hydrolase that suddenly endowed Flavobacterium sp . K172 with the capacity to live off nylon by-products arose by the above mechanism . A corollary to the above argument is the expectation that those viral base sequences that are known to use two of the three alternative reading frames to encode two different polypeptide chains should invariably contain recognizable remains of the oligomeric tandem repeats, and as a consequence, various oligopeptidic repeats should also be present in the amino acid sequence of each . Furthermore, two polypeptide chains encoded by the same base sequence translated in different reading frames should show segmental homology of the type depicted previously . In the present paper, the base sequence of human hepatitis B virus ayw subtype that encodes an 832 amino acid residue long putative nucleic acid polymerase in one reading frame and a 226 residue long human hepatitis B surface antigen in the other reading frame was examined . All three predictions noted above were satisfied.

Infect Control, 1984 May, 5(5), 237 - 9
Flavobacterium meningosepticum; Ratner H; lavobacterium meningosepticum is an opportunistic pathogen of low virulence found in the hospital environment in water-containing equipment . Of primary importance is its role in outbreaks of neonatal meningitis which tends to be severe with a high mortality rate and serious sequelae . Changing all equipment concerned with humidifying or administering gases every 24 hours can help prevent these outbreaks in neonatal nurseries . Treatment is difficult because of the resistance of F . meningosepticum to most antimicrobial agents used to treat meningitis . Sensitivity tests, using a MIC method, are mandatory for infections caused by F . meningosepticum and, pending these results, vancomycin, given intravenously and, if necessary, intrathecally, appears to be the drug of choice for initial therapy.

J Bacteriol, 1984 May, 158(2), 419 - 24
DNA-DNA hybridization analysis of nylon oligomer-degradative plasmid pOAD2: identification of the DNA region analogous to the nylon oligomer degradation gene; Negoro S et al.; Fine structure of the gene of 6-aminohexanoic acid cyclic dimer hydrolase, one of the enzymes responsible for the degradation of the nylon oligomer (6-aminohexanoic acid cyclic dimer), on the plasmid pOAD2 harbored in Flavobacterium sp . KI72 was determined by constructing miniplasmids from plasmid pNDH5 (a hybrid plasmid consisting of pBR322 and a 9.1-kilobase-pair HindIII fragment of pOAD2 ) . The 6-aminohexanoic acid cyclic dimer hydrolase produced by cells of Escherichia coli C600 harboring pNDH5 or its miniplasmid was examined immunologically and electrophoretically and was found to be identical to that of Flavobacterium sp . KI72 . A fragment of pOAD2 (17.2- to 19.1-kilobase-pair region on pOAD2 ) was detected as hybridized fragment by Southern blotting experiments, indicating the presence of the DNA region analogous to the 6-aminohexanoic acid cyclic dimer hydrolase gene on the plasmid.

Carbohydr Res, 1984 Apr 2, 127(1), 75 - 94
Structural studies on heparin . Tetrasaccharides obtained by heparinase degradation; Linker A et al.; Three tetrasaccharides representing major structural sequences of heparin were isolated in good yield and characterized after degradation of heparin by purified flavobacterial heparinase . N-Desulfation was necessary to achieve good separation of these closely related compounds from each other . One of the tetrasaccharides was shown to be derived from the fully sulfated repeating segments; to contain L-iduronic acid and six sulfate groups, and have the structure delta 4,5- HexpA -(2-SO4)-(1----4)-alpha-D- GlcpN -(N-SO4)-(6-SO4)-(1- ---4)-alpha -L- IdopA -(2-SO4)-(1----4)-D- GlcN -(N-SO4)-(6-SO4) . The second contained a D-glucuronic acid unit that was nonsulfated instead of the L-iduronic acid, and the third, obtained in a fairly low yield, contained five sulfate groups, three of which being located on the disaccharide at the nonreducing end, and having the structure delta 4,5- HexpA -(2-SO4)-(1----4)-alpha-D- GlcpN -(N-SO4)-(6-SO4)-(1- ---4)-alpha -L- IdopA -(2-SO4)-(1----4)-D- GlcN -(N-SO4) . All tetrasaccharides had a sulfated, unsaturated uronic acid unit at the nonreducing end, confirming that the heparinase requires sulfated L-iduronic acid units for activity.

J Clin Microbiol, 1984 Apr, 19(4), 568 - 9
Flavobacterium multivorum septicemia in a hemodialyzed patient; Potvliege C et al.; A case of Flavobacterium multivorum septicemia in a hemodialyzed patient is reported . Two blood cultures were positive at 48 h, and the patient became afebrile only after antimicrobial therapy . The origin of the septicemia could not be determined.

Proc Natl Acad Sci U S A, 1984 Apr, 81(8), 2421 - 5
Birth of a unique enzyme from an alternative reading frame of the preexisted, internally repetitious coding sequence; Ohno S; The mechanism of gene duplication as the means to acquire new genes with previously nonexistent functions is inherently self limiting in that the function possessed by a new protein, in reality, is but a mere variation of the preexisted theme . As the source of a truly unique protein, I suggest an unused open reading frame of the existing coding sequence . Only those coding sequences that started from oligomeric repeats are likely to retain alternative long open reading frames . Analysis of the published base sequence residing in the pOAD2 plasmid of Flavobacterium Sp . K172 indicated that the 392-amino acid-residue-long bacterial enzyme 6-aminohexanoic acid linear oligomer hydrolase involved in degradation of nylon oligomers is specified by an alternative open reading frame of the preexisted coding sequence that originally specified a 472-residue-long arginine-rich protein.

EMBO J, 1984 Mar, 3(3), 581 - 4
Integrity of the pericellular fibronectin matrix of fibroblasts is independent of sulfated glycosaminoglycans; Hedman K et al.; The pericellular matrix fibers of cultured human fibroblasts contain fibronectin, other glycoproteins, and heparan and chondroitin sulfate proteoglycans . In the present study, cell-free pericellular matrices were isolated from metabolically labeled fibroblast cultures . The isolated matrices were digested with heparinase from Flavobacterium heparinum, and then analyzed for sulfated glycosaminoglycans (GAGs) . Nitrous acid degradation was used to distinguish the N-sulfated GAGs (heparan sulfate) from chondroitin sulfate . Fibronectin and the other major matrix polypeptides were studied using gel electrophoresis, enzyme immunoassay and immunofluorescence . Upon heparinase digestion, greater than 95% of sulfated GAGs were degraded in the matrix without detectable release of fibronectin or other matrix polypeptides or alteration of the fibrillar matrix structure . We conclude that in fibroblast cultures the integrity of the fibrillar matrix is independent of sulfated GAGs . Together with earlier observations, this suggests that filamentous polymerization of fibronectin forms the backbone of early connective tissue matrix.

Appl Biochem Biotechnol, 1984 Feb, 9(1), 41 - 55
An immobilized microbial heparinase for blood deheparinization; Linhardt RJ et al.; A new medical application of an immobilized microbial enzyme is described . Extracorporeal devices require systemic heparin administration to prevent thrombus formation; however, the use of heparin often leads to serious hemorrhagic complications . Heparinase isolated from Flavobacterium has been immobilized and used in a fluidized bed reactor to eliminate heparin from blood passing through an extracorporeal circuit both in vitro and in vivo . This paper discusses the stepwise development of this heparinase reactor including: (1) improvements in the fermentation resulting in an inexpensive large-scale source of heparinase without the addition of the previously required inducer, heparin; (2) the use of batch processes to adapt previous purification schemes to large-scale heparinase production and the subsequent purification of heparinase to a single SDS-PAGE banding protein; (3) the immobilization of heparinase with a 91% activity recovery and good stability, (4) the design and successful testing of a fluidized bed reactor containing immobilized heparinase in the removal of clinically used quantities of heparin from both human blood in vitro and canine blood in vivo; and (5) the initiation of animal studies focusing on the toxicology of heparinase-derived heparin degradation products and the short and long term effects of exposure to these products and to heparinase.

Mol Biochem Parasitol, 1984 Jan, 10(1), 99 - 109
Characterization of polysaccharides of the eggs and adults of Hymenolepis diminuta; Robertson NP et al.; Polysaccharides and other complex carbohydrates were released by proteolysis of the chloroform-methanol insoluble residue of 10 day-old worms and eggs of Hymenolepis diminuta . Gas-liquid chromatographic analysis of alditol acetate derivatives of monosaccharides released from the polysaccharides by hydrolysis revealed that in the 10 day-old worm, glucose was the most abundant sugar, followed by galactose, glucosamine, galactosamine, fucose and possibly rhamnose . Mannose was least abundant and xylose was absent . In the egg, glucose and galactose were equally abundant, followed by the same sugars found in 10 day-old worms, and xylose was present . Uronic acid was detected in both fractions by specific chemical tests . None of the saccharide material from eggs and worms was susceptible to degradation by Streptomyces hyaluronidase, chondroitinase AC, and slightly susceptible to chondroitinase ABC, as shown by electrophoretic analysis on composite 2.2% acrylamide-agarose slab gels and 4.5/12.5% polyacrylamide gels before and after enzymatic treatment . One of the gel-separable bands, however, was degradable by both nitrous acid and Flavobacterium heparinase . Both bands from eggs were degradable by nitrous acid . These results suggest that eggs contain heparin and/or heparan sulfate and perhaps dermatan sulfate and that 10 day-old worms also have these polyglycans but possibly not chondroitin sulfate or hyaluronic acid.

Nouv Rev Fr Hematol, 1984, 26(4), 255 - 60
Heparin-like molecules as regulators of atherogenesis; Rosenberg RD et al.; Bovine aortic endothelial cells release a heparin-like substance in the presence of 0.4% fetal calf serum . This substance inhibited the growth of smooth muscle cells in vitro by about 70% . Substitution of platelet-poor plasma for serum resulted in minimal liberation of inhibitory activity from the cells unless at least 10-fold higher concentrations of platelet-poor plasma were utilized . This suggested that a platelet product was involved in the release process . Therefore, we examined the ability of the platelet heparitinase recently isolated in our laboratory to release heparin-like species from cultured endothelial cells . Our results show that when endothelial cells were exposed to serum-free medium containing 1 ng/ml of the purified platelet endoglycosidase, at least as much inhibitory activity was released as was obtained with 0.4% serum . Dose response experiments indicated that only 10 pg/ml of the enzyme were necessary to liberate 50% of the inhibitory activity from endothelial cells . The heparin-like nature of the inhibitory substance was demonstrated by its sensitivity of Flavobacterium heparinase . Utilizing appropriate controls, the release of heparin-like material by the endoglycosidase was shown to be enzyme-specific and was not due to artifacts of experimental manipulations . In addition, this enzyme did not convert prereleased material to an active component, but directly liberated the active heparin-like species from endothelial cells . A simple model describing the possible role of heparin-like components and the endoglycosidase in the normal and injured wall will be presented.

Microbiol Immunol, 1984, 28(10), 1083 - 92
Biological activities of partially purified elastase produced by Flavobacterium meningosepticum; Miyazaki S; Injection of viable cells of Flavobacterium meningosepticum at doses of 10(6) colony-forming units/eye caused ophthalmia in rabbits' corneas . Elastase, free of protease activity, was partially purified from broth cultures of F . meningosepticum ATCC12535 and TMS516 . The optimum pH of the elastase activity was about 8.0 . The activity was suppressed by Ni, Zn, Co, EDTA, and o-phenanthroline, and accelerated by Cu and Fe . The minimum dose of the elastase causing ophthalmia in the eyes of rabbits was 5 micrograms/eye (0.065 elastase units) . The minimum lethal dose of the elastase for suckling mice by intracerebral injection was 0.4 micrograms (0.0052 elastase units)/suckling mouse, and its minimum lethal dose for adult mice by intraperitoneal injection was 740 micrograms/mouse (9.52 elastase units).

Appl Environ Microbiol, 1983 Dec, 46(6), 1447 - 9
Satellite growth of Legionella pneumophila with an environmental isolate of Flavobacterium breve; Wadowsky RM et al.; Legionella pneumophila serogroup 1 was observed to satellite around colonies of Flavobacterium breve on an L-cysteine-deficient medium which did not support growth of legionellae . Both isolates were recovered from the hot water tanks of hospitals . Ferric PPi stimulated satellite growth between 0.01 and 0.1%.

Nature, 1983 Nov 10-16, 306(5939), 203 - 6
Evolutionary adaptation of plasmid-encoded enzymes for degrading nylon oligomers; Okada H et al.; Flavobacterium sp . KI72 metabolizes 6-aminohexanoic acid cyclic dimer, a by-product of nylon manufacture, through two newly evolved enzymes, 6-aminohexanoic acid cyclic dimer hydrolase (EI) and 6-aminohexanoic acid linear oligomer hydrolase (EII) . These enzymes are active towards man-made compounds, the cyclic dimer and linear oligomers of 6-aminohexanoic acid respectively, but not towards any of the natural amide bonds tested . The structural genes of EI (nylA) and EII (nylB) are encoded on pOAD2, one of three plasmids harboured in Flavobacterium sp . KI72 . This plasmid contains two kinds of repeated sequence (RS-I and RS-II); one of the two RS-II sequences, RS-IIA, contains the nylB gene, while the other, RS-IIB, contains a homologous nylB' gene . From comparisons of the nucleotide sequences and gene products of the nylB and nylB' genes, we now conclude that EII enzyme is newly evolved by gene duplication followed by base substitutions on the same plasmid.

J Lab Clin Med, 1983 Nov, 102(5), 828 - 37
Heparinase: in vivo activity and immunogenicity in rabbits; Klein MD et al.; Anticoagulation with heparin is required during extracorporeal circulation for hemodialysis and cardiopulmonary bypass as well as during vascular surgery . Reversal of anticoagulation with protamine may be associated with hypotension and rebound anticoagulation and requires stoichiometric doses . Heparinase from Flavobacterium heparinum catalytically degrades heparin and reverses its anticoagulant effect . Heparin was administered to New Zealand White rabbits and plasma levels were assayed with the APTT anticoagulant assay and the azure A chemical assay . Heparinase actively degraded heparin both in vitro in rabbit plasma and in vivo in rabbit blood as determined by both the anticoagulant and chemical assays when compared to control heparin disappearance curves . Antibodies to heparinase were demonstrated by the ELISA technique in rabbits receiving i.v . heparinase . These antibodies, however, did not effect the activity of the enzyme in vitro or in vivo . No toxic effects of heparinase were noted in observations of the animals or in blood and histologic studies . Heparinase, either free or immobilized, may be a useful heparin-reversing agent without the drawbacks of protamine.

Am J Physiol, 1983 Nov, 245(5 Pt 1), H725 - 33
Microvascular heparin-like species with anticoagulant activity; Marcum JA et al.; Calf microvasculature was isolated from retina and cerebral gray matter . These preparations contained 0.048-0.060 U of heparin-like anticoagulant activity per gram of wet tissue . The retinal microvascular material contained no detectable mast cells . The anticoagulant potency of this product was associated solely with endothelial cells . This property appears to be due to a heparinlike proteoglycan since molecular species with biologic activity are precipitated with 10% (wt/vol) trichloroacetic acid and are destroyed by incubation with Flavobacterium heparinase . Furthermore, the above component functions in a manner virtually identical to heparin since approximately 60% of these species with anticoagulant activity bind to antithrombin-concanavalin A-Sepharose 4B, and only 15% of their biologic potency is expressed in the presence of antithrombin modified near the mucopolysaccharide binding domain . The cerebral microvascular tissue contained a trace subpopulation of mast cells (approximately 0.3%) . The anticoagulant activity of this preparation is most probably associated with both endothelial cells and mast cells . However, complete separation of these two cellular elements has proven difficult with current methodology.

Hoppe Seylers Z Physiol Chem, 1983 Oct, 364(10), 1467 - 73
{Biosynthesis of phenylalanine and tyrosine in Flavobacteria}; Waldner-Sander S et al.; The enzymes of the terminal steps of phenylalanine and tyrosine biosynthesis, chorismate mutase, prephenate dehydratase, arogenate dehydratase, prephenate dehydrogenase and arogenate dehydrogenase, were studied in 11 different species of the genus Flavobacteria . A comparison of the specific activities, cofactor specificity and regulation of the enzymes, allows a differentiation within the Flavobacteria . All strains studied utilize both arogenate and p-hydroxyphenylpyruvate as an intermediate in L-tyrosine synthesis . Phenylpyruvate was found to be the precursor of phenylalanine in most bacteria . No feedback inhibition of arogenate dehydrogenase by phenylalanine and tyrosine was observed . The diverse strains of the flavobacteria were found to possess different regulatory patterns with respect to the action of phenylalanine and tyrosine on the other enzymes . On the basis of these results a tentative classification of the Flavobacteria within the two groups formed by the different DNA base ratios is proposed.

Surg Neurol, 1983 Oct, 20(4), 294 - 6
Meningitis caused by Flavobacterium meningosepticum after transsphenoidal hypophysectomy with recovery; Chan KH et al.; Adult meningitis caused by Flavobacterium meningosepticum is rare . Five cases have been reported in the literature . The case reported herein developed in a woman after a transsphenoidal hypophysectomy . The isolate was resistant to erythromycin . Treatment with oral rifampicin combined with intravenous chloramphenicol and cefoperazone resulted in complete recovery.

Appl Environ Microbiol, 1983 Oct, 46(4), 917 - 24
Bacteriophages of methanotrophs isolated from fish; Tyutikov FM et al.; Bacteriophages of methanotrophic bacteria were isolated from 67 fish . Only two phages isolated from two fish species specifically lysed Methylocystis sp . and Flavobacterium gasotypicum . The phages lysing these species were designated 63-F and CMF-1-F, respectively . The isolated phages differed greatly in the fine structure of the virion, plaque morphology, spectrum of lytic action, serological properties, and UV sensitivity . At the same time, they had identical one-step growth characteristics: their latent period equalled 5 h, lysis time was 3 to 4 h, and burst size was about 240 virions . The phages had guanine- and cytosine-rich double-stranded DNAs consisting of common nitrogen bases . The molecular masses of the DNAs as determined by the sums of restriction endonuclease cleavage fragments were 28 X 10(6) daltons for phage 63-F and 31 X 10(6) daltons for phage CMF-1-F.

Biochemistry, 1983 Sep 13, 22(19), 4480 - 5
Evidence showing that a proline-specific endopeptidase has an absolute requirement for a trans peptide bond immediately preceding the active bond; Lin LN et al.; The proline-specific endopeptidase (EC 3.4.21.26) from Flavobacterium meningosepticum is specific for the cleavage of peptide bonds on the C-terminal side of prolyl residues . Such bonds will normally exist in the all-trans configuration . However, the preceding peptide bond in the sequence (i.e., on the N-terminal side of the prolyl residue) will exist as a mixture of cis and trans forms in solution . In this study, the activity of the proline-specific endopeptidase toward the substrates N-Cbz-Gly-Pro-MCA (where MCA = 4-methylcoumarinyl-7-amine) and N-Cbz-Gly-Pro-Leu-Gly has been examined . At a high ratio of enzyme activity/substrate concentration, the hydrolysis pattern for each substrate shows two well-separated kinetic phases . It is concluded that the fast kinetic phase, whose velocity depends on enzyme concentration, results from the direct hydrolysis of the active substrate bond (i.e., either the Pro-MCA or Pro-Leu bond, respectively) in molecules where the preceding Gly-Pro bond is trans . The slow phase, whose velocity is independent of enzyme concentration, is rate-limited by the cis-to-trans isomerization of those substrate molecules which initially have the preceding Gly-Pro bond in the cis configuration . That is, substrate molecules having the cis form of the Gly-Pro bond which precedes the active bond cannot be hydrolyzed directly but must first isomerize to the trans form before cleavage can occur . The amplitude, relaxation time, and activation energy for the slow phase are consistent with this interpretation . Thus, the proline-specific endopeptidase from Flavobacterium has an absolute requirement for a trans peptide bond at the position immediately preceding the active bond.

Eur J Pediatr, 1983 Sep, 140(4), 337 - 8
Successful treatment of neonatal Flavobacterium meningosepticum infection; Kaplan M et al.; An 8-day-old, 2.48-kg, 35-week gestation infant developed neonatal sepsis and meningitis due to Flavobacterium meningosepticum serotype F . Treatment with a new antibiotic, azlocillin, in combination with chloramphenicol, led to complete recovery.

Ann Trop Paediatr, 1983 Sep, 3(3), 125 - 8
Antimicrobial treatment of Flavobacterium meningosepticum infection; Johny M et al.; A six-month-old boy in hospital developed septicaemia and meningitis due to Flavobacterium meningosepticum type A . Although the organism was susceptible to amikacin, chloramphenicol and sulphonamides when tested by the disc diffusion method, therapy with these drugs was ineffective . Intravenous therapy with cotrimoxazole gave complete recovery . Various strains of flavobacteria including F . meningosepticum type B were isolated from the environment . Antimicrobial susceptibilities of patients and environmental strains against chloramphenicol, sulphonamides and rifampicin showed no correlation between disc diffusion results and minimum inhibitory concentration (MIC) . On the other hand, all the strains were found by both methods to be susceptible to amikacin, erythromycin, clindamycin and trimethoprim . Disc diffusion method should be supplemented with determination of MIC when testing antimicrobial susceptibility of Flavobacterium sp.

Rev Infect Dis, 1983 Jul-Aug, 5 Suppl 3, S600 - 5
Rifampin therapy for brucellosis, flavobacterium meningitis, and cutaneous leishmaniasis; Conti R et al.; Rifampin has a broad antibacterial spectrum . At high concentrations it also is active in vitro against protozoa, i.e., different species of Leishmania . Rifampin has been used against bacterial and occasionally protozoal infections . Addition of rifampin to a tetracycline regimen was found to reduce the number of relapses in patients with acute and chronic brucellosis . In a few cases of meningitis due to Flavobacterium meningosepticum, which failed to respond to other drugs, patients were treated successfully with rifampin administered either orally or intravenously . Rifampin at doses of greater than or equal to 600 mg daily has been administered to patients with cutaneous leishmaniasis . Healing of skin lesions was observed in the majority of treated patients . Controlled studies are needed to assess the usefulness of rifampin in this disease.

Anal Biochem, 1983 Jul 1, 132(1), 41 - 9
Purification and characterization of an extracellular protease from Flavobacterium arborescens; Boguslawski G et al.; An extracellular protease from Flavobacterium arborescens has been purified to an apparent homogeneity and characterized . The enzyme is most active at pH 8-10.5, requires no metal cofactor, and is inhibited by diisopropyl fluorophosphate . The protease is nonspecific, is active at temperatures up to 60 degrees C, and is completely free of nucleases . The ease of purification and freedom from nucleolytic contaminants make the protease a useful deproteinizing agent in DNA and RNA manipulations.

J Bacteriol, 1983 Jul, 155(1), 22 - 31
Plasmid-determined enzymatic degradation of nylon oligomers; Negoro S et al.; The nylon oligomer (6-aminohexanoic acid cyclic dimer) degradation genes on plasmid pOAD2 of Flavobacterium sp . KI72 were cloned into Escherichia coli vector pBR322 . The locus of one of the genes, the structural gene of 6-aminohexanoic acid linear oligomer hydrolase, was determined by constructing various deletion plasmids and inserting the lacUV5 promoter fragment of E . coli into the deletion plasmid . Two kinds of repeated sequences (RS-I and RS-II) were detected on pOAD2 by DNA-DNA hybridization experiments . These repeated sequences appeared five times (RS-I) or twice (RS-II) on pOAD2 . One of the RS-II regions and the structural gene of the hydrolase overlapped.

Exp Cell Res, 1983 Jun, 146(1), 29 - 42
Adhesive responses of fibroblast and neuroblastoma cells to substrata coated with polyvalent or monoclonal antibody to fibronectin; Harrison DD et al.; Both polyvalent and hybridoma-produced antibodies to fibronectin (Fn) were used to 'map' the immunoaccessible subsets of cell surface fibronectin on virus-transformed murine fibroblast SVT2 and rat neuroblastoma B104 cells . As one approach to this end, attachment and spreading responses of cells were measured on tissue culture substrata coated with antibody or with plasma fibronectin to compare their adhesive responses . Both SVT2 and B104 cells adhere poorly to polyvalent anti-Fn-coated substrata over short time intervals, but within several hours changes occur which permit cells to attach and spread as well on anti-Fn as on Fn (post-adsorption of the anti-Fn with Fn also generates a maximal response) . This adhesive response could be completely prevented by predigesting the cells with Flavobacterium heparanase, but not with chondroitinase ABC, indicating that the cell surface Fn responsible for antibody-mediated adhesion is associated with heparan sulfate proteoglycans on the cell surface . The compositions of the substratum-attached material (left bound after EGTA-mediated detachment of cells) from cells attaching to anti-Fn or Fn were analysed by SDS-PAGE and found to be identical within the same cell type for the two different substrata . Three hybridoma-produced antibodies, which recognize different determinants on Fn, generated different adhesive responses for SVT2 or B104 cells when adsorbed to the substratum . SVT2 cells adhered well to antibody no . 32-coated substrata but poorly to antibodies 92 or 136; on the other hand, B104 cells responded similarly to all three antibodies over short times of attachment but much better to no . 32 after a several hour incubation . These experiments indicate that (1) much of the cell surface fibronectin is complexed with heparan sulfate proteoglycan and is initially inaccessible to bind to polyvalent antibody on the substratum to promote adhesion; (2) the surface of neuroblastoma cells contains a fibronectin-like molecule which is important in their substratum adhesion; and (3) monoclonal antibodies are valuable tools in 'mapping' the orientation of cell surface molecules like fibronectin by measuring adhesive responses to antibody-coated substrata.

J Clin Microbiol, 1983 Jun, 17(6), 970 - 4
Patterns of extracellular proline-specific endopeptidases in Legionella and Flavobacterium spp . demonstrated by use of chromogenic peptides; Berdal BP et al.; Some Legionella strains possess a strong extracellular proline-specific endopeptidase (PSE) activity . Using an enlarged selection of chromogenic peptides representing a variety of N-terminal amino-acids binding to a -prolyl-proline, paranitroanilide chain, PSE activity of Legionella and Flavobacterium strains was examined . Differences in PSE activity emphasized the importance of the chemical structure at the nonchromogenic end of the peptide substrates . There seem to be distinct patterns of N-terminal specificity of PSE in the two bacterial groups.

J Antibiot (Tokyo), 1983 May, 36(5), 471 - 7
Marinactan, antitumor polysaccharide produced by marine bacteria; Umezawa H et al.; Extracellular polysaccharides of marine bacteria were screened for their antitumor activity against sarcoma-180 solid tumor in mice . An active polysaccharide was purified and named marinactan . The producing microorganism has a typical marine bacterial nature requiring sea water for growth and was identified as Flavobacterium uliginosum . Marinactan is a novel heteroglycan consisting of glucose, mannose and fucose in a ratio of approximately 7:2:1 . Marinactan, 10-50 mg mg/kg daily for 10 days i.p., produced 70-90% inhibition of the growth of solid sarcoma 180 . Complete regression of the tumor was observed in some treated mice . Its administrations before and after tumor transplantation showed almost the same inhibitory effect . Marinactan prolonged markedly the survival period of mice bearing ascites sarcoma 180.

Can J Microbiol, 1983 May, 29(5), 619 - 21
Lectin-binding activity and antibody response with Pseudomonas paucimobilis and Flavobacterium multivorum; Smalley DL et al.; Pseudomonas paucimobilis and Flavobacterium multivorum (formerly Group IIK biotype 1 and biotype 2, respectively) showed lectin-binding activity with Helix aspersa and no activity with eight other well-characterized lectins . Microagglutination titrations and immunodiffusion precipitation revealed specific antibody activity to immunizing antigens . However, no major antigens were found to be common from crude antigen preparations of P . paucimobilis and F . multivorum when tested against opposing antisera.

Biochem Biophys Res Commun, 1983 Mar 29, 111(3), 865 - 71
Structural differences of heparan sulfates according to the tissue and species of origin; Dietrich CP et al.; Some structural features of thirteen heparan sulfates isolated from different mammalian tissues and species are reported . Two N-acetylated disaccharides, one of then O-sulfated and two N-sulfated disaccharides, one of then 6-sulfated are formed from these compounds by the combined action of heparitinases I and II from Flavobacterium heparinum . The relative proportions of the four disaccharide units vary quite significantly among the thirteen heparan sulfates indicating that the structure of these polymers are tissue and species specific . Based on the frequency of appearance of each one of the disaccharides it was calculated that 10(36) types of heparan sulfates might theoretically be found . The possible role of these polyanions in cell-cell recognition is discussed in view of the present findings.

Appl Environ Microbiol, 1983 Mar, 45(3), 804 - 10
Nutritional versatility of a starch-utilizing Flavobacterium at low substrate concentrations; van der Kooij D et al.; A starch-utilizing yellow-pigmented bacterium, isolated from tap water, was tested for the utilization of 64 natural compounds at a concentration of 1 g/liter by measuring colony growth on agar media . Only 12 carbohydrates and glycerol promoted growth . Growth experiments with the organism in pasteurized tap water supplied with mixtures of substrates at concentrations of 1 or 10 micrograms of C of each substrate per liter, followed by separate experiments with a number of carbohydrates at 10 micrograms of C per liter showed that of these 64 natural compounds only sucrose, maltose, raffinose, starch, and glycerol promoted growth at very low concentrations . Also maltotriose, -tetraose, -pentaose, -hexaose, and stachyose, which were not included in the mixtures, enhanced growth, and generation times of 3 to 5 h at 10 micrograms of C per liter were observed . The organism, which was tentatively identified as a Flavobacterium species, thus appeared to be highly specialized in the utilization of glycerol and a number of oligo- and polysaccharides at very low concentrations.

J Bacteriol, 1983 Mar, 153(3), 1238 - 46
Unusual sulfonolipids are characteristic of the Cytophaga-Flexibacter group; Godchaux W 3rd et al.; Capnocytophaga spp . contain a group of unusual sulfonolipids, called capnoids (W . Godchaux III and E . R . Leadbetter, J . Bacteriol . 144:592-602, 1980) . One of these lipids, capnine, is 2-amino-3-hydroxy-15-methylhexadecane-1-sulfonic acid; the others are, apparently, N-acylated versions of capnine . The lipids were found, in amounts ranging from 2.5 to 16 mumol of capnoid sulfur per g of cells (wet weight), in two Cytophaga spp . and also in several closely related organisms: several Capnocytophaga spp., Sporocytophaga myxococcoides, two Flexibacter spp., and two Flavobacterium spp . With the exception of the flavobacteria, all of these bacteria have been shown to exhibit gliding motility . The two Flavobacterium spp . belong to a subset of that genus that shares many other characteristics with the cytophagas . Only the Capnocytophaga spp . contained large quantities of capnine as such; in all of the others, most (and possibly all) of the capnoids were present as N-acylcapnines . Capnoid-negative bacteria included some gliding organisms that may not be closely related to the cytophagas: two fruiting myxobacters, a gliding cyanobacterium (Plectonema sp.), Beggiatoa alba, Vitreoscilla stercoraria, Herpetosiphon aurantiacus, and Lysobacter enzymogenes . Nongliding bacteria representing nine genera were also tested, and all of these fell into the capnoid-negative group.

S Afr Med J, 1983 Feb 12, 63(7), 247 - 50
Cutaneous flavobacteriosis--polymorphous skin granulomas from Flavobacterium capsulatum . A case report; Findlay GH et al.; A case of multiple eruptive skin granulomas caused by Flavobacterium capsulatum is described . The organism was resistant or poorly sensitive to all antibiotics except carbenicillin . Cure was brought about by using maximal doses of this drug . The source of the infection could not be proved, but it dated from an orthopaedic procedure to the elbow which was followed by a chronic cellulitis at the operation site . Since this is an organism known to occur in stored water, it was presumed that the flavobacterium was introduced into the wound from bottles of boiled and cooled water used in the operating theatre.

Acta Pathol Microbiol Immunol Scand {B}, 1983 Feb, 91(1), 35 - 41
Studies on a collection of strains of the genus Flavobacterium . 2 . Nutritional studies; Bruun B; An attempt has been made to further characterize the Flavobacterium groups found in an earlier study by determining utilizable carbon-energy sources . As a preliminary, it was initially tried to define minimal growth factor requirements . It was found likely that most of the Flavobacterium groups have amino acid requirements, but a requirement for vitamins was also established for some strains . Testing Flavobacteria for carbon-energy sources by the replica plating method was found sufficiently technically difficult as to result in dubious readings at times . However, the overall results of the carbon-energy tests are in good agreement with the subdivision resulting from the previously performed biochemical tests, and the validity of the groups delimited in the earlier study is confirmed.

J Appl Bacteriol, 1983 Feb, 54(1), 45 - 56
The microbial flora of smoked pork loin and frankfurter sausage stored in different gas atmospheres at 4 degrees C; Blickstad E et al.; The development of the microflora of smoked pork loin and frankfurter sausage was followed during storage in vacuum, N2 and CO2 atmospheres at 4 degrees C . The total aerobic count on the smoked pork loin reached 10(7) organisms/g after 37 d in vacuum, 43 d in N2 and 49 d in CO2 . The corresponding value for the sausage was 77 d in vacuum, while the growth stopped at 6 x 10(40 organisms/g after 98 d in N2, and at 4 x 10(2) organisms/g after 48 d in CO2 . The predominant organisms on the fresh products were Bacillus spp., coryneform bacteria, Flavobacterium spp . and Pseudomonas spp . At the end of the storage time the microflora on both products in the three gas atmospheres, consisted mainly of Lactobacillus spp . and two large groups of organisms that could not be identified as any described genus . Some of the unidentified strains could be classified as a Lactobacillus sp . after subsequent subculturing on laboratory media . The numbers of Lactobacillus spp . at the end of storage decreased in the order, CO2 greater than N2 greater than vacuum . Lactobacillus viridescens generally constituted a substantial part of the Lactobacillus flora (5-72%) . On the sausages two large uniform groups of unidentifiable homofermentative Lactobacillus spp . were also found.

Jpn J Antibiot, 1983 Jan, 36(1), 111 - 6
{An in vitro study of susceptibility of Flavobacterium meningosepticum to antibiotics}; Igari J et al.; The present study concern in vitro observation on the susceptibility to 20 antibacterial agents of the strains of Flavobacterium meningosepticum from clinical specimens at Juntendo University Hospital during the 1 year period of 1981 . The tests for susceptibility of the 144 strains to the drugs were all performed by the serial 2-fold agar plate dilution method on Mueller Hinton agar (Difco), standardized by the Japan Society of Chemotherapy using the Microplanter apparatus with one loop of approximately 10(8) cells/ml . The antibacterial agents tested were as follows: ampicillin, sulbenicillin, piperacillin, mezlocillin, cefazolin, cefmetazole, cefoperazone, cefotaxime, ceftizoxime, kanamycin, gentamicin, tobramycin, amikacin, tetracycline, doxycycline, minocycline, nalidixic acid, pipemidic acid, erythromycin and clindamycin . 1 . Thirty-seven per cent of the strains were isolated from sputum, 23% from urine (greater than or equal to 10(5) ml), and 18% from pus and exudate . Only 3 and 2 strains were isolated from cerebrospinal fluid and blood, respectively . 2 . A large number of the strains tested were highly resistant to ABPC, SBPC, CEZ, KM, TOB, AMK, PPA and EM, and moderately resistant to PIPC, MZPC, CMZ, CPZ, CTX, CZX, GM, TC and NA . The most of the strains were highly sensitive to MINO, CLDM and DOXY in this order . There was no strain resistant to MINO in this study . 3 . The yearly change of the antibacterial activities of DOXY, MINO, CLDM was not significant and 9% of the strains in 1981 were highly resistant to CLDM . The strains resistant to EM were more frequently seen in 1981 than in 1974--1976.

Toxicol Lett, 1983 Jan, 15(1), 57 - 60
Reversion by vitamin K of aflatoxin B1 (AFB)-induced inhibition of oxygen uptake in three AFB-susceptible bacteria; Uwaifo OA; The effect of Vitamin K, on the inhibition of oxygen uptake in Bacillus brevis (2611), Bacillus megaterium (1368) and Flavobacterium aurantiacum by aflatoxin B1 (AFB) has been investigated . At 1.5 mM, vitamin K completely reversed oxygen inhibition by 10 micrograms/ml AFB in the three bacteria and by 50 micrograms/ml AFB in B . brevis, the most susceptible of the three bacteria to AFB . Vitamin K did not reverse the inhibitory effect of 100 micrograms/ml AFB in any of the three bacteria except B . megaterium, neither did 1.5 mM 2,4-dinitrophenol (DNP) reverse the inhibition at 10 micrograms/ml and 100 micrograms/ml levels of AFB in any of the three bacteria.

Ann Biol Clin (Paris), 1983, 41(3), 187 - 98
{Isolation, identification and clinical significance of species of the Flavobacterium genus}; Richard C et al.; Strictly aerobic Gram negative bacteria are found more and more often in human pathological specimens and in the environment . Amongst these bacteria, Flavobacterium poses problems of bacteriological diagnosis as their precise taxonomy is still recent . In this study, conducted on 321 strains of Flavobacterium from various sources, but essentially from patients in the intensive care unit, the authors define the methods and features of identification and the clinical significance of the species of the Flavobacterium genus . One species is particularly important in medical bacteriology, Flavobacterium meningosepticum which can cause septicaemia and neonatal meningitis which are difficult to treat.

J Gen Microbiol, 1982 Dec, 128 (pt 12), 2945 - 54
The similarities between Pseudomonas paucimobilis and allied bacteria derived from analysis of deoxyribonucleic acids and electrophoretic protein patterns; Owen RJ et al.; The chromosomal DNA was isolated and purified from 17 strains of Pseudomonas paucimobilis, and from the type or reference strains of Flavobacterium capsulatum, F . devorans, F . multivorum, 'Chromobacterium lividum', Xanthomonas campestris and seven species of Pseudomonas . The DNA base compositions (mol% G + C) of P . paucimobilis strains were between 62.2 and 68.6%, and typical strains had a mean value of 65.3 +/- 0.4 mol%, determined from thermal denaturation temperature . DNA-DNA molecular hybridization with 3H-labelled probe DNA from NCTC 11030 P . paucimobilis (the type strain) indicated that the species comprised a core of 13 closely related strains (74 to 96%), which included F . devorans NCIB 8195 (= ATCC 10829) . Four P . paucimobilis strains displayed lower levels of hybridization (less than or equal to 38%) . The hybridization results showed that P . paucimobilis was not closely related to allied yellow-pigmented bacteria or to other reference pseudomonads . The electrophoretic protein patterns of representative strains were analysed by computer-assisted techniques and similarity coefficients were calculated . A high degree of congruence was obtained with the DNA hybridization data, and the protein analyses indicated that the four atypical P . paucimobilis strains were heterogeneous and not a single group within the species . The generic affinities of P . paucimobilis, F . capsulatum, 'P . azotocolligans' and P . echinoides are uncertain, but available chemotaxonomic data indicate these species could provide the basis for a new genus.

Acta Pathol Microbiol Immunol Scand {B}, 1982 Dec, 90(6), 415 - 21
Studies on a collection of strains of the genus Flavobacterium . 1 . Biochemical studies; Bruun B; The genus Flavobacterium has undergone a number of changes during the last years resulting in a revised description of the genus in which restriction of Flavobacterium to certain low G + C content species is proposed . The purpose of the present study has been to see how a collection of 184 clinical and environmental strains of Flavobacterium-like organisms isolated in Denmark would fit into this latest revision . It was found that most of the different biochemical patterns represented gradually merged into each other, with certain patterns, however, occurring more frequently than others and corresponding to the already described taxa: F . meningosepticum, Flavobacterium group IIb, F . breve, F . odoratum, F . multivorum and F . spiritivorum . The lately proposed subdivision of the genus will need either revision or extension or both to cope satisfactorily with the situation revealed in this study.

Hoppe Seylers Z Physiol Chem, 1982 Nov, 363(11), 1365 - 75
{Properties of aromatic-amino-acid aminotransferases from two chloramphenicol-resistant Flavobacteria}; Beschle HG et al.; Two enzymes of chloramphenicol-resistant Flavobacterium strain CB 60 and strain CB 6 which catalyse the transamination of tyrosine, phenylalanine and tryptophan were enriched 43-fold and 31-fold . The molecular mass for the aromatic-amino-acid aminotransferases of both strains was found to be 120000 Da and the isoelectric point was at about pH 4.2-4.3 . Both enzymes are not influenced by EDTA . A ping pong bi-bi-mechanism was obtained for the kinetic mechanism of the reaction . The aminotransferases of strain CB 60 and CB 6 differ in the pH optimum (pH 8.4-9.0 and 7.8-9.0), in the optimum of temperature (40.0 degrees C and 57.5 degrees C) and in a higher heat stability of the enzyme of strain CB 60 in comparison to that of strain CB 6 . The catalytic activity of the enzyme of strain CB 60 is not influenced by cycloserine, contrary to the enzyme of CB 6 which is inhibited to 89% . Isonicotinohydrazide does not inhibit the enzyme of strain CB 6, but reduces the catalytic activity of the enzyme of strain CB 60 to 64% . Both enzymes are inhibited differently by phenylhydrazine {78% (CB 60) and 56% (CB 6)} . Data in percentages are related to 0.15 mM inhibitor . For the substrates tyrosine, phenylalanine, tryptophan and 2-oxoglutarate, the aminotransferase of strain CB 60 has Km values of 15.0, 16.6, 16.6 and 1.7mM, and for the enzyme of strain CB 6 Km values of 1.40, 1.33, 1.37 and 0.97mM were obtained.

J Biol Chem, 1982 Oct 10, 257(19), 11256 - 60
Inhibition of vascular smooth muscle cell growth by endothelial cell-derived heparin . Possible role of a platelet endoglycosidase; Castellot JJ Jr et al.; Bovine aortic endothelial cells release a heparin-like substance in the presence of 0.4% fetal calf serum . This substance inhibited the growth of smooth muscle cells in vitro by about 70% . Substitution of platelet-poor plasma for serum resulted in minimal liberation of inhibitory activity from the cells unless at least 10-fold higher concentrations of platelet-poor plasma were utilized . This suggested that a platelet product was involved in the release process . Therefore, we examined the ability of the platelet heparitinase described in the preceding communication to release heparin-like species from cultured endothelial cells . Our results show that when endothelial cells were exposed to serum-free medium containing 1 ng/ml of the purified platelet endoglycosidase, at least as much inhibitory activity was released as was obtained with 0.4% serum . Dose response experiments indicated that only 10 pg/ml of the enzyme were necessary to liberate 50% of the inhibitory activity from endothelial cells . The heparin-like nature of the inhibitory substance was demonstrated by its sensitivity to Flavobacterium heparinase . Utilizing appropriate controls, the release of heparin-like material by the endoglycosidase was shown to be enzyme-specific and was not due to artifacts of experimental manipulations . In addition, this enzyme did not convert prereleased material to an active component, but directly liberated the active heparin-like species from endothelial cells . A simple model describing the possible role of heparin-like components and the endoglycosidase in the normal and injured wall is presented.

J Clin Microbiol, 1982 Sep, 16(3), 564 - 5
Enzymatic profile of Pseudomonas paucimobilis; Smalley DL; Pseudomonas paucimobilis, previously known as group IIK biotype 1, was found to have a significantly different enzyme profile from that of Flavobacterium multivorum, formerly group IIK biotype 2 . P . paucimobilis produces several esterases and phosphatases which may contribute to its virulence.

J Pharmacobiodyn, 1982 Sep, 5(9), 734 - 40
An inhibitor for post-proline cleaving enzyme; distribution and partial purification from porcine pancreas; Yoshimoto T et al.; An inhibitor(s) for post-proline cleaving enzyme was checked by using a fluorogenic substrate, Z-Gly-Pro-4-methyl coumarinamide, and was found to be distributed widely in rat and porcine organs . The highest inhibitory activity on the enzyme was observed in pancreas and the inhibitor was partially purified from porcine pancreas extract by heat treatment, chromatographies on DEAE-Sephadex and Sephadex G-50 and affinity chromatography on trypsin-Sepharose . This inhibitor was very stable against temperature, pH and trichloroacetic acid treatment . The molecular weight was estimated to be 6500 by gel filtration . This inhibitor was highly specific for prolyl endopeptidases from mammals and Flavobacterium and inhibited the enzyme competitively . It acted neither on proline specific exopeptidases such as dipeptidyl aminopeptidase IV, proline aminopeptidase, prolidase, nor usual endopeptidases such as trypsin and alpha-chymotrypsin.

Proc Natl Acad Sci U S A, 1982 Aug, 79(15), 4540 - 4
endo-beta-N-acetylglucosaminidase F: endoglycosidase from Flavobacterium meningosepticum that cleaves both high-mannose and complex glycoproteins; Elder JH et al.; We have detected an endoglycosidase activity produced by Flavobacterium meningosepticum . This enzyme, named endo F, cleaves glycans of both the high-mannose and the complex type linked through asparagine to the protein backbone . The data indicate that cleavage occurs via hydrolysis of the glycosidic bond of the N,N'-diacetylchitobiose core structure adjacent to asparagine, similar to that due to endo H and endo D . Extreme variability was noted in the availability of this cleavage site among N-linked glycoproteins . Glycoproteins of retrovirus, lymphocytic choriomeningitis virus, Pichinde virus, and HLA-A and -B antigens were readily cleaved in the presence of nonionic detergent . Others, such as ovalbumin, fetuin, bromelain, ovomucoid, alpha 1-acid glycoprotein, immunoglobulin G, and influenza virus hemagglutinin became susceptible only after reduction and alkylation or when cleavage was performed in the presence of 1% 2-mercaptoethanol . Endo F should prove useful in the study of glycans and protein backbones as discrete entities and for defining the nature of the glycan-protein interface.

Jpn J Antibiot, 1982 Jun, 35(6), 1579 - 84
{Infections during induction chemotherapy of acute leukemia and their control V . Clinical evaluation of a large dose of amikacin injected intravenously}; Moriyama Y et al.; In this study, we treated severe infections (21 cases) accompanied with induction chemotherapy in 20 patients with acute leukemia by the combination of a large dose (600 approximately 1,200 mg/day) of amikacin with other antibiotics . Infections during induction chemotherapy of acute leukemia consisted of sepsis (8 cases), pneumonia (7) and others (6), and most of causative organisms were Gram-negative bacteria, such as Ps . aeruginosa (7 strains), Flavobacterium (5), Serratia (3), Ps . cepacia (2), E . coli (2) and others . The combination chemotherapy of a large dose of amikacin with other antibiotics was found to be effective (71.4%) for such infections . Side effects were negligible except for drug eruption . Therefore, a large dose of amikacin should be given for the treatment of severe infections accompanied with induction chemotherapy of acute leukemia.

J Biochem (Tokyo), 1982 Jun, 91(6), 1899 - 906
Proline-specific dipeptidyl aminopeptidase from Flavobacterium meningosepticum; Yoshimoto T et al.; A proline-specific dipeptidyl aminopeptidase was highly purified from cell-free extract of Flavobacterium meningosepticum by a series of column chromatographies on DEAE-Sephadex A-50, Sephadex G-150, hydroxyapatite, and a second gel filtration on Sephadex G-150 . The enzyme was most active at pH 7.4-7.8 for both Gly-Pro-beta-naphthylamide (Gly-Pro-2-NNap) and Gly-Pro-p-nitroanilide (Gly-Pro-pNA) and was stable between pH 7 and 9.5 . The enzyme was markedly inhibited by diisopropylphosphofluoridate (DFP) and mercury ion but not by sulfhydryl-blocking reagents and metal chelators . The molecular weight of the enzyme was about 160,000 as judged by the gel filtration method and the subunit molecular weight was estimated to be 75,000 by sodium dodecyl sulfate (SDS)-gel electrophoresis, suggesting a dimeric form of the native enzyme . The isoelectric point was at pH 9.5 . The enzyme hydrolyzed peptides and peptide amides at the carboxyl side of a proline residue penultimate to the amino-terminal amino acid, as did post-proline dipeptidyl aminopeptidases from various mammals . However, antiserum raised against post-proline dipeptidyl aminopeptidase from porcine kidney did not cross-react with the Flavobacterium dipeptidyl aminopeptidase.

Eur J Clin Microbiol, 1982 Jun, 1(3), 138 - 43
Flavobacterium meningosepticum ventriculitis: in vivo and in vitro results with the combinations rifampicin-erythromycin and mezlocillin-cefoxitin; Kelsey MC et al.; A case of Flavobacterium meningosepticum ventriculitis is described in which the failure of therapy with a combination of rifampicin and erythromycin is attributed to inadequate antibiotic levels in cerebrospinal fluid . The successful eradication of the organism was achieved with the use of mezlocillin and cefoxitin given by the intravenous and intraventricular route . In vitro sensitivity tests of recently isolated strains of Flavobacterium meningosepticum suggested that the combination mezlocillin and cefoxitin is more often synergistic than rifampicin and erythromycin.

Biochim Biophys Acta, 1982 Apr 3, 702(2), 197 - 203
Mode of action of heparin lyase on heparin; Linhardt RJ et al.; Heparinase (heparin lyase, EC 4.2.2.7) prepared from Flavobacterium heparinum was used to digest heparin . The products of digestion were examined with a viscosometric assay at various stages of the reaction to measure their average molecular weight . By comparison with computer simulations of various models, heparinase was shown to act in a random endolytic mode . The relative abundance of intermediates in heparin degradation catalyzed by heparinase immobilized on Sepharose 4B was measured by high pressure liquid chromatography (HPLC) at various time points . The results obtained using HPLC were consistent with a random endolytic mechanism . The heparin digestion products were separated and identified using gel permeation chromatography . The final distributions of heparin degradation products for free and immobilized heparinase were identical . Contaminating sulfatases and glycuronidases which could have subsequently acted on heparin degradation products were not found in significant amounts in the heparinase preparation studied.

Hoppe Seylers Z Physiol Chem, 1982 Apr, 363(4), 439 - 44
Conversion of chloramphenicol degradation products by tyrosine aminotransferase from Flavobacteria; Beschle HG et al.; The tyrosine aminotransferase of Flavobacterium strain CB 60, strain CB 6 and F . devorans r - a partially purified enzyme was used - is able to deaminate oxidatively p-aminophenylalanine and the intermediate products of chloramphenicol degradation p-nitrophenylserine and p-aminophenylserine . The aminotransferases of the strains CB 6 and CB 60 also convert p-aminophenylserinol . p-Nitrophenylserinol only reacts with the enzyme from strain CB 6 . Determination of substrate specificity from strain CB 6 shows that an alcoholic group in C3 position (ring proximal) and to a lower degree an alcoholic group in C1 position (ring distal) decrease the turnover rate . Based on its broad substrate specificity the tyrosine aminotransferase has the ability not only to metabolize physiological compounds but also degradation products of chloramphenicol.

Biochim Biophys Acta, 1982 Feb 15, 710(2), 117 - 21
Stabilization of lipoprotein lipase by endothelial cells; Shimada K et al.; Lipoprotein lipase, purified from bovine milk, lost 90% of its activity when incubated in Hanks' balanced salt solution for 5 min at 37 degrees C . Bovine pulmonary artery endothelial cells, maintained in culture, markedly stabilized this enzyme . The stabilizing factor of endothelial cells was non-dialyzable, resistant to heating at 100 degrees C and to changes in pH, and unaffected by treatments of cells with proteolytic enzymes or with heparinase (Flavobacterium heparinum enzyme) . However, the stabilizing effect on lipoprotein lipase was reduced by 60-70% by the extraction of cells with chloroform/methanol (2:1) . The lipid extract of the cells stabilized the enzyme, suggesting that lipid component(s) of the endothelial cells account for their stabilizing effect . Since the endothelial cell is thought to be the site of action of lipoprotein lipase, stabilization of the enzyme by this cell may play a role in its preservation and function in vivo.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1981 Dec, 174(4), 314 - 24
{Identification of bacteria from use-surface disinfectant solutions and their sensibility against disinfectants (author's transl)}; Dott W et al.; The investigations are concerned with bacteria which had been isolated from use-surface disinfectant solutions originated from 19 disinfection dosage apparatus of different hospitals and from 8 taps of a centralised disinfection dosage apparatus in an other hospital . The greatest part of the isolated bacteria belonged to the genus Pseudomonas . Beyond it there were yellow-pigmented bacteria like Xanthomonas, Flavobacterium and Micrococcus . After subculture on solid peptone-containing media most of the isolated bacteria turned out to be sensitive to the disinfection solutions in a concentration range, that had been applied in the dosage apparatus . The examination of 6 disinfectants showed, that there were considerable differences in the efficiency to kill the bacteria . Even one preparation pointed out to be absolutely ineffective at applied concentrations.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1981 Dec, 174(4), 299 - 313
{Microbiological investigations with decentral dosing apparatuses for disinfectants . Part 1: investigations of cell number (author's transl)}; Heinzel M et al.; Cell numbers have been investigated for several months in disinfectant solutions which were measured out with tap water by decentral automatically dosing apparatuses . They differed not only quantitatively but also qualitatively with respect to the products used: With one product on the bases of aldehydes + organic tin compound 3 h after samples have been collected up to 100 bacteria/100 ml could be cultivated . Thus, cell numbers in disinfectant solutions were in the same range of cell numbers of tap water . Sometimes the cell numbers in both substrates seem to go parallel (Fig . 1) . The bacterial compositions in both were similar (Table 3); gramnegative, yellow bacteria of the genera Pseudomonas and Flavobacterium dominated . After the disinfectant in one apparatus has been substituted by one which is composed of aldehydes + a quaternary ammonium compound cell numbers rapidly and permanently decreased to the limit of detectability . Only coryneform bacteria survived for a longer time which were characterized (Tab . 4) but not determined taxonomically . Their facultatively methylotrophic growth was conspicuously.

J Clin Microbiol, 1981 Dec, 14(6), 612 - 6
Cellular fatty acid composition of organisms frequently associated with human infections resulting from dog bites: Pasteurella multocida and groups of EF-4, IIj, M-5, and DF-2; Dees SB et al.; The cellular fatty acid composition of Pasteurella multocida and four unclassified groups of gram-negative bacteria (EF-4, M-5, IIj, and DF-2) which are frequently associated with human dog-bite infections was determined by gas-liquid chromatography . Strains of P . multocida were readily distinguished from the unclassified groups by the presence of 3-hydroxy myristic acid (3-OH- 14:0) . Groups M-5 and EF-4 were characterized by the presence of 3-hydroxy lauric (3-OH-12:0) acid . Only group EF-4 organisms contained 2-OH-16:0, a-17:0, and 17:0 cyclopropane acids . Groups IIj and DF-2 differed from the other groups by the presence of large amounts of a branched-chain 15-carbon acid (i-15:0); they differed from each other by the presence of i-2-OH-15:0 and i-17:1 acids in IIj, which were absent in DF-2 . The data indicate that gas-liquid chromatographic analysis for cellular fatty acids provides an additional test for rapid differentiation of these gram-negative organisms associated with dog-bite infections . Similarities observed in the fatty acid compositions of Flavobacterium, IIj, and DF-2 suggest that these two unclassified groups may be additional species of Flavobacterium.

Gene, 1981 Dec, 16(1-3), 73 - 8
New restriction endonucleases from Flavobacterium okeanokoites (FokI) and Micrococcus luteus (MluI); Sugisaki H et al.; Two new restriction endonucleases have been isolated from Flavobacterium okeanokoites IFO12536 and Micrococcus luteus IFO12992 and named FokI and MluI, respectively . Based on analysis of the sequences around the restriction sites, the recognition sequences and cleavage sites of these endonucleases were deduced as below: FokI: (formula: see text) . MluI introduces double-strand cleavages at unique sequences that are completely two-fold rotationally symmetric like most type II restriction endonucleases . FokI belongs to a class of restriction endonucleases that recognize specific but asymmetric nucleotide sequences and introduce staggered cleavages at appointed positions away from the recognition sequences.

J Cell Biol, 1981 Nov, 91(2 Pt 1), 427 - 37
Structural features of alveolar wall basement membrane in the adult rat lung; Vaccaro CA et al.; The ultrastructural characteristics of alveolar (ABM) and capillary (CBM) basement membranes in the adult rat lung have been defined using tannic acid fixation, ruthenium red staining, or incubation in guanidine HCl . ABM is dense and amorphous, has 3- to 5-nm filaments in the lamina rara externa (facing the alveolus) that run between the lamina densa and the basal cell surface of the epithelium, has an orderly array of ruthenium red-positive anionic sites that appear predominantly (79%) on the lamina rara externa, and has discontinuities beneath alveolar type II cells but not type I cells that allow penetration of type II cytoplasmic processes into the interstitium of the alveolar wall . The CBM is fibrillar and less compact than ABM, has no lamina rara filaments, and has one fifth the number of ruthenium red-positive anionic sites of ABM that appear predominantly (64%) overlying the lamina densa . Incubation of lung tissue with Flavobacterium heparinum enzyme or with chondroitinase has shown that ABM anionic sites represent heparan sulfate proteoglycans, whereas CBM anionic sites contain this and other sulfated proteoglycans . The CBM fuses in a local fashion with ABM, compartmentalizing the alveolar wall into a thick and thin side and establishing a thin, single, basement-membrane gas-exchange surface between alveolar air, and capillary blood . The potential implications of ABM and CBM ultrastructure for permeability, cell differentiation, and repair and morphogenesis of the lung are discussed.

Biochim Biophys Acta, 1981 Oct 12, 677(2), 194 - 9
Bacterial metabolism of ethylene glycol; Willetts A; Metabolism of ethylene glycol as the sole source of carbon by a species of Flavobacterium was affected by the dissolved oxygen tension of the growth medium . Under strongly aerobic conditions the diol was exclusively metabolised to glycollate by an initial oxidase, subsequently metabolised to acetyl-CoA with no net change in ATP, and then oxidised to CO2 by the tricarboxylic acid cycle yielding large amounts of reduced nicotinamide nucleotides which were used to generate a net gain in ATP by oxidative phosphorylation . Under microaerophilic conditions, some ethylene glycol after initial metabolism to acetyl-CoA by the oxidase-inhibited pathway, was subsequently catabolised to acetyl phosphate and then acetate, yielding a net gain in ATP by substrate-level phosphorylation: additionally some diol was catabolised by an inducible diol dehydratase to acetaldehyde and subsequently reduced to ethanol as a terminal metabolite.

J Clin Invest, 1981 Oct, 68(4), 995 - 1002
Involvement of cell surface heparin sulfate in the binding of lipoprotein lipase to cultured bovine endothelial cells; Shimada K et al.; It has been postulated that lipoprotein lipase, an enzyme important in the uptake of fatty acids into tissues, is bound to the vascular endothelial cell surface and that this binding occurs through attachment to heparinlike glycosaminoglycans . Furthermore, it is thought that heparin releases the enzyme from its attachment to the endothelium into the circulation . These hypotheses have never been tested directly in cell systems in vitro . In the present study we have directly evaluated the interaction of lipoprotein lipase, purified from bovine skim milk with monolayer cultures of endothelial cells, isolated from bovine pulmonary artery . Endothelial cells in primary culture had no intrinsic lipoprotein lipase activity but were able to bind lipoprotein lipase quantitatively . The binding reached equilibrium and was saturable at 0.24 nmol of lipoprotein lipase/mg of cell protein . The concentration of lipoprotein lipase at half-maximal binding was 0.52 microM . Bound lipoprotein lipase could be detached from cultured cells by increasing concentrations of heparin, and at and above 0.6 microgram/ml of heparin, 90% of the cell-bound lipoprotein lipase activity was released . Heparan sulfate and dermatan sulfate released the enzyme to a lesser extent and chondroitin sulfate caused little, if any, release of lipoprotein lipase . The release of lipoprotein lipase with heparin was not associated with a release of {3S}glycosaminoglycans from 35S-prelabeled cells . Reductions of lipoprotein lipase binding to endothelial cells and of cell surface-associated {3S}glycosaminoglycans in 35S-prelabeled cells occurred in parallel both when cells were pretreated with crude Flavobacterium heparinum enzyme before lipoprotein lipase binding and when cells were treated with this enzyme after lipoprotein lipase binding . The removal of heparan sulfate from the cell surface by purified heparinase totally inhibited the binding of lipoprotein lipase by endothelial cells, but the removal of chondroitin sulfate by chondroitin ABC lyase had no effect on this binding . These results provide direct evidence for lipoprotein lipase attachment to endothelial cells through heparan sulfate on the cell surface, and provide evidence for the release of lipoprotein lipase by heparin through a detachment from this binding site.

P N G Med J, 1981 Sep, 24(3), 169 - 73
The laboratory diagnosis of opportunistic infections caused by uncommon bacteria in Papua New Guinea; Gratten M; Flavobacterium meningosepticum, Chromobacterium violaceum and Listeria monocytogenes are opportunistic pathogens of man and are occasional but important causes of infections in neonates, infants and adults in Papua New Guinea . Mortality is high . Flavobacterium meningosepticum should be suspected when a weak indole producing oxidase positive Gram-negative bacillus producing a discrete yellow non-diffusing pigment at room temperature and showing a resistant antibiotic sensitivity pattern is isolated from the blood and/or C.S.F . of a neonate . Chromobacterium violaceum can be presumptively identified if a Gram-negative bacillus producing an intense non-diffusing violet pigment is cultured, commonly in associated with multiple abscess formation in the host . The isolation of a slender Gram-positive bacillus from C.S.F . or blood which produces catalase and acetylmethylcarbinol is consistent with Listeria monocytogenes . This organism forms discretely beta haemolytic colonies on blood agar and attacks the carbohydrates salicin and trehalose.

Eur J Biochem, 1981 Jun 1, 116(3), 547 - 51
Purification and characterization of 6-aminohexanoic-acid-oligomer hydrolase of Flavobacterium sp . Ki72; Kinoshita S et al.; 6-Aminohexanoic-oligomer hydrolase of Flavobacterium sp . KI72 was purified to homogeneity by column chromatography three times, and by preparation polyacrylamide gel electrophoresis twice . The purified enzyme had the following characteristics . 1 . The molecular weight was estimated to be 84000 by Sephadex G-200 molecular-sieve chromatography . The enzyme consisted of two homologous subunits of 42000, judged from sodium dodecylsulfate/polyacrylamide gel electrophoresis . 2 . The optimum pH for activity was between 8 and 9, the optimum temperature was 40 degrees C for a 1-h reaction . The Michaelis-Menten constants and turnover numbers for the 6-aminohexanoic acid dimer and trimer were 5.9 mM and 2.4 s-1, and 6.2 mM and 2.0 s-1 respectively . 3 . The enzyme was inhibited by 0.37 mM diisopropylfluorophosphate and by 0.013 mM p-chloromercuribenzoate . 4 . The enzyme was active on 6-aminohexanoic acid oligomers from dimer to hexamer and icosamer but not on hectamer, and the activity decreased with the increase of the polymerization number of the oligomer . The oligomers were hydrolyzed so as to remove the 6-aminohexanoic acid residue successively from the amino terminus . The enzyme could not hydrolyze other linear amides, cyclic amides, dipeptides, tripeptides or casein . 5 . 6-aminohexanoic-acid-oligomer hydrolase was classified as a new member of the linear amidases (EC 3.5.1.-).

South Med J, 1981 Jun, 74(6), 764 - 6
Flavobacterium meningosepticum sepsis: disease due to bacteria with unusual antibiotic susceptibility; Harrington SP et al.; The three cases we have presented implicate F meningosepticum as a significant pathogen causing disease in immunocompromised adult patients . Since this organism is of low pathogenicity but may be found in the hospital environment, its identification as a pathogen raises the suspicion of a nosocomial source of infection, and a search for the source should be made . Furthermore, with regard to the choice of antimicrobial therapy, it must be remembered that this organism is resistant to most antibiotics commonly used to treat gram-negative bacilli, and disk diffusion technics may not reliably predict actual antibiotic sensitivity.

South Med J, 1981 Jun, 74(6), 757 - 9
Flavobacterium meningosepticum in the neonatal period; Ferlauto JJ et al.; We have described a term newborn infant who developed Flavobacterium meningosepticum meningitis at 9 days of age . The case demonstrates the tenacity and changing antibiotic sensitivity of this organism . We used trimethoprim-sulfamethoxazole (Bactrim) intravenously to obtain CSF sterilization . On follow-up this child was neurologically intact.

J Biol Chem, 1981 May 10, 256(9), 4310 - 20
Cells selected for high tumorigenicity or transformed by simian virus 40 synthesize heparan sulfate with reduced degree of sulfation; Winterbourne DJ et al.; Cell lines, selected from two independent clones of an established mouse embryo cell line by their ability to grow as solid tumors in immunocompetent syngeneic hosts, were found to have the same alteration in anion exchange properties as was previously reported for simian virus 40 (SV40)-transformed subclones . One tumor cell line (219CT) and one SV40-transformed subclone (215CSC) were selected for further detailed comparison with their common parent clone (210C) . Cellulose acetate electrophoresis at pH 1.0 showed that 215CSC heparan sulfate had a slight overall decrease in sulfation compared with heparan sulfate from 210C; however, no gross difference in sulfation could be detected between heparan sulfate from 219CT and 210C . Analysis of the products of deaminative cleavage of heparan sulfate by nitrous acid under conditions where cleavage occurs quantitatively at N-sulfated glucosamine residues showed that, although heparan sulfate from the three cell lines gave similar yields of O-sulfated disaccharides, both 215CSC and 219CT had only about half as many O-sulfate residues in higher molecular weight oligosaccharides compared to heparan sulfate from 210C . Enzymatic degradation of heparan sulfate with a mixture of enzymes from Flavobacterium heparinum showed that this common alteration in heparan sulfate from both 215CSC and 219CT resulted from a 30% decrease in glucosamine residues bearing 6-O-sulfate groups . As this decrease in 6-O-sulfate glucosamine residues occurs in regions of the chain containing relatively few sulfate groups, it is clear that certain sequences of charged groups present in heparan sulfate frm 210C will be found only rarely in heparan sulfate from 215CSC and 219CT . It is suggested that this will result in alterations of the interaction of heparan sulfate with other molecules in the microenvironment at the cell surface which may be important in the control of such phenomena as cell growth and adhesion.

J Clin Pathol, 1981 Apr, 34(4), 429 - 33
Flavobacterium meningosepticum infection: an epidemiological study in a newborn nursery; Thong ML et al.; After an outbreak of flavobacterium meningitis in the newborn nursery of the University Hospital, an investigation was carried out to determine the possible sources of this organisms in the nursery . Various serotypes of the organism were recovered from a variety of sources such as wash basins, sinks, suction apparatus, and disinfectants in the nursery as well as the neighbouring wards . Colonisation of the pharynx with this organism was demonstrated in several clinically healthy babies in the nursery . The possible role of fomites, hospital staff, newborn babies, and their mothers in the transmission of the organism is considered together with the measures taken to reduce environmental contamination.

Eur J Biochem, 1981 Apr, 115(2), 287 - 91
A bacterial glucoamylase degrading cyclodextrins . Partial purification and properties of the enzyme from a Flavobacterium species; Bender H; A Flavobacterium species has been isolated which produces a cyclodextrin-degrading glucoamylase . The inducible, cell-bound enzyme was purified about 10-fold to 75% purity in 57% yield . The action of the enzyme was studied with the main cyclodextrins (cyclohexaamylose, cycloheptaamylose and cyclooctaamylose) and with the typical glucoamylase substrates, respectively . The final degradation product with all the substrates was glucose . Small amounts of maltose, which could be detected in the course of cyclodextrin degradation, were hydrolyzed at a lower rate . V for cyclohexaamylase was found to be about 14-15 mumol glucose min-1 (mg pure protein)-1, the Km for cyclohexaamylose was 0.142 mM . Apparently the enzyme preferred shorter alpha-D-glucopyranosyl chains . Besides maltose, amylopectin and glycogen proved to be very poor substrates . Some properties of the enzyme have been described.

Carbohydr Res, 1981 Feb 2, 88(2), 291 - 303
Purification of heparinase and heparitinase by affinity chromatography on glycosaminoglycan-bound AH-Sepharose 4B; Ototani N et al.; Heparinase and heparitinase were separated from an extract of Flavobacterium heparinum, induced with heparin by using column chromatography on hydroxylapatite . As the heparinase preparation contained chondroitinases B and C, chondroitinase B was removed by rechromatography on a hydroxylapatite column . Chondroitinase C was then eliminated by column chromatography on O-phosphono("phospho")-cellulose . The heparinase preparation thus obtained was free from sulfoamidase for 2-deoxy-2-sulfoamino-D-glucose (GlcN-2S), sulfatase for 2-amino-2-deoxy-6-O-sulfo-D-glucose (GlcN-6S), as well as delta 4,5glycosiduronase for the unsaturated disaccharides obtained from heparin . The remaining sulfatase for 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid 2-sulfate (delta UA-2S) in the heparinase preparation was removed by affinity chromatography with dermatan sulfate-bound AH-Sepharose 4B coated with dermatan sulfate . The heparitinase preparation separated by column chromatography on hydroxylapatite was purified by affinity chromatography with heparin-bound AH-Sepharose 4B coated with heparin . Sulfatase for 2-amino-2-deoxy-6-O-sulfo-D-glucose (GlcN-6S) and delta 4,5glycosiduronase for the unsaturated disaccharides obtained from heparin were removed by this chromatography . Sulfatase for 4-deoxy-alpha-L-threo-hex-4-enopyranosyluronic acid 2-sulfate (delta UA-2S) remaining in the heparitinase preparation was finally removed by column chromatography on hydroxylapatite . The recoveries of the purified preparations of heparinase and heparitinase were estimated to be 39 and 50%, respectively, from the crude enzyme fractions obtained by the first column chromatography on hydroxylapatite . The purified heparinase and heparitinase were free from all enzymes that could degrade the sulfated unsaturated disaccharides produced from heparin with heparinase.

Appl Environ Microbiol, 1981 Feb, 41(2), 360 - 5
Heparinase production by Flavobacterium heparinum; Galliher PM et al.; Heparinase production by Flavobacterium heparinum in complex protein digest medium, with heparin employed as the inducer, has been studied and improved . The maximum productivity of heparinase has been increased 156-fold over that achieved by previously published methods to 375 U/liter per h in the complex medium . Rapid deactivation of heparinase activity, both specific and total, was observed at the onset of the stationary phase . Nutritional studies on growth and heparinase production showed an obligate requirement for L-histidine and no vitamin requirement . L-Methionine partially relieved the L-histidine requirement . A defined medium containing glucose, ammonium sulfate, basal salts, L-methionine, and L-histidine was developed for growth and heparinase production . The growth rate in this medium was 0.21 h-1, which is 40%, higher than that in complex medium . The maximum volumetric productivity of heparinase in the defined medium was increased to 1,475 U/liter per h, providing a 640-fold increase over that achieved by previously published methods . No rapid deactivation was observed . An examination of alternate inducers for heparinase showed that heparin degradation products, hyaluronic acid, heparin monosulfate, N-acetyl-D-glucosamine, and maltose, induce heparinase in complex medium . An Azure A assay was modified and fully developed to measure the heparin concentration during fermentation and the heparinase specific activity of crude extracts of F . heparinum obtained from sonication, thus negating the need for further purification to measure activity."

J Gynecol Obstet Biol Reprod (Paris), 1981, 10(2), 119 - 25
{The bacteriostatic and bactericidal effect of amniotic fluid (author's transl)}; Rouquet Y et al.; The bacteriostatic and bactericidal effect of 100 samples of amniotic fluid (LA) was studied against 5 bacterial species that are responsible for neonatal infections . These results show that there is a bacteriostatic activity in liquor . 52 specimens of liquor amnii (52%) were shown to be active against at least one of the 5 bacterial specimens studied . 32 samples of liquor (32%) were active against Listeria monocytogenes, 21 (21%) against Flavobacterium meningosepticum, 18 (18%) against Escherichia coli, 17 (17%) against group B streptococci and 7 (7%) against Bacteroides fragilis . A simultaneous study of the 6 types of germs show a separate characteristics of this activity . In this way the number of samples of liquor that were active against one or at the same time against 2, 3, 4 or 5 bacteria were respectively 24, 19, 5, 2 and 2 . The bacteriostatic effect was more frequently active in those samples of liquor which were studied near term (57.5%) than in the samples studied nearer the beginning of pregnancy (31.5%) . All the same, this difference is not statistically significant . There was no difference in the antibacterial activity of samples of liquor from normal and from abnormal pregnancies . The bacterial effect was found only in 12% of samples of liquor, particularly against streptococcus B (8%) and against Listeria monocytogenes (4%) . This bactericidal effect was only found after the 31st week of amenorrhoea.

J Biochem (Tokyo), 1981 Jan, 89(1), 161 - 7
Degradative removal of heparan sulfate from the surface of an ascites hepatoma, AH 66, by heparitinase; Ohkubo Y et al.; Heparitinase {EC 4.2.2.8, heparitin sulfate lyase} was prepared from an extract of cultured cells of Flavobacterium heparinum . Purification of the enzyme was achieved by repeating the hydroxyapatite column chromatography . The enzyme was used to degrade heparan sulfate occurring on the surfaces of ascites hepatoma cells, AH 66 . From the supernatant of the enzyme-treated cells, breakdown products from heparan sulfate could be detected by paper chromatography . The heparitinase was found to be more effective than trypsin in removing heparan sulfate from the cells . Furthermore, on analyzing glycosaminoglycans and glycopeptides from the enzyme-treated cells and control cells, it was concluded that heparan sulfate was exclusively present on the cell surface and accessible to the heparitinase whereas other cell surface complex carbohydrates remained intact.

Acta Microbiol Pol, 1981, 30(3), 283 - 94
Studies on psychrophilic bacteria in two lakes of different trophy; Stopinski M; The number of bacteria capable of growth at low temperatures in two lakes was found to be subject to considerable variation throughout the year although the changes were not correlated with changes in ambient temperature . A correlation was, however, observed between temperature and ratio of number of psychrophiles to "total" count . All of the psychrophilic bacteria isolated from the studied lakes were able to grow at temperatures below 0 degrees C . Microorganisms isolated from eutrophic lakes were characterized by more pronounced stenothermy than those isolated from lake with low tropic level . These bacteria represented a type resembling a type resembling obligatory or ,,true" psychrophiles . Most of the isolates belonged to the genus Pseudomonas and to the Flavobacterium--Cytophaga group.

J Bacteriol, 1980 Oct, 144(1), 375 - 81
Bacteriophages of methanotrophic bacteria; Tyutikov FM et al.; Bacteriophages of methanotrophic bacteria have been found in 16 out of 88 studied samples (underground waters, pond water, soil, gas and oil installation waters, fermentor cultural fluids, bacterial paste, and rumen of cattle) taken in different geographic zones of the Soviet Union . Altogether, 23 phage strains were isolated: 10 strains that specifically lysed only Methylosinus sporium strains, 2 strains that each lysed 1 of 5 Methylosinus trichosporium strains studied, and 11 strains that lysed Flavobacterium gasotypicum and, at the same time, 1 M . sporium strain . By fine structure, the phages were divided into two types (with very short or long noncontractile tails); by host range and serological properties, they fell into three types . One-step growth characteristics of the phages differed only slightly; the latent period varied from 6 to 8 h, the rise period varied from 4 to 6 h, and the average burst size was 100 . All phages had guanine- and cytosine-rich double-stranded deoxyribonucleic acid consisting of common nitrogen bases . The molecular mass of the deoxyribonucleic acid as determined by restriction endonuclease analysis was 29.4 X 10(6) for M . sporium phages and 44 X 10(6) for F . gasotypicum phages . By all of the above-mentioned properties, all phages within each of the groups were completely identical to one another, but differed from phages of other groups . Bacteriophages lysing M . sporium and M . trichosporium GB2 were identical to phages M1 and M4, respectively, which were isolated earlier in the German Democratic Republic on the same methanotrophic species.

Carbohydr Res, 1980 Aug 15, 83(2), 303 - 13
Dextran alpha-(1 yields 2)-debranching enzyme from Flavobacterium Sp . M-73 . Properties and mode of action; Mitsuishi Y et al.; The general properties and specificity of a dextran alpha-(1 yields 2)-debranching enzyme from Flavobacterium have been examined in order to apply this enzyme to the structural analysis of highly branched dextrans . The optimum pH range and temperature were pH 5.5-6.5, and 45 degrees, respectively . The enzyme was stable up to 40 degrees on heating for 10 min, and over a pH range of 6.5-9.0 on incubation at 4 degrees for 24 h . The effects of various metal ions and chemical reagents have also been examined . The debranching enzyme has a strict specificity for the (1 yields 2)-alpha-D-glucosidic linkage at branch points of dextrans and related branched oligosaccharides, and produces D-glucose as the only reducing sugar . The degree of hydrolysis of the dextrans by this enzyme and the Km value (mg/mL) were as follows: B-1298 soluble, 25.2%, 0.21; B-1299 soluble, 31.5%, 0.27; and B-1397, 11.8%, 0.91 . the debranching enzyme thus has a novel type of specificity as a dextranhydrolase . We have termed this enzyme as dextran alpha-(1 yields 2)-debranching enzyme, and its systematic name is also discussed.

J Bacteriol, 1980 Aug, 143(2), 772 - 80
Cellular location of enzymes involved in chondroitin sulfate breakdown by Bacteroides thetaiotaomicron; Salyers AA et al.; Bacteroides thetaiotaomicron, a gram-negative anaerobe found in human colons, could utilize chondroitin sulfate, a tissue mucopolysaccharide, as its sole source of carbohydrate . The enzymes responsible for the breakdown of chondroitin sulfate by B . thetaiotaomicron were similar to those produced by Proteus vulgaris and Flavobacterium heparinum and included a lyase (EC 4.2.2.4), which degraded chondroitin sulfate into sulfated disaccharides, sulfatases (EC 3.1.6.4), which removed the sulfate residues, and a glucuronidase, which broke the unsulfated disaccharides into monosaccharide components . Chondroitin sulfate lyase, the first enzyme in the breakdown sequence, was not extracellular . It appeared to be located in the periplasmic space since lyase activity was released by treatment with ethylenediaminetetraacetate and lysozyme . Moreover, sodium polyanethole sulfonate, a high-molecular-weight inhibitor of chondroitin lyase, did not inhibit breakdown of chondroitin sulfate by intact bacteria . The sulfatase and glucuronidase appeared to be intracellular . None of these enzymes was strongly bound to membranes, and none of the steps in the breakdown of chondroitin sulfate was sensitive to oxygen.

Biochim Biophys Acta, 1980 Jul 10, 614(1), 63 - 70
A novel purification procedure of L-lysine 6-aminotransferase from Flavobacterium lutescence; Yagi T et al.; A new method for the purification of L-lysine 6-aminotransferase (L-lysine: 2-oxoglutarate 6-aminotransferase, EC 2.6.1.36) was devised, in which affinity chromatography with L-lysylacetamidododecyl-Sepharose 6B, the most effective affinity adsorbent, was substituted for the heat treatment . The yield of the enzyme with the present procedure was approx . twice as high as that with the previous procedure (Soda, K . and Misono, H . (1968) Biochemistry 7, 4110-4119) . The enzyme purified by this method was activated 2-fold by heat treatment (65 degrees C for 5 min) . The enzyme has absorption maxima at 340 and 415 nm, derived from the bound pyridoxal 5'-phosphate, which are identical with those of the enzyme obtained with the procedure including heat treatment . These results rule out the possibility that the formation of the 340-nm pyridoxal 5'-phosphate of the enzyme is an artifact of heat treatment.

Antimicrob Agents Chemother, 1980 Apr, 17(4), 679 - 85
L-lysine epsilon-aminotransferase involved in cephamycin C synthesis in Streptomyces lactamdurans; Kern BA et al.; In Streptomyces lactamdurans, the precursor of the alpha-aminoadipoyl side-chain of cephamycin C is L-lysine . In this regard, streptomycetes differ strikingly from the fungi, which produce alpha-aminoadipic acid during the synthesis, rather than the breakdown, of L-lysine . Studies using a cell-free system showed that an aminoadipic acid . The product of this reaction was trapped and subsequently purified by ion-exchange chromatography . Thin-layer chromatography, spectrophotometry, and amino acid oxidase digestion studies identified the reaction product as L-1-piperideine-6-carboxylate, implying enzymatic removal of the epsilon amino group of L-lysine . This enzymatic activity (E.C . 2.6.1.36; L-lysine: 2-oxoglutarate 6-aminotransferase) is highly unusual and was previously conclusively demonstrated only in the genus Flavobacterium . In S . lactamdurans, the specific activity of this enzyme reaches a peak early in the fermentation (approximately 20 h) and decreases as the antibiotic begins to appear.

Hoppe Seylers Z Physiol Chem, 1980, 361(6), 809 - 18
{A prephenate dehydratase from Flavobacterium devorans stimulated by aromatic amino acids (author's transl)}; Krauss G et al.; Isolation and properties of prephenate dehydratase from Flavobacterium devorans r are described . The enzyme was enriched 43-fold by ammonium sulfate precipitation, by gel chromatography on ultrogel ACA-34 and by hydrophobic chromatography on decylagarose . The molecular weight was estimated to be 1.35 x 10(5) . The enzyme was activated in 1mM solutions of L-tyrosine 4,4-fold, of L-phenylalanine 3.9fold and by L-tryptophan 1.9-fold in 0.02M Tris/HCl buffer at pH 7.7 . The Km values were, when adding the activator, at L-tyrosine 1.5 x 10(-5) M, at L-phenylalanine 1.3 x 10(-5) M and at L-tryptophan 1 x 10(-5) M . Regulation of aromatic amino acid is discussed in connection with the prephenate dehydratase.

Hoppe Seylers Z Physiol Chem, 1980, 361(6), 801 - 7
{Growth inhibition by phenylalanine and tyrosine--metabolism of phenylalanine in Flavobacterium devorans (author's transl)}; Krauss G et al.; Growth of a chloramphenicol-resistant mutant of Flavobacterium devorans (F . devorans r) was inhibited by L-phenylalanine and L-tyrosine and furthermore by L-tryptophan, 4-amino-benzoate, anthranilate and homogentisate . The inhibitionwas reversed partially by simultaneous addition of aromatic amino acids and 4-amino-benzoate . After cultivating F . devorans r in a glucose-minimal medium containing L-phenylalanine, the metabolites L-tyrosine, 4-hydroxyphenylpyruvate, 4-hydroxyphenylacetate, 4-hydroxybenzaldehyd, 4-hydroxybenzoate, homogentisate and anthranilate were found . Growth inhibition and a hypothetical degrading pathway of phenylalanine are discussed.

Antonie Van Leeuwenhoek, 1980, 46(1), 41 - 9
Deoxyribonucleic acid relatedness of some menaquinone-producing Flavobacterium and Cytophaga strains; Callies E et al.; Nine menaquinone-forming strains of the Flavobacterium--Cytophaga complex with DNA base compositions between 35 and 45 moles percent guanine-plus-cytosine were investigated for genome sizes and DNA relatedness by DNA:DNA hybridization in vitro, using the optically recorded initial reassociation kinetics . Two strains representing C . hutchinsonii and C . marinoflava proved to be related on the 50 percent binding level, i.e . on a level of DNA relatedness commonly found within well-classified conventional genera of bacteria . Strains of C . johnsonae, F . heparinum, F . meningosepticum, F . odoratum, F . pectinovorum, and an unnamed Flavobacterium--Cytophaga strain were found to be interrelated, and linked to the genus Cytophaga, on the 30, or 20 percent binding levels, respectively . These findings indicate that the organisms in question are related to Cytophaga . They therefore should be transferred into the family Cytophagaceae.

Zentralbl Bakteriol {B}, 1980, 170(5-6), 469 - 78
{On the occurrence of staphylococci and pseudomonas in swimming-pool water (author's transl)}; Jentsch F et al.; 1 . Analyses made at a bath with an additional ozone processing stage showed that the bathers caused a marked bacterial contaminatin on busy days and that this contamination did not originate from the purification plant . 2 . This finding and numerous individual analyses of water from various swimming pools yielded a spectrum of 14 micro-organisms, 4 genera being of frequent incidence: bacillus, staphylococcus, pseudomonas and flavobacterium . In general, bacterial contamination exists not only in the pool itself but also after filtering and even subsequent to the addition of chlorine . Staphylococci were found to be particularly invulnerable even to high chlorine contents, which is likely to be attributable to their enclosure in organic material . 5 . Tenfold sampling demonstrated that the identification of Staphylococcus aureus in individual samples is unreliable despite its ubiquitous occurrence, probably due to their uneven distribution in the water, while the identification or exclusion of Pseudomonas aeruginosa proves sufficiently dependable even with only individual samples . 4 . The general routine analysis of swimming pool water for the presence of staphylococcus aureus appears not justifiable, whilst testing for Pseudomonas aeruginosa appears justifiable, especially for identifying potentially dangerous filter contaminations.

Med J Aust, 1979 Dec 29, 2(13), 703 - 4
Colonization of an amputation site by Flavobacterium odoratum after gentamicin therapy; Davis JM et al.; Flavobacterium odoratum was isolated from the stumps of the amputated toes of a debilitated patient . Gentamicin therapy and the poor state of health of the patient provided suitable conditions for this gentamicin-resistant species to multiply . When the gentamicin therapy was stopped, and as the patient's heal improved, F . odoratum gradually disappeared to be replaced by Staphylococcus aureus . This appears to be the first report of the isolation of F . odoratum in Australia, and it indicates the potential of this resistant species to act as a source of infection for groups at risk.

Biochim Biophys Acta, 1979 Dec 11, 588(3), 302 - 9
Bacterial metabolism of propane-1,2-diol; Willetts A; The pathway of propane-1,2-diol metabolism by a species of Flavobacterium able to grow on the diol as the sole source of carbon was influenced by the degree of aeration of the growth medium . Under strongly aerobic conditions the diol was exclusively catabolised to lactaldehyde by an initial diol oxidase, subsequently metabolised to pyruvate and then oxidised to CO2 by the tricarboxylic acid cycle . Under microaerophilic conditions some propane-1,2-diol was catabolised by the oxidase-initiated pathway, but some diol was alternatively catabolised by an inducible diol dehydrase to propionaldehyde and subsequently reduced to n-propanol as an end product of metabolism.

J Antibiot (Tokyo), 1979 Dec, 32(12), 1293 - 302
Chloramphenicol resistance of three different flavobacteria; Sussmuth R et al.; The chloramphenicol resistance of some flavobacteria was investigated comparatively . This resistance can be explained either by acetylation of chloramphenicol to O-acetylchloramphenicol via constitutively formed acetyltransferases, followed by cometabolic degradation (strain CB 60), or by limited uptake and total degradation (strain CB 6) by inducible enzymes or by other mechanisms (F . devorans) . The mechanisms of resistance, CM-acetylation, CM-degradation and limited uptake are discussed.

Biochim Biophys Acta, 1979 Nov 15, 588(1), 20 - 5
Kinetic control of phosphoglycerate mutase from Flavobacterium sp . grown on ethylene glycol; Willetts A; A species of Flavobacterium able to oxidise ethylene glycol to pyruvate via glyoxylate, glycerate, 2-phosphoglycerate and phosphoenolpyruvate exploits phosphoglycerate mutase to initiate gluconeogenesis . Partially purified phosphoglycerate mutase from this bacterium is independent of adenylate charge control but is activated by phosphoenolpyruvate . The possible significance of this regulation is discussed.

Carbohydr Res, 1979 Nov, 76, 203 - 13
Partial purification and properties of a bacterial isoamylase; Evans RM et al.; Isoamylase has been prepared by affinity chromatography of a commercial enzyme-preparation from a strain of Cytophaga (also known as a Flavobacterium or Polyangium) . The enzyme was not very stable, but the stability could be improved by calcium ions . The enzyme had a very low but significant activity on pullulan and on alpha-dextrins having maltosyl side-chains . This observation, which is contrary to previous reports, has been related to the specificity of isoamylase and other bacterial debranching-enzymes.

J Clin Pathol, 1979 Sep, 32(9), 953 - 5
Meningitis caused by Pseudomonas paucimobilis; Hajiroussou V et al.; This appears to be the first report of meningitis due to Pseudomonas paucimobilis and the first report of a clinically significant isolate of this species in the UK . Characteristics by which the species may be recognised are given, and attention is drawn to the possible confusion of Ps . paucimobilis with other yellow-pigmented pseudomonads and Flavobacterium species.

J Clin Microbiol, 1979 Aug, 10(2), 206 - 9
Cellular fatty acid composition of Pseudomonas paucimobilis and groups IIk-2, Ve-1, and Ve-2; Dees SB et al.; The cellular fatty acid composition of Pseudomonas paucimobilis (IIk-1) and group IIk-2, Ve-1, and Ve-2 was determined by gas-liquid chromatography . The unnamed groups were readily distinguished from P . paucimobilis by cellular fatty acids . The data strongly suggest that these bacteria may be additional species of Pseudomonas and Flavobacterium.

J Clin Microbiol, 1979 Aug, 10(2), 155 - 60
Airway colonization by Flavobacterium in an intensive care unit; du Moulin GC; A total of 195 patients admitted to a respiratory-surgical intensive care unit became colonized with species of Flavobacterium during a 70-month prospective study . By biochemical, cultural, and morphological criteria and a comparison of antibiotic susceptibilities, all patient isolates of Flavobacterium were apparently related . The origin of these organisms was sought . Flavobacterium were recovered from different water-associated areas of the hospital and from the hands of respiratory-surgical intensive care care unit staff . The organisms were also found in university dormitory sinks . The isolation of these organisms from tap water led to their recovery from reservoirs supplying drinking water to the city of Boston and surrounding communities . These organisms are resistant to chlorine concentrations found in municipal water . There was no proven case of pneumonia caused by Flavobacterium in 2,329 consecutive patients studied in our respiratory-surgical intensive care unit.

Ann Microbiol (Paris), 1979 Jul, 130B(1), 141 - 4
{Six new serotypes of "Flavobacterium meningosepticum" (author's transl)}; Richard C et al.; Six new serotypes (G to L) of Flavobacterium meningosepticum are described herein . Cross-reactions between the 12 serotypes, A to L (including the 6 serotypes A to F of Owen and Lapage, 1974) are described . Sixty-six strains isolated in France (61 from Strasbourg) belong to the following serotypes (with number of strains): A (1), F (4), G (51), H (1), I (1), J (6), K (1) and L (1) . Use of meningosepticum serotyping is recommended as an epidemiological tool.

Appl Environ Microbiol, 1979 Jun, 37(6), 1063 - 6
L-Glutamine formation by Flavobacterium rigense; Yamada S et al.; A penicillin-resistant mutant of Flavobacterium rigense designated as strain 703, FERM-P no . 3628, was obtained after ultraviolet treatment of F . rigense FERM-P no . 3556 . The parent strain produces 0-2-hydroxypropylhomoserine from 1,2-propanediol . The mutant was found to be a good producer of L-glutamine . The physiological characteristics of strain 703 were different from the general group of L-glutamic acid-producing bacteria . Strain 703 required L-tryptophan and thiamine but not biotin for its growth . L-Glutamine formation on a specific basis, however, was independent of L-tryptophan and thiamine . Biotin and penicillin were also not effective . Only ammonium fumarate acted as an effective factor on L-glutamine formation . Accumulation of L-glutamine by strain 703 was 10 mg/ml at 30 degrees C for 48 h in a chemically defined medium containing 3% diammonium fumarate.

Carbohydr Res, 1979 May, 70(2), 295 - 306
Purification of chondroitinase B and chondroitinase C using glycosaminoglycan-bound AH-Sepharose 4B; Ototani N et al.; Chondroitinase B and chondroitinase C were separated from an extract of Flavobacterium heparinum induced with chondroitin 6-sulfate by using column chromatography on hydroxylapatite . Chondroitinase C was eluted together with the activities of hyaluronidase, delta4,5glycosiduronase, and sulfatase . The latter two activities were eliminated exclusively by passing the crude chondroitinase C fraction through a phosphono-cellulose column pre-equilibrated with 0.07M sodium phosphate buffer (pH 6.8) . Chondroitinase C was then purified by affinity chromatography using dermatan sulfate-bound AH-Sepharose 4B coated with the same glycosaminoglycan . Purification of the enzyme was achieved 18-fold and in 73% yield . On the other hand, the activities of delta4,5glycosiduronase and sulfatase were decreased to 50 and 60%, respectively, as compared with those in the crude chondroitinase B fraction, after passing the fraction through a column of phosphono-cellulose pre-equilibrated with 0.1M sodium phosphate buffer (pH 6.8) . The remaining activities of these two enzymes were then eliminated from chondroitinase B by affinity chromatography with heparin-bound AH-Sepharose 4B coated with dermatan sulfate . In the affinity chromatography used in the present study, non-covalent coating of the glycosaminoglycan-bound (covalently) AH-Sepharose 4B with the same or another glycosaminoglycan was found to be important.

Biochim Biophys Acta, 1979 Jan 4, 582(1), 33 - 43
Chemistry of heparitin sulfate and heparin from normal tissues and from patients with Hunter syndrome; Nader HB et al.; Some structural features of heparitin sulfate excreted by patients with Hunter syndrome are described . It is shown, with the aid of heparitinases and heparinase from Flavobacterium heparinum, that the Hunter heparitin sulfate is a very complex structure composed of nine different disaccharide units containing regions akin to normal heparitin sulfate and regions akin the heparin . Two-thirds of the iduronic acid residues of Hunter heparitin sulfate are devoid of sulfate, contrasting with heparin in which most of the iduronic acid residues are sulfated . The isolation and characterization of the non-reducing ends of heparin and of the heparitin sulfates is also described . Based on these results the specificity of the heparinase and heparitinases as well as the biosynthesis of iduronic acid-containing heparin-like compounds is discussed.

Microbiol Immunol, 1979, 23(5), 319 - 28
Chemical composition of Streptococcus mutans cell walls and their susceptibility to Flavobacterium L-11 enzyme; Inoue M et al.; The susceptibility to a cell wall lytic L-11 enzyme from Flavobacterium sp . and the quantitative and/or qualitative composition of the cell walls of some strains of cariogenic Streptococcus mutans and a non-cariogenic strain of Streptococcus mitis were determined . The purified cell walls of S . mutans strains HS-1 (serotype a), BHT (b), NCTC10449 (c), C67-1 (c), C67-25 (c), OMZ 176 (d), MT703 (e), MT557 (f), OMZ65 (g), and AHT (g), and S . mitis CHT contained glutamic acid, alanine, and lysine as well as muramic acid and glucosamine as a peptidoglycan component . Besides these amino acids, significant amounts of threonine were detected in strains HS-1, OMZ65, and AHT cell walls, and considerable amounts of aspartic acid and/or threonine as well as several other amino acids in OMZ176, OMZ65, and CHT cell walls . Rhamnose was a common special component of the cell walls of S . mutans strains BHT, NCTC10449, MT703, B2 (e), MT557, and AHT, and S . mitis CHT . An additional sugar component, glucose, was detected in the cell walls of all of these strains except BHT, and galactose was found in BHT, AHT, and CHT cell walls . Galactosamine was present in S . mitis CHT cell walls . Varying amounts of phosphorus were detected in the cell walls of all the strains examined . The cell walls of all these streptococcal strains except MT703, 6715, and AHT were susceptible to the lytic action of the L-11 enzyme to various extents . No consistent relationship was observed between the amino acid and sugar composition of these cell walls and their susceptibility to the L-11 enzyme . The chemical composition of these cell walls is discussed in terms of the serological classification of S . mutans.

J Clin Pathol, 1979 Jan, 32(1), 73 - 7
Flavobacterium odoratum: a species resistant to a wide range of antimicrobial agents; Holmes B et al.; During the period 1966-77, 24 strains of Flavobacterium odoratum were identified from among strains of Gram-negative, non-fermentative bacteria submitted to the National Collection of Type Cultures for computer-assisted identification . The F . odoratum strains showed resistance to therapeutic levels of gentamicin, tobramycin, amikacin, and carbenicillin as well as to several other antimicrobial agents generally useful in the treatment of infections caused by Gram-negative, non-fermentative bacteria . Two strains isolated from amputation stumps and another three strains isolated in significant numbers from urine specimens were possibly opportunist pathogens . The biochemical characteristics of the 24 strains, the proposed neotype strain of F . odoratum, and three strains representative of a group, referred to at the Center for Disease Control, Atlanta as group M-4f, were compared with those of biochemically similar species which may be isolated from clinical material.

J Clin Microbiol, 1978 Dec, 8(6), 772 - 4
Cellular fatty acids of Flavobacterium meningosepticum and Flavobacterium species group IIb; Moss CW et al.; The cellular fatty acid profiles of Flavobacterium meningosepticum and Flavobacterium species group IIb were markedly different from those of related bacteria . The profiles were characterized by the presence of 13-methyl-tetradecanoate and three uncommon acids: 2-hydroxy-13-methyl-tetradecanoate, 15-methyl-hexadecanoate, and 3-hydroxy-15-methyl-hexadecanoate.

J Biochem (Tokyo), 1978 Oct, 84(4), 1005 - 8
Interaction of mucopolysaccharides with glycosaminoglycans on glycosaminoglycan-bound AH-Sepharose 4B; Ototani N et al.; Chondroitinase C, chondroitinase AC, heparinase, and heparitinase separated from an extract of Flavobacterium heparinum were subjected to affinity chromatography with glycosaminoglycan-bound AH-Sepharose 4B, previously coated non-covalently with glycosaminoglycan, as the matrix . The results suggested the importance of coating the matrix with glycosaminoglycan in the binding of the enzyme protein to the matrix.

Clin Allergy, 1978 Sep, 8(5), 511 - 6
Humidifier fever and endotoxin exposure; Rylander R et al.; Three cases of humidifier fever were detected in an office environment . Flavobacteria were found in the contaminated water in a humidifier . After an experimental exposure, the three persons with previous symptoms suffered from fever and slight respiratory symptoms . A leucocytosis and an increase in the number of segmented white blood cells were found the day following the exposure . General immunoglobulins as well as antibodies to Flavobacterium and endotoxin were slightly elevated in the exposed group . The possibility that endotoxins may be the causative agent by means of an indirect complement activation is discussed.

Appl Environ Microbiol, 1978 Jun, 35(6), 1046 - 51
Extracellular accumulation of a new amino acid, O-2-hydroxypropylhomoserine, from 1,2-propanediol by flavobacterium rigense; Yamada S et al.; During an investigation of microorganisms utilizing petrochemicals, a strain identified as Flavobacterium rigense was found to accumulate a new amino acid in a medium containing 1,2-propanediol as the sole carbon source . Cultural conditions for the accumulation of the product were investigated, and as a result, the yield was increased to 2.8 mg/ml after a 5-day incubation in a medium containing 8% 1,2-propanediol . The pure amino acid was isolated, and its structure was investigated . Elemental analysis and infrared, nuclear magnetic resonance, and mass spectral analyses indicated that the amino acids is O-2-hydroxypropylhomoserine.

J Bacteriol, 1978 May, 134(2), 506 - 13
Carboxypeptidase displaying differential velocity in hydrolysis of methotrexate, 5-methyltetrahydrofolic acid, and leucovorin; Albrecht AM et al.; An enzyme that catalyzes the hydrolysis of folic acid and the antifolate methotrexate nearly 20 times more rapidly than the hydrolysis of 5-methyltetrahydrofolate was extraced from a gram-negative bacterium tentatively identified as a Flavobacterium sp . The enzyme was purified 500-fold and found to have a molecular weight of about 53,000 . Apparently a metallo-enzyme, it is inhibited by citrate and ethylenediaminetetraacetic acid (EDTA) . Ca2+, Co2+, Mg2+, and Zn2+ reverse inhibition by EDTA, whereas Ca2+ and Zn2+ are weak activators in the absence of EDTA . The enzymatic reaction releases the carboxy-terminal glutamyl moiety of derivatives of pteroyl-mono-L-glutamic acid . Substituents on N5 of the pteridine ring decrease the velocity of hydrolysis . Some non-specificity for the terminal amino acid is expressed . The strikingly different rates of hydrolysis of methotrexate and 5-methyltetrahydrofolate have stimulated interest in this enzyme for its potential clinical value in improving the therapeutic index of methotrexate.

J Biochem (Tokyo), 1978 Apr, 83(4), 1213 - 6
Isolation of a novel sphingoglycolipid containing glucuronic acid and 2-hydroxy fatty acid from Flavobacterium devorans ATCC 10829; Yamamoto A et al.; A new acidic sphingoglycolipid has been isolated from a Gram-negative, glucose-non-fermentative (obligatory aerobic) bacterium, Flavobacterium devorans ATCC 10829, by thin-layer chromatography on silica gel after mild alkaline hydrolysis of the cellular lipids . Chemical degradation studies, thin-layer chromatographic behavior, IR and mass-spectrometric analysis of the original and reduced glycolipid with LiA1H4 revealed that the lipid contained glucuronic acid, long-chain bases, and fatty acids in a molar ratio of approximately 1:1:1 . The major long-chain bases were identified by gas chromatography-mass spectrometry as dihydrosphingosine (d-18 :0) and longer homologues, while the N-acyl group was exclusively 2-hydroxy myristic acid . The most probable structure of this glycolipid appeared to be a ceramide glucuronic acid (N-acyl dihydrosphingosine 1-glucuronic acid).

Appl Environ Microbiol, 1978 Apr, 35(4), 679 - 84
Bacterial oxidation of polyethylene glycol; Kawai F et al.; The metabolism of polyethylene glycol (PEG) was investigated with a synergistic, mixed culture of Flavobacterium and Pseudomonas species, which are individually unable to utilize PEGs . The PEG dehydrogenase linked with 2,6-dichlorophenolindophenol was found in the particulate fraction of sonic extracts and catalyzed the formation of a 2,4-dinitrophenylhydrazine-positive compound, possibly an an aldehyde . The enzyme has a wide substrate specificity towards PEGs: from diethylene glycol to PEG 20,000 Km values for tetraethylene glycol (TEG), PEG 400, and PEG 6,000 were 11, 1.7, and 15 mM, respectively . The metabolic products formed from TEG by intact cells were isolated and identified by combined gas chromatography-mass spectrometry as triethylene glycol and TEG-monocarboxylic acid plus small amounts of TEG-dicarboxylic acid, diethylene glycol, and ethylene glycol . From these enzymatic and analytical data, the following metabolic pathway was proposed for PEG: HO(CH2CH2O)nCH2CH2OH leads to HO(CH2CH2O)nCH2CHO leads to HO(CH2CH2O)nCH2COOH leads to HO(CH2CH2O)n-1CH2CH2OH.

J Clin Pathol, 1978 Mar, 31(3), 220 - 2
Flavobacterium meningosepticum as an opportunist; Mani RM et al.; Flavobacterium meningosepticum was isolated from the cerebrospinal fluid of an adult immunodeficient female . In spite of prompt therapy the patient succumbed to the infection . The opportunistic role of the organism is discussed.

An Esp Pediatr, 1978 Feb, 11(2), 147 - 50
{Neonatal meningitis by "flavobacterium meningoseptium" (author's transl)}; Cintado Bueno C et al.; We informed a case of neonatal meningitis caused by "Flavobacterium meningosepticum", the first to be described in our country, taking into consideration its ethioepidemiology and therapy.

Biochim Biophys Acta, 1978 Jan 18, 538(2), 316 - 27
Microbial metabolism of aliphatic glycols . Bacterial metabolism of ethylene glycol; Child J et al.; A species of Flavobacterium isolated from pond water by its ability to grow aerobically on ethylene glycol as the role source of carbon initially oxidised the diol to glyoxylate via glycollate . The glyoxylate was metabolised by the glycerate pathway to acetyl-CoA . The acetyl-CoA was further metabolised by the tricarboxylic acid cycle plus malate synthase acting anaplerotically.

J Nutr Sci Vitaminol (Tokyo), 1978, 24(3), 229 - 35
Vitamin B6 biosynthesis and nucleoside effect in the resting cell system of Flavobacterium sp . 238-7; Tani Y et al.; As one of the attempts to find clues on precursors or intermediates of the vitamin B6 biosynthetic pathway, the resting cell system of Flavobacterium sp . 238-7, a bacterium producing a high amount of the vitamin, was employed . Among various compounds tested as an additive to the reaction mixture, L-glutamate and nucleoside derivatives increased the formation of vitamin B6, respectively . The possible mechanism of the participation of these compounds in vitamin B6 biosynthesis is discussed . The identification of vitamin B6 formed in the reaction mixture was done using column chromatography.

Can J Microbiol, 1977 Dec, 23(12), 1599 - 653
The history, biology, and taxonomy of the Cytophaga group; Christensen PJ; The first section of this review covers the important characteristics of the genera Cytophaga and Sporocytophaga . The topics discussed include vegetative cell structure, the spreading habit, and degradation of macromolecules . A historical account of these two genera follows, together with a discussion on the definition of, and species differentiation with the genus Cytophaga, and on the taxonomy of Sporocytophaga . The third section deals with the relationships of the cytophagas with the flavobacteria and includes a brief history of Flavobacterium, reassignation of some species to Cytophaga, differentiation from Cytophaga, and a discussion on the definition of the genus Flavobacterium . This is followed by a section dealing with the relationship of Cytophaga with the flexibacteria, starting with an introduction to the diversity of flexing organisms and taxonomic developments, and proceeding with the differentiation within the family Cytophagaceae, and species differentiation in Flexibacter . The concluding section includes a proposed redefinition of Cytophaga, a proposal regarding species conservation in this genus, and discussions on the relationship between the cytophagas and the myxobacteria and on the significance of cytophagas in the environment . The characteristics of all described species of Cytophaga, Flexibacter, and relevant flavobacteria are tabulated and a bibliography is presented.

Environ Health Perspect, 1977 Dec, 21, 279 - 83
Biodehalogenation; Castro CE; Haloorganic biocides are widely employed as soil fumigants to combat the destructive action of plant parasitic nematodes and fungi . These substances are dehalogenated by soil organisms, principally species of Pseudomonas and Flavobacteria, to nontoxic metabolities . The paths of metabolism of a vareity of simply alkyl halides are described with emphasis upon the biodehalogenation step.

J Clin Microbiol, 1977 Nov, 6(5), 450 - 55
Two outbreaks of Flavobacterium meningosepticum type E in a neonatal intensive care unit; Hazuka BT et al.; Two separate outbreaks due to Flavobacterium meningosepticum type E occurred in a neonatal intensive care unit in March-April and July 1975 . The first outbreak involved all five infants in the unit . Two infants developed meningitis, one had bacteremia, and two were colonized . During the second outbreak, five of seven infants were colonized but none developed disease . The upper respiratory tract was colonized first in most instances, and the organism persisted at this site for a mean of 17.3 days . Duration of colonization was more prolonged in infants receiving antibiotics than in untreated infants . Extensive environmental surveillance failed to demonstrate a reservoir, however, F . meningosepticum was recovered from three nasoendotracheal tubes and from an aerosol tube before colonization of four infants . The organism was resistant to most antimicrobial colonization of four infants . The organism was resistant to most antimicrobial agents tested and developed resistance to others during the treatment course of one infant . Although F . meningosepticum was not recovered from cultures of transport vehicles, several other gram-negative bacteria were isolated and were also resistant to multiple antibiotics.

Biochem J, 1977 Aug 1, 165(2), 287 - 93
Specificity of flavobacterial glycuronidases acting on disaccharides derived from glycosaminoglycans; Hovingh P et al.; The specificity of the unusual flavobacterial glycuronidases that act on disaccharides containing delta4,5-unsaturated uronic acids was reinvestigated . The results show that the enzyme that hydrolyses the uronidic bond in disaccharides from hyaluronic acid and the chondroitin sulphates appears to be mainly specific for beta-D-(1 leads to 3)-derived linkages . The enzyme that hydrolyses the uronidic bond in a variety of disaccharides obtained from heparan sulphate and heparin appears to be specific for beta-D-(1 leads to 4)- and alpha-L-(1 leads to 4)- derived linkages . Thus the glycuronidases seem to be specific for linkage position rather than anomeric configuration, as had been thought previously . In addition, the data confirm other evidence that the major glucuronidic linkages in heparan sulphate and heparin have the beta-D-configuration, and the iduronidic linkages the alpha-L-configuration.

Arch Microbiol, 1977 May 13, 113(1-2), 33 - 7
The carotenoids of Flavobacterium strain R1560; Britton G et al.; The main carotenoid of Flavobacterium strain R1560 has been identified as (3R,3'R)-zeaxanthin . Also present were small amounts of 15-cis-phytoene, phytofluene, "zeta-carotene" (7,8,7',8'-tetrahydro-psi, psi-carotene plus 7,8,11,12-tetrahydro-psi, psi-carotene), neurosporene, lycopene, beta-zeacarotene, gamma-carotene, beta-carotene, beta-cryptoxanthin, rubixanthin, 3-hydroxy-beta-zeacarotene and several apo-carotenals . Zeaxanthin production was inhibited by nicotine (10 mM), and lycopene and rubixanthin accumulated . The biosynthesis of zeaxanthin is discussed in terms of pathways and also of "half-molecule" reaction sequences . The presence of zeaxanthin may be a characteristic of a group of Flavobacterium species, and may thus be useful in the taxonomic classification of these organisms.

Health Lab Sci, 1977 Apr, 14(2), 107 - 16
Pigment analysis in the identification of "Flavobacteria"; Cohen JR et al.; Spectral properties of the intracellular pigments of flavpbacteroa were studied by "scanning" ultrasonic preparations, methanol extracts, MF preparations, slide cultures and whole cell suspensions . No genus-specific Amax were demonstrated with methanol extracts . In situ analyses of pigment, using ultrasonic preparations or whole cell suspensions, demonstrated that all strains of Flavobacterium spp . surveyed showed a discrete Amax between 418 and 421 nm . This peak was absent from scans of other yellow gram negative rods" . Amplitude of Amax was accentuated by variation in incubation parameters . Good resolution was obtained by repositioning cuvettes in "conventional" spectrophotometers and attaching opal glass plates to the "detector suface" of cuvettes . The presence of a genus-specific Flavobacterium pigment, or pigment fraction, detected through in situ analysis, adds another feature to keys or schema used for characterization and identification of the organisms comprising this presently ill-defined taxon.

Biken J, 1977 Mar, 20(1), 11 - 20
The mode of hydrolysis of a glycan portion of Micrococcus lysodeikticus cell walls by endo-N-acetylglucosaminidase or endo-N-acetylmuramidase isolated from crude barley beta-amylase; Iwata S et al.; Micrococcus lysodeikticus cell walls were digested with a pI 6.8 endo-N-acetylglucosaminidase or a pI 9.5 endo-N-acetylmuramidase . The digests were further treated with a N-acetylmuramyl-L-alanine amidase of Flavobacterium L-11 enzyme to remove the peptide portion . The products were fractionated by gel filtration and ion-exchange chromatography, and the glycan portion of fractions were analyzed for their average amino sugar chain lengths . The following results were obtained . 1 . The glycan portion of the main products in the pI 6.8 enzyme digest consisted of (-N-acetylmuramic acid-N-acetyl-glucosamine-)2-3 . 2 . The glycan moiety of the pI 9.5 enzyme digest was mainly composed of (-N-acetylglucosamine-N-acetylmuramic acid-)3-4 . 3 . The glycosidic linkages around the muramic acid 6-phosphate residues which linked to a special structure through a phosphodiester bond were rather refractory to the glycosidase action of both pI 6.8 and 9.5 enzymes.

Jpn J Antibiot, 1977 Mar, 30(3), 242 - 9
{Clinical and bacteriological study on Flavobacterium meningosepticum (author's transl)}; Igari J et al.; The present study concerns in vitro observations on the susceptibility to antibiotics and chemotherapeutic agents of the strains of F . meningosepticum isolated from various clinical specimens at the Clinical Laboratory of Juntendo University Hospital and the assessment of their clinical significance of the patient, when they are isolated from clinical specimens . 1) Fifty-two % of the strains was isolated from sputum, 20% from urine and 8% from pus and exudate . Only 2 strains were obtained from blood and one strain from cerebro-spinal fluid . 2) The patients with F . meningosepticum from sputum had respiratory distress due to the basic disorder of central nervous system, respiratory system or cardiovascular system . An endotracheal tube or a tracheal canule was placed in most of the patients to assure a patient airway, and they received massive antibiotic therapy . Most of the patients with infected urine had functional and structural abnormality of the urinary tract, as well as a history of instrumentation, and had previously received antibiotic therapy . The bacteremia due to F . meningosepticum occurred after major operation in 2 patients, and one of them suffered from hydrocephalus associated with meningitis, moreover, in whom the organism was isolated from cerebro-spinal fluid . The route of infection with F . meningosepticum cannot be assessed certainly, but the correlation with catheterization or canulation strongly suggests that the instrumentation is required for the organism to become established . 3) In vitro test for sensitivity to 19 chemotherapeutic agents were done with the strains of F . meningosepticum isolated at the laboratory from August 1974 through July 1976 . A large number of the strains were highly resistant to SB-PC, CB-PC, DKB, LCM, CL and NF, and resistant to AB-PC, CEZ and AMK . All strains were highly sensitive to MINO, DOTC and CLDM in this order, and were sensitive to EM, PA and NA . The relative sensitivity was found to TC, CP, GM and PPA.

Acta Pathol Microbiol Scand {B}, 1977 Feb, 85B(1), 27 - 37
Oxidase positive rods from cases of suspected gonorrhoea . A comparison of conventional, gas chromatographic and genetic methods of identification; Bovre K et al.; Genito-urethral specimens from 3260 women and 1170 men, with ailments suggestive of gonorrhoea, were examined for growth of oxidase positive rodshaped bacteria, as well as of gonococci . Moraxella osloensis was identified in 26 cases (0.64 per cent of women and 0.43 per cent of men) . Three patients harboured phenylalanine negative (or weakly reacting) and tryptophan deaminase negative M . phenylpyrouvica and, in three cases, a Flavobacterium species was detected . Among six oropharyngeal specimens from patients suspected of gonorrhoea, two yielded growth of oxidase positive rods, Kingella kingae and Neisseria elongata, respectively, N . gonorrhoeae was isolated from 537 patients, i.e., 12.1 per cent of all cases . The isolates of oxidase positive rods were in most cases completely identified by streptomycin resistance transformation . On this basis, the diagnostic reliability of some morphological and cultural-biochemical tests and gas chromatography was examined . Gas chromatographic analysis of fatty acid and alcohol composition of whole cells proved distinctive of species defined genetically, irrespective of confusing behaviour of some strains in other tests.

Biochim Biophys Acta, 1976 Dec 21, 451(2), 436 - 43
Structure of chondroitin sulfates . Analyses of the products formed from chondroitin sulfates A and C by the action of the chondroitinases C and AC from Flavobacterium heparinum; Michelacci YM et al.; The structures of chondroitin sulfate A from whale cartilage and chondroitin sulfate C from shark cartilage have been examined with the aid of the chondroitinases AC and C from Flavobacterium heparinum . The analyses of the products formed from the chondroitin sulfates by the action of the chondroitinases have shown that three types of oligosaccharides compose the structure of chondroitin sulfate A, namely, a dodeca-, hexa- and a tetra-saccharide, containing five, two and one 4-sulfated disaccharides per 6-sulfated disaccharide residue, respectively . The polymer contains an average of 3 mol of each oligosaccharide per mol of chondroitin sulfate A . Each mol of chondroitin sulfate C contains an average of 5 mol of 4-sulfated disaccharide units . A tetra-saccharide containing one 4-sulfated disaccharide and one 6-sulfated disaccharide C indicating that the 4-sulfated disaccharides are not linked together in one specific region but spaced in the molecule.

J Biochem (Tokyo), 1976 Dec, 80(6), 1209 - 14
Action of chondroitinases . II . Numerical calculation of the degree of multiple attack; Hiyama K; Further investigation was carried out on the action patterns of two chondroitinase-AC {EC 4.2.2.5.} preparations obtained from Arthrobacter aurescens and Flavobacterium heparinum . To infer the action patterns of the chondroitinases, we proposed a new method for the calculation of the degree of multiple attack, based on the concept established by Robyt and French ((1967) Arch . Biochem . Biophys . 122, 8-16) . It was shown that the degree of multiple attack (DM) is represented by the ratio of the initial velocity of number-average degree of scission to that of viscosity-average degree of scission . By this method, DM for A-Chase was estimated to be 3.03 and for F-chase, 1.31.

J Biochem (Tokyo), 1976 Dec, 80(6), 1201 - 7
Action of chondroitinases . I . The mode of action of two chondroitinase-AC preparations of different origin; Hiyama K et al.; The modes of action of two chondroitinases-AC {EC 4.2.2.5} from Arthrobacter aurescens and Flavobacterium heparinum were examined . By comparison of the increase of viscosity and by analyses of digests using paper chromatography and gel filtration, it was shown that the Arthrobacter enzyme (A-Chase) degrades the substrate by a stepwise attack, while the Flavobacterium enzyme (F-Chase) exhibits a more random attack, though the first attack of both enzymes is of endo-type . Further study was carried out of the initial rate and final extent of the enzymic degradation of various mucopolysaccharides . The order of the initial rates at which the native mucopolysaccharides are degraded was similar for both enzymes . For A-Chase, the initial rate and final extent of degradation of modified chondroitin sulfate C or chondroitin methyl ester, however, were low compared with those of F-Chase . These results also suggested that A-Chase degrades the substrate by stepwise attack and F-Chase by random attack.

Mikrobiologiia, 1976 Nov-Dec, 45(6), 1056 - 62
{Isolation of bacteriophages of methane oxidizing bacteria and study of their properties}; Tiutikov FM et al.; Five strains of bacteriophages were isolated for the first time in the USSR from the water of ponds, the paste of methane oxidizing bacteria and the cultural broth of the experimental plant . The strains are specific of the following species: Methylostinus sporium, Methylosinus trichosporium, and Flavobacterium gasotypicum . Bacteriophages lysing Methylocystis impression . Methylomonas agile and Methylococcus capsulatus were not isolated so far . The fine structure of the phages, the shape of negative colonies, the spectrum of lytic activity, and serological properties were studied . The phages can be subdivided into two groups according to the morphology of virions, the shape of negative colonies and serological properties, or into three groups according to the spectrum of lytic activity.

Arch Ophthalmol, 1976 Nov, 94(11), 1907 - 9
Flavobacterium endophthalmitis following keratoplasty . Use of a tissue culture medium-stored cornea; LeFrancois M et al.; This case of Falvobacterium meningosepticum endophthalmitis in the immediate postoperative period following a penetrating keratoplasty represents the first known isolation of F meningosepticum from the eye and, also, our first encounter with bacterial endophthalmitis following keratoplasty . This case occurred soon after conversion from whole eye moist chamber storage to tissue culture storage (McCarey-Kaufman medium) . This storage technique may predispose to development of postoperative endophthalmitis . We offer suggestions to minimize the possibility of such infection.

Infect Immun, 1976 Oct, 14(4), 888 - 93
Serum bactericidal activity in the horseshoe crab, Limulus polyphemus; Pistole TG et al.; Serum from the horseshoe crab, Limulus polyphemus, was examined for bactericidal activity against five species of bacteria . Greatest activity was found against Pseudomonas putida and Flavobacterium sp.; with the former, serum dilutions as high as 1:20 were capable of reducing viable counts by 50% within 2 h . Bactericidal activity of a significantly lesser magnitude was demonstrated against Serratia marcesencs and Salmonella minnesota . No killing was seen when the lobster pathogen Aerococcus viridans (formerly Gaffkya homari) was used . The cidal activity against Flavobacterium sp . remained relatively consistent for 6 months.

Lipids, 1976 Sep, 11(9), 685 - 8
Occurrence of 2- and 3-hydroxy fatty acids in high concentrations in the extractable and bound lipids of Flavobacterium meningosepticum and Flavobacterium IIb; Yano I et al.; The major hydroxy fatty acids of cellular lipids in Flavobacterium meningosepticum and Flavobacterium sp . King's group UUb were identified as 2-hydroxy 13-methyltetradecanoic, 3-hydroxy 13-methyltetradecanoic, 3-hydroxy palmitic, and 3-hydroxy 15-methylhexadecanoic acids using gas chromatography-mass spectrometry and GC-mass fragmentography . The concentration of these hydroxy fatty acids comprised up to 30-40% of the total extractable and 20-30% of the bound lipid fatty acids, respectively . From the stability for mild alkaline hydrolysis, 2-hydroxy fatty acids seemed to be attached with ester linkage, and 3-hydroxy fatty acids with amide linkage.

J Clin Pathol, 1976 Sep, 29(9), 824 - 6
Flavobacterium meningosepticum in the hospital environment; Coyle-Gilchrist MM et al.; Ten strains of Flavobacterium meningosepticum were isolated from routine clinical material during a period of two years . Of a number of antibiotics tested, clindamycin was the only one to which they were fully sensitive . The organism was found in chlorhexidine gluconate (Hibitane) solutions in use on the wards for the storage of thermometers and for routine disinfection.

Appl Environ Microbiol, 1976 Aug, 32(2), 222 - 31
Volatiles produced by microorganisms isolated from refrigerated chicken at spoilage; Freeman LR et al.; Volatile components present at spoilage of refrigerated chicken breasts were identified using high-vacuum-low-temperature distillation techniques followed by analysis with combined temperature-programmed gas chromatography and mass spectrometry . A comparison was made of the compounds detected from both irradiated and non-irradiated muscle stored at 2 and 10 degrees C under both aerobic and anaerobic conditions . Isolates were randomly selected from the spoiled poultry, identified, and evaluated for their ability to produce volatile spoilage noted when grown on radiation-sterilized chicken . Several isolates that produced off-odors on sterile chicken breasts were examined . Twenty-two compounds were associated with spoilage . Some of the compounds found on both irradiated and unirradiated samples were considered to play only a minor role in the spoilage aroma or were present in low concentrations, since the aroma of spoiled irradiated chicken lacked the harsh odor notes typical of spoiled unirradiated chicken . Fifteen of the 22 compounds were considered to be unique to unirradiated, aerobically spoiled samples . Nine of these compounds, hydrogen sulfide, methyl mercaptan, dimethyl sulfide, dimethyl disulfide, methyl acetate, ethyl acetate, heptadiene, methanol, and ethanol, were found on chicken spoiled at both 2 and 10 degrees C . xylene, benzaldehyde, and 2,3-dithiahexane were detected only in samples stored at 2 degrees C and methyl thiolacetate, 2-butanone, and ethyl propionate were associated with 10 degrees C spoilage . Fifty-eight isolates randomly selected from fresh, radiation-pasteurized, and unirradiated spoiled poultry were classified taxonomically, and 10 of them, which produced spoilage odors on sterilized chicken breasts, were selected for subsequent analysis of their volatiles . Isolates identified as Pseudomonas putrefaciens and Pseudomonas species that were members of groups I and II of Shewan's classification, as well as Flavobacterium and oxidative Moraxella, produced a number of the compounds found in the aroma of spoiled chicken . A total of 17 compounds were identified . Whereas no isolate produced all of the aroma compounds found in the aroma of spoiled chicken, together they did produce the nine found in unirradiated samples spoiled at either 2 or 10 degrees C, as well as methyl thiolacetate and xylene . Six compounds were present in the volatiles produced by the isolates but were absent in the volatiles identified from spoiled chicken . These were hydrogen cyanide, methyl isopropyl sulfide, 2-propane thiol, methyl propionate, ethyl benzene, and an unidentified compound.

Arch Microbiol, 1976 Aug, 109(1-2), 127 - 33
A description of glucose uptake in Navicula pelliculosa (Breb) Hilse including a brief comparison with an associated Flavobacterium sp; Jolley ET et al.; Navicula pelliculosa and an associated Flavobacterium sp . were isolated from the epiphyton of Scirpus maritimus, an emergent macrophyte growing in a brackish drainage dyke . Both micro-organisms possessed active transport systems for glucose uptake . In N . pelliculosa the transport system was fully induced in the dark in the absence of glucose, and subsequently inactivated when transferred to the light in the absence of the substrate . The presence of glucose during the dark induction period prevented the achievement of maximum specific activity of the transport system, while incubation at a high light intensity with or without the presence of the substrate resulted in a very marked inhibition of glucose uptake . Inhibition in the light was partially offset by blocking photosynthetic electron flow with 3'(3,4 dichlorophenyl)1'1' dimethyl urea . The transport system accumulated 3-O-methyl glucose against a concentration gradient and was highly specific for glucose as there was no competition by most of the other sugars tested . However, 6-deoxyglucose was taken up instead of glucose and this suggested that glucose was transported in a non-phosphorylated state, whereas inhibition of glucose transport activity with dicyclohexylcarbodimide implicated the involvement of an adenosine triphosphatase on the cell membrane . Inhibitors of oxidative phosphorylation tetrachlorosalicylaniline and carbonylcyanide m-chlorophenylhydrazone also inhibited glucose transport activity . The affinity of the diatom for glucose was greater than that shown by the bacterium, but the Km for glucose transport, 1.5x10-5M was too high to allow effective removal of glucose at in situ concentrations.

Biochim Biophys Acta, 1976 Jun 23, 437(1), 129 - 41
On the structure of heparitin sulfates . Analyses of the products formed from heparitin sulfates by two heparitinases and a heparinase from Flavobacterium heparinum; Silva M et al.; The analyses of the products formed from heparitin sulfates by the action of two heparitinases and a heparinase from Flavorbacterium heparinum is reported . Heparitin sulfates A and B are degraded by heparitinase I yielding two disaccharides, one of them composed of N-acetylucosamine and an unsaturated uronic, joined by alpha(1 lead to 4) linkage, and the other, with the same composition but with an O-sulfate at the hexosamine moiety . A third disaccharide is also formed from heparitin sulfate B, by the action of the same enzyme, composed of glucosamine N-sulfate and an unsaturated uronic acid joined probably by alpha(1 lead to 4) linkage . Besides these three disaccharides, heparitin sulfate B yields, by the action of heparitinase I, an oligosaccharide (with an average molecular weight of 6000) which is completely degraded by the heparitinase II yielding a disaccharide composed of glucosamine 2,6-disulfate and unsaturated uronic acid . All the disaccharides are further degraded by alpha-glycuronidase from Flavobacterium heparinum yielding the respective monosaccharides . Based on these and other analyses the possible structures of the heparitin sulfates are proposed.

J Gen Microbiol, 1976 Mar, 93(1), 89 - 102
Deoxyribonucleic acid reassociation in the classification of flavobacteria; Owen RJ et al.; DNA-DNA reassociation studies showed that Flavobacterium emningosepticum strains had a genetic relatedness of 91 to 100% with inter-strain duplexes having high thermal stabilities . The only exception, strain NCTC10016, had an average relatedness of only 43% to other strains of F . meningosepticum . The apparent divergence in DNA base sequence of this strain was reflected in the structural differences of some enzymes . There was a gradation of DNA relatedness among the Flavobacterium group II-b strains, but three strains were sufficiently related to constitute a species . Low levels of genetic relatedness were confirmed between F . meningosepticum and strains of Flavobacterium group II-b, group II-f, F . aquatile, F . breve, F . heparinum, F . pectinovorum, F . odoratum and Moraxella saccharolytica . All strains had base compositions in the range 32 to 46% guanine plus cytosine . The genome sizes of representative strains of F . meningosepticum and Flavobacterium group II-b were 2-50 X 10(9) to 3-52 X 10(9) daltons . The taxonomic implications of these findings are discussed.

Arch Dis Child, 1976 Mar, 51(3), 209 - 13
Rifamycin in neonatal flavobacteria meningitis; Lee EL et al.; Three newborn infants with meningitis due to Flavobacterium meningosepticum were treated with rifamycin administered parenterally and directly into the cerebral ventricles . Antibiotic concentrations of blood and cerebrospinal fluid (CSF) were monitored during treatment . There was rapid sterilization of the CSF after this antibiotic . Jaundice was the only toxicity noted . All 3 infants developed hydrocephalus and are shunt dependent . Two of them are otherwise free of neurological complications and are developing normally . Rifamycin is a safe and effective antibiotic in this form of neonatal meningitis.

J Biol Chem, 1976 Feb 25, 251(4), 1154 - 8
Chondroitinase C from Flavobacterium heparinum; Michelacci YM et al.; A chondroitinase that acts upon chondroitin sulfate C and hyaluronic acid was isolated from Flavobacterium heparinum . This enzyme was seperated from constitutional chondroitinase AC and an induced chondroitinase B also present in extracts of F . heparinum previously grown in the presence of chondroitin sulfates A, B or C . The enzyme acts upon chondroitin sulfate C producing tetrasaccharide plus an unsaturated 6-sulfated disaccharide (delta Di-6S), and upon hyaluronic acid producing unsaturated nonsulfated disaccharide (delta Di-OS) . Chondroitin sulfate A is also degraded producing oligosaccharides and delta Di-6S but not delta Di-4S . The chondroitinase C is also distinguished from the chondroitinases B and AC by several properties, such as effect of ions, temperature for optimal activity, and susceptibility to increasing salt concentrations . The substrate specificity of the chondroitinase C is different from that of any other chondroitinase or hyaluronidase described so far.

Arch Microbiol, 1976 Feb, 107(1), 1 - 6
Localization of selenium in bacterial cells using TEM and energy dispersive X-Ray analysis; Silverberg BA et al.; Bacteria isolated from lake sediment samples reduced sodium selenite to elemental selenium . Finestructural observations were made on a number of different bacterial species cultured in the presence of sodium selenite . Examination of Escherichia coli and a Pseudomonas species revealed electron-dense deposits of irregular shape, composed of smaller units, within the cytoplasm but not on the cell wall and cell membrane . Cells of Aeromonas and Flavobacterium species exhibited conspicuous intranuclear fibrillary aggregates and different electron-dense inclusions . It appeared that the membrane structures were somewhat more easily stained in some bacterial cells after growth on agar plates containing sodium selenite . The deposits and fibrillary accumulations were interpreted to contain selenium on the basis of energy dispersive X-ray analysis . Control preparations and cells grown in the presence of sodium selenate were void of any fine-structural abnormalities . Alterations in fine structure are discussed in relation to the metabolism of selenium by bacterial cells and possible sites of inhibition.

Health Lab Sci, 1976 Jan, 13(1), 5 - 10
Sources of the slow-growing pigmented water bacteria; Herman LG; The group of organisms often labelled as "slow-growing pigmented water bacteria" may include various species of the Xanthomonas, Cytophoga, Pseudomonas, Aeromonas and Flavobacterium genera . Due to their slow growth, this group is usually overlooked in the average clinical laboratory since the pigmented colonies are most easily noted on membrane filter pads or on surface streaked agar plates, but only after 5-15 days of incubation at room temperature . Although present at low levels in free-flowing domestic water supplies (less than 1 per ml), they can be easily isolated by swabbing sink faucets, drinking fountain heads, surgical scrub sink heads and aerators, dental chair spray units, and from the air water interface in humidifiers, nebulizers, water baths, and reservoirs where the water may remain static for weeks . Although not considered primary pathogens, several strains have been implicated in infections in nurseries and among patients in intensive care units . Contamination control in critical research and patient care areas demands scrupulous sanitation practices, regular cleaning and decontamination of all equipment, and the use of bacteria-free water whenever possible.

J Bacteriol, 1975 Dec, 124(3), 1489 - 501
Isolation and purification of Flavobacterium alpha-1,3-glucanase-hydrolyzing, insoluble, sticky glucan of Streptococcus mutans; Ebisu S et al.; Studies were made on the physical and chemical properties of polysaccharides synthesized by cell-free extracts of Streptococcus mutans, Streptococcus sanguis, and Streptococcus sp . and their susceptibilities to dextranases . Among the polysaccharides examined, insoluble glucans were rather resistant to available dextranase preparations, and the insoluble, sticky glucan produced by S . mutans OMZ 176, which could be important in formation of dental plaques, was the most resistant . By enrichment culture of soil specimens, using OMZ 176 glucans as the sole carbon source, an organism was isolated that produced colonies surrounded by a clear lytic zone on opaque agar plates containing the OMZ 176 glucan . The organism was identified as a strain of Flavobacterium and named the Ek-14 bacterium . EK-14 bacterium was grown in Trypticase soy broth, and an enzyme capable of hydrolyzing the OMZ 176 glucan was concentrated from the culture supernatant and purified by negative adsorption on a diethylaminoethyl-cellulose (DE-32) column and gradient elution chromatography with a carboxymethyl-cellulose (CM-32) column . The enzyme was a basic protein with an isoelectric point of pH 8.5 and molecular weight of 65,000 . Its optimum pH was 6.3 and its optimal temperature was 42 C . The purified enzyme released 11% of the total glucose residues of the OMZ 176 glucan as reducing sugars and solubilized about half of the substrate glucan . The products were found to be isomaltose, nigerose, and nigerotriose, with some oligosaccharides . The purified enzyme split the alpha-1,3-glucan endolytically and was inactive toward glucans containing alpha-1,6, alpha-1,4, beta-1,3, beta-1,4, and/or beta-1,6 bonds as the main linkages.

Health Lab Sci, 1975 Oct, 12(4), 34 - 5
Identification of pigmented gram negative bacilli; Gilardi GL; Identification of chromogenic gram-negative bacilli on the basis of pigment production as a taxonomic criterion is unreliable . Because of this, a wide range of morphological, cultural and biochemical characteristics of bacteria in this category was examined . The salient features found to be useful for identification of pigmented bacilli, including Flavobacterium, Xanthomonas, the fluorescent pseudomonads, Pseudomonas cepacia, P . stutzeri, P . maltophilia, P . putrefaciens, and Group VE, are reviewed.

Health Lab Sci, 1975 Oct, 12(4), 305 - 10
Hospital infections with pigmented water bacteria; Matsen JM; Two types of cultures normally expected to be sterile, i.e., urine and blood, were chosen for review at a large university hospital . The bacteria isolated from urine cultures during 1966-70 and from blood cultures from 1966-73 are listed . Two organisms, Flavobacterium meningosepticum and Pseudomonas cepacia, were selected for more detailed study and review . The antibiotic susceptibility patterns for these two organisms are presented.

Biochem J, 1975 Oct, 151(1), 121 - 9
A comparative study between a chondroitinase B and a chondroitinase AC from Flavobacterium heparinum: Isolation of a chondroitinase AC-susceptible dodecasaccharide from chondroitin sulphate B; Michelacci YM et al.; A chondroitinase that degrades only chondroitin sulphate B was isolated from Flavobacterium heparinum, and separated from a constitutive chondroitinase AC also present in extracts of F . heparinum . The enzyme acts only on chondroitin sulphate B, producing oligo- and tetra-saccharides, plus an unsaturated 4-sulphated disaccharide (deltaDi-4S) . The oligosaccharide fraction (mol . wt . 3000) is susceptible to chondroitinase AC, producing mainly deltaDi-4S . The chondroitinase B is distinguished from chondroitinase AC by several properties, such as the effect of certain metal ions, temperature for optimal activity, and susceptibility to increasing salt concentrations . The enzyme is induced in F . heparinum by all the chondroitin sulphates, as well as by the disaccharides prepared from the chondroitins . The mechanism of induction of the enzyme and the structure of chondroitin sulphate B are discussed in relation to these results.

J Biol Chem, 1975 Sep 10, 250(17), 6841 - 6
Structure of heparin . Characterization of the products formed from heparin by the action of a heparinase and a heparitinase from Flavobacterium heparinum; Silva ME et al.; The total degradation of heparin by the joint action of a purified heparinase and a heparitinase from Flavobacterium heparinum is reported . The heparinase acts directly upon heparin, yielding 52% of a trisulfated disaccharide (O-(alpha-L-ido-4-enepyranosyluronic acid 2-sulfate)-(1leads to 4)-2sulfoamino-2-deoxy-D-glucose 6-sulfate) and 40% of a tetrasaccharide besides small amounts of hexa- and disaccharides . The tetrasaccharide is in turn completely degraded by the heparitinase, forming trisulfated disaccharide and disulfated disaccharide (O-(alpha-D-glyco-4-enepyranosyluronic acid)-(1leads to 4)-2-sulfoamino-2-deoxy-D-glucose 6-sulfate) in equal amounts . These and other results indicate that the tri- and disulfated disaccharides are linked alternately, in a proportion of 3:1, respectively . The primary structure of heparin and the mode of action of the heparinase and the heparitinase are proposed based on the analysis of the different products formed by the action of the enzymes.

J Biol Chem, 1975 Sep 10, 250(17), 6837 - 40
Equilibrium dialysis and cell binding studies on Bandeiraea simplicifolia lectin; Hayes CE et al.; The total degradation of heparin by the joint action of a purified heparinase and a heparitinase from Flavobacterium heparinum is reported . The heparinase acts directly upon heparin, yielding 52% of a trisulfated disaccharide (O-(alpha-L-ido-4-enepyranosyluronic acid 2-sulfate)-(1leads to4)-2-sulfoamino-2-deoxy-D-glucose 6-sulfate) and 40% of a tetrasaccharide besides small amounts of hexa- and disaccharides . The tetrasaccharide is in turn completely degraded by the heparitinase, forming trisulfated disaccharide and disulfated disaccharide (O-(alpha-D-glyco-4-enepyranosyluronic acid)-(1leads to4)-2-sulfoamino-2-deoxy-D0glucose 6-sulfate) in equal amounts . These and other results indicate that the tri- and disulfated disaccharides are linked alternately, in a proportion of 3:1, respectively . The primary structure of heparin and the mode of action of the heparinase and the heparitinase are proposed based on the analysis of the different products formed by the action of the enzymes.

N Engl J Med, 1975 May 22, 292(21), 1099 - 102
Indwelling arterial catheters as a source of nosocomial bacteremia . An outbreak caused by Flavobacterium Species; Stamm WE et al.; Between mid-May and mid-October, 1973, 49 blood cultures from 14 patients in an intensive care unit were positive for flavobacterium species, Group II-b . We conducted an investigation to determine how patients were being infected with this unusual organism . Comparison of the 14 infected patients with 37 controls associated indwelling arterial catheters with subsequent flavobacterium bacteremia (p = 0.005) . Risk of infection was greatest during the period in which blood gas determinations were done most frequently (the first three days of catheterization) and in which infected patients had more blood gas determinations than control patients with arterial catheters (p less than 0.05) . Flavobacterium species was cultured from in-use arterial catheters, from stopcocks, and from ice in the intensive-care unit's ice machine; the catheters were probably contaminated by syringes that were cooled in ice before being used to obtain arterial specimens for blood gas determination . This outbreak calls attention to arterial monitoring systems as a potential source of nosocomial infection.

Biochim Biophys Acta, 1975 Apr 7, 385(2), 324 - 33
Structural studies of heparitin sulfates; Linker A et al.; Heparitin sulfate fractions with a large range in sulfate content were subjected to degradation by Flavobacterium heparinase and by nitrous acid . The products obtained were fractionated by chromatography, characterized, and used to arrive at tentative structures for these complex polysaccharides . The heparitin sulfate chains examined appear to be composed of: 1 . uninterrupted blocks of N-acetylglucosamine containing disaccharides; 2 . larger blocks with a molecular weight range of 5000 to 6000 which include the N-acetyl block but do not contain heparinase sensitive linkages; 3 . segments containing mainly areas where N-acetyl, N-sulfate and some disulfated units alternate in the chain . The size and arrangement of these polymer segments seem to vary with the sulfate content of a particular heparitin sulfate . For instance, the polysaccharides with the highest degree of sulfation do not appear to contain N-acetyl blocks of significant size.

Biochim Biophys Acta, 1975 Mar 14, 385(1), 1 - 10
Enzymatic degradation of heparin-related mucopolysaccharides from the surface of endothelial cell cultures; Buonassisi V et al.; When cultures of endothelial cells prelabeled with H2 -35-SO4 are exposed to a purified preparation from induced Flavobacterium heparinum containing heparinase and heparitinase activities, radioactivity accumulates in the supernatant medium . After further treatment in vitro with crude enzyme this material migrates, in part, as glucosamine (N,O-disulfated glucosamine), a break-down product characteristic of heparin and heparin-related mucopolysaccharides . After exposure of the cultures to the purified enzyme, the amount of acid-insoluble -3 5-S radioactivity that can be removed with EDTA is decreased compared to that that can be removed from control cultures . Since the amount of radioactivity that is released as break-down products is much higher than the amount of radioactivity that is secreted into the supernatant medium as intact (non-dialysable) mucopolysaccharide chains in control plates, the action of the enzyme appears to be on the cell itself . The data presented support previous studies suggesting that chains of heparitin sulfate that are accessible to the action of the enzyme are present at the surface of endothelial cells.

Can J Microbiol, 1975 Mar, 21(3), 392 - 4
A cluster analysis of some bacteria in the water column of Green Lake, Washington; Lighthart B; Mass inoculation and computer clustering techniques were used in an abbreviated procedure to group similar bacteria isolated from four depths in the water column of Green Lake, Washington . Four groups of bacteria were differentiable and were catagorized as orange-yellow Flavobacterium-Cytophaga, yellow Flavobacterium-Cytophaga, Vibrio-Aeromonas-, and Pseudomonas-like . Some of the groups were most prevalent in certain sample depths in the water column.

Appl Microbiol, 1975 Mar, 29(3), 414 - 21
Chondroitinase-producing bacteria in natural habitats; Kitamikado M et al.; A search was undertaken for bacteria which degrade chondroitin sulfate in nature and to find bacteria with a usefully high rate of chondroitinase (ChSase) productivity . First, 253 ChSase-producing bacteria were obtained from aquatic and land environments in Japan by aerobic and anaerobic screening methods . Identification according to Bergey's Manual of Determinative Bacteriology or Bain and Shewan (1968) permitted assignment of the majority of the isolates to seven genera, Aeromonas, Vibrio, Flavobacterium, Beneckea, Proteus, Micrococcus, and Arthrobacter . Next, ChSase productivities of all the isolates were compared with those of two established ChSase-producing stock strains, Proteus vulgaris NCTC 4636 and Flavobacterium heparinum ATCC 13125 . As a result, special attention was given to production by a strain of Aeromonas sp . of large quantities of extracellular ChSase-AC . None of the isolates from the current study displayed significant ChSase-ABC productivity . Finally, ChSase-AC was prepared from the culture fluid of the Aeromonas strain by fractional precipitation with ammonium sulfate, chromatography on phospho-cellulose and diethylaminoethyl-cellulose, and gel filtration on Sephadex G-200 . It was concluded that the Aeromonas strain may represent a profitable source of the enzyme ChSase-AC.

Biochim Biophys Acta, 1975 Feb 19, 377(2), 410 - 20
Purification and properties of a beta-1,6-clucosidase from Flavobacterium; Sano K et al.; An intracellular beta-1,6-glucosidase (beta-D-glucoside glucohydrolase, EC 3.2.1.21) was produced semiconstitutively by Flavobacterium M64 . This enzyme was purified 180-fold by fractionation with ammonium sulfate followed by chromatographies on carboxymethylcellulose, hydroxyapatite and Sephadex G-100 . The final preparation appeared homogeneous on disc electrophoresis on polyacrylamide gel . The molecular weight of the enzyme was determined to be ca . 59 000 by Sephadex G-100 gel filtration and sodium dodecylsulfate-polyacrylamide gel electrophoresis . The optimum pH of the enzyme was 5.8 and the optimum temperature was 40 degrees C . The enzyme readily hydrolyzed oligomers with beta-a,6-glucosidic linkages, converting them to glucose . The Km values for gentio-biose, -triose, -tetraose and -pentaose were 2.8, 3.0, 4.2 and 4.6 times 10- minus 4 M, respectively . The rates of their hydrolyses decreased with increase in their chain lengths . The enzyme was concluded to be a beta-1,6-glucosidase from its substrate specificity, production of glucose, transferring ability and inhibition by glucono-delta-lactone . The enzyme activity was inhibited by Hg-2+, Cu-2+, Ag-+, Fe-3+, p-chloromercuribenzoate, N-ethylmaleimide, glucose and trishydroxyaminomethane (Tris) but not by ethylenediaminetetraacetic acid.

J Biochem (Tokyo), 1975 Jan 1, 77(1?), 171 - 80
Purification and properties of elastolytic enzyme from Flavobacterium immotum; Ozaki H et al.; Elastolytic enzyme was purified and crystallized from culture fluid of Flavobacterium immotum No . 9-35 . The purified enzyme was homogeneous on polyacrylamide gel electrophoresis . The molecular weight was determined by Sephadex G-100 gel filtration to be 13,000 . The isoelectric point was between pH 8.3 and 8.9 . The optimum pH of the enzyme was 7.2 for elastolytic activity . The purified enzyme showed not only elastolytic activity, but also non-specific proteolytic activity against various other proteins . Milk-clotting activity was also observed . The enzyme did not act on keratin, collagen, or fourteen amino acid esters, including N-benzoyl-L-alanine methyl ester, N-benzoyl-L-arginine ethyl ester, and N-acetyl-L-tyrosine ethyl ester, which were typical substrates of pancreatic elastase {EC 3.4.21.11}, trypsin {EC 3.4.21.4}, and chymotrypsin {EC 3.4.21.1}, respectively . However, the enzyme selectively hydrolyzed elastin when both elastin and albumin were present in the reaction mixture . The enzyme was inhibited by o-phenanthroline and various heavy metals such as cadmium, lead, zinc, and mercury . Various inhibitors, such as diisopropyl phosphofluoridate, tosyl-L-lysine chloromethyl ketone, tosyl-L-phenylalanine chloromethyl ketone, trypsin inhibitor, iodoacetamide, etc., had no effect on the elastolytic activity.

Prep Biochem, 1975, 5(4), 281 - 303
Proteoglycans of soluble fraction of mouse mastocytoma; Chandrasekaran EV et al.; Proteoglycans have been isolated from a high speed supernatant fraction of a mouse mastocytoma by procedures which should minimize alteration of the native protein-polysaccharide molecule . The methods used include in vivo labeling proteoglycans with 35S-sulfate, 3H-leucine and 3H-lysine, centrifugation of the tumor homogenate at 105,000 g, cetylpyridinium fractionation of the supernatant, and further purification of some of the fractions obtained by DEAE-cellulose column chromatography, gel filtration on Sepharose 4B and cellulose acetate electrophoresis . Two major sulfated proteoglycans were obtained, one containing keratan sulfate-like material (KSP-S), the other a heparin-like polymer (HP-S) . The presence in HP-S of a compound similar to heparin was confirmed by its digestibility with flavobacterium heparinase . HP-S contained about 4 per cent protein . Glycine was the predominant amino acid, and serine did not appear to be involved in the peptide-carbohydrate linkage . The proteoglycan present in HP-S appeared to be homogeneous when examined using cellulose acetate electrophoresis . KSP-S was found to contain sialic acid and its protein content was significantly higher than that of HP-S . Glutamic and aspartic acids were the most abundant amino acids in KSP-S.

What Is Environmental Microbiology?, What Is Listeria Monocytogenes?, What Is Staphylococcus Aureus?, What Is Botulism?, What Is Functional Genomics?, e, Microorganisms, a, Microorganism, a, Microbe, r, Bacteriology, o, Bacteria, n, Antibiotic prophylaxis, n, Yeasts, c, Antimicrobials, o, Halophilic bacterium, e, Staphylococcus, o, Antibiotic prophylaxis, o, Microorganisms, r, S. cerevisiae, o, Bifidobacterium, i, Achromobacter, r, Prokaryotes, i, Growth media, c, Antimicrobial, c, Enterococci, a, Clostridia, c, Rhizobacter, o, Escherichia coli, n, Escherichia coli, a, Escherichia coli, o, Erwinia, i, Cholera

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005