Expanded diversity among Californian borrelia isolates and description of Borrelia bissettii sp. nov. (formerly Borrelia group DN127)

Up to now, the only species in the complex Borrelia burgdorferi sensu lato known to cause Lyme borreliosis in the United States has been B. burgdorferi sensu stricto. However, some atypical strains closely related to the previously designated genomic group DN127 have been isolated in the United States, mostly in California. To explore the diversity of B. burgdorferi sensu lato group DN127, we analyzed the nucleotide sequences of the rrf-rrl intergenic spacer regions from 19 atypical strains (18 from California and one from New York) and 13 North American B. burgdorferi sensu stricto strains (6 from California). The spacer region sequences from the entire B. burgdorferi sensu lato complex available in data banks were used for comparison. Phylogenetic analysis of sequences shows that the main species of the B. burgdorferi sensu lato complex (B. afzelii, B. garinii, B. andersonii, B. japonica, B. burgdorferi sensu stricto, B. valaisiana, and B. lusitaniae) each form a coherent cluster. A heterogeneous group comprising strains belonging to the previously designated group DN127 clustered separately from B. burgdorferi sensu stricto. Within this cluster, the deep branches expressing the distances between the rrf-rrl sequences reflect a high level of divergence. This unexpected diversity contrasts with the monomorphism exhibited by B. burgdorferi sensu stricto. To clarify the taxonomic status of this highly heterogeneous group, analysis of the rrs sequences of selected strains chosen from deeply separated branches was performed. The results show that these strains significantly diverge at a level that is compatible with several distinct genomic groups. We conclude that the taxonomy and phylogeny of North American B. burgdorferi sensu lato should be reevaluated. For now, we propose that the genomic group DN127 should be referred to as a new species, B. bissettii sp. nov., and that other related but distinct strains, which require further characterization, be referred to as Borrelia spp.     

Growth characteristics and a novel method for identification (the WEE-TAB system) of Porphyromonas species isolated from infected dog and cat bite wounds in humans

Thirty-nine clinical isolates of Porphyromonas species recovered from infected cat and dog bite wounds in humans and eight American Type Culture Collection and National Collection of Type Cultures type strains were characterized by using the API ZYM system, the RapID ANA II system, and conventional biochemical methods. Growth characteristics on various agar media were compared. All strains grew on brucella blood agar supplemented with vitamin K1 and hemin and on brucella laked blood agar supplemented with vitamin K1 and hemin. In contrast, only 34% of strains grew on unsupplemented brucella blood agar, 62% grew on Columbia blood agar, and 70% grew on tryptic soy blood agar (the last three media did not contain vitamin K1 or hemin). The ability of the single-tube, triple-substrate WEE-TAB system to detect the preformed enzymes N-acetyl-beta-D-glucosaminidase, alpha-D-galactosidase, beta-D-galactosidase, alpha-fucosidase, trypsin-like activity, and chymotrypsin was evaluated. The WEE-TAB test results were easy to interpret; the WEE-TAB tests were more sensitive than the comparable tests with the API ZYM and RapID ANA II systems for the detection of alpha-D-galactosidase, beta-D-galactosidase, trypsin, and chymotrypsin, and the WEE-TAB tests accurately identified Porphyromonas species.     

Isolation of Arthrobacter spp. from clinical specimens and description of Arthrobacter cumminsii sp. nov. and Arthrobacter woluwensis sp. nov

Arthrobacter spp. are very widely distributed in the environment (e.g., soil) but have not been described as causing disease in humans. Over a 6-year period, two reference laboratories isolated or received 11 strains which were eventually identified as belonging to the genus Arthrobacter. These strains had been initially identified as Centers for Disease Control and Prevention coryneform group B-1 and B-3 bacteria (whitishgrayish colonies of 2 mm or greater in diameter after 24 h of incubation, respiratory metabolism, absent or weak acid production from sugars, and hydrolysis of gelatin). However, chemotaxonomic investigations revealed lysine as the diamino acid of the cell wall and the presence of branched cellular fatty acids (with anteiso-pentadecanoic acid predominating) which was compatible with an assignment of the 11 isolates to the genus Arthrobacter only. Peptidoglycan and 16S rRNA gene sequence analyses demonstrated that three of the strains studied were representatives of a new Arthrobacter species for which the name Arthrobacter cumminsii sp. nov. is proposed and that one other strain represented a second new Arthrobacter species for which the name Arthrobacter woluwensis sp. nov. is proposed. This report is the first on the isolation of Arthrobacter spp. from clinical specimens.     

Amaricoccus gen. nov., a gram-negative coccus occurring in regular packages or tetrads, isolated from activated sludge biomass, and descriptions of Amaricoccus veronensis sp. nov., Amaricoccus tamworthensis sp. nov., Amaricoccus macauensis sp. nov., and Amaricoccus kaplicensis sp. nov

Three isolates of gram-negative bacteria, strains Ben 102T, Ben 103T, and Ben 104T, were obtained in pure culture by micromanipulation from activated sludge biomass from wastewater treatment plants in Italy, Australia, and Macau, respectively. These isolates all had a distinctive morphology; the cells were cocci that usually were arranged in tetrads. Based on this criterion, they resembled other bacteria from activated sludge previously called "G" bacteria. On the basis of phenotypic characteristics and the results of 16S ribosomal DNA sequence analyses, the three isolates were very similar to each other, but were sufficiently different from their closest phylogenetic relatives (namely, the genera Rhodobacter, Rhodovulum, and Paracoccus in the alpha subdivision of the Proteobacteria) to be placed in a new genus, Amaricoccus gen. nov. Each of the three isolates represents a new species of the genus Amaricoccus; strains Ben 102T, Ben 103T, and Ben 104T are named Amaricoccus veronensis, Amaricoccus tamworthensis, and Amaricoccus macauensis, respectively. An isolate designated Ben 101T, which was isolated independently by Cech and Hartman in Kaplice, Czech Republic, was also characterized and belongs to the same genus. We propose that the isolate of Cech and Hartman should be placed in another new species, Amaricoccus kaplicensis.     

Spiroplasma lineolae sp. nov., from the horsefly Tabanus lineola (Diptera: Tabanidae)

Spiroplasma strain TALS-2T from the viscera of the striped horsefly, Tabanus lineola, collected in Georgia was serologically distinct from other Spiroplasma species, groups, putative groups, and subgroups. Light and electron microscopy of cells of strain TALS-2T revealed helical motile cells surrounded only by a single cytoplasmic membrane. The organism grew in M1D and SP-4 liquid media. Growth also occurred in 1% serum fraction medium and in conventional horse serum medium. Growth in liquid media was serum dependent. The strain passed through 220-nm filter pores, but was retained in filters with 100-nm pores. The optimum temperature for growth was 30 degrees C. Multiplication occurred at temperatures from 20 to 37 degrees C, with a doubling time at the optimum temperature of 5.6 h in M1D broth. Strain TALS-2T catabolized glucose but hydrolyzed neither arginine nor urea. The guanine-plus-cytosine content of the DNA was 25 +/- 1 mol%. The genome size was 1,390 kbp. Six isolates serologically similar to strain TALS-2T were obtained from the same host in coastal Georgia. Three strains closely related to strain TALS-2T were isolated from the horsefly Poeciloderas quadripunctatus in Costa Rica. Strain TALS-2T (= ATCC 51749), a representative of group XXVII, is designated the type strain of a new species, Spiroplasma lineolae (Mollicutes: Entomoplasmatales).     

Massilia timonae gen. nov., sp. nov., isolated from blood of an immunocompromised patient with cerebellar lesions

A fastidious, slowly growing, strictly aerobic, gram-negative bacterium was isolated from a culture of blood from a 25-year-old man with common variable immunodeficiency. The man had been admitted to hospital with febrile progressive cerebellar ataxia. The use of standard phenotypic schemes did not lead to identification, but sequence analysis demonstrated that the 16S rRNA gene of the isolate was most similar to those of the environmental bacteria Duganella zoogloeoides (formerly Zoogloea ramigera 115) and Telluria mixta. Further characterization of the bacterium by biochemical analysis, electron microscopy, G+C content estimation, and fatty acid analysis demonstrated significant differences between the bacterium and D. zoogloeoides and Telluria species; thus, we propose it as a new taxon with the name Massilia timonae gen. nov., sp. nov.     

Description of Gemella sanguinis sp. nov., isolated from human clinical specimens

Six strains of a hitherto undescribed gram-positive, catalase-negative, facultatively anaerobic coccus isolated from human sources were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strains were genealogically identical and constitute a new subline within the genus Gemella. The unknown bacterium was readily distinguished from Gemella haemolysans, Gemella bergeriae, and Gemella morbillorum by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Gemella sanguinis sp. nov. The type strain is CCUG 37820(T).     

Mycoplasma lagogenitalium sp. nov., from the preputial smegma of Afghan pikas (Ochotona rufescens rufescens)

Organisms with characteristics typical of mycoplasmas were isolated from the preputial smegma of Afghan picas (Ochotona rufescens rufescens). The results of growth inhibition tests, metabolic inhibition tests, and immunobinding assays showed that the isolated strains were identical and that they were distinct from previously described Mycoplasma, Entomoplasma, Mesoplasma, and Acholeplasma species. These organisms represent a new species, for which the name Mycoplasma lagogenitalium is proposed. M. lagogenitalium ferments glucose, does not hydrolyze arginine or urea, reduces tetrazolium chloride, possesses phosphatase activity, does not digest gelatin or casein, and does not produce films or spots. It lyses sheep erythrocytes and does not adsorb sheep, rabbit, or horse erythrocytes. Cholesterol or serum is required for growth. The growth temperature is 37 degrees C. The guanine-plus-cytosine content of the DNA is 23.0 +/- 1.0 mol%. The type strain is M. lagogenitalium 12MS (= ATCC 700289T).     

Rileyella petauri gen. nov., sp. nov. (Pentastomida: Cephalobaenida) from the lungs and nasal sinus of Petaurus breviceps (Marsupialia: Petauridae) in Australia

A new cephalobaenid pentastome, Rileyella petauri gen: nov., sp. nov. from the lungs and nasal sinus of the petaurid marsupial, Petaurus breviceps, is described. It is the smallest adult pentastome known to date, represents the first record of a mammal as the definitive host of a cephalobaenid and may represent the only pentastome known to inhabit the lungs of a mammal through all its instars, with the exception of patent females. Adult males, non-gravid females and nymphs moulting to adults occur in the lungs; gravid females occur in the nasal sinus. R. petauri is minute and possesses morphological features primarily of the Cephalobaenida but the glands in the cephalothorax and the morphology of the copulatory spicules are similar to some members of the remaining pentastomid order, the Porocephalida. This unusual combination of features distinguish the new genus from other genera in the Cephalobaenida. The occurrence of only seven fully-formed larvae in eggs in the uterus, each representing about 10% of the length of the patent female, and her presence in the nasal sinus of a dependent juvenile P. breviceps (36 gm) implies a direct life cycle.     

Phenotypic and phylogenetic characterization of some Gemella-like organisms from human infections: description of Dolosigranulum pigrum gen. nov., sp. nov

A phylogenetic analysis was performed on two previously uncharacterized Gram-positive, catalase-negative bacteria from clinical sources. 16S rRNA sequencing studies revealed the isolates represent a new line of descent within the lactic acid group of bacteria. On the basis of the phylogenetic findings and phenotypic distinctiveness of the organisms, it is proposed that they be classified in a new genus Dolosigranulum, as Dolosigranulum pigrum sp. nov. The type strain of Dolosigranulum pigrum is NCFB 2975.     

Oswaldocruzia venezuelensis sp. n. (Nematoda:Trichostrongylina, Molineoidea), a parasite of Bufo marinus from Venezuela

A new species of Oswaldocruzia Travassos, 1917, a parasite of Bufo marinus L. from Venezuela, is described. Like most Neotropical Oswaldocruzia, Oswaldocruzia venezuelensis sp. n. is characterized by spicules with three principal branches: blade, shoe and fork, and by a division of the fork within the distal third of the spicule length. O. vaucheri Ben Slimane et Durette-Desset, 1993 is the most closely related species due to its caudal bursa of type II and its cervical alae of the same shape but it differs in the following characters: the position of the papillae of rays 4 situated nearer the papillae of rays 3 rather than rays 5, a higher percentage of the ridges in the oesophageal region, the cervical alae three times longer and sharp and the spicular fork divided less deeply.     

Hymenobacter roseosalivarius gen. nov., sp. nov. from continental Antartica soils and sandstone: bacteria of the Cytophaga/Flavobacterium/Bacteroides line of phylogenetic descent

Aseptically collected sandstone and soil samples from the antarctic Dry Valleys were inoculated into oligotrophic media and incubated under low light intensities. A total of 41 Gram-negative isolates were obtained with reddish colonies spreading on agar. A sandstone isolate and four soil strains were characterized further. They were nearly identical in morphological, physiological, biochemical and chemotaxonomic properties. They produced large amounts of extracellular polymer and utilized for growth: glucose, saccharose, mannitol, sorbitol, L-aspartate, malate and acetate, but not D-ribose, adonitol, DL-alanine, glutamate, glycolate, lactate or succinate. All strains hydrolyzed gelatin, starch, casein, xylan, Tweens 80 or 60 and dead or living yeast cells, but not cellulose or pectin. Nitrate was not reduced, ethanol was not oxidized and acid was not produced from maltose, mannitol or dulcitol. Ammonia was not produced from peptone. They were strictly aerobic. Major fatty acids were n 16:1 d 9, n 16:1 d 11, n 17:1 d 11, and i 15:0. The strains contained the quinone MK-7 and phosphatidylethanolamine as the main phospholipid. The base ratio ranged from 55 to 61 mol% G+C. A 16S rRNA sequence analysis of strains AA-688 and AA-718 showed these to be identical and to represent a special phylogenetic group within the Cytophaga/Flavobacterium/Bacteroides major line of descent. Three soil strains labeled "Taxeobacter" Txc1, Txg1, and Txo1 (Reichenbach, 1992) belonged to the same group but had lower sequence similarities (<95%). Some of their characteristics were different from those of the antarctic strains: the utilization of C-compounds, hydrolysis of polymers, temperature tolerances, major fatty acids and base ratios. Txc1 and Txg1 may later have to be considered as members of this group, possibly on the species level, while Txo1 could represent a different related genus. It is concluded that the five antarctic strains represent a new genus and species for which the name of Hymenobacter roseosalivarius is proposed. The type strain is AA-718T (DSM 11622T).     

Haloanaerobium lacusroseus sp. nov., an extremely halophilic fermentative bacterium from the sediments of a hypersaline lake

A new extremely halophilic chemoorganotrophic bacterium (strain H200T [T = type strain]) was isolated from the hypersaline sediments of Retba Lake in Senegal. This organism was a sluggishly motile, rod-shaped, non-spore-forming, gram-negative, obligate anaerobe that grew optimally at 40 degrees C in the presence of 180 to 200 g of NaCl per liter. The DNA base composition was 32 mol% guanine plus cytosine. The fermentation products from glucose were ethanol, acetate, H2, and CO2. Yeast extract was required for growth. The fermentable substrates included D-fructose, galactose, D-xylose, cellobiose, lactose, maltose, sucrose, starch, D-mannitol, glycerol, and Casamino Acids. On the basis of the results of a 16S rRNA sequence analysis, strain H200T was found to be related to Haloanaerobium species. The 16S rRNA sequence of strain H200T differed from the sequences of the three previously described Haloanaerobium species, and strain H200T also differed from these organisms in its NaCl range for growth (60 to 340 g/liter); strain H200T grew in the presence of the highest NaCl concentration recorded for any halophilic anaerobic organism, including the three previously described Haloanaerobium species. We propose that strain H200T (= DSM 10165) belongs to a new Haloanaerobium species, Haloanaerobium lacusroseus.     

Phalocrocorax olivaceus (Aves: Phalacrocoracidae), a new vector of Henneguya sp. in aquatic ecosystems from southern Chile

This is the first report of Henneguya sp., a myxozoan parasite of fishes, in faeces of Phalacrocorax olivaceus, a piscivorous bird, from Valdivia river, Chile Phalacrocorax olivaceus would be a mechanical vector of Hennguya sp. and contributes to disseminate the infection in fishes.     

Monoecocestus thomasi sp. n. (Cestoda: Anoplocephalidae) from the northern flying squirrel, Glaucomys sabrinus (Shaw), in Oregon

Monoecocestus thomasi sp. n. (Cestoda: Anoplocephalidae), from the northern flying squirrel, Glaucomys sabrinus (Shaw), in Oregon, is described and distinguished from its congeners. Monoecocestus thomasi is the sixth species of Monoecocestus to be described from nearctic rodents, and the first from a member of the family Sciuridae. Our comparisons of taxonomic characters of the nominal species of Monoecocestus in North American rodents have shown that M. giganticus Buhler 1970 is a synonym of M. americanus (Stiles 1895), both from the porcupine, Erethizon dorsatum (Linnaeus). Insemination in Monoecocestus ssp. is discussed, with the conclusion that it takes place only by way of the vagina in early immature segments.     

Arthrobacter, Brachybacterium and Planococcus isolates identified from antarctic sea ice brine. Description of Planococcus mcmeekinii, sp. nov

Escherichia coli aspartate transcarbamylase (ATCase) catalyzes the first committed step in pyrimidine biosynthesis, the condensation of aspartate and carbamyl phosphate. ATCase is positively allosterically regulated by ATP and negatively regulated by CTP. We have used mild solvent perturbation to gain global molecular information about the mechanism of heterotropic allostery. The [NaCl], temperature, and osmotic pressure dependence of the enzymatic activity of ATCase has been examined in the presence and absence of allosteric effectors. The results indicate that: 1) Regulation of aspartate binding by CTP appears to involve a unique set of electrostatic interactions not involved in enzyme function in the presence of ATP or in the absence of effectors. 2) Aspartate binding is enthalpically driven in the presence and absence of allosteric effectors. 3) The apparent enthalpy and entropy of aspartate binding (delta H, delta S), and activation energy of catalysis (Ea) are substantially altered in the presence of CTP but not ATP. 4) The change in hydration of ATCase upon substrate binding is the same in the presence and absence of allosteric effectors. 5) The linkage between heterotropic and homotropic allostery is different for ATP and CTP.     

Bacillus oleronius sp.nov., a member of the hindgut flora of the termite Reticulitermes santonensis (Feytaud)

A new rod-shaped endospore-forming bacterium is described, which was isolated from the hindgut of the termite Reticulitermes santonensis (Feytaud). The isolate stains Gram negative and its DNA has a guanine-plus-cytosine content of 35 mol%. Despite the Gram-staining reaction, both biochemical and physiological features place the isolate in the genus Bacillus and indicate a phenotypic resemblance to the Bacillus firmus-lentus group of species. On the basis of comparative 16S rRNA analysis and some phenotypic features the isolate clearly represents a new species for which the name Bacillus oleronius is proposed. The type strain is Bacillus oleronius Rt 10 (DSM 9356).     

Sumixam maximus gen.and sp.nov.,a New Pennsylvanian Species of Archaeorthoptera(Neoptera;Indiana,USA)

A new Mazon-Creek-type fossil specimen is described as Sumixam maximus gen. and sp. nov.,fossil relatives): it exhibits one of the diagnostic character states of this taxon, which is the fusion of CuA (emerging from M + CuA) with CuP, or a branch of it. A more precise taxonomic assignment is out of reach. The new taxon exhibits a fusion of the anterior branch of MA with RP, and a point of divergence of MA and MP located near the point of divergence of CuA (from M + CuA), but lack a branching of CuPa. This unique combination of character states is probably derived with respect to a number of contemporaneous species. Sumixam maximus gen. and sp. nov. is most likely the closest known relative of the panorthopterans, which include all the recent Archaeorthoptera.     

Sphaerechinorhynchus ophiograndis n. sp. (Acanthocephala:Plagiorhynchidae:Sphaerechinorhynchinae), described from the intestine of a king cobra, Ophiophagus hannah

BACKGROUND/AIMS: The mechanism of action of recombinant interferon-alpha (rIFN alpha) treatment in chronic hepatitis C is not fully understood, and may include modulation of the immune system as well as a direct antiviral effect. We have therefore evaluated the plasma concentrations of pro- and anti-inflammatory cytokines in patients with chronic hepatitis C before and during treatment with rIFN alpha. METHODS: Twenty-three patients were studied. Plasma concentrations of IL-1 beta, IL-6, TNF, IL-1 receptor antagonist (IL-1RA) and soluble TNF receptors (sTNFRs) type I and type II were determined twice before rIFN alpha treatment (on day -11 and day 1), and on days 11, 32 and 120 of treatment. RESULTS: IL-1 beta, IL-6 and TNF plasma concentrations were rarely increased before treatment (in one, six and seven patients, respectively), and usually declined during treatment. sTNFRs I and II plasma concentrations were not increased either before or during treatment. This was not the case for IL-1RA. In untreated patients, the plasma concentration of IL-1RA was higher than normal in 16 out of 23 patients. When rIFN alpha treatment was initiated, there was a constant and dramatic increase in IL-1RA levels, which reached 8 times the upper limit of the normal range (p < 0.001 as compared to pretreatment values). This increase was sustained up to day 120. CONCLUSIONS: These results indicate that induction of an anti-inflammatory status through modulation of the IL-1/IL-1RA balance may be a key mechanism of action of rIFN alpha treatment in chronic hepatitis C.