Cellular Fatty Acid Composition of Nine Pathovars of Xanthomonas campestris

Cellular fatty acids of 80 strains of Xanthomonas campestris, representing 9 different pathovars, were analyzed by gas-liquid chromatography and mass spectrometry. A total of 48 fatty acids were identified, the most important being the 16:0 (averaging at least 4.5 % of the total), the cis- and trans- 9 16 : 1 (over 14.4 %), and the iso and anteiso 15 : 0 (over 30 %). Other major fatty acids (averaging over 1 % of total) were the saturated 14 : 0 and 15 : 0, the hydroxy-substituted iso 3-OH 11 : 0, 3-OH 12 : 0 and iso 3-OH 13 : 0, and the branch-chained iso 11 : 0, iso 16 : 0, iso 17 : 1, iso 17 : 0 and anteiso 17 : 0. Of 33 minor fatty acids detected and identified, only 7 have been previously reported in the xanthomonads. Significant differences in mean percentages of 5 major fatty acids and 4 (chemical) class totals were detected among pathovars, which statistically segregated into three groups by rank analysis. X. campestris pv. dieffenbachiae was in a group by itself; pvs. campestris, citri (pathotypes A and B), manihotis, phaseoli, pruni and vesicatoria were in a seond group, and pvs. glycines, begonia and citri (pathotype E) were in a third.     

SYNTHESIS OF GLYCOPROTEINS IN BRAIN: IDENTIFICATION, PURIFICATION AND PROPERTIES OF GLYCOSYL TRANSFERASES FROM PURIFIED SYNAPTOSOMES OF GUINEA PIG CEREBRAL CORTEX

Abstract— Four glycoprotein:glycosyl transferases (a fetuin:N-acetylglucosaminyl transferase; a bovine submaxillary mucin: N-acetylgalactosaminyl transferase; a collagen: glucosyl transferase and an orosomucoid: galactosyl transferase) were purified 34-, 45-, 37- and 47-fold, respectively, from synaptosomes prepared from guinea pig cerebral cortex. Purifications were achieved by centrifugation and by column chromatography on Sephadex G-100 and G-150 of 0 , 1% (w/v) Triton X-100 extractsof the purified cerebral cortical synaptosomes. The enzymes were separated from endogenous acceptors and were highly specific for specific macromolecular acceptors; small molecules were ineffective as acceptors. The fetuin: N-acetylglucosaminyl transferase functioned only with fetuin minus N-acetylneuraminic acid, galactose and N-acetylglucosamine; the bovine submaxillary mucin: N- acetylgalactosaminyl transferase with bovine submaxillary much minus N-acetylneuraminic acid and N-acetylgalactosamine; the collagen: glucosyl transferase with collagen minus glucose; and the orosomucoid: galactosyl transferase with either orosomucoid minus N-acetylneuraminic acid and galactose or fetuin minus N-acetylneuraminic acid and galactose. Each transferase required a specific (XDP)-monosaccharide for transfer. The transferases were entirely dependent on either Mn²⁺ or Mg²⁺ for activation and Fe²⁺ and Hg²⁺ inhibited each of the four enzymes. The optimum pH's for the enzymes were: for fetuin: N-acetylglucosaminyl transferase, 7 , 4–8.0; for bovine submaxillary mucin: N-acetylgalactosaminyl transferase, 7 , 7; for collagen: glucosyl transferase, 7 , 7 and for orosomucoid: galactosyl transferase, 6 , 6. The enzymes were distributed subsynaptosomally primarily in the synaptosomal plasma membrane and in the mitochondria of the synaptosome. The respective values for K_m (μM) and V_mex (pmoles/h/mg of protein) for the transferases were: fetuin: N-acetylglucosaminyl transferase, 12 and 143; for bovine submaxillary mucin: N-acetylgalactosaminyl transferase, 25 and 166; for collagen: glucosyl transferase, 4 and 10 and for orosomucoid:galactosyl transferase, 8 and 111.     

Specificity of mutations induced by riboflavin mediated photosensitization in the supF gene of Escherichia coli

Riboflavin-mediated photosensitization has been shown to produce 8-hydroxyguanine (oh⁸Gua) in DNA. We investigated the specificity of mutation of photosensitized supF gene induced in Escherichia coli. The oh⁸Gua repair deficient E. coli mutant mutM and mutY were transformed with plasmid pUB3 carrying the supF gene irradiated with white light in the presence of riboflavin. Under these conditions, riboflavin photosensitization increased the amounts of oh⁸Gua in pUB3 DNA. Three types of a single base substitution occurring at G:C pairs were detected in both wild-type and mutM mutant strains. Almost all base substitutions were transversions to T:A or C:G pairs occurring at a similar extent in both wild-type and mutM strains. Mutations derived from mutY strain transformed with photosensitized DNA were only G:C to T:A transversions. These G:C to T:A transversions observed in the mutY strain were suggested to be the result of mispairing of oh⁸Gua with adenine. Riboflavin-mediated photosensitization may also produce lesions on DNA causing G:C to C:G changes by unknown mechanisms.     

Changes of urinary phospholipids in the chronic kidney disease patients

Abstract

Objective: To evaluate whether urinary phospholipids could be regarded as biomarkers of chronic kidney disease.

Materials and methods: Thirteen healthy volunteers and 26 consecutive chronic kidney disease patients were included. Urinary phospholipids were quantified by high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry.

Results: Urinary phosphatidylcholines concentrations (PC 16:0/16:0, 16:0/22:3, 16:0/18:1 and 16:0/18:2) were significantly higher both in glomerulonephritis group (all p?<?0.001) and in tubulointerstitial injury group (all p?<?0.05) than in healthy control group. Meanwhile, sphingomyelin concentrations (SM 18:1/16:0 and 18:1/18:0) in glomerulonephritis group were significantly higher than those in healthy control group (all p?<?0.001). Urinary PCs and SMs were positively correlated with proteinuria but negatively correlated with serum albumin. Meanwhile, PCs were positively correlated with serum creatinine.

Conclusion: Our work first demonstrated that urinary phospholipids might be biomarkers for the chronic kidney disease patients. Increased urinary phospholipids in chronic kidney disease patients might result from proteinuria, damaged kidney function or proteinuria induced hypoalbuminemia or lipotoxicity.     

Characterization of Shiga toxin‐producing Escherichia coli isolated from dairy cows in Argentina

Aims: To feno‐genotypically characterize the Shiga toxin‐producing Escherichia coli (STEC) population in Argentinean dairy cows. Methods and Results: From 540 STEC positive samples, 170 isolates were analyzed by multiplex PCR and serotyping. Of these, 11% carried stx1, 52%stx2 and 37%stx1/stx2. The ehxA, saa and eae were detected in 77%, 66% and 3%, respectively. Thirty‐five per cent of strains harboured the profile stx1, stx2, saa, ehxA and 29%stx2, saa, ehxA. One hundred and fifty‐six strains were associated with 29 different O serogroups, and 19 H antigens were distributed among 157 strains. STEC O113:H21, O130:H11 and O178:H19 were the most frequently found serotypes. The STEC O157:H7 were detected in low rate and corresponded to the stx2⁺, eae⁺, ehxA⁺ virulence pattern. Conclusions: We detected a diversity of STEC strains in dairy cattle from Argentina, most of them carrying genes linked to human disease. Significance and Impact of the study: The non‐O157 STEC serotypes described in this study are associated worldwide with disease in humans and represent a risk for the public health. For this, any microbiological control in dairy farms should be targeted not only to the search of O157:H7 serotype.     

Reliable identification of Prevotella and Butyrivibrio spp. from rumen by fatty acid methyl ester profiles

Data for bacterial identification were provided by culturing anaerobic bacteria under standardized conditions followed by extraction and methylation of cellular long-chain fatty acids and gas chromatographic analysis. The databases of fatty acid methyl ester (FAMEs) profiles for two predominant ruminal genera,Prevotella andButyrivibrio, were created. Major long-chain cellular fatty acids found in the 23 analyzedPrevotella strains were 15:0 (anteiso), 15:0, 15:0 (iso) and 16:0. The strains ofPrevotella could be well identified on species level by the characteristic ratios among major fatty acids and by acids unique fatty for each species. The 45Butyrivibrio strains were grouped into 4 major and 2 minor groups according to FAMEs profiles. The major fatty acids for the bulk of theButyrivibrio strains were 14:0, 15:1, 16:0 and 16:0 (iso). This groups corresponded to those based on 16S rDNA sequences.     

In Vivo Cloning of Arylsulfatase-Tyramine Oxidase Genes in rec Strains of Klebsiella aerogenes

A general, genetic technique for in vivo cloning of bacterial genes is presented. We previously introduced the Mu phage into various genera of bacteria including Klebsiella aerogenes with RP4 : : Mu. Using these strains carrying RP4 : : Mu cts and thermo-inducible Mu prophage in the chromosome, we cloned in vivo the arylsulfatase (ats) and tyramine oxidase (tyn) genes by partial thermo-induction. The donor strains carrying the recombinant plasmids were conjugated with K. aerogenes rec strains, which were isolated as UV-sensitive mutants. The resultant recombinant plasmids, pAT1 and pAT2, were purified and used for the transformation of mutant strains deficient in the ats and tyn genes. The ats-tyn genes seemed to be transposed into the RP4::Mu cts plasmid together with other chromosomal DNA fragments. This in vivo cloning method is applicable to a wide variety of gram-negative bacteria.     

滇藏热泉arxA基因型厌氧砷氧化原核微生物多样性及其影响因素

【目的】探究滇藏地区热泉沉积物微生物及arxA基因的多样性及其影响因素。【方法】采用Illumina MiSeq高通量测序技术和统计学的分析方法,针对3个滇藏地热区的22个样点,进行地球化学分析及微生物多样性分析,并分析其与环境因子之间的相关性。【结果】沉积物微生物群落主要门类及其丰度占比为Chloroflexi(21.27%)、Deinococcus-Thermus(17.25%)、Aquificae(13.39%)、Proteobacteria(9.27%)、Acetothermia(8.3%)、Bacteroidetes(4.96%)与Crenarchaeota(4.57%);具有arxA基因的微生物主要门类及其丰度占比分别为Proteobacteria(64.87%)、Bipolaricaulota (9.55%)、Deinococcus-Thermus (6.42%)与Crenarchaeota (4.05%);不同地热区优势种群有所差异,主要受温度、pH值、砷浓度、总溶解固体(TDS)及海拔等因素的影响,Mantel检验呈显著相关性(P<0.001)。【结论】沉积物中...     

HIGH POLYUNSATURATED FATTY ACID LEVELS IN TWO SUBTROPICAL MACROALGAE,CLADOSIPHON OKAMURANUS AND CAULERPA LENTILLIFERA1

The lipid and fatty acid compositions in two edible subtropical algae (the brown alga Cladosiphon okamuranus Tokida and the green alga Caulerpa lentillifera J. Agardh) were determined to clarify their lipid characteristics and nutritional values. Glycolipids and phospholipids were the major lipid classes, with significant levels of triacylglycerols. Polyunsaturated fatty acids (PUFA) were the major fatty acids of both algae. The lipid class composition and major fatty acids were similar in both the algal species, irrespective of wild and cultured specimens. Typical n‐6 PUFA, such as 18:2n‐6 (linoleic acid) and 20:4n‐6 (arachidonic acid), occurred in characteristically high levels in both of the algae. High levels of n‐3 PUFA were measured in all lipid classes of both species without 22:6n‐3 (docosahexaenoic acid), 18:3n‐3, 18:4n‐3, and 20:5n‐3 (eicosapentaenoic acid) for Cl. okamuranus; and 16:3n‐3, 18:3n‐3, and 20:5n‐3 for Ca. lentillifera. The finding suggests that the green algal species, which mainly biosynthesizes short‐chain (C₁₆ and C₁₈) PUFA, differs from that of the brown alga, which is capable of biosynthesizing high 20:5n‐3 levels. The PUFA levels in glycolipids of the two algal species comprised up to 60%, even though they are subtropical marine species. High n‐6 PUFA levels in the algal lipids probably influence the significant levels of n‐6 PUFA in herbivorous fishes, because the n‐6 PUFA levels in marine fish lipids are generally undetectable or negligible.     

Use of lac Gene Fusions to Study Regulation of Tyramine Oxidase,Which is Involved in Derepression of Latent Arylsulfatase in Escherichia coli

Strains with lac fused to each of the arylsulfatase (ats) and tyramine oxidase (tyn) operons in Escherichia coli were isolated. Synthesis of β-galactosidase in strains with tyn:: lac fusions was induced by tyramine, histamine, tryptamine, dopamine and octopamine, and the induction of the tyn operon was subject to catabolite and ammonium repressions. These repressions were relieved when the cells were grown with a poor carbon or nitrogen source. No arylsulfatase activity is detected in E. coli strains. Synthesis of β-galactosidase in strains with ats:: lac fusions was repressed by sulfur compounds. The repression was relieved by monoamine compounds, which induced tyramine oxidase synthesis. The inhibition of tyramine oxidase activity by cysteine resulted in a decrease of the derepressed synthesis of β-galactosidase in the ats:: lac fusion. Repressing and derepressing conditions for the tyn operon prevented and stimulated, respectively, expression of the ats operon. Thus, the expression of latent arylsulfatase in E. coli seems to be regulated by expression of the tyn operon.     

Lasioseius Fimetorum: a soil-dwelling predator of glasshouse pests?

Female lifespan and reproduction, in terms of numberof larvae produced, of the soil-dwelling predatorymite Lasioseius fimetorum Karg (Acari:Podocinidae) fed on mould mites (Tyrophagusputrescentiae [Schrank] [Acarina: Acaridae]) wereinvestigated by laboratory experiments at 20 °C,as were the mite's consumption rates of various prey.After a preoviposition period of 10.7 days, L.fimetorum produced progeny at a daily rate of 0.7.The oviposition period lasted 23.6 days and a total of19.4 progeny were produced per female. Females livedfor 38.6 days. Eggs of the Collembola Isotomurusspp. (Collembola: Isotomidae) were consumed in thelargest amount by L. fimetorum followed by mouldmite nymphs, larvae and pupae of thrips (Frankliniella occidentalis [Pergande] [Thysanoptera:Thripidae]), eggs of the Collembola Micrisotomaspp. (Collembola: Isotomidae), Isotomurus spp.nymphs and sciarid larvae (Bradysia pauperaTuomikoski and B. tritici (Coquillet) [Diptera:Sciaridae]). Immature drain flies (Psychoda spp.[Diptera: Psychodidae]) were not consumed by L.fimetorum. The suitability of L. fimetorum forbiological control of glasshouse pests withsoil-dwelling stages is discussed in comparison withanother predatory mite Hypoaspis miles Berlese(Acarina: Hypoaspididae).     

Germinal virus vector WDV (wheat dwarf virus)-mediated multiple insertions of a maize transposon,Ds (dissociation), in rice

Wheat dwarf virus (WDV) is a monocot-infecting geminivirus that replicates in infected tissue as double-stranded DIMA. We evaluated whether the WDV vector system bearingDs could be used as an effective insertional mutagen in rice. Molecular data showed thatDs was excised from WDV vectors once the WDV-carryingDs (WDV::Ds) and the genomicAc vector were co-introduced into rice calli. Mature TO and T1 transgenic plants were analyzed for the distribution and inheritance ofDs inserts. Southern analysis indicated that theDs elements excised from WDV vectors were stably inserted into genomes. The number of transposedDs ranged from zero to three copies, among independent transformants. Meanwhile, untransposedDs (WDV::Ds) were present in multiple-copies in genomes. Southern analysis of the selfed progeny of T0 plants demonstrated that most WDV::Ds were co-segregated among siblings. This indicated that these elements were integrated into the same single loci. However, a fewDs were found to segregate independently from the majority ofDs. In this report, we discuss the efficiency of WDV vectors in generating multicopyDs in rice genomes.     

Undergraduate teaching in gerodontology in Austria, Switzerland and Germany

Objective: To survey the present state of undergraduate teaching in the domain of gerodontology in Germany, Switzerland and Austria. Study participants: All universities of Austria (A), Germany (D) and Switzerland (CH). Protocol: A questionnaire on undergraduate teaching in gerodontology was mailed to all Deans (A: n = 3; CH: n = 4; D: n = 31) and all independent departments except paediatric dentistry and orthodontics (A: n = 11; CH: n = 15; D: n = 111). Results: The questionnaires were completed and returned by 29 Deans (A: n = 2; CH: n = 4; D: n = 23) and 102 departments (A: n = 7; CH: n = 8; D: n = 87). In Austria, gerodontology is a very small component of the dental curriculum and the Deans did not want this to be increased. Most German universities claimed to teach some aspects of gerodontology to undergraduate students and 87.4% of the Deans voted for separate lectures in gerodontology. In Switzerland, gerodontology seems well established. The results of questionnaires from the independent departments revealed that in all three countries lectures were more prevalent (A: n = 0; CH: n = 4; D: n = 6) than practical training in nursing homes (A: n = 0; CH: n = 3; D: n = 6). Conclusion: Considering the demographical shift which is leading to an increasing proportion of elderly in the population, the weighting of gerodontology in the undergraduate dental curriculum should be considered for revision in Austria and Germany.     

EFFECT OF NUTRIENT LIMITATION ON FATTY ACID AND LIPID CONTENT OF MARINE MICROALGAE1

Phaeodactylum tricornutum and Chaetoceros sp. (Badllariophyceae), Isochrysis galbana (clone T-Iso) and Pavlova lutheri (Prymnesiophyceae), Nannochloris atomus (Chlorophyceae), Tetraselmis sp. (Prasinophyceae), and Gymnodinum sp. (Dinophyceae) were cultured at different extents of nutrient-limited growth: 50 and 5% of μ_max. The lipid content of the algae was in the range 8.3–29.5% of dry matter and was generally higher in the Prymnesiophyceae than in the Prasinophyceae and the Chlorophyceae. Increasing extent of phosphorus limitation resulted in increased lipid content in the Bacillariophyceae and Prymnesiophyceae and decreased lipid content in the green flagellates N. atomus and Tetraselmis sp. The fatty acid composition of the algae showed taxonomic conformity, especially for the Bacillariophyceae, where the major fatty adds were 14:0, 16:0, 16:1, and 20:5n-3. These fatty acids were dominant also in the Prymnesiophyceae together with 22:6n-3. An exception was I. galbana, in which 18:1 was the major monounsaturated fatty add and 20:5n-3 was absent. The fatty acids of N. atomus and Tetraselmis sp. varied somewhat, but 16:0, 16:1, 18:1, 18:3n-3, and 20:5n-3 were most abundant. Gymnodinum sp. contained mainly 16:0, 18:4n-3, 20: 5n-3, and 22:6n-3. An increased level of nutrient limitation (probably phosphorus) resulted in a higher relative content of 16:0 and 18:1 and a lower relative content of 18:4n-3, 20:5n-3, and 22:6n-3. The nutrient limitation probably reduced the synthesis of n-3 polyunsaturated fatty acids.     

A general method for isolation of Mu d1-8(Aprlac) operon fusions in Salmonella typhimurium LT2 from Tn10 insertion strains: chlC::Mu d1-8

Summary A general method was developed for the isolation of Salmonella thyphimurium LT2 Mu d1–8 (Ap^r lac) operon fusions in a gene displacing a Tn10 insertion. Random Mu d1–8 fusion pools were prepared to grow phage P22 lysates which transduced chlC::Tn10 to Ap^rTet^s on fusaric acidampicillin plates. Among these Ap^rTet^s potential chlC::Mu d1–8 fusions, a simple spot test identified the fusions that were closely linked to the Tn10 insertion in chlC. Out of 68 Ap^rTet^s isolates 7 chlC::Mu d1–8 fusions with a nitrate-induced Lac⁺ phenotype were obtained. When oxrA::Tn10 was transduced into these chlC::Mu d1–8 fusions, they became Lac^- even in the presence of nitrate, confirming that they were chlC::Mu d1–8 fusions.     

Chewing lice from high‐altitude and migrating birds in Yunnan,China, with descriptions of two new species of Guimaraesiella

In total, 366 birds representing 55 species in 24 families and eight orders, were examined for chewing lice (Phthiraptera: Amblycera, Ischnocera) in two high‐altitude localities in Yunnan Province, China. In Ailaoshan, almost all of the birds examined were resident passeriforms, of which 36% were parasitized by chewing lice. In Jinshanyakou, most birds were on migration, and included both passerine and non‐passerine birds. Of the passerine birds caught in Jinshanyakou, only one bird (0.7%) was parasitized by chewing lice. The prevalence of Myrsidea and Brueelia‐complex lice on birds caught in Ailaoshan was higher than in previous reports. Of the chewing lice identifiable to species level, three represent new records for China: Actornithophilus hoplopteri (Mjöberg, 1910), Maculinirmus ljosalfar Gustafsson & Bush, 2017 and Quadraceps sinensis Timmermann, 1954. In total, 17 new host records are included, of which we describe two as new species in the Brueelia‐complex: Guimaraesiella (Cicchinella) ailaoshanensis sp. nov. ex Schoeniparus dubius dubius (Hume, 1874) and G. (C.) montisodalis sp. nov. ex Fulvetta manipurensis tonkinensis Delacour & Jabouille, 1930. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:9FC3D8EE‐2CED‐4DBE‐A1DB‐471B71260D27 .     

Response of bioluminescent bacteria to sixteen azo dyes

Recombinant bioluminescent bacteria were used to monitor and classify the toxicity of azo dyes. Two constitutive bioluminescent bacteria,Photobacterium phosphoreum andEscherichia coli, E. coli GC2 (lac::luxCDABE), were used to detect the cellular toxicity of the azo dyes. In addition, four stress-inducible bioluminescentE. coli, DPD2794 (recA::luxCDABE), a DNA damage sensitive strain; DPD2540 (fabA::luxCDABE), a membrane damage sensitive strain; DPD2511 (katG::luxCDABE), an oxidative damage sensitive strain; and TV1061 (grpE::luxCDABE), a protein damage sensitive strain, were used to provide information about the type of toxicity caused by crystal violet, the most toxic dye of the 16 azo dyes tested. These results suggest that azo dyes result in serious cellular toxicity in bacteria, and that toxicity monitoring and classification of some azo dyes, in the field, may be possible using these recombinant bioluminescent bacteria.     

Phospholipid fatty acid composition of gorgonians of the genus Pseudopterogorgia: Identification of tetracosapolyenoic acids

The phospholipid fatty acid composition of the Caribbean gorgonians Pseudopterogorgia acerosa (Pallas), Pseudopterogorgia americana (Gmelin), Pseudopterogorgia bipinnata (Verrill) and Pseudopterogorgia rigida (Bielschowsky) is described for the first time. The main phospholipids identified were phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine. All four gorgonians presented a similar phospholipid fatty acid composition. The main fatty acids were 16:0, 18:3(n-6), 18:4(n-3), 20:4(n-6), 22:6(n-3), 24:5(n-6) and 24:6(n-3). In all of the studied Pseudopterogorgia gorgonians, high amounts of the tetracosapolyenoic fatty acids 24:5(n-6) and 24:6(n-3) were identified. In the four gorgonians studied, n-6 polyunsaturated fatty acids predominated. These results suggest that the occurrence of tetracosapolyenoic fatty acids in the Gorgoniidae is more general than previously recognized.