Competitive adsorption of binary mixture of <Emphasis Type="Italic">Leptospirillum ferriphilum</Emphasis> and <Emphasis Type="Italic">Acidithiobacillus caldus</Emphasis> onto pyrite

Leptospirillum ferriphilum and Acidithiobacillus caldus are two important acidophilic microorganisms involved in iron and sulfur oxidation during bioleaching. Cell adsorption to mineral surfaces is important for the direct leaching or contact leaching of minerals. In this study, we report the competitive adsorption of binary mixtures of L. ferriphilum LF-104 and A. caldus MTH-04 onto pyrite surfaces. The Langmuir adsorption parameter (C_Am) indicated that these two bacteria underwent competitive adsorption to pyrite. Real-time quantitive PCR was used to quantify the relative amounts of L. ferriphilum and A. caldus adsorbed onto the surfaces of pyrite following exposure to a mixture of these two organisms. The adsorption of L. ferriphilum was not affected by A. caldus. However, adsorption of A. caldus was greatly affected by the presence of L. ferriphilum. Zeta-potential measurements and FT-IR spectroscopic studies showed that L. ferriphilum had a higher electrostatic attraction towards pyrite when compared to A. caldus. Based on the above results, we propose a competitive adsorption model to explain the mechanism by which L. ferriphilum and A. caldus compete in their adsorption to pyrite, although L. ferriphilum dominated in the competitive adsorption process. This work provides a better understanding of the adsorption behavior of mixed microbial populations onto mineral surfaces.     

Isolation of a strain of <Emphasis Type="Italic">Acidithiobacillus caldus</Emphasis> and its role in bioleaching of chalcopyrite

A moderately thermophilic and acidophilic sulfur-oxidizing bacterium named S₂, was isolated from coal heap drainage. The bacterium was motile, Gram-negative, rod-shaped, measured 0.4 to 0.6 by 1 to 2 μm, and grew optimally at 42–45°C and an initial pH of 2.5. The strain S₂ grew autotrophically by using elemental sulfur, sodium thiosulfate and potassium tetrathionate as energy sources. The strain did not use organic matter and inorganic minerals including ferrous sulfate, pyrite and chalcopyrite as energy sources. The morphological, biochemical, physiological characterization and analysis based on 16S rRNA gene sequence indicated that the strain S₂ is most closely related to Acidithiobacillus caldus (>99% similarity in gene sequence). The combination of the strain S₂ with Leptospirillum ferriphilum or Acidithiobacillus ferrooxidans in chalcopyrite bioleaching improved the copper-leaching efficiency. Scanning electron microscope (SEM) analysis revealed that the chalcopyrite surface in a mixed culture of Leptospirillum ferriphilum and Acidithiobacillus caldus was heavily etched. The energy dispersive X-ray (EDX) analysis indicated that Acidithiobacillus caldus has the potential role to enhance the recovery of copper from chalcopyrite by oxidizing the sulfur formed during the bioleaching progress.     

Physiological and Morphological Characteristics of Acidophilic Bacteria <Emphasis Type="Italic">Leptospirillum ferriphilum</Emphasis> and <Emphasis Type="Italic">Acidithiobacillus thiooxidans</Emphasis>, Members of a Chemolithotrophic Microbial Consortium

A thermoacidophilic consortium of chemolithotrophic microorganisms oxidizing the concentrate of high-pyrrhotite pyrite?arsenopyrite ore at 38–40°C was isolated. The most active members of the consortium were identified as Leptospirillum ferriphilum, Acidithiobacillus thiooxidans, Ferroplasma acidiphilum, and Sulfobacillus thermotolerans. Leptospirillum and Thiobacillus species were the most numerous members of the consortium and had the highest activity of ferrous iron and sulfur oxidation, respectively. The optimal temperature values for the growth of both isolates were within 35–38°C. The optimal ranges of initial pH were 1.0–1.2 and 1.75–1.85 for leptospirilla and 1.7–3.3 for thiobacilli with the pH optimum of 1.9. Significant polymorphism and specific cyclic growth with formation of vibrios, spirilla, rods with different end shape, spiral filaments, numerous “pseudococci,” and densely packed spiral filaments surrounded by a sheath were revealed for the Leptospirillum isolate. Two latter morphoforms of L. ferriphilum were not previously described. Differences in ability of the morphoforms to oxidize Fe²⁺ were revealed. For the first time, the possibility of growth in the presence of organic substances was demonstrated for A. thiooxidans. The rates of growth and substrate oxidation, cell size, and the maximal cell yield decreased insignificantly in comparison with the lithoautotrophic strain.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

Bio-adsorption and Bio-transformation of Arsenic by <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis> BY3

Arsenic, a toxic element in the environment, has seriously threatened the health of hundreds of millions of people in the world. Meanwhile, microorganisms play an important role in the adsorption and bio-transformation of arsenic. Here, we compared the biological characteristics of Acidithiobacillus ferrooxidans BY3 in different media systems, such as arsenic bio-adsorption and bio-transformation capacities. We show that arsenic stress significantly affected the pH and Eh of the culture systems, as well as the oxidation rates of Fe²⁺ and bacteria numbers. Furthermore, arsenic influenced bacterial structure and composition of the cell membrane, caused volume decreased and changed the vibration conditions of characteristic peaks of surface groups (-CH₂, -NH, and –OH) on cell membranes. In addition, At.f-BY3 shows high bio-adsorption abilities and certain bio-transformation abilities for iAs^III. Bio-adsorption and conversion efficiency was also shown to be significantly affected by Fe²⁺ concentrations in the reaction systems. Statistic analysis revealed 10.07-fold increase of the transformation ability of iAs^III into iAs^V in the 9 K growth media containing 1600 mg/L NaAsO₂ compared with that in the 1 K growth media. Our findings contribute to understand the applications and microbiological mechanisms of Acidithiobacillus ferrooxidans in arsenic pollution.

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Graphical Abstract Bio-adsorption and bio-transformation are used as a biological method of heavy metals pollution, such as Cu²⁺, Ni²⁺, Pb²⁺, Cr²⁺, Zn²⁺, Cd²⁺, As³⁺ and As⁵⁺ in acid mine water. The aim of this investigation was to assess the performance of arsenite (iAsIII) to adsorption and transformation by Acidithiobacillus ferrooxidans BY3, and application of Acidithiobacillus ferrooxidans BY3 on the aspect of arsenic pollution has great potential of exploration.

     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

Phylogenetic analyses of <Emphasis Type="Italic">Uromyces viciae-fabae</Emphasis> and its varieties on <Emphasis Type="Italic">Vicia</Emphasis>, <Emphasis Type="Italic">Lathyrus</Emphasis>, and <Emphasis Type="Italic">Pisum</Emphasis> in Japan

A pea rust fungus, Uromyces viciae-fabae, has been classified into two varieties, var. viciae-fabae and var. orobi, based on differences in urediniospore wall thickness and putative host specificity in Japan. In principal component analyses, morphological features of urediniospores and teliospores of 94 rust specimens from Vicia, Lathyrus, and Pisum did not show definite host-specific morphological groups. In molecular analyses, 23 Uromyces specimens from Vicia, Lathyrus, and Pisum formed a single genetic clade based on D1/D2 and ITS regions. Four isolates of U. viciae-fabae from V. cracca and V. unijuga could infect and sporulate on P. sativum. These results suggest that U. viciae-fabae populations on different host plants are not biologically differentiated into groups that can be recognized as varieties.Contribution no. 184, Laboratory of Plant Parasitic Mycology, Institute of Agriculture and Forestry, University of Tsukuba, Japan     

Genes and pathways for CO<Subscript>2</Subscript> fixation in the obligate,chemolithoautotrophic acidophile, <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis>, Carbon fixation in <Emphasis Type="Italic">A. ferrooxidans</Emphasis>

Background  

Acidithiobacillus ferrooxidans is chemolithoautotrophic γ-proteobacterium that thrives at extremely low pH (pH 1-2). Although a substantial amount of information is available regarding CO₂ uptake and fixation in a variety of facultative autotrophs, less is known about the processes in obligate autotrophs, especially those living in extremely acidic conditions, prompting the present study.     

Bio-resolution of glycidyl (<Emphasis Type="Italic">o</Emphasis>, <Emphasis Type="Italic">m</Emphasis>, <Emphasis Type="Italic">p</Emphasis>)-methylphenyl ethers by <Emphasis Type="Italic">Bacillus megaterium</Emphasis>

A newly isolated Bacillus megaterium with epoxide hydrolase activity resolved racemic glycidyl (o, m, p)-methylphenyl ethers to give enantiopure epoxides in 84–99% enantiomeric excess and with 21–73 enantiomeric ratios. The (S)-enantiomer was obtained from rac-glycidyl (o or m)-methylphenyl ether while the (R)-epoxides was obtained from glycidyl p-methylphenyl ether. The observations are explained at the level by enzyme-substrate docking studies.     

<Emphasis Type="Italic">KRAS</Emphasis>, <Emphasis Type="Italic">BRAF</Emphasis>, <Emphasis Type="Italic">EGFR</Emphasis> and <Emphasis Type="Italic">HER2</Emphasis> gene status in a Spanish population of colorectal cancer

To evaluate the KRAS, BRAF, EGFR, and HER2 gene status in colorectal cancer by novel techniques and evaluate whether anti-HER2 therapies could be offered in the treatment of these patients. There are conflicting data on the prevalence of BRAF mutations and EGFR and HER2 gene amplification in colorectal KRAS wild type patients. In our study we tried to evaluate these expressions and their relationship to future treatment assays. Clinical–pathological data and paraffin-embedded specimens were collected from 186 patients who underwent colorectal resections at General Yagüe Hospital in Burgos, Spain. KRAS and BRAF status was analyzed by real-time PCR in all patients. EGFR and HER2/NEU gene amplification was detected using fluorescent in situ hybridisation technique (FISH) in 38 KRAS and BRAF wild type patients. KRAS mutations were present in 48% of the colorectal cancer patients. BRAF mutations were present in 6.25% of the KRAS wild type patients. EGFR and HER2 gene amplification was observed in 5.3% and 26.3%, respectively, of KRAS and BRAF wild type colorectal cancer patients. HER2, but not EGFR gene amplification, was frequently observed in KRAS and BRAF wild type colorectal cancer patients. These data indicate that HER2 amplification could be one of the genes to be considered in the therapeutic management of colorectal cancer.     

Redescriptions of the Indo-Pacific atherinid fishes <Emphasis Type="Italic">Atherinomorus forskalii</Emphasis>, <Emphasis Type="Italic">Atherinomorus lacunosus</Emphasis>, and <Emphasis Type="Italic">Atherinomorus pinguis</Emphasis>

The Indo-Pacific marine atherinid fishes Atherinomorus forskalii (Rüppell, 1838), Atherinomorus lacunosus (Forster, 1801), and Atherinomorus pinguis (Lacepède, 1803) are redescribed as valid species based on the types and non-type specimens collected throughout the Indo-Pacific. They are similar to each other chiefly in having a wide midlateral band (almost the same or greater than the midlateral scale width), large mouth (posterior tip of upper jaw reaching to or beyond a vertical through anterior margin of pupil), and no distinct tubercle at the posterior end of the dentary. All three species are distinguishable from congeners by those characters. The three species have long been confused with each other or synonymized erroneously as a single species. Atherinomorus forskalii, known from the Red Sea and eastern Mediterranean, differs from Atherinomorus lacunosus and Atherinomorus pinguis in having conspicuous, large endopterygoid teeth, forming obvious tooth ridges. Atherinomorus lacunosus, widely distributed in almost the entire Indo-Pacific, from East Africa to Tonga, north to southern Japan, and south to northern Australia, differs from Atherinomorus pinguis in having a wider midlateral band (the lower margin reaching to almost the center of the fourth scale row at level of the anal fin origin vs. the lower margin reaching to the ventral end of the third scale row in Atherinomorus pinguis) and more numerous midlateral scales (40–44 vs. 38–41 in Atherinomorus pinguis). Atherina morrisi Jordan and Starks, 1906, Hepsetia pinguis mineri Nichols and Roemhild, 1951, Pranesus capricornensis Woodland, 1961, Pranesus maculatus Taylor, 1964, and Pranesus pinguis ruppelli Smith, 1965, are regarded as junior synonyms of Atherinomorus lacunosus. Atherinomorus pinguis is also widely distributed in the Indo-West Pacific, from East Africa to northern Australia and north to southern Japan. Atherina pectoralis Valenciennes, 1835, is considered a junior synonym of Atherinomorus pinguis. Supplementary material to this paper is available in electronic format at     

Association between <Emphasis Type="Italic">cagA</Emphasis>, <Emphasis Type="Italic">vacAi</Emphasis>, and <Emphasis Type="Italic">dupA</Emphasis> genes of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> and gastroduodenal pathologies in Chilean patients

In addition to the already known cagA gene, novel genetic markers have been associated with Helicobacter pylori (H. pylori) virulence: the dupA and vacAi genes. These genes might play an important role as specific markers to determine the clinical outcome of the disease, especially the vacAi gene, which has been expected to be a good marker of severe pathologies like gastric adenocarcinoma. In the present study, the association of cagA, dupA, and vacAi genes with gastroduodenal pathologies in Chilean patients was studied. One hundred and thirty-two patients positive for H. pylori were divided into two groups—non-severe and severe gastric pathologies—and investigated for the presence of cagA, dupA, and vacAi H. pylori virulence genes by PCR. The cagA gene was detected in 20/132 patients (15.2%), the vacAi1 gene was detected in 54/132 patients (40.9%), the vacAi2 gene was detected in 26/132 patients (19.7%), and the dupA gene was detected in 50/132 (37.9%) patients. Logistic regression model analysis showed that the vacAi1 isoform gene in the infected strains and the severity of the diseases outcome were highly associated, causing severe gastric damage that may lead to gastric cancer (p < 0.0001; OR = 8.75; 95% CI 3.54–21.64). Conversely, cagA (p = 0.3507; OR = 1.62; 95% CI 0.59–4.45) and vacAi2 (p = 0.0114; OR = 3.09; 95% CI 1.26–7.60) genes were not associated with damage, while the dupA gene was associated significantly with non-severe clinical outcome (p = 0.0032; OR = 0.25; 95% CI 0.09–0.65). In addition, dupA gene exerts protection against severe gastric pathologies induced by vacAi1 by delaying the outcome of the disease by approximately 20 years.     

Molecular analysis of the genus <Emphasis Type="Italic">Anoxybacillus</Emphasis> based on sequence similarity of the genes <Emphasis Type="Italic">recN</Emphasis>, <Emphasis Type="Italic">flaA</Emphasis>, and <Emphasis Type="Italic">ftsY</Emphasis>

Genome predictions based on selected genes would be a very welcome approach for taxonomic studies. We analyzed three genes, recN, flaA, and ftsY, for determining if these genes are useful tools for systematic analyses in the genus Anoxybacillus. The genes encoding a DNA repair and genetic recombination protein (recN), the flagellin protein (flaA), and GTPase signal docking protein (ftsY) were sequenced for ten Anoxybacillus species. The sequence comparisons revealed that recN sequence similarities range between 61% and 99% in the genus Anoxybacillus. Comparisons to other bacterial recN genes indicated that levels of similarity did not differ from the levels within genus Anoxybacillus. These data showed that recN is not a useful marker for the genus Anoxybacillus. A 550–600-bp region of the flagellin gene was amplified for all Anoxybacillus strains except for Anoxybacillus contaminans. The sequence similarity of flaA gene varies between 61% and 76%. Comparisons to other bacterial flagellin genes obtained from GenBank (Bacillus, Pectinatus, Proteus, and Vibrio) indicated that the levels of similarity were lower (3–42%). Based on these data, we concluded that the variability in this single gene makes it a particularly useful marker. Another housekeeping gene ftsY suggested to reflect the G+C (mol/mol) content of whole genome was analyzed for Anoxybacillus strains. A mean difference of 1.4% was observed between the G+C content of the gene ftsY and the G+C content of the whole genome. These results showed that the gene ftsY can be used to represent whole G+C content of the Anoxybacillus species.     

Bioleaching of uranium from low grade black schists by <Emphasis Type="Italic">Acidithiobacillus ferrooxidans</Emphasis>

Summary The feasibility of bacterial recovery of uranium from the low grade black schists occurring in the Okcheon district, South Korea, was investigated. Following the introduction of Acidithiobacillus ferrooxidans, 80% of the uranium could be extracted from the schists, which contain 0.01% U₃O₈ by weight, within 60 h at a pulp density of 100 g-ore/l. Only 18% of the uranium was extracted without microbial activity. The uranium-leaching efficiency was not greatly affected by the addition of Fe²⁺ in the range of 5–9 g/l, and the leaching efficiency of uranium from the schists by A. ferrooxidanscould be efficiently maintained at high pulp densities (up to 500 g-ore/l).     

Forecasting model of <Emphasis Type="Italic">Corylus</Emphasis>, <Emphasis Type="Italic">Alnus</Emphasis>, and <Emphasis Type="Italic">Betula</Emphasis> pollen concentration levels using spatiotemporal correlation properties of pollen count

The aim of the study was to create and evaluate models for predicting high levels of daily pollen concentration of Corylus, Alnus, and Betula using a spatiotemporal correlation of pollen count. For each taxon, a high pollen count level was established according to the first allergy symptoms during exposure. The dataset was divided into a training set and a test set, using a stratified random split. For each taxon and city, the model was built using a random forest method. Corylus models performed poorly. However, the study revealed the possibility of predicting with substantial accuracy the occurrence of days with high pollen concentrations of Alnus and Betula using past pollen count data from monitoring sites. These results can be used for building (1) simpler models, which require data only from aerobiological monitoring sites, and (2) combined meteorological and aerobiological models for predicting high levels of pollen concentration.     

A PCR-RFLP method on faecal samples to distinguish <Emphasis Type="Italic">Martes martes</Emphasis>, <Emphasis Type="Italic">Martes foina</Emphasis>, <Emphasis Type="Italic">Mustela putorius</Emphasis> and <Emphasis Type="Italic">Vulpes vulpes</Emphasis>

A non-invasive genetic approach has been recently employed in the population genetics of wild species, using faeces that could be easily collected, particularly of elusive or endangered species. However, faeces of pine (Martes martes) and beech marten (M. foina) can be morphologically similar and could be confused with those of polecat (Mustela putorius) and red fox (Vulpes vulpes). On this basis, a rapid, simple and inexpensive RFLP protocol using a cytochrome b (Cyt b) gene fragment of mtDNA, isolated from faecal samples, was performed to distinguish the above-mentioned species.