Efficient separation and purification of allophycocyanin from <Emphasis Type=Italic>Spirulina</Emphasis> (<Emphasis Type=Italic>Arthrospira</Emphasis>) <Emphasis Type=Italic>platensis</Emphasis>

Allophycocyanin (APC) is a minor component of phycobiliproteins in cyanobacteria and red algae. This paper describes a simple and inexpensive extracting method for isolating APC from Spirulina (Arthrospira) platensis with high efficiency. The crude phycobiliprotein extract was pretreated by ammonium sulfate fractionation. Then, by adding hydroxylapatite into crude phycobiliprotein extract dissolved in 20 mM phosphate buffer (pH 7.0), APC was selectively adsorbed by hydroxylapatite but C-phycocyanin (C-PC) was not. The hydroxylapatite was collected and APC was extracted from the crude phycobiliprotein extract. Then, the enriched APC was washed off from the hydroxylapatite using 100 mM phosphate buffer (pH 7.0). In this simple extracting method it was easy to remove C-PC and isolate APC in large amounts. The absorbance ratio A ₆₅₀/A ₂₈₀ of extracted APC reached 2.0. The recovery yield was 70%, representing 4.61 mg · g⁻¹ wet weight. The extracted APC could be further purified by a simple anion-exchange chromatography with a pH gradient from 5.6 to 4.0. The absorbance ratio A ₆₅₀/A ₂₈₀ of the purified APC reached 5.0, and the overall recovery yield was 43%, representing 2.83 mg · g⁻¹ wet weight. Its purity was confirmed by native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate-PAGE.     

Habitat,density, and spatial distribution of <Emphasis Type=Italic>Rivulus giarettai</Emphasis> (<Emphasis Type=Italic>Actinopterygii</Emphasis>, <Emphasis Type=Italic>Cyprinodontiformes</Emphasis>) in southeastern Brazil

We present data on the habitat, density, and spatial distribution of Rivulus giarettai, and discuss some biotic and abiotic variables related to its abundance in Free Flowing Waters (FFW) and Dam Reservoirs (DR) in palm grove (Mauritia flexuosa) marshes (Veredas) in Uberlandia, Minas Gerais, southeastern Brazil. The mean density (individuals/plot) of R. giarettai was about 13 times higher in FFW than in DR. In FFW, the density of R. giarettai was highest at intermediate amounts of substrate (plant mass) and it was positively rank-correlated with the depth, and the number of arthropods. Individuals occurred in an aggregated distribution. The aggregated pattern could be related to a concentration of individuals in microhabitats neither too much exposed nor completely saturated by plants. R. giarettai was relatively abundant and tolerant to slight man-made habitat modifications. Damming appeared to be especially problematic by negatively affecting its density.     

<Emphasis Type=Italic>Pseudomonas</Emphasis> bacteria and phosphorous fertilization,affecting wheat (<Emphasis Type=Italic>Triticum</Emphasis><Emphasis Type=Italic>aestivum</Emphasis> L.) yield and P uptake under greenhouse and field conditions

We have recently indicated the plant growth promoting activities of Pseudomonas sp. as well as their alleviating effects on some soil stressors such as salinity. This is because in recent years, biological fertilizers have received special attention by scientists in sustainable agriculture. Accordingly, it is pertinent to specify the beneficiary level of such soil bacteria on plant growth including phosphorous (P) uptake. Hence, the objectives were to determine: (1) the plant growth promoting effects of the tested Pseudomonas sp., and (2) its combined effects with different P fertilization rates on the nutrient uptake (N, P, and K) and yield of wheat (Triticum aestivum L.) under greenhouse and field conditions. The experiments were factorially arranged on the basis of a completely randomized block design with three replicates and were conducted at the Research Farm of Agriculture and Natural Resources Research Center of Khorasan, Mashhad, Iran. P was fertilized at three levels including 0, 25 and 50 kg/ha P₂O₅. Pseudomonas sp. including Pseudomonas fluorescens 153, P. fluorescens 169, P. putida 4, and P. putida 108 were tested. Activities such as production of ACC deaminase and IAA-like products, as well as P solubilization were among the most important activities of the tested Pseudomonas sp. Such bacterial effects greatly enhanced wheat growth and yield under greenhouse and field conditions. The results also showed that the effects of Pseudomonas sp. on wheat nutrient uptake and the effects of bacteria as well as P fertilization on wheat yield were significant. P. putida 108 was the most effective strain enhancing wheat P uptake and grain yield under greenhouse (96 and 58%) and field (80 and 37%) conditions, respectively. Hence, although Pseudomonas sp. could be a suitable replacement for high P fertilization, however, the optimum wheat yield resulted when the bioinoculants are combined with 50% (25 kg/ha P₂O₅) P fertilization. This finding has great agricultural and environmental implications.     

Light respiration in <Emphasis Type=Italic>Chlorella sorokiniana</Emphasis>

Respiration and photosynthesis are two important processes in microalgal growth that occur simultaneously in the light. To know the rates of both processes, at least one of them has to be measured. To be able to measure the rate of light respiration of Chlorella sorokiniana, the measurement of oxygen uptake must be fast, preferably in the order of minutes. We measured the immediate post-illumination respiratory O₂ uptake rate (OUR) in situ, using fiber-optic oxygen microsensors, and a small and simple extension of the cultivation system. This method enables rapid and frequent measurements without disturbing the cultivation and growth of the microalgae. Two batch experiments were performed with C. sorokiniana in a short light-path photobioreactor, and the OUR was measured at different time points. The net oxygen production rate (net OPR) was measured online. Adding the OUR and net OPR gives the gross oxygen production rate (gross OPR), which is a measure for the oxygen evolution by photosynthesis. The gross OPR was 35–40% higher than the net OPR for both experiments. The respiration rate is known to be related to the growth rate, and it is suggested that faster algal growth leads to a higher energy (ATP) requirement, and as such, respiratory activity increases. This hypothesis is supported by our results, as the specific OUR is highest in the beginning of the batch culture when the specific growth rate is highest. In addition, the specific OUR decreases toward the end of the experiments until it reaches a stable value of around 0.3 mmol O₂ h⁻¹ g⁻¹. This value for the specific OUR is equal to the maintenance requirement of C. sorokiniana as determined in an independent study of (Zijffers et al. 2010 (in press)). This suggests that respiration could fulfill the maintenance requirements of the microalgal cells.     

Presence of <Emphasis Type=Italic>C. albidus</Emphasis>, <Emphasis Type=Italic>C. laurentii</Emphasis> and <Emphasis Type=Italic>C. uniguttulatus</Emphasis> in Crop and Droppings of Pigeon Lofts (<Emphasis Type=Italic>Columba livia</Emphasis>)

Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this disease.     

<Emphasis Type=Italic>Boletus kermesinus</Emphasis>, a new species of <Emphasis Type=Italic>Boletus</Emphasis> section <Emphasis Type=Italic>Luridi</Emphasis> from central Honshu,Japan

Boletus kermesinus, a new species of Boletus section Luridi, is fully described and illustrated based on the materials collected in subalpine coniferous forests of central Honshu, Japan. It has distinctive features of dark-red basidiomata having distinct viscidity in the pileus surface, usually unchanging flesh, discolorous red pores, and an entirely reticulate stipe becoming coarsely lacerate-rimose with age.     

Attenuation of fibrosis <Emphasis Type=Italic>in vitro</Emphasis> and <Emphasis Type=Italic>in vivo</Emphasis> with <Emphasis Type=Italic>SPARC</Emphasis> siRNA

Introduction  

SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.     

Multiple host-ant use by the predatory social parasite <Emphasis Type=Italic>Phengaris</Emphasis> (=<Emphasis Type=Italic>Maculinea</Emphasis>) <Emphasis Type=Italic>arion</Emphasis> (Lepidoptera,Lycaenidae)

Phengaris (=Maculinea) arion is an endangered social parasite of Myrmica ants, and for a very long time was considered as specific to Myrmica sabuleti. Previous studies carried out in Poland suggested some discrepancies within this assumption, and therefore a much more intensive survey was undertaken. The host ant use of P. arion was studied at five sites in different types of biotopes in Poland, i.e. xerothermal grasslands where Thymus pulegioides was used as a larval food plant by the butterfly, and more or less sandy biotopes with Thymus serpyllum. Altogether nine Myrmica species were recorded, and considerable variation in species composition and density of nests was recorded. At four localities M. sabuleti proved to be the most common ant. A total of 529 Myrmica nests were examined, and only 20 of them contained larvae and pupae of P. arion. Host ants belonged to five different species, i.e. M. sabuleti, Myrmica scabrinodis, Myrmica schencki, Myrmica lobicornis and Myrmica hellenica. Only at one site (NE Poland) was a significant heterogeneity in parasitation rates among Myrmica species detected. M. lobicornis was the most often infested ant there, which may suggest local specialisation of the butterfly. Overall low parasitism rates may explain the vulnerability of P. arion in Central Europe but further studies are also necessary.     

Interaction of <Emphasis Type=Italic>Bdellovibrio bacteriovorus</Emphasis> with bacteria <Emphasis Type=Italic>Campylobacter jejuni</Emphasis> and <Emphasis Type=Italic>Helicobacter pylori</Emphasis>

Interaction of Bdellovibrio bacteriovorus 100NCJB with bacteria Campylobacter jejuni (strains 1, 2, 3, 4, and 5) and Helicobacter pylori, strain TX30a, was confirmed. The results indicate that lytic activity of bdellovibrios both in liquid media and cells attached to a surface was observed. The potential use of the antimicrobial activity of predatory bacteria for environmental bioprotection and public health is discussed.     

Manifestation of salt tolerance of <Emphasis Type=Italic>Spirulina platensis</Emphasis> and <Emphasis Type=Italic>Spirulina maxima</Emphasis> cyanobacteria of the genus <Emphasis Type=Italic>Arthrospira (Spirulina)</Emphasis>

The salt tolerance of two representatives of genus Spirulina (Arthrospira) Spirulina platensis and Spirulina maxima has been investigated. They both are the wide-spread objects of photobiotechnology and it has been shown that the content of 5–15 % sea-water in medium has not caused the decreasing of biomass yield more than 15–20% as compared with control. The further decreasing of biomass was proportionate to sea-water content in medium. The investigation of reactivity of native (intravital) exometabolites secreted into cultural medium has showed that the sea-water content influence the oxidative activity (OA) of exometabolites and hour’s rhythmics.     

<Emphasis Type=Italic>In vitro</Emphasis> organogenesis of <Emphasis Type=Italic>Passiflora alata</Emphasis>

Passiflora alata in vitro organogenesis was studied based on explant type, culture medium composition, and incubation conditions. The results indicated that the morphogenic process occurred more efficiently when hypocotyl segment-derived explants were cultured in media supplemented with cytokinin and AgNO₃ incubated under a 16-h photoperiod. The shoot bud elongation and plant development were obtained by transferring the material to MSM culture medium supplemented with GA₃ and incubated in flasks with vented lids. Histological analyses of the process revealed that the difficulties in obtaining plants could be related to the development of protuberances and leaf primordia structures, which did not contain shoot apical meristem. Roots developed easily by transferring elongated shoots to 1/2 MSM culture medium. Plant acclimatization occurred successfully, and somaclonal variation was not visually detected. The efficiency of this organogenesis protocol will be evaluated for genetic transformation of this species to obtain transgenic plants expressing genes that can influence the resistance to Cowpea aphid borne mosaic virus.