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.     

Validation of <Emphasis Type=Italic>Christianella</Emphasis> Guiart, 1931 (Cestoda: Trypanorhyncha) and its taxonomic relationship with <Emphasis Type=Italic>Grillotia</Emphasis> Guiart, 1927

Christianella Guiart, 1931 (Cestoda: Trypanorhyncha) is redefined as a subgenus of Grillotia Guiart, 1927 based on the type-species, G. (C.) minuta (van Beneden, 1849), from the elasmobranch Squatina squatina (Linnaeus). Grillotia smarisgora (Wagener, 1854) is treated as a synonym of G. (C.) minuta, as are G. angeli Dollfus, 1969 and G. bothridiopunctata Dollfus, 1969. Other species included in the subgenus are G. (C.) carvajalregorum Menoret & Ivanov, 2009 (formerly Progrillotia dollfusi Carvajal & Rego, 1983), G. (C.) australis Beveridge & Campbell, 2001, G. (C.) longispinis (Linton, 1890) n. comb. (formerly Rhynchobothrium longispine Linton, 1890) and G. (C.) yuniariae Palm, 2004. The subgenus is similar to Grillotia Guiart, 1927 (sensu stricto), having two bothria and an atypical heteroacanthous armature, but differs in having a single row of intercalary hooks, fewer, elongate segments with testes often in longitudinal columns, a distinctive basal armature, an internal seminal vesicle which extends beyond the cirrus or hermaphroditic sac and no uterine pore. The adults of three species are known, all parasitising members of Squatina Duméril.     

<Emphasis Type=Italic>Barleria compacta</Emphasis>: a new species in <Emphasis Type=Italic>Barleria</Emphasis> sect. <Emphasis Type=Italic>Prionitis</Emphasis> (Acanthaceae) from Somalia

Following a re-examination of the material treated under Barleria brevispina (Fiori) Hedrén in the recent Flora of Somalia account of the Acanthaceae, it is concluded that two distinct species are involved and Barleria compacta Malombe & I. Darbysh. is described here from north-eastern Somalia. Its affinities and conservation status are discussed.     

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.     

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.     

<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.     

Demographic characteristics of the copepod <Emphasis Type=Italic>Acanthocyclops americanus</Emphasis> (Sars, 1863) (Copepoda: Cyclopoida) fed mixed algal (<Emphasis Type=Italic>Scenedesmus acutus</Emphasis>)-rotifer (<Emphasis Type=Italic>Brachionus havanaensis</Emphasis>) diet

The predatory copepod Acanthocyclops americanus is commonly found in shallow ponds and lakes in Mexico. We tested the survivorship and reproduction-related variables of this copepod, isolated from Lake Huetzalin (Xochimilco, Mexico City), on four mixed diets comprising two algal concentrations (0.8 × 10⁶ and 1.6 × 10⁶ cells ml⁻¹ of Scenedesmus acutus) and two rotifer densities (1 and 8 ind. ml⁻¹ of Brachionus havanaensis). Survivorship patterns of naupliar and adult stages of A. americanus were evaluated at 24 ± 1°C. The demography experiments were initiated with five females and five males (1:1 ratio, 5th copepodites) into each of the 16 test [= 4 diet combinations × 4 replicates (cohorts)] jars containing 50 ml medium and with one of the chosen algal-rotifer diets. Data on the survival of naupliar and copepodites (up to V) showed a high death rate (80%) in A. americanus by the time nauplii reached the adult stage. The duration of developmental time for nauplii and the copepodite V (C-V) was about 28 days by which time A. americanus became adults. Food combination had a significant effect on both survival and duration of the naupliar and C-V but not on the development time in C-I, C-II, C-III, and C-IV. Naupliar stages showed highest life expectancy at low food levels (rotifers or alga). Adult male and female copepods showed better survival under higher availability of rotifers in the medium, while higher algal density resulted in steep mortality during the first days. Average adult lifespan and life expectancy of A. americanus varied from 19 to 23 days, depending on the diet combination. Age-specific reproductive output (m _x ) of female A. americanus showed low reproduction (<10 offspring per female on any given day) when raised on low rotifer density, regardless of the algal concentration. On the other hand, enhanced offspring production (average of about 50 nauplii per female) was recorded when the rotifer density was higher (i.e., 8 ind. ml⁻¹). Gross and net reproductive rates varied from 48 to 438 and 27 to 318 offspring per female, respectively. In general, higher availability of rotifers in the medium resulted in higher (8–10 times as compared to those in low rotifer density) gross and net reproductive rates. There was no significant difference in the generation time (13–14 days) in relation to the diet combination. The rate of population increase (r) (range: 0.29–0.69 per day) increased with greater abundance of B. havanaensis in the medium. Thus, our study showed that inclusion of animal protein in the diet is necessary for high population growth rates of A. americanus.     

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.