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 overlap between sympatric European hares (<Emphasis Type=Italic>Lepus europaeus</Emphasis>) and Eastern cottontails (<Emphasis Type=Italic>Sylvilagus floridanus</Emphasis>) in northern Italy

In northern Italy, the range of the Eastern cottontail (Sylvilagus floridanus) largely overlaps with that of the native European hare (Lepus europaeus) on the Po Plain. Both species appear to have similar habitat requirements. We studied habitat selection by hares and cottontails during feeding activity from September 2006 to August 2007 in two areas where they occur alone (allopatry) and in one area where they occur together (sympatry). The three areas were basically similar, so that shifts in habitat use observed in sympatry should reflect the response to interspecific competition. Habitat selection was examined at micro- and macro-habitat levels throughout seasons. Habitat breadth of both species followed the change of resource availability through seasons in allopatry as well as in sympatry. No shifts in habitat use were evident at macro-habitat level, even during autumn which was the limiting season. Exploitation of shared habitats by the two species seems to be promoted by differential micro-habitat use within macro-habitat types. Cottontails used woods with dense understory in greater proportion than hares, and their present sites were concentrated within the maximum distance of 20 m of the nearest shelter site. Hares were more likely than cottontails to exploit crops, and their sites were distributed even greater than 80 m away from permanent cover patches. The habitat heterogeneity of agricultural ecosystems within the sympatry range could buffer the negative effects of external factors (climate, human disturbance and predation) on hares, and enhance the chances of exploitation of shared habitats by both species.     

On the distribution of seed-eating <Emphasis Type=Italic>Dicraeus nartshukae</Emphasis> (Diptera,Chloropidae) and its host plant <Emphasis Type=Italic>Leymus mollis</Emphasis>

The distribution ranges of Dicraeus nartshukae Kanmiya (Diptera, Chloropidae) and its host plant Leymus mollis (Trin.) Pilger are discussed. Larvae of D. nartshukae develop in unripe seeds of Leymus mollis but not in the seeds of the closely related L. villosissimus and L. interior. The spikes, spikelets, and glumes of L. mollis are larger than in other species, and usually softly hirsute. The glumes of L. villosissimus and L. interior possess dense hirsute pubescence which appears to be unfavorable for oviposition of Dicraeus nartshukae. The absence of seedeating Dicraeus on L. villosissimus and L. interior is an additional argument in support of distinctness of these species. The distribution range of Dicraeus nartshukae is meridional, or Pacific littoral.     

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.     

The notoriously destructive potato stalk borer (<Emphasis Type=Italic>Trichobaris trinotata</Emphasis>) has negligible impact on its native host, <Emphasis Type=Italic>Solanum carolinense</Emphasis> (horsenettle)

While herbivores can have a wide range of impacts on their host plants, instances in which herbivore damage is truly beneficial are rare. Thus, it was surprising that damage by the potato stalk borer (PSB) was associated with increased fitness of its native host plant (horsenettle) in a recent field study, particularly because this insect can have such severe negative effects on potato crops. A controlled experiment with potted horsenettle was conducted to investigate the impact of damage by PSB to see if it has any negative impacts on the plants, or whether horsenettle truly overcompensates for its damage. Damage by PSB had negligible impact on sexual reproduction and potential future vegetative propagation through root production in this experiment. Additionally, PSB damage did not exacerbate the negative fitness impact of heavy leaf damage by the eggplant lace bug. The lack of positive impacts in the potted horsenettle study suggests that selection for decreased resistance to PSB in the field study was due to correlated selection for increased plant vigor, rather than to overcompensation. That is, when given options, PSB females may preferentially oviposit on larger, healthier plants that are likely to be better hosts for their larvae. An important implication of these results is that a wild weed’s high tolerance of a natural herbivore can compound the weed’s potential to serve as a reservoir from which the herbivore can switch to closely related agricultural plants that are not tolerant of its damage.     

Food web overlap among native axolotl (<Emphasis Type=Italic>Ambystoma mexicanum</Emphasis>) and two exotic fishes: carp (<Emphasis Type=Italic>Cyprinus carpio</Emphasis>) and tilapia (<Emphasis Type=Italic>Oreochromis niloticus</Emphasis>) in Xochimilco,Mexico City

Two exotic fishes, common carp (Cyprinus carpio) and tilapia (Oreochromis niloticus), were introduced more than 20 years ago into Xochimilco for aquaculture, and now dominate the system in terms of biomass and numbers. Over this same period, wild populations of the microendemic axolotl salamander (Ambystoma mexicanum) have been dramatically reduced, and it currently persists in isolated areas of this aquatic system, which is situated at the edge of Mexico City. In this study, we examine potential trophic interactions and niche overlap among two exotic fishes: carp and tilapia, and the native axolotl. Axolotl had more diverse diets and a higher trophic position compared to the exotics. Stable isotope analysis revealed substantial trophic niche overlap among axolotl and the exotics. The two exotics occupied a larger niche area than the axolotl, suggesting higher levels of omnivory and diet specialization. Current exotic fish removal efforts will further our understanding of interactions between the axolotl and exotic species, and are expected to benefit axolotl recovery efforts.     

Cell-free culture medium of <Emphasis Type=Italic>Burkholderia cepacia</Emphasis> improves seed germination and seedling growth in maize (<Emphasis Type=Italic>Zea mays</Emphasis>) and rice (<Emphasis Type=Italic>Oryza sativa</Emphasis>)

The saprophytic bacterium Burkholderia cepacia has been shown to play an active role as plant growth promoting bacteria (PGPB). In this study, the ability of cell-free culture medium (CFCM) of B. cepacia to improve early developmental stages of plants has been assessed on two agronomically important crops, maize (Zea mays) and rice (Oryza sativa). Treating maize and rice seeds for 45 min before germination significantly improved seed germination and consequent seedling growth. The effect of CFCM was confirmed by the increased biomass of the shoot and, mainly, the root systems of treated seedlings. Chromatographic characterization of the CFCM revealed that the spent culture medium of B. cepacia is a complex mix of different classes of metabolites including, among others, salicylic acid, indole-3-acetic acid (IAA) and several unidentified phenolic compounds. Fractionation of the CFCM components revealed that the impressive development of the root system of CFCM-treated seedlings is due to the synergistic action of several groups of components rather than IAA alone. The data presented here suggest that a CFCM of B. cepacia can be used to improve crop germination.     

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.     

<Emphasis Type=Italic>Asthenes dorbignyi</Emphasis> (Passeriformes: Furnariidae) host of <Emphasis Type=Italic>Argas neghmei</Emphasis> (Acari: Argasidae)

Argas neghmei Kohls & Hoogstraal is a Neotropical tick species parasite of poultry and man in the arid western of the southern cone of America in Argentina and Chile. Males, females and nymphs of an argasid were collected from 20 to 22 of June, 2007 in nests of Creamy-breasted Canastero, Asthenes dorbignyi (Reinchenbach) (Passeriformes: Furnariidae) at about 7 km S of La Poma (24°46′S, 66°12′W) and a male tick at 24°48′S, 66°10′W on Nov 27, 2008, Salta province, Argentina. Scanning electron microscope micrographies and sequences of 16S rDNA confirmed that the ticks found were A. neghmei. Development in breeding areas of A. dorbignyi is probably a source of infestation for animal premises and houses. However, the role of other unknown wild birds may also contribute to the invasion of A. neghmei in chicken houses or residences.     

Positioning of the major locus for <Emphasis Type=Italic>Puccinia psidii</Emphasis> rust resistance (<Emphasis Type=Italic>Ppr1</Emphasis>) on the <Emphasis Type=Italic>Eucalyptus</Emphasis> reference map and its validation across unrelated pedigrees

In this report the major locus for Puccinia psidii rust resistance, Ppr1, was positioned on the reference genetic map for Eucalyptus. Additionally, its position was validated by association genetics in a related and two unrelated pedigrees involving different Eucalyptus grandis resistant trees crossed to individuals of two other species, Eucalyptus tereticornis and Eucalyptus camaldulensis. These results are consistent with the hypothesis that Ppr1 controls a large proportion of the variation for rust resistance, strengthening its role as a major locus in Eucalyptus and providing its unequivocal genomic position on linkage group 3. A localized map with 19 microsatellite loci was built around Ppr1. Multiallelic profiles were observed at several mapped microsatellites suggesting recent tandem duplications in the genomic landscape surrounding Ppr1. Markers EMBRA125 and EMBRA1071 flank Ppr1 at 9.5% and 7% recombination, respectively, and were found to be in linkage equilibrium in a E. grandis breeding population, consistent with the expectations in outcrossed Eucalyptus. Their potential use for MAS will specifically be directed to identifying resistant offspring of P. psidii resistant parent trees that are heterozygous at Ppr1. In these circumstances, a significant amount of LD is expected to occur between specific alleles at flanking microsatellites and the resistance allele at Ppr1. Moreover, the positional information of Ppr1 paves the way for prospective undertakings in this genomic region with the upcoming availability of a draft genome for E. grandis.     

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.     

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.