Genetic variation assessed with microsatellites in mass selection lines of the Pacific oyster (<Emphasis Type="Italic">Crassostrea gigas</Emphasis>) in China

Four successive mass selection lines of the Pacific oyster, Crassostrea gigas, selected for faster growth in breeding programs in China were examined at ten polymorphic microsatellite loci to assess the level of allelic diversity and estimate the effective population size. These data were compared with those of their base population. The results showed that the genetic variation of the four generations were maintained at high levels with an average allelic richness of 18.8–20.6, and a mean expected heterozygosity of 0.902–0.921. They were not reduced compared with those of their base population. Estimated effective population sizes based on temporal variances in microsatellite frequencies were smaller to that of sex ratio-corrected broodstock count estimates. Using a relatively large number of broodstock and keeping an equal sex ratio in the broodstock each generation may have contributed to retaining the original genetic diversity and maintaining relatively large effective population size. The results obtained in this study showed that the genetic variation was not affected greatly by mass selection progress and high genetic variation still existed in the mass selection lines, suggesting that there is still potential for increasing the gains in future generations of C. gigas. The present study provided important information for future genetic improvement by selective breeding, and for the design of suitable management guidelines for genetic breeding of C. gigas.     

Genetic transformation of <Emphasis Type="Italic">Nannochloropsis oculata</Emphasis> with a bacterial phleomycin resistance gene as dominant selective marker

The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2×10⁷ cells mL^?1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL^?1 zeocin. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.     

Spatial pattern analysis and associations of different growth stages of populations of <Emphasis Type="Italic">Abies georgei</Emphasis> var. <Emphasis Type="Italic">smithii</Emphasis> in Southeast Tibet,China

Abies georgei var. smithii is an important plant species in Southeast Tibet, China. It has high ecological value in terms of biodiversity protection, as well as soil and water conservation. We analyzed the spatial pattern and associations of A. georgei var. smithii populations at different growth stages by using Ripley’s L function for point pattern analysis. The diameter structure was a nearly reverse ‘J’ shape. The amount of saplings and medium-sized trees accounts for a large part of the entire population, suggesting a high regeneration rate and an expanding population. In the transition from saplings to medium trees or to large trees, saplings show a significant aggregation distribution at small scales, while medium trees and large trees show a random distribution. There are significant inverse associations between saplings and medium trees and large trees at small scales, while there are no obvious associations between medium trees and large trees. The natural regeneration was affected by interspecific competition, and it was also affected by intraspecific competition. The joint effects of biological characteristics and environmental factors contribute to the spatial distribution pattern and associations of this A. georgei var. smithii population.     

Significance of different microalgal species for growth of moon jellyfish ephyrae, <Emphasis Type="Italic">Aurelia</Emphasis> sp.1

The scyphozoan Aurelia aurita (Linnaeus) sp. l., is a cosmopolitan species-complex which blooms seasonally in a variety of coastal and shelf sea environments around the world. The effects of different microalgal species on the growth of newly-released Aurelia sp.1 ephyrae were studied under laboratory conditions. We fed ephyrae with four different microalgal species (diatom, autotrophic dinoflagellate, heterotrophic dinoflagellate, and chlorophyta) plus Artemia nauplii for 12-24 d at 18°C. Results showed that the growth rate diverged significantly for Artemia nauplii compared to other food types. In addition, there was no significant variation between the growth rates for Skeletonema costatum and Prorocentrum donghaiense, and no significant variation was found in the growth rates for N. scintillans and P. subcordiformis. Artemia nauplii could support the energy requirement for the newly-released ephyrae to develop to meduase, and the ephyrae with Artemia nauplii showed a significant average growth rate of 25.85% d^?1. Newly-released ephyrae could grow slightly with some species of microalgae in the earliest development stage. Chain diatom Skeletonema costatum and autotrophic dinoflagellate Prorocentrum donghaiense, could not support the growth of the ephyrae, while heterotrophic dinoflagellate Noctiluca scintillans and chlorophyta Platymonas subcordiformis could support the growth of the ephyrae. However, none of the ephyrae fed with the tested phytoplankton could mature to medusae.     

Variations in leaf functional traits and physiological characteristics of <Emphasis Type="Italic">Abies georgei</Emphasis> var. <Emphasis Type="Italic">smithii</Emphasis> along the altitude gradient in the Southeastern Tibetan Plateau

Variations in leaf functional traits of Abies georgei var. smithii at 3700, 3900, 4100, 4300, and 4390 m altitude were investigated in 15 typical plots in the Southeastern Tibetan Plateau. In each plot, three seedlings were selected, of which functional leaves in current-year sunny branches were chosen for the measurement of morphological, photosynthetic, and physiological and biochemical characteristics, and their variations were analyzed. Results showed that significant variations existed among the leaf functional traits of A. georgei var. smithii along the altitudinal gradient, as well as their physiological adaption indicators. Leaf area decreased, while the mass per area and thickness of leaf increased at an altitude above 4,100 m. The maxima of pigment, total nitrogen concentration, net photosynthesis rate during light-saturated, and when water use efficiency appeared at 4100 m altitude. In addition, A. georgei var. smithii seedlings regulated the activities of superoxide dismutase and ascorbate peroxidase to resist abiotic stress under 4100 m altitude. Meanwhile, malondialdehyde concentration and the dark respiration rate rapidly increased, which indicates that A. georgei var. smithii seedlings suffered from heavy abiotic stress from 4100 m to 4390 m altitude. Basing on variations in leaf functional traits along the altitude gradient, we inferred that 4100 m altitude was the suitable region for A. georgei var. smithii growth in the Sygera Mountain. Moreover, the harsh environment was the main limiting factor for A. georgei var. smithii population expansion to high altitude.     

Genotype and Phylogenetic Diversity of <Emphasis Type="Italic">Symbiodinium</Emphasis> ITS2 Sequences Within Clade C in Three Typical Coral Species from Luhuitou Fringing Reef of the South China Sea

Dinoflagellates in the genus Symbiodinium, including nine clades (A–I), mainly form mutualistic symbioses with corals. More than 100 Symbiodinium molecular types have been identified by the ITS2-based genotype method within any given clade, and specifically within Symbiodinium clade C. However, the genotype identification method using the ITS2 sequence is likely to lead to high diversity estimates due to the intra-genomic variations in the ITS2 space; thus, further validation is essential for a correct identification. In this study, the molecular diversity of Symbiodinium ITS2 sequences cloned from two stone corals, Acropora sp. SY-01 and Pocillopora sp. SY-05, and one soft coral, Sarcophyton sp. SY-07, living in the northern part of South China Sea (SCS), were analyzed and compared using the ITS2-based genotype identification method, coupled with ITS2-based secondary structural and phylogenetic analyses. As the result, 12 Symbiodinium ITS2 genotypes were identified, while only six and three Symbiodinium ITS2 genotypes were supported by ITS2-based secondary structural and phylogenetic analyses, respectively. In addition, no shared Symbiodinium ITS2 genotypes were observed among the three coral species, suggesting coral species-dependent Symbiodinium genotypes were within clade C. In summary, the present study provides a theoretical basis for validating the molecular diversity of Symbiodinium ITS2 genotypes in corals.     

Trophic Interaction in a <Emphasis Type="Italic">Portunus rituberculatus</Emphasis> Polyculture Ecosystem Based on Carbon and Nitrogen Stable Isotope Analysis

Trophic interaction among various biomass groups in a swimming crab Portunus trituberculatus polyculture pond was investigated using carbon and nitrogen stable isotope analysis. The polycultured animal species also included white shrimp Litopenaeus vannamei, short-necked clam Ruditapes philippinarum, and redlip mullet Liza haematochila. The mean δ¹³C value for all the biomass groups in polyculture ecosystem ranged from ?25.61‰ to ?16.60‰, and the mean δ¹⁵N value ranged from 6.80‰ to 13.09‰. Significant difference in the δ¹³C value was found between particulate organic matter (POM) and sediment organic matter (SOM) (P < 0.05), indicating that these two organic matter pools have different material sources. Assuming that a ¹³C-enrichment factor of 1.00‰ and a ¹⁵N-enrichment factor of 2.70‰ existed between consumer and prey, diets of the four cultured animals were estimated using a stable isotope mixing model. The estimated model results indicated that P. trituberculatus mainly feed on Aloidis laevis; L. vannamei mainly feed on shrimp feed; while A. laevis, R. philippinarum and L. haematochelia mainly feed on POM. Shrimp feed was also an important food source of R. philippinarum and L. haematochelia. The diets of P. trituberculatus, L. vannamei, R. philippinarum, and L. haematochila showed complementary effects in this polyculture ecosystem. Our finding indicated that the polyculture of these four organisms with suitable farming density could make an effective use of most of the food sources, which can make a highly efficient polyculture ecosystem.     

Fatty acid composition of <Emphasis Type="Italic">Euphausia superba</Emphasis>, <Emphasis Type="Italic">Thysanoessa macrura</Emphasis> and <Emphasis Type="Italic">Euphausia crystallorophias</Emphasis> collected from Prydz Bay,Antarctica

The information of trophic relationship is important for studying the Southern Ocean ecosystems. In this study, three dominant krill species, Euphausia superba, Thysanoessa macrura and Euphausia crystallorophias, were collected from Prydz Bay, Antarctica, during austral summer of 2009/2010. The composition of fatty acids in these species was studied. E. superba and T. macrura showed a similar fatty acid composition which was dominated by C14:0, C16:0, EPA (eicosapentenoic acid) and DHA (decosahexenoic acid) while E. crystallorophias showed higher contents of C18:1(n-9), C18:1(n-7), DHA and EPA than the former two. Higher fatty acid ratios of C18:1(n-9)/18:1(n-7), PUFA (polyunsaturated fatty acid)/SFA (saturated fatty acid), and 18PUFA/16PUFA indicated that E. crystallorophias should be classified as a typical omnivore with a higher trophic position compared with E. superba and T. macrura.     

Development of three multiplex PCR primer sets for ark shell (<Emphasis Type="Italic">Scapharca broughtonii</Emphasis>) and their validation in parentage assignment

Scapharca broughtonii is a commercially important and over-exploited species. In order to investigate its genetic diversity and population structure, 43 novel polymorphic microsatellites were isolated and characterized. The number of alleles per locus ranged from 3 to 22 with an average of 6.93, and the observed and expected heterozygosities varied between 0.233 and 1.000, and 0.250 and 0.953, with an average of 0.614 and 0.707, respectively. Three highly informative multiplex PCRs were developed from nine of those microsatellites for S. broughtonii. We evaluated and validated these multiplex PCRs in 8 full-sib families. The average polymorphism information content (PIC) was 0.539. The frequency of null alleles was estimated as 3.13% of all the alleles segregation based on a within-family analysis of Mendelian segregation patterns. Parentage analysis of real offspring demonstrated that 100% of all offspring were unambiguously allocated to a pair of parents based on 3 multiplex sets. Those 43 microsatellite loci with high variability will be helpful for the analysis of population genetics and conservation of wild stock of S. broughtonii. The 3 sets of multiplex PCRs could be an important tool of pedigree reconstruction, population genetic analysis and brood stock management.     

Effects of different phosphorus concentrations and N/P ratios on the growth and photosynthetic characteristics of <Emphasis Type="Italic">Skeletonema costatum</Emphasis> and <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis>

The effects of different phosphorus (P) concentrations (0.36, 3.6, and 36 μmol/L corresponding to low-, middle-, and high-P concentration groups, respectively) and nitrogen (N)/P ratios on the growth and photosynthetic characteristics of Skeletonema costatum and Prorocentrum donghaiense were studied. For both species, the high-P (HP) concentration group showed the greatest algal density and highest specific growth rate. Changes in the maximum efficiency of photosystem II (F _v /F _m ) were monitored under the various P and N/P conditions. The largest decrease in F _v /F _m was in the low-P (LP) group in S. costatum and in the HP group in P. donghaiense. There were high rapid light curves and photochemical quantum yields (Φ _PSII) for S. costatum in the HP group, while the actual photosynthetic capacity was higher in P. donghaiense than in S. costatum in the MP group. Under eutrophic but relatively P-restricted conditions, P. donghaiense had higher photosynthetic activity and potential, which could cause this dinoflagellate to increasingly dominate the phytoplankton community in these conditions. Under the same P concentration and N/P ratio, P. donghaiense had a larger relative maximum rate of electron transport and higher Φ _{PS II} values than those of S. costatum. These differences between P. donghaiense and S. costatum may explain the interaction and succession patterns of these two species in the Changjiang (Yangtze) River estuary from a photosynthesis perspective.     

Effects of <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> YC459 and soil types on seed germination and seedling growth in rock slope restoration

We carried out experiments with various concentrations of Trichoderma harzianum YC459 in different soil types (forest soil, mixed soil, merchantable soil, and leaf mold soil) to evaluate its effect on seed germination and seedling establishment of four species (Festuca arundinacea Schreb., Dianthus barbatus var. asiaticus Nakai, Lespedeza cyrtobotrya Miq., and Parthenocissus tricuspidata Planch) for rock slope restoration. We also investigated the use of drilled slanted holes on the rock slopes for seedling establishment. The results showed that T. harzianum concentration had significant effects on seed germination, seedling growth, and seedling survival for all the species with different soil types. Seed germination and survival rates peaked at 5% T. harzianum concentration with leaf mold soil and decreased as T. harzianum concentration increased from 5% to 10%. Seedling survival rates of all four species were generally lowest at 0% T. harzianum concentration in all soil types. The height of F. arundinacea and L. cyrtobotrya peaked at 5% T. harzianum concentration whereas that of D. barbatus and P. tricuspidata peaked at 10% T. harzianum concentration. We concluded that 5% T. harzianum concentration with leaf mold soil is appropriate for seed germination and seedling survival rates of most species, thus enhancing seedling establishment. Practical application of the findings of this study will contribute in the vegetation restoration of steep rocks in mountain environments     

Allometric equations of select tree species of the Tibetan Plateau,China

The Tibetan forest is one of the most important national forest zones in China. Despite the potentially important role that Tibetan forest will play in the Earth?s future carbon balance and climate regulation, few allometric equations exist for accurately estimating biomass and carbon budgets of this forest. In the present study, allometric equations,both species-specific and generic, were developed relating component biomass(DW) to diameter at breast height(DBH) and tree height(H) for six most common tree species in Tibetan forest. The 6 species were Abies georgei Orr., Picea spinulosa(Griff.)Henry, Pinus densata Mast., Pinus yunnanensis Franch., Cypresses funebris Endl. and Quercus semecarpifilia Smith.. The results showed that, both DBH-only and DBH2 H based species-specific equations showed a significant fit(P0.05) for all tree species and biomass components. The DBH-only equations explained more than 80% variability of the component biomass and total biomass, adding H as a second independent variable increased the goodness of fit, while incorporating H into the term DBH2 H decreased the goodness of fit. However, not all DBH-H combined equations showed a significant fit(P0.05) for all tree species and biomass components. Hence, the suggested species-specific allometric equations for the six most common tree species are of the form ln(DW) = c + αln(DBH). The generalized equations of mixed coniferous component biomass against DBH, DBH2 H and DBH-H also showed a significant fit(P0.05) for all biomass components. However, due to significant species effect, the relative errors of the estimates were very high. Hence, generalized equations should only be used when there are too many different tree species, or there is no species-specific model of the same species or similar growth form in adjacent area.     

Local records of long-term dynamics of bamboo gregarious flowering in northern Laos and regional synchronicity of <Emphasis Type="Italic">Dendrocalamus membranaceus</Emphasis> in two flowering sites

Bamboos are widely distributed in mainland Southeast Asia and have abundant biomass. They are characterized by prolonged vegetative growth and semelparity. Where bamboos are dominant, their synchronous flowering and death has a major impact on forest vegetation. Although the small-scale dynamics of this process have become clearer in recent years, the history, geographical scale and synchronicity of bamboo flowering over broad areas remains unknown. This study focused on the flowering history of six bamboo species, Bambusa tulda, Cephalostachyum virgatum, Dendrocalamus hamiltonii, Dendrocalamus membranaceus, Indosasa sinica and Oxytenanthera parvifolia, over 40 years across a broad area of northern Laos. We also examined the synchronicity of flowering in D. membranaceus. We visited 49 villages in northern Laos and surveyed knowledgeable inhabitants about bamboo flowering history. The timing, scale and synchronicity of gregarious flowering varied among species. D. hamiltonii and D. membranaceus showed higher flowering synchronicity than other species. All the species except I. sinica had both sporadic and gregarious flowering traits, and showed conspicuous variability in their flowering scale. The flowering bamboo population at two gregarious flowering sites for D. membranaceus was surveyed. While this species had the highest synchronicity in this study, its synchronicity was lower than other species in previous studies worldwide. We found that the gregarious flowering of bamboos in northern Laos over the last 40 years showed lower synchronicity than bamboo flowering reported in other areas of the world. The historical dynamics and scale of bamboo flowering must be further clarified to understand the vegetation composition of this area.     

Effects of a Potential Autochthonous Probiotic <Emphasis Type="Italic">Bacillus subtilis</Emphasis> 2-1 on the Growth and Intestinal Microbiota of Juvenile Sea Cucumber, <Emphasis Type="Italic">Apostichopus japonicus</Emphasis> Selenka

The effects of Bacillus subtilis 2-1 from the intestine of healthy sea cucumber on the growth, digestive enzyme activities and intestinal microbiota of juvenile sea cucumber (Apostichopus japonicus) were determined in the present study. Sea cucumber was fed with Sargassum thunbergii powder supplemented with B. subtilis 2-1 at different concentrations varying among 0 (control), 10⁵, 10⁷, and 10⁹ CFU g^?1 for 8 weeks. Results showed that the growth performance and intestinal amylase and trypsin activities were significantly increased by dietary B. subtilis 2-1 at 10⁹ CFU g^?1 (P < 0.05). However, dietary B. subtilis 2-1 had no significant influence on the lipase activity in sea cucumber (P > 0.05). The polymerase chain reaction denaturing gradient gel electrophoresis and 16S rRNA gene sequencing analysis indicated that dietary B. subtilis 2-1 at 10⁵ and 10⁷ CFU g^?1 inhibited most of the Proteobacteria including those in genus Vibrio. Dietary B. subtilis 2-1 at 10⁹ CFU g^?1 not only decreased the abundance and species of genus Vibrio, but also increased the intensity of genera Psychrobacter and Bacillus. A specific dosage of dietary B. subtilis 2-1 could increase the growth and modulate the intestinal microbiota of sea cucumber; thus it might be a novel probiotic for keeping the health of sea cucumber.     

Immunogenicity and protective role of antigenic regions from five outer membrane proteins of <Emphasis Type="Italic">Flavobacterium columnare</Emphasis> in grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.     

Inbreeding depression on growth and survival of full-sib family of Manila clam (<Emphasis Type="Italic">Ruditapes philippinarum</Emphasis>)

In present study, the inbreeding depression (ID) of growth and survival of Manila clam (Ruditapes philippinarum) was investigated at larval and juvenile stages. Nine inbred families (A ₂, B ₂, C ₂, D ₂, E ₂, F ₂, G ₂, H ₂ and I ₂) were established by mating within nine full-sib families with expected inbreeding coefficient of 0.25. Inbred families showed significant differences in shell length and hatching rate of D-larvae (straight-hinged larvae). The larvae of the nine inbred families grew slower than those of control group (CG), and their ID value ranged from 0.81% ± 6.09% to 16.10% ± 1.49%. The ID value of larval survival rate varied between 27.47% ± 9.36% and 70.50% ± 13.66%. The ID was also detected for juvenile growth in A ₂, B ₂, C ₂, and D ₂, which ranged from 4.60 ± 2.21 to 17.71 ± 7.73. The A ₂ family maintained the highest juvenile survival rate, whereas the other inbred families exhibited ID values varying between 62.79% ± 4.54% and 96.14% ± 0.87%. The linear relationship of estimated ID between growth and survival was negatively correlated (R = ?0.434, P < 0.05). The results of this study suggested that the ID of growth was common at the larval stage but was less prevalent at juvenile stage. In contrast, the ID of survival increased from larval to juvenile stage. A better understanding of the effect of inbreeding may aid to selective breeding of Manila clam.     

Screening and isolation of the algicidal compounds from marine green alga <Emphasis Type="Italic">Ulva intestinalis</Emphasis>

Twenty species of seaweed were collected from the coast of Zhejiang, China, extracted with ethanol, and screened for algicidal activity against red tide microalgae Heterosigma akashiwo and Prorocentrum micans. Inhibitory effects of fresh and dried tißsues of green alga Ulva intestinalis were assessed and the main algicidal compounds were isolated, purified, and identified. Five seaweed species, U. intestinalis, U. fasciata, Grateloupia romosissima, Chondria crassicaulis, and Gracilariopsis lemaneiformis, were investigated for their algicidal activities. Fresh tissues of 8.0 and 16.0 mg/mL of U. intestinalis dissolved in media significantly inhibited growth of H. akashiwo and P. micans, respectively. Dried tissue and ethyl acetate (EtOAc) extracts of U. intestinalis at greater than 1.2 and 0.04 mg/mL, respectively, were fatal to H. akashiwo, while its water and EtOAc extracts in excess of 0.96 and 0.32 mg/mL, respectively, were lethal to P. micans. Three algicidal compounds in the EtOAc extracts were identified as 15-ethoxy-(6z,9z,12z)-hexadecatrienoic acid (I), (6E,9E,12E)-(2-acetoxy-β-D-glucose)-octadecatrienoic acid ester (II) and hexadecanoic acid (III). Of these, compound II displayed the most potent algicidal activity with IC₅₀ values of 4.9 and 14.1 µg/mL for H. akashiwo and P. micans, respectively. Compound I showed moderate algicidal activity with IC₅₀ values of 13.4 and 24.7 µg/mL for H. akashiwo and P. micans, respectively. These findings suggested that certain macroalgae or products therefrom could be used as effective biological control agents against red tide algae.     

The mycobiota of discoloured needles of the Swiss stone pine (<Emphasis Type="Italic">Pinus cembra</Emphasis> L.) in the Tatra Mountains (Poland)

The Swiss stone Pine (Pinus cembra L.) is an alpine species, fairly commonly occurring in the Alps and the Carpathians, close to the timberline. Natural sites of the Swiss stone pine in Poland are found exclusively in the Tatra Mountains, within the area of the Tatra National Park (TNP). In 2017, the health status of the needles of P. cembra was determined and their mycological analysis was carried out. As a result, 11 species of fungi were isolated from the needles showing disease symptoms, manifested by various shades of discoloration. The most frequently observed species was Fusarium oxysporum. It probably does not bring on disease symptoms, but may affect the development of other frequently isolated species (such as Sydowia polyspora, Lophodermium sp. and Lophodermium conigenum) that are pathogenic to Pinus spp.     

Effects of dietary genistein on GH/IGF-I axis of Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis>

There is considerable concern that isoflavones, such as genistein in fish feed composed of soybean protein, aff ects somatic growth in fish. Our previous works demonstrated that 30 and 300 μg/g dietary genistein had no significant eff ect on growth performance in Nile tilapia (Oreochromis niloticus), but the higher level of genistein (3 000 μg/g) significantly depressed growth. This study was conducted to further examine the eff ects of dietary genistein on the endocrine disruption on growth hormone/insulin-like growth factor-I (GH/IGF-I) axis in Nile tilapia (O. niloticus). Juvenile fish were fed by hand twice daily to satiation with one of four isonitrogenous and isoenergetic diets, each containing either 0, 30, 300 or 3 000 μg/g genistein. Following an 8-week feeding period, plasma GH and IGF-I levels were investigated by radioimmunoassay and gene expression levels of gh, ghrelin, gnrhs, ghr, npy, npyrs, pacap, ghrs, igf-I, igf-Ir, and igfbp3 were examined by real-time PCR. The results show that no significant change in plasma GH and IGF-I levels in fish fed with diets containing 30 μg/g and 300 μg/g genistein. mRNA expression of genes along the GH/IGF-I axis remained unaff ected, except for igf-Ir, which was stimulated by the 300 μg/g genistein diet. While in fish fed the 3 000 μg/g genistein diet, the plasma GH and IGF-I levels decreased, and mRNA expression of gh, ghr2, npyr1, igf-I, and igf-Ir were also significantly depressed. In contrast, npy and igfbp3 mRNA expression were enhanced. This study provides convincing evidence for growth impediment by genistein by disturbing the GH/IGF-I axis in Nile tilapia O. niloticus.