A genetic and ultrastructural study of three clones of Porphyridium purpureum (Bory de Saint-Vincent, 1797) Drew et Ross, 1965 (Rhodophyta) from the marine microalgae collection of the Zhirmunsky institute of marine biology

In this paper, we present the results of a comparative genetic and ultrastructural study of three clones of the microalga Porphyridium purpureum (Rhodophyta) from the culture collection of marine microalgae of the Zhirmunsky Institute of Marine Biology. All clones, which have different geographical origins, showed a high similarity in terms of the ultramicroscopic structure and the nucleotide sequences of the nuclear ribosomal DNA genes (18S rDNA, ITS1-5.8S rDNA-ITS2, D1-D2 region of 28S rDNA). The obtained data are very helpful for the certification of two strains of P. purpureum that were isolated for the first time in the practice of Russian algological research.     

Feeding Ecology of Padogobius nigricans (Canestrini, 1867) and P. bonelli (Bonaparte, 1846) in Aggia River (Umbria,Italy) and Their Diet Overlap

The Arno goby Padogobius nigricans is an endemic species to Central Italy. The populations of this species are in sharp decline due to habitat modifications and potentially competition with Padanian goby Padogobius bonelli, an endemic species to Northern Italy, introduced in some watercourses of Central Italy. The purposes of this paper were to analyze dietary composition of the two species, determine the degree of overlap in their diets and investigate the feeding ecology of P. nigricans in terms of absence and presence of the alien species. Specimens were collected from the Aggia River, a tributary of Tiber River. Gut contents were analyzed and prey items were identified to the lowest recognizable taxa and counted. All the obtained results showed that P. nigricans and P. bonelli in the Aggia River share a similar diet. Both species fed extensively on Chironomids and no statistically significant ontogenetic changes in feeding habits were found. A high degree of diet overlap was observed. Diet of P. nigricans and P. bonelli also included fish and where the species live in syntopy, individuals belonging to Padogobius genus were found in stomach, suggesting that mutual predation may occur.     

Effects of salt stress on nitrogenase activity and growth of four legumes

The effects of salt stress on nitrogenase (N2-ase) activity, growth and nitrogen content ofVicia faba (L.),Medicago sativa (L.) Merrill,Glycine max andVigna sinensis (L.) were investigated. Four levels of salinity were applied and salt treatments were imposed on inoculated and N-fertilized plants.M. sativa tolerated mild levels of salinity but higher salt concentrations depressed N₂-ase activity of this species. The other three legumes were considerably affected by salt treatments, and N₂-ase activity was significantly reduced by salinity. Vicia faba, carrying elongate nodules, could restore a partial N₂-ase activity upon recovery from salt stress whereasG. max andV. sinesis, both with spherical nodules, could not regain significant activity when salinity was removed. Salt stress retarded growth of both inoculated and N-fertilized plants. The nitrogen content of both treatments was also affected by salinity and the effect was more severe for inoculated than N-fertilized plants.     

Regulation of sodium in the shore crabCarcinus maenas,adapted to environments of constant and changing salinities

The activity of Na-K-ATPase was determined in the posterior gills of the shore crabCarcinus maenas during a period following transfer from 35 to 10 ‰ salinity and vice versa at 15 °C. After transfer from high to low salinity, Na-K-ATPase activity increased from 3.2 to 7.0 μmoles P_i mg protein^?1 h^?1 within a period of 2 to 3 weeks. Transfer of crabs from low to high salinity resulted in reduction of activity from 7.4 to 4.5 μmoles P_i mg protein^?1 h^?1 within about the same period. The relatively slow response following salinity change indicates that the amounts of Na-K-ATPase in the gills may play a role in hyperionic Na regulation in relatively constant brackish-water environments. Instant responses to salinity result from activation and inhibition of Na-K-ATPase activity by Na. Gill Na-K-ATPase is activated by the Na concentration of the incubation medium to attain a steep maximum at about 75 mM Na, which corresponds to the lowest environmental Na levels tolerated byC. maenas equivalent to a salinity of ca 6 ‰. Activity greatly decreased towards higher Na levels, equivalent to the salinity of normal sea water, at which hyperregulation no longer occurs. Selective addition of either Na or Cl to brackish water of 9 ‰ S resulted in effective hyperregulation of the non-increased ion, and passive distribution between medium and blood of the increased ion. These data indicate that under appropriate conditions the normally coupled transport of Na and Cl may be uncoupled and take place independently of each other.     

Picophytoplankton predominance in hypersaline lakes (Transylvanian Basin,Romania)

The occurrence and importance of photoautotrophic picoplankton (PPP, cells with a diameter <2 μm) was studied along a trophic and salinity gradient in hypersaline lakes of the Transylvanian Basin (Romania). The studied lakes were found to be rich in PPP, with abundances (maximum 7.6 × 10⁶ cells mL^?1) higher than in freshwater and marine environments of similar trophic conditions. The contribution of PPP to the total phytoplankton biovolume did not decrease with increasing trophic state as it was generally found in other aquatic environments. Regardless of the trophic conditions, the contribution of PPP could reach 90–100 % in these hypersaline lakes. We hypothesized that the PPP predominance might be the result of the low grazing pressure, since heterotrophic nanoflagellates (the main grazers of PPP) were absent in the studied samples. There were significant differences in community composition among the lakes along the salinity gradient. CyPPP predominated in less saline waters (mainly below 5 %), while EuPPP were present along the entire salinity range (up to 18.7 %), dominating the phytoplankton between 3 and 13 % salinity. Above 13 % salinity, the phytoplankton was composed mainly of Dunaliella species.     

Freshwater Crustacea (Cladocera,Copepoda) of Iceland: taxonomy,ecology, and biogeography

Of the many types of freshwater bodies across Iceland (e.g., lakes, springs, rivers, and ponds), greater effort has been invested in researching the freshwater Crustacea (Cladocera, Copepoda) of more permanent, larger, and deeper water bodies than on smaller or more ephemeral sites. To address this, we described the fauna of 12 shallow freshwater bodies distributed from the coast to the central highlands of Iceland. We identified 16 species in 11 genera belonging to six families of Cladocera, and 12 species in eight genera belonging to four families of Copepoda. Five of the species we identified (Alona werestschagini, Cyclops vicinus, Daphnia cristata, Diacyclops abyssicola, and Heterocope borealis) represent new distribution records for Iceland, increasing the number of species now known from this region to 34 Cladocera and 38 Copepoda taxa. Additionally, we discuss the taxonomy of some species with doubtful taxonomical status. Six crustacean taxocenes are described, each characterized by the dominance of different taxa, with bottom sediment characteristics best explaining species assemblage structure. We evaluated the relationships between Icelandic Cladocera and Copepoda faunas and those of the adjacent Svalbard, Faroe, and Shetland Islands, Greenland, and the mainland Norway. The taxonomic composition of the Icelandic fauna is dominated by widely distributed Holarctic, and secondarily Palaearctic and Nearctic species, and is more similar to the faunas of the major North Atlantic islands (64.2–71.8 % similarity) than it is to that of continental Norway.     

Transcription and processing of rRNA in higher plant cells (Daucus carota,Petroselinum crispum,Acer pseudoplatanus)

Actively dividing cells from parsley (Petroselinum crispum) and carrot (Daucus carota) (bothApiaceae) andAcer pseudoplatanus (Aceraceae) were used to detect the primary gene product of the rDNA in plant cells. Parsley and carrot cells were labelled with [³²P] orthophosphate. In both cases only one high molecular weight rRNA precursor was present on polyacrylamide gels under non-denaturing conditions. Its molecular weight did not exceed 2.5 × 10⁶ daltons. The component emerged from the heterogenous material after a labelling period of 5–10 min. In parsley cells 45 min after onset of incubation labelled mature rRNA (25S and 18S) arrived in the cytoplasm. InAcer pseudoplatanus (incubation period 60 min) two rapidly labelled components did emerge from polyacrylamide gels; their molecular weights were 2.3 and 3.2^? 3.4 × 10⁶ daltons. After electrophoresis under denaturing conditions the larger component was no longer present, thus indicating that it was an aggregate of different RNA molecules. The molecular weights of the rRNA precursors ofD. carota andP. crispum determined after electrophoresis in formamide gels were about 2.1 × 10⁶ daltons. From these results we have no evidence for the existence of rRNA precursors exceeding the molecular weight of 2.5 × 10⁶ daltons.     

A Laboratory Case Study of Efficient Polyhydoxyalkonates Production by Bacillus cereus,a Contaminant in Saccharophagus degradans ATCC 43961 in Minimal Sea Salt Media

A contaminating bacterium growing along with the stock culture of Saccharophagus degradans ATCC 43961 (Sde 2-40) on marine agar plate was isolated and investigated for its ability to produce polyhydoxyalkonates (PHA). Preliminary screening by Sudan black B and Nile blue A staining indicated positive characteristic of the isolate to produce PHA. The isolate was able to grow and produce PHA in minimal sea salt medium broth. PHA quantification studies with gas chromatographic analyses of the dry cells derived from culture broths revealed accumulation of PHA in bacterial cells. PHA production started after 20 h and increased with cell growth and attained maximum values of 61 % of dry cell weight at 70 h of cultivation. After 70 h, a slight decrease in the level of PHA content was observed. The nature/type of PHA was found to be poly(3-hydroxybutyraye) by Fourier transform-infrared spectroscopy. Microbiological and 16S rRNA gene sequencing analyses suggested that the PHA producing bacterial isolate belongs to Bacillus genera and shows 100 % nucleotide sequence similarity with Bacillus cereus species in GenBank. This study is a first report for ability of Bacillus species to grow in marine sea salt media and produce PHA. The media used for the polymer production was novel in the context of the genus Bacillus and the production of PHA was three-fold higher than Sde 2-40 using same growth medium. This study shows that the contaminant bacteria once properly investigated can be used for advantageous characteristic of metabolites production in place of original cultures.     

Molecular cloning,characterisation and expression of the translationally controlled tumor protein gene in rock bream (Oplegnathus fasciatus)

Translationally controlled tumor protein (TCTP) is an important immune regulator that has been implicated in a number of cellular processes, including cell growth, cell cycle progression, apoptosis regulation and protection of cells against various environmental stresses. In this study, we cloned and characterised TCTP from rock bream (Oplegnathus fasciatus), which is an economically important species in the Korean aquaculture industry. The full-length rock bream TCTP (RbTCTP) cDNA was of 1,041 bp and contained an open reading frame of 513 bp, which encoded 170 amino acids. The 5′ untranslated region (UTR) was 90 bp, while the 3′ UTR was 438 bp, containing a polyadenylation signal. RbTCTP showed 76, 75 and 74 % amino acid sequence identities to those of tilapia (Oreochromis niloticus), orange-spotted grouper (Epinephelus coioides) and Japanese sea perch (Lateolabrax japonicus), respectively. The positions of microtubule binding region, Ca⁺ binding region and TCTP signature regions in RbTCTP were similar to other fish species and mammals. RbTCTP mRNA expression level was highest in the gill compared to other tissues. The level of RbTCTP mRNA expression was significantly regulated by injection of red seabream iridovirus, Streptococcus iniae and Edwardsiella tarda.     

Predicted effects of increasing salinity on the crustacean zooplankton community of Pyramid Lake,Nevada

Since 1933 the salinity of Pyramid Lake, Nevada, U.S.A., has increased 32% to nearly 5.5‰. We tested the hypothesis that further increases of 1.5 to 2 times (1.5× to 2×) its present salinity would significantly reduce species richness and alter population structures of the existing crustacean zooplankton community. Three strategies were applied: in addition to monitoring zooplankton in semicontrolled indoor microcosms at 1×, 1.5× and 2× and conducting range-finding, acute, and chronic salinity bioassays, the present zooplankton community of Walker Lake (2×) was compared with that existing in Pyramid Lake (1×). Ceriodaphnia quadrangula and Diaphanosoma leuchtenbergianum, both collected from Pyramid Lake, were lacking in Walker Lake. Populations of Cyclops vernalis were significantly lower and those of Diaptomus sicilis and Moina hutchinsoni were significantly higher in Walker Lake than in Pyramid Lake. Densities of Ceriodaphnia and Cyclops were low in microcosms at salinities > 1×. Diaphanosoma could not be maintained in microcosms, regardless of salinity. Numbers of Diaptomus and Moina in microcosms were proportional to salinity level. Short-term LC50 salinities (‰) were as follows: Diaphanosoma, 6.5; Ceriodaphnia, 7.1; Diaptomus, 13.3; Cyclops, 14.8; and Moina, 17.8. Multiple-generation, chronic bioassays were run only on Cyclops and Diaptomus. Three generations of Cyclops were produced at salinities of 4.0 to 8.5‰, but not at 9.8‰ or higher. Diaptomus was unable to complete three generations at salinities ?9.6‰. We speculate that high salinity in Walker Lake may indirectly benefit Diaptomus by negatively affecting predatory Cyclops, and benefit Moina by causing extinction of competing salinity-intolerant Diaphanosoma and Ceriodaphnia. Except for the response of Diaptomus, results from bioassays were in general agreement with results from microcosms and with field data. Untested predator-prey interactions could be responsible for the apparent discrepancy.     

Feral pigs as hosts for Amblyomma sculptum (Acari: Ixodidae) populations in the Pantanal,Mato Grosso do Sul,Brazil

The Pantanal in Brazil is the largest floodplain of the world. This ecosystem, rich in wildlife, has a large feral pig population. Such a large host biomass must have a strong influence on the parasite fauna. In this work, we evaluated the role of feral pigs in the maintenance of Amblyomma sculptum (formerly Amblyomma cajennense), the most prevalent tick species in the Pantanal. Tick infestations were evaluated on 243 feral pigs and their environment. The suitability of domestic pigs, representing their feral relatives, to A. sculptum adults and nymphs was assessed experimentally. Tick infestation of feral pigs was strongly associated with that of the environment: 96 and 97 % of the ticks, respectively, were A. sculptum. The infestation prevalence on this host species was close to 90 % in the dry season and 100 % in the wet season and mean infestation intensity was above 30 ticks in both seasons. Suitability of pigs as hosts for A. sculptum was shown by the high proportion of nymphs and female ticks found engorging on captured feral pigs and adequate biological parameters displayed by ticks from experimental infestations of domestic pigs. Other tick species on feral pigs, albeit in much lower numbers, were Amblyomma parvum and Ornithodorus rostratus. Results show that feral pigs feed a high proportion of the A. sculptum adults and nymphs in their territories and should be a target for tick-borne diseases studies. This is particularly relevant to public health because all the main tick species found on feral pigs are aggressive to humans as well.     

Cytotaxonomy ofCalliphoridae (Diptera)

A modification of the general system of classification ofCalliphoridae is proposed in this report. Information about the chromosome complements of about 90 species is provided including references to the earlier literature on chromosomes of this family. With one exception all species studied by us or by earlier investigators have 2n=12. The total complement length of 556 complements from 83 species averaged 66.4 μ. Most species have a short heteromorphic sex pair but the size and morphology vary considerably and one species,Hemipyrellia fernandica, seems to have a quadrivalent sex-chromosome complex in a 2n=14 complement. The autosomal pairs vary in both length and arm ratios between the species but seem to be more stable than the sex chromosomes. Only minor variations were found between different collections of the same species except for two collections ofCalliphora croceipalpis. Although presently available data on the chromosomes of Diptera species suggest that increases in total complement length accompany evolutionary progress, no such general trends were obvious in either the total complement lengths or chromosome morphology, either within or between the tribes ofCalliphoridae. Thus the karyotypical reorganizations in the family have apparently not been directly associated with changes in the general morphological features of the adults which may have responded in these respects more extensively to genetic mutations.     

High salinity induced expression profiling of differentially expressed genes in shrimp (Penaeus monodon)

Four suppression subtractive hybridization (SSH) cDNA libraries were constructed to identify differentially expressed salinity stress responsive genes of black tiger shrimp, Penaeus monodon exposed to high (55 ppt) salinity conditions. One each of the forward and reverse SSH cDNA libraries were developed from the gill and gut tissues of shrimp and clones having inserts larger than 300 bp were unidirectionally sequenced. Based on the sequence homology search, the identified genes were categorized for their putative functions related to a wide range of biological roles, such as nucleic acid regulation and replication, immune response, energy and metabolism, signal transduction, cellular process, structural and membrane proteins, stress and osmoregulation. Gene expression levels in response to high salinity conditions at 2 weeks post salinity stress for some of the differentially expressed genes (Na⁺/K⁺-ATPase α-subunit, glutathione peroxidase, intracellular fatty acid binding protein, elongation factor 2, 14-3-3 like protein, penaeidin, translationally controlled tumor protein, transglutaminase and serine proteinase inhibitor B3) identified from SSH cDNA libraries were analysed by real-time RT-PCR. The highest gene expression levels was observed for Na⁺/K⁺-ATPase α-subunit in gill tissues (15.23-folds) and antennal glands (12.01-folds) and intracellular fatty acid binding protein in gut tissues (14.05-folds) respectively. The differential and significant levels of gene expression indicate the functional role of these genes in shrimp salinity stress adaptive mechanisms.     

Spittlebugs (Hemiptera: Cercopidae) and their host plants: a strategy for pasture diversification

Information about the biology of spittlebugs on different species of forage is necessary for the development of tactics for their control, and is also essential for evaluating, selecting and launching new forage cultivars. The aims of this study were to compare the nutritional suitability of different forage species–Brachiaria ruziziensis (Congo grass), Pennisetum purpureum (elephant grass), Panicum maximum (Guinea grass), and Cynodon dactylon (Bermuda grass)–for the spittlebug species Mahanarva spectabilis (Distant) (Hemiptera: Cercopidae), Deois schach (Fabricius) (Hemiptera: Cercopidae) and Notozulia entreriana (Berg) (Hemiptera: Cercopidae), and to evaluate the performance of each spittlebug species on forage monocultures. B. ruziziensis was the most suitable host plant for D. schach. Since N. entreriana did not show differences in the evaluated parameters, it was not possible to determine its ideal forage species. For M. spectabilis, P. purpureum was the most suitable forage species. The evaluation of the performance of the different spittlebugs on a single forage species also showed which spittlebugs are a problem. M. spectabilis and D. schach developed better on a monoculture of B. ruziziensis. M. spectabilis was a major problem when it fed on P. purpureum, but on C. dactylon and P. maximum, none of the spittlebugs developed properly, indicating that these plants are less suitable for them. These results suggest that the forage diversification of pastures may be a strategy for the integrated management of spittlebugs.     

Molecular evidence to reconcile taxonomic instability in mahseer species (Pisces: Cyprinidae) of India

The mahseers are an important group of fishes endemic to Asia with most species considered threatened. Conservation plans to save declining wild populations are hindered by unstable taxonomy, and detailed systematic review could form a solid platform for future management and conservation. D-loop and cytochrome c oxidase I (COI) mtDNA sequences were examined in nine mahseer species of Tor, Neolissochilus, and Naziritor. Pseudogenes amplified in a portion of the species limited the utility of the D-loop region. ABGD analysis, NJ, ML, and MP methods and genetic distance (TrN?+?I?+?G) using COI data revealed concordant species delimiting patterns. The three genera were monophyletic, separated as distinct clades (TrN?+?I?+?G 0.064 to 0.106), and Naziritor was flagged as a separate genus, distinct from Puntius (TrN?+?I?+?G 0.196). Out of seven nominal species known for Tor cogeners from India, only five were recovered with mtDNA data (TrN?+?I?+?G 0.000 to 0.037) and two species could not be distinguished with the molecular data set employed. Tor mosal, synonymized as Tor putitora, was rediscovered as a distinct species (TrN?+?I?+?G 0.031) based on its type locality. Tor mussulah was confirmed as a separate species (TrN?+?I?+?G 0.019 to 0.026). Two valid species, Tor macrolepis and T. mosal mahanadicus, were not distinct from T. putitora (TrN?+?I?+?G 0.00). The high divergence with mtDNA data failed to validate T. mosal mahanadicus as a subspecies of T. mosal (TrN?+?I?+?G 0.031). Morphological outliers discovered within the distribution range of Tor tor (TrN?+?I?+?G 0.022 to 0.025) shared the same lineage with T. putitora (TrN?+?I?+?G 0.002 to 0.005), indicating a new extended distribution of the Himalayan mahseer T. putitora in the rivers of the Indian central plateau. The findings indicate the need for integrating molecular and morphological tools for taxonomic revision of the Tor and Naziritor genera, so that taxa are precisely defined for accurate in situ and ex situ conservation decisions.