Partial 16S rDNA Sequencing Research Articles

Description of Lelenchus brevislitus n. sp. (Nematoda: Tylenchidae), an example of a cryptic species from Iran and its phylogenetic relationships with other species in the family

Lelenchus brevislitusn. sp. is described and illustrated based on morphological, morphometric and molecular data. The new species is characterised by having females with slender, 452-488 μm long, body lateral field vestigial as a plain simple band under scanning electron microscopy (SEM) in vulval region and absent in proximal and distal body ends, cephalic region short,ca2 μm high, conical and truncate in anterior end in lateral view under SEM, dorso-ventrally flattened, amphidial pouches pocket-shaped and large under light microscopy (LM), their apertures short (ca1 μm long in SEM) longitudinal sinuous slits remarkably wide near the small oral disc and significantly narrowing posteriad stylet, slender. 6.5-8.0 μm long, developed median bulb with distinct valvular plates, monodelphic-prodelphic reproductive system having a short post-vulval uterine sac (PUS), transverse vulval slit (V = 47.5-51.7) bearing small lateral membranes on each side, long filiform female tail (154-170 μm, c = 2.8-3.1, c′ = 21.5-28.3), and males unknown. The new species is considered to be a cryptic species inside the Tylenchidae. Morphological comparisons with all species of the genus are discussed. Phylogenetic studies were performed for further characterisation of the new species, and the results, using nearly full length 18S rDNA data, revealed it formed a clade with an isolate ofL. leptosoma, both of which are in a sister clade with an isolate ofSakia. In a reconstructed phylogenetic tree using partial 28S rDNA sequences, the new species formed a monophyletic group, again withL. leptosoma, a sister clade to someMalenchusspp.

Description of three species of Isorchis (Digenea: Atractotrematidae) from Australia.

Three species of Isorchis Durio and Manter, 1969 are described from Australian waters. Isorchis megas sp. nov. is described from the spotbanded scat, Selenotoca multifasciata (Richardson), off Western Australia (WA) and Northern Territory (NT); Isorchis currani sp. nov. is described from S. multifasciata off NT; and Isorchis anomalus sp. nov. is described from the milkfish, Chanos chanos Forsskål, off WA. Isorchis megas sp. nov. can be differentiated from the other species of Isorchis by possessing a single, large egg that is greater than 20% of the body length; having a shorter body (the largest specimen is less than 500 μm); and utilizing a scatophagid rather than a chanid host. Isorchis currani sp. nov. can be differentiated from species of Isorchis other than I. megas sp. nov. by utilizing a scatophagid rather than a chanid host; it is differentiated from I. megas sp. nov. in having eggs that are 11-15% of the body length. Isorchis anomalus sp. nov. can be differentiated from all other species of Isorchis in possessing an irregular shaped genital pore rather than one that is circular to oblong. A Bayesian inference analysis of partial 28S rDNA sequences of the three new species of Isorchis and 30 other haploporoids revealed 1) the monophyly of the Atractotrematidae Yamaguti, 1939, 2) the two species of Isorchis infecting S. multifasciata were each other's closest relative, and 3) that Isorchis was most closely related to Pseudomegasolena Machida and Komiya, 1976 rather than Atractotrema Goto and Ozaki, 1929 although sequence data are not yet available for a member of Pseudisorchis Ahmad, 1985.

<i>Ex Situ</i> Biodegradation of Crude Oil Using Bacterial Isolates From Palm Oil Mill Effluent

Bioremediation of crude oil is quite unique, complex and relatively a slow process. They study area for this research work was a cottage industry located in Orumba North local government area of Anambra state, Nigeria. Evaluating the degradation potential of bacterial isolates from palm oil mill effluent on crude oil was conducted using standard methods, which includes screening, inoculums development and degradation studies to assess the most promising hydrocarbon degraders. Pooled samples collected from different points on the palm oil mill effluent disposal channels were inoculated mineral salt medium containing crude oil to assess degradation abilities of different bacterial species. Data indicated that these bacterial isolates were capable of thriving in the mineral salt medium between the pH range of 6.0-7.0 and bacterial proliferation were quantified by assessing the total viable count which ranged between 2 x 105-9 x 105 cfu/ml based on individual abilities to utilize crude oil as a sole carbon source. Cultural, morphological and biochemical characterizations were conducted on these isolates and Bergey’s manual of determinative bacteriology was used to suggest possible isolates, while molecular identification was done Centre for Agriculture and biosciences International (CABI), UK using partial 16S rDNA sequencing analysis and FASTA algorithm with prokaryote database from EBI. Escherichia fergosonii, Klebsiella variicola and Microccocus luteus were identified. Gas chromatographic analysis was applied to quantify the extent of degradation of crude oil by the isolates. All the isolated bacterial species displayed varying degree of crude hydrocarbon biodegradation.

Morphology and molecular analysis of life-cycle stages of Proctoeces maculatus (Looss, 1901) (Digenea: Fellodistomidae) in the Bizerte Lagoon, Tunisia.

The life cycle of Proctoeces maculatus (Looss, 1901) (Digenea, Fellodistomidae) was studied in Bizerte Lagoon (Tunisia). Three sequential hosts appear to be involved: the Mediterranean mussel Mytilus galloprovincialis Lamarck, 1819 (Mytilidae) as the first intermediate host; the polychaete Sabella pavonina Savigny, 1822 (Sabellidae), as the second intermediate host; and fishes (Lithognathus mormyrus (Linnaeus, 1758) (Sparidae), Trachinotus ovatus (Linnaeus, 1758) (Carangidae) and Sparus aurata Linnaeus, 1758 (Sparidae) as the definitive hosts. It should be noted that S. pavonina was recorded as second intermediate host for P. maculatus for the first time. Molecular confirmation of the morphological identification of the life-cycle stages of this digenean was obtained using partial 28S rDNA sequences. Comparative sequences revealed that the sporocysts and the metacercariae are conspecific but they diverged by 0.3% from the adults. The present results raised the possibility of the existence of cryptic species within the different developmental stages. However, all the present isolates differed from material from Archosargus probatocephalus in the Gulf of Mexico identified as P. maculatus.

Identification and characterization of psychrotolerant coliform bacteria isolated from pasteurized fluid milk

The presence of coliform bacteria in pasteurized fluid milk typically indicates that product contamination occurred downstream of the pasteurizer, but it may also indicate pasteurization failure. Although coliform detection is frequently used as a hygiene indicator for dairy products, our understanding of the taxonomic and phenotypic coliform diversity associated with dairy products is surprisingly limited. Therefore, using Petrifilm Coliform Count plates (3M, St. Paul, MN), we isolated coliforms from high-temperature, short-time (HTST)-pasteurized fluid milk samples from 21 fluid milk processing plants in the northeast United States. Based on source information and initial characterization using partial 16S rDNA sequencing, 240 nonredundant isolates were obtained. The majority of these isolates were identified as belonging to the genera Enterobacter (42% of isolates), Hafnia (13%), Citrobacter (12%), Serratia (10%), and Raoultella (9%); additional isolates were classified into the genera Buttiauxella, Cedecea, Kluyvera, Leclercia, Pantoea, and Rahnella. A subset of 104 representative isolates was subsequently characterized phenotypically. Cold growth analysis in skim milk broth showed that all isolates displayed at least a 2-log increase over 10 d at 6°C; the majority of isolates (n=74) displayed more than a 5-log increase. In total, 43% of the representative isolates displayed lipolysis when incubated on spirit blue agar at 6°C for 14 d, whereas 71% of isolates displayed proteolysis when incubated on skim milk agar at 6°C for 14 d. Our data indicate that a considerable diversity of coliforms is found in HTST-pasteurized fluid milk and that a considerable proportion of these coliforms have phenotypic characteristics that will allow them to cause fluid milk spoilage.

Data on some members of the family Tylenchidae (Nematoda: Tylenchina) from Iran

AbstractThree species belonging to the family Tylenchidae, Malenchus nanellus, M. undulatus and Tylenchus naranensis, are reported for the first time for Iran. These species are characterized based on morphological and morphometric data. The Iranian population of M. nanellus is characterized by its body length ranging from 410-485 μm, cuticle annuli 1.1- 1.5 μm wide at mid-body and lateral field with two crenate lines, starting at the mid-region of procorpus and ending near 1/3 of tail length. The population of M. undulatus is characterized by its 458-526 μm body length, cuticle coarsely annulated, annuli 1.8-2.4 μm wide at mid-body, lateral field with crenate incisures, beginning at about half of the stylet length, ending at middle of tail, head narrower than adjacent body, median bulb well developed with prominent valve plates and functional males in population. Iranian population of T. naranensis, is characterized by having a 631-774 μm body length, lateral field with four lines, outer lines crenate, a stylet length ranging from 10-11 μm and a tail of 102-131 μm long. Molecular phylogenetic analyses based on partial 28S rDNA sequences of T. naranensis revealed its close affinity with the genus Filenchus. Other Tylenchidae genera, such as Aglenchus (including the newly sequenced isolate from Iran) and Coslenchus were sister taxa and formed a well-supported clade. Malenchus exiguus, a previously reported species from Iran and sequenced in the present study, formed a monophyletic clade with other species of Malenchus and Lelenchus leptosoma.

Impact of Zinc Orthophosphate on Simulated Drinking Water Biofilms Influenced by Lead and Copper

AbstractPhosphate, a key nutrient for bacterial growth, is also a key component of many corrosion-control programs to manage lead and copper corrosion in premise plumbing. Bench-scale stagnant water galvanic macrocells with lead and copper components were fed with drinking water containing three levels of zinc orthophosphate [0 (control), 1, and 3 mg l−1-PO4]. Suspended polycarbonate coupons, representing benign downstream fixtures, were placed in the macrocells, thus enabling biofilm formation on this material. Community profiling using denaturing gradient gel electrophoresis (16S rDNA PCR-DGGE) revealed that phosphate dose (primarily) and metal type (to a lesser extent) influenced biofilm community diversity. Generally, community diversity increased with increasing heterotrophic plate counts that in turn rose in response to elevated phosphate. Partial 16s rDNA sequences obtained from DGGE gel bands identified the dominant bacterial taxa as the phyla Verrumicrobia, Firmicutes, Bacteroidetes, and α-Prote...

The systematics of carnivorous sponges.

Carnivorous sponges are characterized by their unique method of capturing mesoplanktonic prey coupled with the complete or partial reduction of the aquiferous system characteristic of the phylum Porifera. Current systematics place the vast majority of carnivorous sponges within Cladorhizidae, with certain species assigned to Guitarridae and Esperiopsidae. Morphological characters have not been able to show whether this classification is evolutionary accurate, and whether carnivory has evolved once or in several lineages. In the present paper we present the first comprehensive molecular phylogeny of the carnivorous sponges, interpret these results in conjunction with morphological characters, and propose a revised classification of the group. Molecular phylogenies were inferred using 18S rDNA and a combined dataset of partial 28S rDNA, COI and ALG11 sequences. The results recovered carnivorous sponges as a clade closely related to the families Mycalidae and Guitarridae, showing family Cladorhizidae to be monophyletic and also including carnivorous species currently placed in other families. The genus Lycopodina is resurrected for species currently placed in the paraphyletic subgenus Asbestopluma (Asbestopluma) featuring forceps spicules and lacking sigmas or sigmancistras. The genera Chondrocladia and Cladorhiza are found to be monophyletic. However, results indicate that the subgenus Chondrocladia is polyphyletic with respect to the subgenera Meliiderma and Symmetrocladia. Euchelipluma, formerly Guitarridae, is retained, but transferred to Cladorhizidae. The four known carnivorous species currently in Esperiopsis are transferred to Abyssocladia. Neocladia is a junior homonym and is here renamed Koltunicladia. Our results provide strong evidence in support of the hypothesis that carnivory in sponges has evolved only once. While spicule characters mostly reflect monophyletic groups at the generic level, differences between genera represent evolution within family Cladorhizidae rather than evolution of carnivory in separate lineages. Conflicting spicule characters can be reinterpreted to support the inclusion of all carnivorous sponges within Cladorhizidae, and a carnivorous habit should thus be considered the main diagnostic character in systematic classification.

Isolation and Characterization of Salt Tolerant Endophytic and Rhizospheric Plant Growth-Promoting Bacteria (PGPB) Associated with the Halophyte Plant (Sesuvium Verrucosum) Grown in KSA

This study was designed to isolate and characterize endophytic and rhizospheric bacteria associated with the halophyte plant Sesuvium verrucosum, grown under extreme salinity soil in Jeddah, Saudi Arabia. The plant growth promotion activities of isolated bacterial were evaluated in vitro. A total of 19 salt tolerant endophytic and rhizospheric bacterial isolates were obtained and grouped into six according to genetic similarity based on RAPD data. These six isolates were identified by amplification and partial sequences of 16S rDNA as Enterobacter cancerogenus,Vibrio cholerae, Bacillus subtilis, Escherichia coli and two Enterobacter sp. Isolates were then grown until exponential growth phase to evaluate the atmospheric nitrogen fixation, phosphate solubilization, and production of phytohormones such as indole-3-acetic acid, as well as 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. While, All of the six strains were negative for ACC deaminaseactivity, two isolates showed Nitrogen fixation activity, three isolates produce the plant hormone (Indole acetic acid) and two isolates have the activity of solubiliztion of organic phosphate. Among the six isolates, the isolate (R3) from the soil around the roots is able to perform the three previous growth promoting possibilities together and it is ideal for use in promoting the growth of plants under the high salinity conditions. This isolate is candidate to prepare a friendly biofertelizer that can be used for the improvement of the crops performance under salinity conditions.Int J Appl Sci Biotechnol, Vol 3(3): 552-560

Genetic diversity of cultured populations of giant freshwater prawn (Macrobrachium rosenbergii) in China using mtDNA COI and 16S rDNA markers

Partial sequences of mitochondrial DNA 16S rDNA and COI genes (395 bp and 498 bp respectively) were sequenced from samples of ten cultured populations of Macrobrachium rosenbergii (Giant Freshwater Prawn – GFP) in Zhejiang, Guangdong and Guangxi Provinces in China and two wild populations of GFP from Mekong River and Dongnai River in Viet Nam. Five haplotypes of 16S rDNA were identified in the 360 samples. The wild populations displayed nucleotide diversity (π) of 0.0008 and 0.0003, and genetic diversity (h) of 0.3030 and 0.1310 in the Mekong River and Dongnai River respectively. The cultured populations displayed no significant genetic diversity. COI sequences identified 17 haplotypes based on 21 polymorphic sites. At this marker, the 12 populations showed a range of h from 0.1290 to 0.6940 and π from 0.0003 to 0.0073. The largest genetic distance (Da) among the 12 populations was 0.0065 (between ZJB and BT/DN populations) and the lowest Da was 0.0003 (between GDD and GDA populations). The wild populations had higher genetic diversity than the cultured populations, but three of cultured populations from Zhejiang (ZJA, ZJB and ZJC) had π higher than wild populations, because they originated from Thailand, Bangladesh and the Mekong River in Viet Nam.

Morphology and molecular analysis of Mizelleus indicus Jain () and M. longicirrus (Tripathi, ) Venkatanarasaiah & Kulkarni 1981 (Monogenea, Dactylogyridae) from the freshwater shark Wallago attu in the Ganga River, India

Species of the genus Mizelleus Jain (1957) have always been controversial regarding identification and validity. Members of this group of species differ from each other in the morphology of their hard parts, which can be misleading and subject to differing interpretation among scientists. Therefore, the main objective of present study was to identify Mizelleus worms by morphological methods and molecular analysis on the basis of 18S ribosomal DNA to clarify their phylogenetic status. In this study, specimens were isolated from the gill filaments of Wallago attu (Siluriformes) and studied morphologically. In accordance with morphological characters, the specimens were found to be Mizelleus indicus and Mizelleus longicirrus. Partial sequences of nuclear 18S rDNA of these two species were amplified. The results confirm the phylogenetic relationships and taxonomic validation of M. indicus and M. longicirrus in India.

Three new species of Myxobolus Bütschli, 1882 (Myxozoa: Myxobolidae) infecting the common nase Chondrostoma nasus (L.) in the River Danube.

The common nase Chondrostoma nasus (L.) is a frequent cyprinid fish in the River Danube. In a survey on its infection with myxosporeans, eight different Myxobolus spp. spore types were found in the gills, swim bladder, fins and intestinal wall. Of these, spore types representing three species were studied in detail by morphological and molecular methods. Based on the differences in 18S rDNA sequences, two new species of Myxobolus Bütschli, 1882 from the gills and one from the swim bladder are described: M. arrabonensis n. sp., M. szentendrensis n. sp. and M. paksensis n. sp. The new species resembled M. muelleri Bütschli, 1882, M. intimus Zaika, 1965 and M. cycloides Gurley, 1893, all parasitic in leuciscine cyprinids, in spore size and location in the host, but exhibited differences in partial 18S rDNA sequences as follows: M. arrabonensis - M. muelleri (1.4%), M. szentendrensis - M. intimus (2.8%), M. paksensis - M. cycloides (2.4%). Based on the significant differences in rDNA sequences, the three forms are considered to represent new, hitherto undescribed species in spite of their morphological similarities to some Myxobolus spp. forming spores in identical locations in genetically closely related cyprinids of the subfamily Leuciscinae.

Molecular phylogenetics of eimeriid coccidia (Eimeriidae, Eimeriorina, Apicomplexa, Alveolata): A preliminary multi-gene and multi-genome approach.

Coccidia possess three distinct genomes: nuclear, mitochondrial, and plastid. Sequences from five genes located on these three genomes were used to reconstruct the phylogenetic relationships of members of the phylum Apicomplexa: 18S rDNA sequences from the nuclear (nu) genome, partial cytochrome c oxidase subunit I sequences from the mitochondrial (mt) genome, and partial 16S and 23S rDNA sequences and RNA polymerase B sequences from plastid (pl) genomes. Maximum parsimony, maximum likelihood, and Bayesian inference were used in conjunction with nuclear substitution models generated from data subsets in the analyses. Major groups within the Apicomplexa were well supported with the mitochondrial, nuclear, and a combination of mitochondrial, nuclear and concatenated plastid gene sequences. However, the genus Eimeria was paraphyletic in phylogenetic trees based on the nuclear gene. Analyses using the individual genes (18S rDNA and cytochrome c oxidase subunit I) resolved the various apicomplexan groups with high Bayesian posterior probabilities. The multi-gene, multi-genome analyses based on concatenated nu 18S rDNA, pl 16S, pl 23S, pl rPoB, pl rPoB1, and mt COI sequences appeared useful in resolving phylogenetic relationships within the phylum Apicomplexa. Genus-level relationships, or higher, appear best supported by 18S rDNA analyses, and species-level analyses are best investigated using mt COI sequences; for parasites for which both loci are available, nuclear 18S rDNA sequences combined with mitochondrial COI sequences provide a compact and informative molecular dataset for inferring the evolutionary relationships taxa in the Apicomplexa.

Atrokylindriopsis, a new genus of hyphomycetes from Hainan, China, with relationship to Chaetothyriales

Atrokylindriopsis setulosa gen. et sp. nov. produces pigmented, septate, setulate conidia that are attached to the monophialidic conidiogenous cells of unbranched, brown, macronematous conidiophores at the midpoint of their long side during conidiogenesis, giving the appearance of a ‘T’. Internal transcribed spacer region (ITS) and partial 28S rDNA sequences indicate placement of Atrokylindriopsis in the Chaetothyriales.

Isolation and Characterization of Multi-Metal-Resistant Halomonas sp. MG from Tamil Nadu Magnesite Ore Soil in India.

The aim of the study was to isolate and characterize potential multi-metal-resistant bacteria from ore soils. A total of three bacteria were isolated and assayed for resistance to arsenic (As), copper (Cu), and lead (Pb). Isolate Halomonas sp. MG exhibited maximum resistance to 1000 mg Pb/L, 800 mg As/L, and 500 mg Cu/L and it was identified as Halomonas sp. based on the partial 16S rDNA sequences. The metal(loid)s resistance mechanisms were further confirmed by amplification of arsC (As) copAU (Cu), and pbrT (Pb) genes. Biological transmission electron micrographs and XRD studies showed that the isolate Halomonas sp. MG transformed and/or biomineralized the metals either intracellularly or extracellularly. These results suggest that the isolate could be used as a potential candidate for the bioremediation of As, Cu, and Pb.

Morphological and molecular-biological characteristics of two gregarine species (Eugregarinida: Gregarinidae) associated to Phaedon brassicae and Phyllotreta striolata (Coleoptera: Chrysomelidae)

A new gregarine species (Eugregarinida: Gregarinidae) was previously reported from daikon leaf beetle, Phaedon brassicae at Daegwallyeong in Korea. Here, we reported another novel gregarine species from the striped flea beetle, Phyllotreta striolata (Coleoptera: Chrysomelidae). The beetles were collected from the three locations; Pyeongchang (760m), Hoengseong (466m), and Inje (290m), Gangwon province during July to September. In this paper, we present the morphological and molecular-biological characteristics of Gregarine species associated to P. brassicae and P. striolata. The size and body shape at mature trophozoite stage were the main morphological diagnostic characteristics of those species. Gregarines from P. brassicae were bigger in size (100μm) with square shaped body while Gregarines from P. striolata were slightly smaller in size (80μm) with egg shaped body. Based on the partial 18S rDNA sequence, this gregarine species was also grouped in eugregarine, and showed 85% homology to Gregarina sp. associated to P. brassicae. However, histological section result showed that each species had their own gregarine parasite beetle association. Species-specific parasitism was confirmed in local populations' beetles. There was positive relationship between infection rates, and the elevation of locations for gregarine related to P. striolata. Therefore, we suggested that the gregarines and their coleopteran hosts have species-specific interaction via co-evolution.

Systematic revision of the freshwater snailMargarya Nevill, 1877 (Mollusca: Viviparidae) endemic to the ancient lakes of Yunnan, China, with description of new taxa

The systematics of the viviparid freshwater snail genus Margarya endemic to the ancient lakes of Yunnan, China, is revised based on comparative analyses of morphological features, including shell, operculum, radula, and genital anatomy, and molecular phylogenetic analyses of partial sequences of the mitochondrial 16S rDNA (16S) and cytochrome c oxidase subunit I (COI) genes, as well as the nuclear Internal Transcribed Spacer 2 (ITS2). The taxonomic utility of key anatomical and morphological features in this group is evaluated. The genus Margarya as delimited previously is split into three genera in order to retain monophyletic taxa: (1) Margaryas.s., consisting of four species, i.e. the type species Margarya melanioides plus Margaryafrancheti, Margaryaoxytropoides, and Margaryamonodi; (2) the previously introduced subgenus Tchangmargarya is elevated to an independent genus containing two species, Tchangmargarya yangtsunghaiensis and the new species Tchangmargarya multilabiata sp. nov.; and (3) a new genus, Anularya gen. nov., is described, also containing two species, i.e. Anularya mansuyi and Anularya bicostata. Molecular phylogenies based on analyses of three gene fragments have identical topologies, supporting the monophyly of these genera. The sister group of Margarya s.s. is Cipangopaludina, whereas the sister group of Anularya is Sinotaia; Tchangmargarya is sister to a clade containing all the aforementioned groups. Features of the operculum and the right male tentacle (penis) are particularly informative on the generic level, whereas shell and radular characters are especially useful to differentiate species. The phylogenetic relationships recovered here are consistent with orogenic patterns of the Yunnan Mountains. Changes in the river system and water area of ancient lakes caused by tectonic activities probably play an important role in speciation and shaping the current pattern of species distribution in Yunnan.

Quantification of the potential biocontrol and direct plant growth promotion abilities based on multiple biological traits distinguish different groups of Pseudomonas spp. isolates

Members of the Pseudomonas genus colonize the rhizosphere of different plant species and display plant-probiotic traits. Therefore, they are interesting candidates for biological agricultural inputs to stimulate plant development and/or promote crop health. We have generated a collection of 19 isolates of pseudomonads obtained from either bulk soil or the rhizosphere of healthy individuals of major extensive crops from different plots under no-till management located in Argentina. Isolates were selected for their ability to antagonize several fungal pathogens recovered from infected soybean and maize plants. Partial sequencing of 16S rDNA, oprF and rpoB genes positioned isolates within all major pseudomonads groups. Most isolates colonized the rhizosphere of soybean and maize seedlings without affecting germination or development. Members of the Pseudomonas chlororaphis subgroup, and one isolate of the Pseudomonas putida subgroup, displayed the widest antifungal spectrum and strongest antagonistic potential. A principal component analysis based on the outcome of quantitative and qualitative tests related to biocontrol of fungal pathogens, direct plant growth promotion, and other root colonization-related traits, showed that isolates with the highest antagonistic potential and mostly of soil origin were grouped together, whereas rhizospheric isolates and those with strong flagellar-dependent motility, exoprotease production and biofilm development, were clustered in a separate group. Interestingly, quantitative comparison of newly developed biocontrol and direct growth promotion indices revealed that these two plant probiotic traits are rather associated to different set of isolates, and rarely present in the same pseudomonad strains. These indices could be employed to screen and categorize isolates by their potential to act as a biocontrol or a biofertilizer agent.

Genetic Diversity of Filamentous Cyanobacteria from Shore Regions of Okinawa

Benthic marine cyanobacteria are ubiquitous oxygenic photosynthetic bacteria and they form major components of various communities, including epiphytic, epilithic and microbial mats, in coral reef environments. Filamentous groups may form almost monospecific macroscopic colonies, and sometimes make potentially harmful algal blooms (HABs). HABs bring serious problems for marine tourism but marine filamentous cyanobacteria are good resource for drug discovery. However, cultivation and identification of such filamentous cyanobacteria are difficult and there are taxonomic confusions. In this study, we investigate 17 samples from coral reef area of 10 localities in Okinawa. Macroscopic mats or cushions of filamentous cyanobacteria were collected. The samples were divided into three parts: (1) for morphological observation, (2) for molecular work, and (3) cultivation for further study. Dominant cyanobacteira of each sample identified morphologically as Anabaena, Hydrocoleum, Lyngbya, Oscillatoria, Phormidium, Schizothrix and Symploca. Molecular results of partial 16S rDNA sequences revealed six groups that composed of heterocystous group, Leptolyngbya group, Symploca group, Moorea group, Oscillatoria group and unknown group, respectively. Some samples corroborated between morphology and molecular results but some were not. In addition, four out of six groups closely related with natural product producers. These results suggest that there are high biodiversity of marine benthic filamentous cyanobacteria in Okinawa that may become seeds of HABs as well as providing promising resources for drug discovery.

Molecular and morphological identification of Cardicola (Trematoda: Aporocotylidae) eggs in hatchery-reared and migratory Atlantic bluefin tuna (Thunnus thynnus L.)

A microscopic and a molecular approach was used to investigate blood fluke infection in the very first specimens of Atlantic bluefin tuna (Thunnus thynnus, Scombridae) that were born in captivity and that were never moved from their specific cage culture sites in the Mediterranean. Data were compared with infection in wild, migratory bluefin tuna which were captured and fattened along the Spanish Mediterranean coast. Wild and cultured populations sampled in the present study harbored the same Cardicola spp. richness and showed statistically insignificant differences in overall parasite prevalence. Three morphotypes, crescent-shaped, fusiform and oval eggs, were observed in both non-migrant and migrant tuna. According to partial ITS2 and 28S rDNA sequences they belong to Cardicola opisthorchis, Cardicola forsteri, and to Cardicola sp., a recently described species, phylogenetically closely related to Cardicola orientalis. All three species were simultaneously present in some specimens. No significant difference in overall parasite prevalence was observed, though the prevalence of eggs of individual species, i.e. C. opisthorchis and C. forsteri, was significantly higher in fattened bluefin tuna. Our results strongly support a Mediterranean origin of the three Cardicola species encountered in cultured specimens and suggest that the intermediate hosts of these Cardicola species occur along the Mediterranean coast. Thus, infection with these parasites is not exclusively a result of infections happening along the migratory routes of wild bluefin tuna, in the Atlantic Ocean, as previously suggested. Once transferred to the sea for fattening, Atlantic bluefin tuna are at risk of infection with up to three different species of the genus Cardicola in along the Mediterranean coast in southeast Spain. On the contrary, C. orientalis infections might be associated to long-distance migration and an overlapping Thunnus maccoyii and Thunnus orientalis distribution.Statement of relevanceThis is the very first parasitological study conducted in T. thynnus born in captivity and cultured local in the Mediterranean. The results show that once transferred to the sea for fattening, they are at risk of infection with up to three different species of the genus Cardicola. These results strongly imply a Mediterranean distribution of these species.