Determination of fluoride and its effects on flavonoids in some legumes

The effects of fluoride (Fl) emission, from an aluminum (Al) reduction plant in Iran, on the concentration of Fl in six legume species were studied. In addition, the influence of Fl on leaf flavonoids was investigated. Determination of Fl in collected plants (Alhagi camelorum Fisch., Cercis siliquastrum L., Glycyrrhiza glabra L. Medicago sativa L., Robinia peseudoacacia L., and Sophora alopecuroides L.) from the factory area was performed using a Fl ion-selective electrode. Identification of flavonoids in the samples was obtained by two-dimensional paper chromatography (2-D PC) and thin layer chromatography (TLC). Controls were obtained from plants located 10 km distance from the factory. Results showed high Fl concentration in all polluted samples. Phytochemical changes in polluted leaves were observed with appearance or disappearance of some flavonoids.     

Phytoremediation of petroleum-contaminated soils: Pre-screening for suitable plants and rhizospheral fungi

A field study was conducted in a petroleum-contaminated site in Kermanshah refinery (Iran) to find the petroleum-resistant plant species and the rhizospheral fungi for being used in bioremediation. Results showed that the amounts of petroleum pollution in non-vegetated areas is 6.8% and in vegetated areas is 2.2%. Plant samples were collected from petroleum-polluted areas and determined using morphological characters. It was found that eight plant species were growing on the contaminated sites: Polygonum aviculare, Amaranthus retroflexus, Noea mucronata, Alhaji cameleron, Crozophora heirosolymitrana, Poa sp., Convolvulus arvensis and Trifolium repense. The rhizospheral fungi of the plants were collected and determined using microscopic studies and taxonomical keys. The results determined the presence of 21 species in the rhizosphere of the plants growing in the polluted areas; three of these species were common in all of the plants and the others have species-specific distribution within the plants. The highest number of rhizospheral fungi (11 species) were determined for P. aviculare in non-polluted areas and nine species in polluted areas. It seems that the plant is the best candidate for using phytoremediation. However, the variation of fungi in petroleum-polluted areas was more than non-polluted zones. The culture of fungi in oil-contaminated media showed that, although all the studied fungi were resistant to low petroleum pollution (1% v/v), a few species, especially Fusarium species, showed higher resistance to petroleum pollution (10% v/v) and hence they may be suitable for mycoremediation in highly polluted areas.     

Genetic population structure of a species' complex of blue mussels (Mytilus spp.)

Blue mussels representing two nominal species (Mytilus trossulus Gould, 1850 and Mytilus galloprovincialis Lamarck, 1819 were collected from 28 intertidal locations along the Pacific coast of the USA in 1990–1991 (total N=1255) and examined for variation at 15 allozyme loci. Twelve samples, mostly from a region of suspected hybridization, were analyzed for variation in seven shell characters. Principal-components analysis of allozyme data revealed three groups based on first principal-component scores, which were identified as M. trossulus, M. galloprovincialis, and hybrids. Canonical discriminant analysis of shell characters was less successful in separating mussels into discrete groups. Each location was characterized for four environmental variables: (1) temperature, (2) salinity, (3) tidal height and (4) degree of exposure to wave action, which were then used as independent variables in a series of multiple-regression analyses, with the proportions of the two species as dependent variables. Temperature and salinity had significant (P<0.05) effects on the macrogeographic distribution of the two species, whereas the effects of height in the tidal zone and degree of wave exposure were not statistically significant. Salinity was found to have a greater influence than temperature on the microgeographic distribution of the two species. M. trossulus was more abundant at locations with lower temperatures and greater salinity variation than M. galloprovincialis. The two species appear to be ecologically distinct, and their genetic integrity is at least partly the result of environmental heterogeneity.     

Sequestration of phorbol esters by aposematic larvae of Hyles euphorbiae (Lepidoptera: Sphingidae)?

Summary. The larvae of the hawkmoth species Hyles euphorbiae have a conspicuous aposematic colouration and show gregarious behaviour. It has thus been suggested that they sequester phorbol esters from their food plants which include different species of the genus Euphorbia (Euphorbiaceae) for chemical protection against predators. To test this hypothesis in more detail, we fed larvae an artificial diet with three doses of 12-tetradecanoyl-phorbol-13-acetate (TPA), then examined the faeces and the larval tissues, such as integument, haemolymph and gut of the caterpillars for the presence of TPA. In order to determine the ability of the larvae to detoxify phorbol esters, other larvae were directly injected with a TPA solution and analysed in the same manner. Our study indicates that the larvae of Hyles euphorbiae do not sequester phorbol esters. Upon oral application TPA was not found in the larval integument or the haemolymph. Instead, it was mostly metabolised (about 70–90%). Nevertheless, about 10-30% were retained and recovered in the faeces. The larvae were also able to metabolise and thus detoxify the phorbol ester when TPA was injected directly into the body. These hawkmoth caterpillars are relatively large and have a gut full of plant material, which they regurgitate into the direction of the predator when attacked in nature. Since phorbol esters are very potent toxins and irritants, we postulate that the gut content (and especially the plant slurry disgorged as regurgitant from the anterior gut) alone could be aversive for a potential predator, even if some metabolism has taken place. Thus, although H. euphorbiae caterpillars do not actively sequester phorbol esters, their aposematic colouration appears to be based on chemical defence through phorbol esters retained in the gut.     

Escape responses of four Chilean intertidal limpets to seastars

The Chilean intertidal Lottiidae limpet fauna consists of at least nine species, grouped into the monophyletic taxon Scurria and a single taxon preliminarily designated as `Lottia' orbignyi (Dall). Within the Scurria clade, the most basal species are Scurria variabilis (Sowerby), S. viridula (Lamarck), and S.zebrina (Lesson), and among the most crown groups are S. ceciliana (Orbigny) and S. araucana (Orbigny). This phylogenetic information, obtained by the analysis of molecular characters (16S mtDNA), provides a framework for a comparative study of behavioral characters. In this study, we analyze the escape responses of two basal limpet species (`L.' orbignyi and S. viridula) and two derived limpet species (S. araucana and S. ceciliana) in the presence of the limpet predatory seastar Heliaster helianthus (Lamarck) and the limpet non-predatory seastars Stichaster striatus Muller and Troschel and Patiria chilensis Lutken. Neither P.chilensis nor Stichaster striatus induced escape responses such as those observed with the predatory H. helianthus. Moreover, in the presence of H. helianthus, basal and derived limpet species differed significantly in the percentage of individuals responding, reaction time, and duration of the response. The basal species `L.' orbignyi and Scurriaviridula exhibited instantaneous and vigorous locomotor responses, whereas the derived species S. araucana and S. ceciliana displayed no locomotor responses. The same pattern was found in responses to H. helianthus homogenates. The results strongly suggest a correlation between phylogeny and anti-predator escape responses to the seastar H. helianthus. Alternative escape responses have evolved for basal and derived species within the Chilean clade. Received: 27 April 2000 / Accepted: 16 August 2000     

Enzymatic and morphological criteria for distinguishing between Cardium edule and C. glaucum of the Portuguese coast

Cardium edule L. and C. glaucum Bruguière are two sibling species of cockles commonly found in littoral systems along the Portuguese coast, where they sometimes occur as mixed populations. The problem of morphological separation of these two cockle species has not been completely solved. Indeed, the distinctive morphological criterion proposed by Brock after identifying the specimens by means of electrophoretic patterns of hepatopancreatic malate dehydrogenase was not effective when applied to Portuguese Cardium spp. The object of the present study was to establish morphological discrimination criteria for distinguishing Portuguese C. edule and C. glaucum on the basis of initial identification of individuals through the enzymatic criterion established by Brock. Cockles were collected from 1988 to 1991 at 16 estuarine and lagoonal systems along the Portuguese coast. Hepatopancreatic extracts were submitted to horizontal starch gel electrophoresis following Brock's procedure. The malate dehydrogenase patterns obtained were identical to those found by Brock. Clear interspecific differences in the leucine aminopeptidase and esterase isozyme systems were also detected. The morphological study of the shells involved 7 morphometric and 9 qualitative and semiquantitative characters. Canonical variate analysis of the morphometric characters showed that these could not be used for identification purposes. However, a clear separation between the two species was obtained by a principal-component analysis of qualitative and semiquantitative characters. The characters mainly useful for distinguishing between the two species were valve profile, shape of ventral valve junction, type of calcareous scales and shell colour.     

Molecular systematics of six Calanus and three Metridia species (Calanoida: Copepoda)

The discrimination of species of the copepod genus, Calanus (Copepoda; Calanoida), is problematical-especially in regions of sympatry. Although the species of Calanus exhibit exceptional morphological similarity, they are quite distinct in genetic character. The DNA base sequences of the mitochondrial large subunit (16S) ribosomal RNA (rRNA) gene unambiguously discriminated C. finmarchicus (Gunnerus 1765), C. glacialis (Jaschnov 1955), C. marshallae (Frost 1974), C. helgolandicus (Claus 1863), C. pacificus (Brodsky 1948), C. sinicus (Brodsky 1965), and C. hyperboreus (Kroyer 1838). Sequence differences among Calanus species for this gene portion range from 7.3% (between C. glacialis and C. marshallae) to 23.9% (between C. glacialis and C. sinicus). Differences among conspecific individuals were approximately 1 to 2%. [These sequence data were determined between April and November 1993; the sequenced domain is similar to that published previously in Bucklin et al. (1992) but are derived from analysis of additional individuals.] Statistical analysis of the sequence data using a variety of tree-building algorithms separated the taxa into one group of species corresponding to the C. finmarchicus group (C. finmarchicus, C. marshallae, and C. glacialis) and another ungrouped set of species corresponding to the C. helgolandicus group (C. helgolandicus, C. pacificus, and C. sinicus). The C. helgolandicus group may be older than the C. finmarchicus group, making the tree topology less reliable in this area. Calanus hyperboreus was an outlier; Nannocalanus minor (Claus 1863) was the outgroup. Similar analysis of Metridia species confirmed that M. lucens (Boeck 1864) and M. pacifica (Brodsky 1948) are distinct species; M. longa (Lubbock 1854) was still more divergent. These sequence data will allow the design of simple, molecular tools for taxonomic identifications. Diagnostic characters, assayed by rapid molecular protocols, will enable biological oceanographers to answer important questions about the distribution and abundance of all life stages (as well as patterns of reproduction) of morphologically similar species, such as those of Calanus.     

Genetic evidence for two sibling species within Polydora cf. ciliata (Polychaeta: Spionidae) from the Sea of Japan

Two morphologically indistinguishable but genetically very distinct polychaete species of the family Spionidae are found in the Peter the Great Bay of the Sea of Japan. Both species bore into molluscan shells and occur sympatrically. They are close in morphology to Polydora ciliata (Johnston, 1838) and some other members of the redefined P. ciliata species complex. The systematics of the two species is still under consideration by one of us (V.I.R.). A total of nine enzyme systems comprising 12 isozyme loci were examined in both the species by means of starch gel electrophoresis. Nei's genetic distance between them proved surprisingly high (D= 1.942) being the highest among estimates so far obtained for pairs of polydorid species. The levels of intraspecific genetic variation in both species (Polydora sp. 1, H _e= 0.287 ± 0.064; Polydora sp. 2, H _e= 0.111 ± 0.045) are comparable to those of other electrophoretically studied polydorids. The problem of taxonomic integrity of cosmopolitan marine invertebrate species is briefly discussed as well as the usefulness of isozymes as species-specific characters suitable for unequivocal discrimination between sibling polydorid species. A list of enzymes characterized by interspecifically variable numbers of expressed isozymes within polydorids is given and recommended for such a discrimination. Received: 22 July 1997 / Accepted: 19 February 1998     

Bioassay-guided isolation of allelochemicals from Avena sativa L.: allelopathic potential of flavone C-glycosides

The allelopathic potential of oat (Avena sativa L., var. Argentina, Poaceae, Cyperales) was investigated under field and laboratory conditions. In field trials, oat plants provided an effective control of weeds, showing a species-specific impact: the most abundant weed species, Picris echioides was reduced by 94% in number of individuals. Aerial parts of oat plants, harvested immediately before soil incorporation, were utilized in a bioassay-guided isolation, which was aimed at identifying the phytotoxic compounds in a methanol/water extract of the aerial parts of the plants. Further partitions of extract gave an active n-butanol portion composed of flavonoids and saponins. Phytotoxic activity was detected for the flavonoid fraction, whereas no activity was found for the saponin mixture. Germination of an indicator species, lettuce (Lactuca sativa L.), was completely inhibited at flavonoid concentrations of 6.7, 10.0, and 20.0 mg/mL, and conversely the number of abnormal seedlings was greatly increased from 2% of control to over 96% at concentrations equal to 6.7 and 10.0 mg/mL. Analysis of the flavonoid fraction by ESI-MS techniques identified two components of the mixture as isoorientin 2-O-glucoside and isovitexin 2″-O-arabinoside. Saponins were characterized as avenacoside A, avenacoside B, 26-desglucoavenacoside A, and 26-desglucoavenacoside B. Both flavone C-glycosides and saponins were isolated previously from oat.     

A stereoscan electron microscope study of male reproductive characters in the Jaera albifrons group of species (Crustacea: Isopoda)

The male secondary sex characters of Jaera albifrons Leach, Jaera ischiosetosa Forsman, Jaera praehirsuta Forsman and Jaera forsmani Bocquet are described with the Stereoscan electron microscope (S.E.M.). These characters, which are present on the peraeopods, are the only means of identification for these Jaera species which together form the Jaera albifrons group (Isopoda, Asellota). The male praeoperculum (modified, fused, first pair of pleopods) and appendix masculinae (modified endites of the second pair of pleopods) have also been studied with the S.E.M. and possible functions of the various sex characters are discussed. Both the male operculum and appendix masculinae are similar for each species, suggesting that copulation with females of a different species is anatomically possible. The fact that hybrids are rarely collected in ecological investigations is discussed with reference to the present known information. Two methods of copulation for the members of this group have been proposed and these are discussed. A method of sperm transfer along the male operculum is outlined.     

Concentrations of Cd,Ni, Pb,and Cr in the two edible fish species Liza klunzingeri and Sillago sihama collected from Hara biosphere in Iran

The concentrations of four metals (Cd, Ni, Pb, and Cr) were determined in the muscles, gills, and livers of two edible fish species (Liza klunzingeri and Sillago sihama) caught from the Hara biosphere of Southern Iran. In both fish species, metal concentrations and bioaccumulation factors were in the sequence liver?>?gill?>?muscle. Bioaccumulation factors were found to be highest in S. sihama. The metal concentrations were descending in the order of Ni?>?Cr?>?Pb?>?Cd, except for muscle samples from S. sihama showing an inversion of Pb and Cr. There is a significant negative correlation between the concentrations of the metals in each tissue with length, weight, and age, except for muscle in L. klunzingeri. Some metal levels in the muscle exceeded the limits recommended by FAO, WHO, and FEPA.     

First genetic validation and diagnosis of the short-finned squid species of the genus <Emphasis Type="Italic">Illex</Emphasis> (Cephalopoda: Ommastrephidae)

Squids of the genus Illex are representative of the family Ommastrephidae and account for 65% of the world’s cephalopod captures. Illex is formed by four taxa distributed throughout the Atlantic Ocean (I. argentinus, I. coindetii, I. illecebrosus and I. oxygonius), whose identification and phylogenetic relationships based on morphological characters remain controversial. Thirty-seven enzyme-coding loci were analysed in 230 individuals from seven populations of Illex and ten specimens of Todaropsis eblanae, which were used as the outgroup. Two to four enzyme loci (ALPDH*, IDHP-1*, MEP* and SOD*) were diagnostic among Illex species depending on the species-pair comparison. Individuals morphologically identified as I. oxygonius were also found genetically distinct, which proves the taxonomic validity of this species. No significant intraspecific genetic heterogeneity was detected within Illex argentinus, I. coindetii and I. illecebrosus (Mean G _ST= 0.011, 0.003, 0.017, respectively). I. illecebrosus and I. oxygonius were shown as sister species with a close relationship to I. argentinus, whereas I. coindetii formed a different lineage.     

Low-temperature adaptation of deep-sea bacteria isolated from the Japan Trench

Montastraea annularis, M. faveolata, and M.␣franksi are three recently separated species that together dominate reefs of the tropical western Atlantic. Despite morphological, life-history, ecological and genetic differences, the legitimacy of their status as separate species has been questioned. This controversy stems from both the scarcity of unambiguous, diagnostic differences among them, and from the possibility of extensive hybridization associated with their approximately synchronous reproduction in sympatry. Here we report on fertilization trials and the timing of spawning, both of which suggest that the potential for hybridization may be limited. Crosses between M. faveolata (the most genetically distinctive taxon) and the other two species were largely unsuccessful, as were selfed matings for all three species. M. annularis and M. franksi showed no evidence of fertilization barriers, but the timing of spawning typically differed between them by 1 to 2 h. We also found that spawning times in the field of M. annularis and M. faveolata were non-overlapping in 1995, and that the timing of spawning in M. annularis could be experimentally shifted forward by simulating earlier than natural sunsets. These findings from Panamá and Honduras, particularly given their consistency with comparable observations and experiments elsewhere, provide evidence of reproductive isolation and support the separate species status of these three taxa. Some hybridization may occur under natural conditions at these sites, particularly between M. annularis and M.␣franksi. Rampant genetic interchange among these taxa in Central America seems unlikely, however, in the light of the concordant associations between morphology, reproductive biology and other characters which are observed. Received: 22 July 1996 / Accepted: 2 October 1996     

Molecular phylogenies help resolve taxonomic confusion with Asian <Emphasis Type="Italic">Crassostrea</Emphasis> oyster species

Identification of oyster species is still largely based on phenotypic characters that are highly plastic. Prompted by the proposed introduction of the Asian oyster species Crassostrea ariakensis into the Chesapeake Bay region of the U.S.A., this study uses molecular genetic information to understand the taxonomic framework surrounding C. ariakensis and to confidently distinguish among various sympatric oyster species. Putative samples of C. ariakensis and other species of cupped oysters from across Asia were collected and phylogenetic analyses were conducted on DNA sequences of both nuclear (ITS-1) and mitochondrial (COI) regions. Trees generated based on the two independent molecular datasets were highly congruent, and indicate that many oysters collected for this study as C. ariakensis were originally misidentified. Results also indicate that C. ariakensis, C. hongkongensis and C. nippona are distinct, but closely related species. There is strong support in both analyses for a close relationship between C. gigas and C. sikamea, as well as between C. belcheri and C. gryphoides, and between C. iredalei and C. madrasensis. The parsimony analyses based on these DNA markers, however, did not provide evidence to support C. angulata as a distinct species from C. gigas. Overall, the results emphasize the need for rigorous species identification, and additional extensive and intensive sampling to more accurately determine relationships among Crassostrea species, define their geographic distributions, and establish existing sympatry patterns.     

Reproductive,growth and life-history contrasts between two species of grapsid crabs,Sesarma cinereum and S. reticulatum

Two grapsid crabs, Sesarma cinereum (Bosc) and S. reticulatum (Say), were studied in the region of Beaufort, North Carolina, USA, during the summers of 1977, 1983, and 1984. The two grapsid species showed differences in social organization, habitat preferences, and number and size of first-stage larvae. We analyzed several other characters (growth rates, size at sexual maturity, reproductive effort per clutch) to determine if these differences could be related to a coherent pattern of life-history trends. While some relationships between characters were consistent with those found among other crab species, few seemed peculiarly matched to the ecological contrasts represented by our species. Our results suggest that other factors (phyletic predisposition; decoupling of selection between larval and adult stages; equally adaptive alternatives) may obscure the rules governing relationships between life-history characters and evolutionary trends.     

Contamination of lead (Pb) in the coastal sediments of north and south of Iran: a review study

Coastal areas of Iran are heavily affected by urbanisation, industrialisation, and maritime activities. One consequence of this environmental pressure is the contaminants accumulation, as heavy metals, into the marine ecosystem. In this review, the coastal areas in the north and south of the country were assessed for lead (Pb) contamination of, one of the most toxic metals found in the environment. All studies conducted during 2006–2016 with at least 10 specimens that reported the mean and standard deviation of Pb were considered in this review. Heterogeneity between studies was assessed using the Q and I² statistics. The Pb mean concentration overall studies was estimated to be 21.88?µg/g (95% CIs: 16.25–27.50). Random effect model showed no statistical difference in mean Pb concentration levels between south and north coasts. However, the variability in Pb mean concentrations within southern coasts was considerable and statistically significant. Moreover, the Pb concentrations in the northern coasts of Persian Gulf and Oman Sea, in south of Iran, decreased in the following order Hormozgan > Khuzestan > Sistan-VA-Balluchestan > Bushehr while in the southern coasts of Caspian Sea, in north of Iran, decreased in the order Guilan > Mazandaran > Golestan.     

Pathological effects of sublethal diazinon on the blood,gill, liver and kidney of the freshwater fish Capoeta damascina

Capoeta damascina has a large distribution in the Middle East and is one of the most abundant fishes in inland waters of Iran. Histopathological symptoms of the blood, gill, liver and kidney of C. damascina were investigated over 1, 5 and 9-day exposure to 0 (control), 0.5, 1 and 1.5?mg?L^?1 diazinon. The data were analysed using analysis of variance and non-parametric multivariate analysis of variance. Diazinon concentration was more important than the exposure time on changes of the tissues. Diazinon decreased red blood cell (RBC), white blood cell (WBC), haematocrit (Hct) and haemoglobin (Hb). The symptoms of the gill were mainly desquamation, epithelial hyperplasia, hyperplasia at the base of secondary lamella, epithelial lifting and curling. The prevalent symptoms of the liver were degeneration of nuclei, alternation in size and shape of hepatocytes, focal necrosis and pyknosis. Degeneration of Bowman's capsule, necrosis in renal tubule and haematopoietic tissue were the most common symptoms found in the kidney. In conclusion, the blood, gill, liver and kidney of C. damascina are sensitive enough to respond to low concentrations of diazinon over a short period and this species can thus be a bioindicator of diazinon.     

The prevalence of heavy metals in Cladophora glomerata L. from Farahabad Region of Caspian Sea – Iran

The presence of toxic heavy metals in the environment is considered as a risk factor for adverse human and environmental health effects. Farahabad Region is a tourist center in the southern coast of Caspian Sea in Mazandaran Province of Iran. Environmental monitoring of this site is important for public health for individuals visiting and residing in this region. Although numerous biomonitoring data are available globally, very few if any apparent investigations have been conducted in this region. In this study, concentration of three heavy metals chromium (Cr), lead (Pb), and cadmium (Cd) was determined for one year in Cladophora glomerata, the predominant macroalga species present in this region. Detection of heavy metals was performed with atomic absorption spectrophotometer using standard methods. Results showed that the range of Cr metal in various algal samples was 29–55 ppm/g dry weight. The levels of Pb in algal samples (Cladophora) ranged from 2 to 8 ppm/g dry weight. The Cd concentrations in C. glomerata biomass ranged from 1.5 to 8.2 ppm/g dry biomass. In view of potential threats of such high metal concentrations in coastal waters and in algal tissues, it is necessary to adopt conservation measures to ensure public health safety.     

Morphological and molecular data suggest a cosmopolitan distribution of the polychaete <Emphasis Type="Italic">Proscoloplos cygnochaetus</Emphasis> Day, 1954 (Annelida,Orbiniidae)

The discovery of a possibly invasive Proscoloplos species on the French Atlantic coast led to a detailed morphological and molecular investigation. Proscoloplos consists of three nominal species, but molecular analyses of the variable ITS1 and ITS2 region revealed no clade support for samples from South Africa, France and Australia, representing at least two of these nominal species. We found no unambiguous diagnostic characters for the three different species even with scanning electron microscopy. Two main characters—first appearance of branchiae and hooks—that were used for species delineation show considerable intrapopulational variation and do not withstand critical evaluation. The obtained data and the observed regenerative capabilities point to a cautious use of the position of branchiae and hooked chaetae as taxonomic marker in Proscoloplos. After experimental bisection of animals, the recovery led to a phenotype without recognizable signs of regeneration, but neither branchiae nor hooked chaetae reoccurred at the same initial segmental position. Summarising the molecular and morphological data we suggest the synonymy of the known Proscoloplos species under the senior synonym Proscoloplos cygnochaetus Day, 1967. The enormous geographic range of this species comprises the temperate waters of the whole southern hemisphere and includes the French population. We discuss a dispersal enabled through the effective anchorage on vessels using mucous glands and chaetae possibly combined with architomic reproduction.