Morphometric and genetic analysis as proof of the existence of two sturgeon species in the Guadalquivir river

Morphometric and genetic methods were used to identify two sturgeon species, Acipenser naccarii Bonaparte, 1836, and A. sturio Linnaeus, 1758, captured in some of the principal rivers of the Iberian Peninsula, including the Guadalquivir. After measuring 25 Iberian specimens from a fishery and several Spanish and Portuguese museums and applying stepwise discriminant analysis (SDA), four specimens preserved in different museums [two specimens from the Guadalquivir river (EBD-8173 and EBD-8174), one specimen from the Tagus river (MUC1) and one specimen from the Mondego river (MUC46B)], as well as five specimens captured in the Guadalquivir river in the 1940s but not preserved (CM1, CM2, CM3, CM4 and CM5), were identified as A. naccarii. After cloning and characterisation of a satellite-DNA family, HindIII, from A.␣naccarii genome, its absence from the genome of A.␣sturio was determined. Using this satellite-DNA as a genetic marker and by means of dot-blotting, we demonstrate that the DNA of the two specimens captured during the mid-1970s in the Guadalquivir river cross-hybridised with HindIII satellite-DNA sequences of A.␣naccarii. We conclude that A. naccarii is autochthonous to the Iberian Peninsula and is not, as was previously believed, endemic to the Adriatic Sea. Received: 28 November 1996 / Accepted: 10 March 1997     

Atypical plant–herbivore association of algal food and a kleptoplastic sea slug (Elysia clarki) revealed by DNA barcoding and field surveys

The identity of food sources and feeding preferences of specialist herbivores have been commonly inferred from spatial associations between consumer and food items. However, such basic information for well-known marine herbivores, sacoglossans (sea slugs), and their algal diets remains disappointingly lacking, especially from field studies. The sacoglossan, Elysia clarki (Pierce et al. in Molluscan Res 26:23–38, 2006), is kleptoplastic and sequesters chloroplasts from algal food to photosynthesize, so DNA identification of sequestered chloroplasts was employed to verify the algal species fed upon by the slug across its geographic range. The molecular information on the algae consumed by E. clarki was combined with field surveys of slugs and algae in slug habitats in the Florida Keys in July and August of 2008 in order to evaluate whether the diet of this herbivore could be predicted based on its spatial association with algae in the field. A considerable mismatch between food availability and kleptoplast identity was recorded. E. clarki commonly occupied areas devoid of potential food and often contained symbiotic plastids from algal species different from those most frequently found in the surveyed habitats. In three of the four study sites, algal species present were poor predictors of slug diet. These findings suggest that the photosynthetic capability of E. clarki may release the slug from the constraint of requiring proximity to its food sources and may allow for the potential lack of spatial coupling between this herbivore and its algal food. This combination of field surveys and DNA barcoding provided critical and previously unavailable information on herbivore feeding in this marine system.     

Resistance and re-organization of an ecosystem in response to biological invasion: Some hypotheses

Using a dynamic model of Lake Chozas developed by Marchi et al. (2011), we tested three hypotheses about recovery of the indigenous community and water quality after radical changes caused by introduction of an invasive allochthonous crayfish, Procambarus clarkii:

1.

Can the lake resist the pressure of an invasive species, like P. clarkii, by adaptation?

2.

Can the ecosystem recover when all the crayfish are removed and low phosphorus concentrations persist in inflow water?

3.

Does the simulated recovery of submerged vegetation occur at a total phosphorus concentration below 100 mg TP m⁻³, as estimated by Scheffer et al. (1993), Scheffer (1997), Jeppesen et al. (1998) and Zhang et al. (2003)?

We obtained the following answers:

1.

Lake Chozas can at least partly resist by adaptation. A combination of possible parameter changes could lead to a significant increase in eco-exergy.

2.

Removal of the phosphorus represented by crayfish (by harvesting) implies complete recovery of the lake and its eco-exergy, albeit not necessarily with the same organisms having the same properties.

3.

The expected hysteresis created by introduction and harvesting of crayfish is observed under the following conditions: phytoplankton dominance at total phosphorus ≥ about 200-250 mg TP m⁻³ and submerged vegetation returns at total phosphorus < 100 mg TP m⁻³.

     

Underlying reasons of the controversy over adverse effects of Bt toxins on lady beetle and lacewing larvae

In their recent study, Hilbeck et al. (2012) report that Cry1Ab causes lethal effects on larvae of the ladybird beetle Adalia bipunctata when fed directly to the predator. Such toxic effects were not previously observed in a direct feeding study conducted by us (álvarez-Alfageme et al. 2011). Because Hilbeck et al. (2012) claim that our study design did not allow us to detect any adverse effects we provide arguments for the value and relevance of our study in this commentary. Furthermore we discuss two additional published studies that have not revealed any direct effects of Cry1Ab on larvae of A. bipunctata and are not mentioned by Hilbeck et al. (2012). One of the studies was conducted in our laboratory under more realistic exposure conditions (álvarez-Alfageme et al. 2011). Feeding A. bipunctata larvae with spider mites reared on Bt maize did not reveal any adverse effects on lethal and sublethal parameters of the predator. This was despite the fact that the larvae had ingested high amounts of biologically-active Cry1Ab protein. Thus, we do not see verified evidence that A. bipunctata larvae are sensitive to Cry1Ab at realistic worst-case exposure concentrations. This, together with the fact that A. bipunctata will be little exposed to Cry1Ab under field conditions, allows us to conclude that the risk of Bt maize to this predator is negligible. Support for this comes from the results of many Bt maize field studies that have not revealed evidence for direct Cry1Ab-effects on non-Lepidoptera species.     

n-Octyl esters of long-chain fatty acids are not anthropogenic pollution markers

n-Octyl esters of higher fatty acids have been reported as markers of urban wastewater in sediments, polychaeta, fish, crabs and oysters (Chaler et al. in J Chromatogr A 1046:203–210, 2004)). However, up to date, there were no subsequent studies to confirm this claim. Likewise, Chaler and co-workers did not consider that the mentioned compounds might occur naturally in the environment. Here we found seven n-octyl straight-chain alkanoates, from C-20 to C-26, in the wild-growing plant Heracleum sphondylium L., Apiaceae, from 14 locations. Those plant metabolites were unambiguously identified by gas chromatographic co-injection of the synthesized esters with the inflorescence washings. All identified octyl esters represent new natural compounds, except for octyl docosanoate. Since we have demonstrated that n-octyl esters occur naturally, and in abundance, they cannot be recognized any longer as wastewater markers. Additionally, here we provide evidences that the compounds identified by Chaler et al. (2004) are in fact 2-ethylhexyl esters, mistakenly identified as n-octyl esters.     

Metals in Cancer irroratus (Crustacea: Decapoda): Concentrations,concentration factors,discrimination factors,correlations

Nine metals were analysed in whole specimens of the decapod crustancean Cancer irroratus, and three in the exoskeleton of Carcinus maenas. In terms of concentrations (expressed as parts per millions) and concentration factors, 2 groups of metals could be distinguished in Cancer irroratus: Zn, Cu, Fe and Mn in one group, Ca, Mg, Sr, Na and K in the other group. Na excepted, all the metals had concentration factors significantly higher (P<0.01) than 1. The concentration factor of Na was significantly lower (P<0.01) than 1, demonstrating the existence in C. irroratus of a barrier to the movement of Na into the crustacean from its environment. Studies of coefficients of discrimination (D), in C. irroratus, for metals considered in pairs, showed, in each case that D was significantly different (Plt;0.01) from 1. Only the discrimination factor between, Zn and Mn was less significantly different (0.05lt;Plt;0.1) from 1. Correlations were made between the concentrations of metals and other parameters: water content, size, and ash content. Significant correlations were found, especially in the cases of the pairs Ca?Sr, Na?K, Cu?Zn, and Mn-animal size.     

Recent strandings of the giant jellyfish Rhizostoma luteum Quoy and Gaimard, 1827 (Cnidaria: Scyphozoa: Rhizostomeae) on the Atlantic and Mediterranean coasts

We present reports of sightings of living and stranded specimens of Rhizostoma luteum on the Atlantic coast of Morocco and along the south shore of the Iberian Peninsula in June–July 2012 and in January–February 2013. During summer 2012 and following the dominant currents, the jellyfish first appeared in the Gulf of Cadiz west of the Strait of Gibraltar. Subsequently, seven additional sightings were reported east of the Strait, in the Alboran Sea. In winter 2013, another event of stranded individuals of this species occurred in the Gulf of Cadiz. A phylogenetic analysis performed on the mitochondrial cytochrome c oxidase I (COI) gene sequence in specimens from both stranding events confirmed the morphological classification, ratifying that R. luteum differs from Rhizostoma octopus and Rhizostoma pulmo. This study records the presence of this species for the first time in 60 years.     

Component-specific toxic concerns of the inhalable fraction of urban road dust

Continuous global urbanisation causes an ever-growing ecological footprint of pollution. Road dust (RD), one of these pollutants, poses a health concern due to carcinogenic and toxic components potentially present in the micron-sized fractions. The literature reports on the concentrations of trace, toxic metals and metalloids present in RD (Hooker and Nathanail in Chem Geol 226:340–351, 2006), but the literature on its molecular composition is limited. Recent reports on the bioaccessibility of platinum group metals are also reported (Colombo et al. in Chem Geol 226:340–351, 2008). In vitro and animal toxicological studies confirmed that the chemical composition of inhaled particles plays a major role in its toxic, genotoxic and carcinogenic mechanisms, but the component-specific toxic effects are still not understood. Particle-bound airborne transition metals can also lead to the production of reactive oxygen species in lung tissue; a special concern amongst particularly susceptible cohorts (children and elderly). The characterisation of the molecular composition of the fine fraction is evidently of importance for public health. During a pilot study, partially characterised size-fractioned RD samples (Barrett et al. in Eviron Sci Technol 44:2940–2946, 2010) were analysed for their elemental concentration using X-ray fluorescence spectrometry and inductively coupled plasma mass spectrometry. In addition, separately dispersed particles (200 particles per size fraction) were analysed individually by means of computer-controlled electron probe X-ray micro-analysis (CC-EPXMA) and their molecular structure probed by studying elemental associations. These were correlated with micro-Raman spectroscopy (MRS) results. It was found that the fine fraction (<38?μm) had the highest Pb (238?ppm) and Cr (171?ppm) concentrations. The CC-EPXMA data showed >50?% association of Cr-rich particles with Pb, and the MRS data showed that the Cr was mostly present as lead chromate and therefore in the Cr(VI) oxidation state. Concentrations of both Pb and Cr decreased substantially (279 (<38?μm)–13?ppm (<1?mm); 171 (<38?μm)–91?ppm (<?1?mm), respectively) in the larger fractions. Apart from rather alarmingly high concentrations of oxidative stressors (Cu, Fe, Mn), the carcinogenic and toxic potential of the inhalable fraction is evident. Preliminary bioaccessibility data indicated that both Cr and Pb are readily mobilised in artificial lysosomal liquid and up to 19?% of Cr and 47?% of Pb were released.     

Third-stage larva shifts host fish from teleost to elasmobranch in the temporary parasitic isopod, Gnathia trimaculata (Crustacea; Gnathiidae)

The life cycle of the fish ectoparasitic isopod Gnathia trimaculata is described based on both field samplings and laboratory observations. Species identification of the larvae was based on morphological observation and supported by molecular analysis. As the results of field samplings in several sites of southwestern and central Japan (24–34°N, 124–139°E) from 2005–2011, approximately 900 third-stage larvae of G. trimaculata were found on 25 elasmobranch species, and 220 first- and second-stage larvae were found on three teleost species. No third-stage larvae were found on the teleosts, and the larvae of younger stages never infested elasmobranchs. Therefore, G. trimaculata is supposed to shift its host from teleosts to elasmobranchs as it develops. We discuss the developmental periods, life span, distribution, and predation risk of the present species compared with other gnathiid species.     

Fatty acid compositions of neutral glycerides from the ovaries of the asteroids Ctenodiscus crispatus,Asterias lincki and Pteraster militaris from Balsfjorden,northern Norway

Fatty acid analyses of lipids in the asteroids Ctenodiscus crispatus (Retzius), Asterias lincki (Müller and Troschel) and Pteraster militaris (O. F. Müller) collected in Balsfjorden, near Tromsø, northern Norway in 1978–1980, were performed to further our understanding of the asteroids' reproductive biology. Fatty acid compositions were determined in phospholipids, triacylglycerols and alkyldiacylglycerols isolated from ovaries of frozen C. crispatus. The major polyunsaturates in phospholipids were 20:4(n-6) followed by 20:5(n-3). Both the triacylglycerols and alkyldiacylglycerols were relatively deficient in polyunsaturates but had 20:1(n-13) and 18:1(n-7) as major components. Both these liqid classes had additionally 15% of their fatty acids as a variety of odd-numbered and branched-chain fatty acids. The fatty acid composition of neutral glycerides, largely alkyldiacylglycerols, from heat-dried ovaries of C. crispatus was not materially different from that of alkyldiacylglycerols isolated from the ovaries of frozen individuals. The fatty acid composition of neutral glycerides, largely alkyldiacylglycerols, from heat-dried ovaries of P. militaris was similar to that for C. crispatus in that 20:1(n-13) and 18:1(n-7) were major components, and odd-numbered together with branched-chain fatty acids accounted for about 15% of the total fatty acids. The fatty acid composition of neutral glycerides, consisting of equal amounts of triacylglycerols and alkyldiacylglycerols, from heat-dried ovaries of A. lincki differed from those of the previous two species in that 20:1(n-9) and 18:1(n-9) were major components and odd-numbered together with branched-chain fatty acids accounted for less than 5% of the total fatty acids. The results are discussed mainly in terms of lipid biosynthetic mechanisms within the asteroids and the interplay between internal biosynthetic activities and dietary fatty acids, particularly those from bacterial sources, is considered.     

Four genes,morphology and ecology: distinguishing a new species of <Emphasis Type="Italic">Acesta</Emphasis> (Mollusca; Bivalvia) from the Gulf of Mexico

There is currently an active scientific debate about the most correct and efficient way to identify species. To date, few studies in the marine realm have combined the available taxonomic methods. In this study we have used morphology, ecology and molecular analyses to identify a new species within the bivalve genus Acesta. All four genes studied (12S, 16S, Cytb, COI) suggested that a common cold-seep species in the Gulf of Mexico, A. bullisi, should be divided into two distinct species. This conclusion was supported by morphological traits and by observations of ecological distribution. A. oophaga Järnegren, Schander and Young n. sp. is described here, and A. bullisi Vokes (Tulane Stud Geol 2:75–92, 1963) is re-described. This study shows that DNA barcoding in combination with traditional morphological and ecological analyses may be an important tools to identify hidden biodiversity among deep-water organisms such as bivalves.     

The free energy and information embodied in the amino acid chains of organisms

It is generally accepted as a useful and workable hypothesis that when an ecosystem receives an inflow of exergy (energy that can do work) it will utilize this flow of exergy to move as far away from thermodynamic equilibrium as possible after the exergy (energy) for maintenance has been covered. If more combinations of system components including organisms are offered, the combination of components and processes that will bring the system most away from thermodynamic equilibrium will win.The amino acid sequences of the proteins e.g. enzymes determine and control the life processes of the organisms and may be viewed as information sensu lato. The free energy of oxidation of the amino acids and the peptide bonds of the cell enzymes expresses therefore the exergy content, eco-exergy or work capacity that the information contributes to “moving further away from thermodynamic equilibrium”. In this paper eco-exergy is calculated and plotted versus the β-values (a measure of the information contained in the genome) for different organisms. The eco-exergy density was previously (see [J?rgensen et al., 1995] and [J?rgensen et al., 2005]) proposed to be calculated as the summation of the product of the β-values representing the information of the genome multiplied by the concentrations of the respective ecosystem components. This analysis shows a strong correlation between the β-values and free energy released when oxidizing the enzymes. The β-values can therefore be assumed to represent the free energy that the organisms have invested in genetic information.     

Low variation in partial cytochrome oxidase subunit I (COI) mitochondrial sequences in the coralline demosponge Astrosclera willeyana across the Indo-Pacific

Partial sequences of the mitochondrial DNA (mtDNA) gene cytochrome oxidase subunit 1 (COI) were analysed from individuals of the coralline demosponge Astrosclera willeyana sensu lato out of ten Indo-Pacific populations from the Red Sea to the central Pacific. This taxon is widely distributed in cryptic coral reef habitats of the Indo-Pacific and is regarded as a modern representative of long-extinct, formerly reef-building stromatoporoid-type sponges. The aims were to clarify phylogeographic and taxonomic relationships in this “living fossil” and to explore mitochondrial DNA sequence variation over a wide geographic range. Very low variability was observed across the Indo-Pacific, as only three COI haplotypes were identified, with a maximum p-distance of 0.418% and low nucleotide diversity (π=0.00049). Very low genetic structure was revealed among populations: Haplotype 1 was found in all specimens from nine Pacific populations (N=45), separated by distances of more than 7,000 km; haplotype 2 was restricted to the Red Sea population (N=4); haplotype 3 was only found in the Tuamoto specimens (N=7). COI data presented here do not support the hypothesis of at least two sibling species belonging to genus Astrosclera in the Pacific. Considering the maximum geographic distance of more than 20,000 km between sampled populations, mtDNA COI sequence variation observed here is among the lowest reported to date for a diploblastic taxon and adds to the growing evidence of a general mtDNA conservation in sponges. It is argued that this low mtDNA variation in A. willeyana s.l. is due to a low rate of mtDNA evolution caused by a combination of long generation time and low metabolic rate.     

Response to Kruse-Plass et al. (2017) regarding the risk to non-target lepidopteran larvae exposed to pollen from one or more of three Bt maize events (MON810, Bt11 and 1507)

We respond to the paper of Kruse-Plass et al. (Environ Sci Eur 29:12, 2017), published in this journal, regarding the risk to non-target lepidopteran larvae exposed to pollen from one or more of three Bt maize events (MON810, Bt11 and 1507). We emphasise that what is important for environmental risk assessment is not the number of pollen grains per se, but the degree of exposure of a NT lepidopteran larva to Bt protein contained in maize pollen. The main text of this response deals with general issues which Kruse-Plass et al. have failed to understand; more detailed refutations of each of their claims are given in Additional file 1. Valid environmental risk assessment requires direct measurement of pollen on leaves at varying distances outside a source field(s); such measurements reflect the potential exposure experienced by an individual larva on a host plant. There are no new data in the Kruse-Plass et al. paper, or indeed any data directly quantifying pollen on actual host-plant leaves outside a maize field; only data gathered within or at the edge of maize crops were reported. Values quoted by Kruse-Plass et al. for deposition on host plants outside the field were estimates only. We reiterate the severe methodological criticisms made by EFSA [Relevance of a new scientific publication (Hofmann et al. 2016) for previous environmental risk assessment conclusions and risk management recommendations on the cultivation of Bt-maize events MON810, Bt11 and 1507. EFSA Supp Publ; EN-1070, 2016], which render this estimation procedure unreliable. Furthermore, criticisms of EFSA (EFSA J 2015(13):4127, 2015) and of EFSA [Relevance of a new scientific publication (Hofmann et al. 2016) for previous environmental risk assessment conclusions and risk management recommendations on the cultivation of Bt-maize events MON810, Bt11 and 1507. EFSA Supp Publ; EN-1070, 2016] made by Kruse-Plass et al. are shown in Additional file 1 to be without foundation. We therefore consider that there is no valid evidence presented by Kruse-Plass et al. to justify their conclusions.     

Shearwater diet during migration along the coast of Rio Grande do Sul,Brazil

The coastline of the state of Rio Grande do Sul (Brazil) is used annually as a migratory route by a number of seabird species. Thirteen surveys were carried out between July 1997 and July 1998 to obtain information on the diet of Puffinus gravis, Puffinus griseus, and Puffinus puffinus along the coast of Rio Grande do Sul. Sampling efforts focused on dead beach-cast specimens. One hundred twenty-one stomach content analyses were performed on P. gravis from a total of 238 specimens; 17 stomach content analyses were performed on P. griseus from a total of 54 specimens; and 25 stomach content analyses were performed on 25 P. puffinus from a total of 61 dead specimens. Cephalopods were the main prey found in the stomachs of the three species and were present in 96% of the P. gravis stomachs. The following cephalopods were observed: Ommastrephidae, Gonatus antarcticus, Gonatus sp., Lycoteuthis lorigera, Lycoteuthis sp., Histioteuthis sp., Cycloteuthis sp., Loligo plei, Loligo sanpaulensis, Sepiidae, and Argonauta nodosa. The latter was the most abundant species in the three species of Puffinus. The following fish were identified from otoliths: Porichthys porosissimus, Ctenosciaena gracilicirrhus, Paralonchurus brasiliensis, Cynoscion guatucupa, and Trichiurus lepturus. There was also a high incidence of synthetic materials such as plastic, polyethylene and rubber.     

A study of osteological and molecular differences in populations of Aphanius fasciatus Nardo 1827, from the central Mediterranean (Teleostei, Cyprinodontidae)

Nine populations of Aphanius fasciatus Nardo, 1827 from the central Mediterranean were analysed by examining the mitochondrial control region and the morphology of the bony elements of the skull and vertebral column, to study the degree of intraspecific differentiation of A. fasciatus considering the level of isolation of the different populations and the palaeogeographic history of the central Mediterranean area. Both the molecular and morphological analyses differentiate between the populations, even if the topologies of the two trees are different. Molecular and osteological investigations have consistently demonstrated a well-supported differentiation of the south-eastern Sicilian populations both within the same group (Tigano et al. in Ital J Zool 71:1124–1133, 2004a; Tigano et al. in Abstract volume XI European Congress of Ichthyology, Tallin, Estonia, 2004b), and from the populations from western Sicily, Tunisia and the island of Malta. The molecular results show that the nine populations are characterised by haplotypes that are well defined in relation to a probably limited gene flow; while, as regards the morphological data the differentiation found could be explained in terms of the geographic isolation of the various populations, although the influence of environmental factors, which differ greatly between the various sites where the populations live, cannot be ruled out.     

Increased seawater temperature and light during early springs accelerate receptacle growth of Fucus vesiculosus in the northern Baltic proper

In many temperate estuarine areas, ongoing climatic changes are expected to lead to higher seawater temperatures, to increased inflow of freshwater and nutrients (due to increased rainfall) and to altered light conditions. During the last two decades, several winters have been unusually mild in southern Finland, and the subsequent springs have been early. This may have consequences for the seasonality of many species, including the bladder-wrack, Fucus vesiculosus, whose reproductive cycle may be especially sensitive. In the present study, F. vesiculosus receptacles from the field were weighed regularly during three warm (early) springs and compared with receptacles from three cold (late) springs. On average, the first sign of receptacle growth in the field occurred 5–6?weeks earlier and receptacles probably matured 2–3?weeks earlier during the warm springs than during the cold springs. This may be due to differences in seawater temperatures among years, but significantly heavier receptacles at 0.8?m compared to at 3.1?m further suggests the importance of light for receptacle growth as a measure of maturation, since there were no depth-related differences in seawater temperatures. Also, when plotting receptacle wet weights against seawater temperatures, the importance of light can be seen as different regression relationships (slopes and intercepts). In an outdoor aquarium experiment, the effects of temperature and light on receptacle growth were also tested, and there were significant differences between shallow and deep specimens, but temperature and light had no effects, except on initial receptacle growth. Since oogonian development, and thereby the time for zygote release, followed the receptacle growth curve, this could mean a mismatch between the timing of F. vesiculosus zygote settlement and the cover of competing filamentous algae, but such species interactions require further investigations in order to better understand their possible consequences.