Study of the Nucleolar Organizer Regions in Palinurus elephas (Crustacea: Decapoda)

Using fluorescence in situ hybridization (FISH), chromomycin (CMA₃) staining and silver staining, we studied the nucleolar organizer regions in the spiny lobster Palinurus elephas in order to extend our knowledge on the karyology of this commercially important species. Multiple NORs have been detected by FISH, and CMA₃ showed a good correspondence between the localization of GC-rich heterochromatin and the ribosomal genes mapped by FISH. In contrast, the number of Ag-positive regions was higher than the number of FISH and CMA₃ signals, which may be explained by silver staining of the kinetochores. A variability in the number of FISH and CMA₃ signals has been detected in metaphases I and II which is probably due to the occurrence of rDNA cistrons on B chromosomes.     

Detecting immigrants in a highly genetically homogeneous spiny lobster population (Palinurus elephas) in the northwest Mediterranean Sea

We investigated the genetic structure of early benthic juveniles of the spiny lobster Palinurus elephas in the northwest Mediterranean Sea by means of ten polymorphic microsatellite markers. Non‐metric Multidimensional Scaling coupled with assignment tests were used as a new approach to further delimit a reference population inside a genetically homogeneous pool of individuals and test for the presence of long distance immigrants. From this approach, we found that most early benthic juveniles collected while settling in the northwest Mediterranean Sea originated from a common larval pool. However, 4.2% of the individuals were classified as immigrants from other genetically differentiated populations, with more immigrants in the south than in the north of the sampled basin. Given currents in the northwest Mediterranean Sea and the long pelagic larval phase of P. elephas that lasts several months, this result suggest a restricted homogenized zone in the studied basin with some individuals probably coming from more differentiated populations through the Almeria‐Oran Front or the Strait of Sicily.     

Main phospholipids and their fatty acid composition in muscle and cephalothorax of the edible Mediterranean crustacean Palinurus vulgaris (spiny lobster)

The total lipids of muscle and cephalothorax of Mediterranean lobster Palinurus vulgaris were found to be 1.0% and 2.4% of the wet tissue of which the phospholipids represented 66.5% and 47.5%, respectively. The main PhL saturated, monounsaturated and polyunsaturated fatty acids in muscle and cephalothorax were C16:0, C18:0, C18:1ω-9, C18:1ω-7, C20:4ω-6, C20:5ω-3 and C22:6ω-3. 2-OH C14:0 and cyclo-17:0 fatty acids were also identified though in low percentages. The main individual PhL in muscle were found to be phosphatidylcholine (53.5%), 72.0% of which corresponded to the structure of 1,2-diacyl-glycerocholine while the rest 28.0% to 1-O-alkyl-2-acyl-glycerocholine or 1-O-(1-alkenyl)-2-acyl-glycerocholine and phosphatidylethanolamine (19.3%), 75.0% of which corresponded to the structure of 1,2-diacyl-glyceroethanolamine and 25% to 1-O-alkyl-2-acyl-glyceroethanolamine or 1-O-(1-alkenyl)-2-acyl-glyceroethanolamine. Cephalothorax main PhL were found to be PC and PE (66.4% and 18.8%, respectively). In muscle and cephalothorax PC ω-3 fatty acids amounted 7.78% and 8.60%, while in PE amounted 30.77% and 23.65% respectively. Furthermore, in both tissues PhL, cardiolipine phosphatidylserine, phosphatidylinositol, sphingomyelin and lysophosphatidylcholine, were also found.     

Behavioral thermoregulation in the spiny lobster Panulirus Argus (latreille)

Ten spiny lobsters (Panulirus argus) were allowed to thermoregulate individually for 3-day periods in an electronic thermoregulatory shuttlebox which allowed them to control water temperatures (and thereby their own body temperatures) by their movements. The range of preferred (voluntarily occupied) temperatures was 25–35°C (mean 29.9°C; mode 30.0°C; median 30.0°C; midpoint 30.0°C; S_k (skewness, Pearson's coefficient) –0.04; s.e.m. 0.19°C; S.D. 2.32°C). The final thermal preferendum (by the gravitation method) in this species is 30°C.     

Boat anchoring on Posidonia oceanica beds in a marine protected area (Italy, western Mediterranean): effect of anchor types in different anchoring stages

Seagrasses worldwide are noted for suffering from mechanical damage caused by boat anchoring. This is particularly so in sites highly frequented by boaters (marine protected areas or coastal urbanised areas). In the last decades, different strategies have been put into practice to reduce such impacts on seagrasses (i.e. by anchoring bans or by deploying boat moorings). More recently, in consideration that few marine protected area (MPA) management bodies or local administrations have the resources to enforce their anchorage regulations, the self-regulatory approach based on education and information of boaters has been preferred in several cases. At present, however, very little is known on the correct anchoring practices to ensure the safeguarding of seagrasses. The aim of the present study was to experimentally quantify in the field the damage caused to Posidonia oceanica shoot density by anchoring. A multifactorial experiment was designed to test whether the damage is dependent on (1) different anchor types (Hall, Danforth and Folding grapnel), (2) the use of a chain vs. a rope, (3) the three anchoring stages (anchor fall, dragging/lock-in and weighing), and finally (4) whether the pattern is consistent among different locations of the meadow.As expected, the three anchor types employed in the present study differed in the levels of damage inflicted on the P. oceanica meadows of the Ustica Island MPA. In particular, the use of the Hall type anchor seems to be preferable to minimise this impact in comparison with the other two anchor types. Moreover, the effect on the meadow of the three anchor types is greatly dependent on the anchoring stage. These results confirm that the weighing stage is the critical stage of the anchoring process. The number of damaged shoots of P. oceanica was not affected by the presence of the chain. These patterns were consistent between locations.In the long term, even anchoring on P. oceanica by small boats using low-impact anchors may potentially have detrimental consequences. For this reason, we suggest that in vulnerable sites, it is preferable to implement an educational program based on information of boaters on correct anchoring practices and anchor typology to use, rather than adopting strong restrictions to boat anchoring or deploying mooring buoys. Although the use of these management strategies is still recommended in the case of anchorage frequented by bigger vessels using heavier anchors and chains.     

Rosette-type tegumental glands associated with aesthetasc sensilla in the olfactory organ of the Caribbean spiny lobster, Panulirus argus

The lateral antennular flagellum of decapod crustaceans bears unique olfactory sensilla, namely the aesthetascs, and other sensilla types. In this study, we identify a new major tissue in the lateral flagellum of the Caribbean spiny lobster, Panulirus argus, namely “aesthetasc tegumental glands” (ATGs), based on immunostaining with antibodies against CUB serine protease (Csp), in situ hybridization with csp-specific probes, labeling with the F-actin marker phalloidin, labeling with the nuclear marker Hoechst 33258, and staining with methylene blue. Each ATG has 12–20 secretory cells arranged in a rosette. Each secretory cell has a Csp-immunoreactive basal portion and an apical portion containing granular material (metachromatic staining indicative of acid mucopolysaccharides). At the center of each secretory rosette is a phalloidin-positive common locus that gives rise to a main drainage duct projecting toward the cuticle. Scanning electron and light microscopy show that thin ducts traverse the cuticle and connect to “peg pores” proximal to the bases of the aesthetascs, with 3.4 peg pores per aesthetasc. Since the number of common loci is correlated with the number of peg pores, we conclude that each pore represents the outlet of one ATG, and that the secretions are released from them. We conclude further that ATGs and aesthetascs are functionally linked. We hypothesize that ATG secretions have antifouling and/or friction-reducing properties, and that they are spread over the surface of the aesthetascs by antennular grooming. A review of the literature suggests that ATGs are common in decapod crustacean antennules, and that rosette glands and grooming might be functionally coupled in other body areas.This study was supported by NSF IBN 0077474 and NIH DC00312.     

Planktonic chlorophyll a and eutrophication in two Mediterranean littoral systems (Mallorca Island,Spain)

Two Mediterranean littoral systems of Mallorca Island have been studied using planktonic chlorophyll a (total concentration and seasonal variations) as trophic indicator. The studied systems are quite different in geomorphology, physicochemical trends and anthropic incidence. S'Albufera-Alcudia Bay is the drainage outlet of a coastal marsh, with a salt wedge estuarine regime. Andratx Harbour is a narrow bay affected by a high anthropic incidence due to the presence of a small fishing port and mainly a large leisure harbour. Both systems are affected by a typical Mediterranean climate and a negligible tide incidence. The present work is based on several year monitoring of physical, chemical and biological water parameters. In S'Albufera-Alcudia Bay, Chl a concentration ranged from 0.01 to 17.10 mg m^–3, the values dropping from the inland site to the sea. The phytoplankton biomass gradient takes its origin in physical and chemical differences. The variance is mainly related to the inland-sea water axis. Nitrogen and silica enriched inland waters are the main source of eutrophication, hence their relation with Chl a. These nutrients entering the Albufera are mainly derived from leaching from farmed soils. The transition from an inland eutrophic system to an oligotrophic marine one show the filter character of these coastal marshes. In Andratx Harbour, Chl a ranged from a non detectable concentration to 6.36 mg m^–3. The ultraplankton (<5 m) was the most important contributor to total concentration. The ultraplankton in this system is related to influence of more oligotrophic open seawater. A greater proportion of microplankton (>20 m) is related both to higher nutrient concentrations and a higher water column stability. In Andratx Harbour, the enclosure state seems very important for development of phytoplankton blooms.     

Diagnostics of nocturnal calls of Sciaena umbra (L., fam. Sciaenidae) in a nearshore Mediterranean marine reserve

Calls emitted by the brown meagre Sciaena umbra (L., fam. Sciaenidae) were recorded at the Natural Marine Reserve of Miramare (Trieste, Italy) in seven nocturnal surveys (12-h continuous sampling) during the summer of 2009. Calls consist of pulses, with the main energy content below 2 kHz and mean peak frequency of c. 270 Hz. Pulses were short, with an average duration of 20 ms and a pulse period of 100 ms. Sounds lasted approximately 500 ms. Three types of sound patterns were recognized: irregular (I), regular (R) and the chorus (C). Their acoustic parameters are described showing that I, R and C differ in pulse duration, pulse peak frequency and pulse period. Occurrence of the three call types changes throughout the night: the R pattern occurred mainly at dawn and dusk, C predominated after nightfall, while I calls were produced sporadically during the whole nocturnal period. Our results indicate that S. umbra has a pronounced nocturnal rhythm in vocalizing behaviour and highlight how the diagnostic time–frequency pattern of S. umbra calls can be used to identify the species in the field. Considering that the abundance of S. umbra is currently declining, the information presented here will be relevant in developing non-invasive and low-cost monitoring acoustic systems for managing S. umbra conservation and fishery along the Mediterranean Sea.     

TheLecanactis grumulosa group (Opegraphaceae) in the Mediterranean region

The morphology, anatomy, and chemistry of five taxa belonging to theLecanactis grumulosa group from several localities of the Mediterranean area have been analysed in detail. One new saxicolous species,L. subgrumulosa, is described from Spain and Morocco. The new combinationL. farinosa is made.L. nothiza, L. monstrosa, L. ramosus, andOpegrapha alboatra are proposed as synonyms ofL. grumulosa. L. werneri andL. farinosa are cited for the first time in Europe and northern Africa, respectively. Anthraquinones are reported as new forLecanactis.     

Fishery of the Deep-water Rose Shrimp Parapenaeus longirostris (Lucas, 1846) (Crustacea: Decapoda) in the Northern Tyrrhenian Sea (Western Mediterranean)

The aim of this study is to provide information on the fishing pattern of Parapenaeus longirostris in the northern Tyrrhenian Sea (western Mediterranean), in order to improve the management of the resource in the area. Data were obtained from commercial fishery, covering the period 1991–2002. Catch rates revealed notable inter-annual fluctuation of the P. longirostris landing, probably due to the life cycle of the species. The highest yields (kg per hour of trawl) of the commercial fishery were obtained at depths ranging between 180 and 300 m, the lower ones at depths between 100 and 180 m, where small specimens are more abundant. Commercial catches were characterised by a very low presence of specimens smaller than 20 mm carapace length (CL), mainly due to the deeper localisation of the fishing grounds when compared to the recruitment areas of the species. The size at which 50% of the specimens were discarded by the fishermen was 15 mm CL, while the estimated size at first capture of the individuals caught by the commercial net was 12.4 mm CL.     

Importance of scrub–pastureland mosaics for wild-living cats occurrence in a Mediterranean area: implications for the conservation of the wildcat (Felis silvestris)

The European wildcat (Felis silvestris) is a threatened species in Europe. Suitable management of forests has been considered crucial for its conservation in Europe. However, this recommendation may not be general due to the lack of studies that test these hypotheses in the Mediterranean area, where landscapes are very different from those of central-north Europe. In this study, wild-living cat habitat associations were analyzed by means of scat surveys in 78 areas distributed in the four main vegetation types of the Mediterranean area of central Spain, where feral cat populations are probably scarce and restricted. Results show higher occurrences of wild-living cats in landscapes covered by scrub–pastureland mosaics rather than forests. Several applied recommendations are given: (1) to include the scrub–pastureland mosaics as protected habitats for wildcats; (2) to encourage further studies about the importance of this habitat in other areas; (3) to avoid the extensive scrubland removal associated with management practices against fires or infrastructure development; and (4) to promote land management practices that enhance these mosaics, and to use shrub species in the reforestation programmes founded by the European Agricultural Policy.     

Feeding habits of grey gurnard,Eutrigla gurnardus (L., 1758), along the Catalan coast (northwestern Mediterranean)

The stomach content of 589 specimens of Eutrigla gurnardus (collected at quarterly intervals) have been analyzed to determine diet according to fish size and season. The results show that crustaceans (% No = 93%, %Weight = 81%) are the basic food and that teleostean fishes are quite important in the adult diet. The importance of nycthemeral migrators decreases significantly as size increases, whilst the importance of benthic prey increases inversely. In every size, the feeding intensity is greater at midday than in the morning. Larger prey are eaten in winter than in summer although feeding intensity is lower in winter.     

Searching for the early benthic phase (EBP) of the European lobster: a trans-European study of cobble fauna

The re-stocking or enhancement of Homarid populations has been the focus of several research projects in recent years. However, despite being able to successfully rear lobsters in captivity, there are few reports of newly settled European lobsters (Homarus gammarus) in the wild. This is of serious concern for fishery managers when trying to ascertain if restocking or enhancement programmes are successful at the fishery level. Based on published work from the western Atlantic seaboard, cobble and boulder substrata have been identified as an important nursery habitat for early benthic phase (EBP) American lobsters (Homarus americanus). This paper, based on an E.U. funded research project (LEAR), describes the results of quantitative airlift suction sampling from cobble habitat in Norway, Ireland the U.K., and Italy. In total, 67 m² of cobble was sampled, but no EBP lobsters were found. Overall, crustaceans and molluscs were the most abundant species in all countries. While the collective densities of animals per m²of cobble were similar to that of the United States, the species diversity in Europe was noticeably higher. Among the crustacea, the reptant decapods dominated at all sites. These findings tend to support the modern day `exclusion hypothesis' that competition and predation from other species limits the successful recruitment of EBP European lobster to the benthos. The results highlight the need to go beyond the single species approach to understanding the factors that influence the recruitment of Homarus gammarus.     

Phenetic,biogeographical, and evolutionary study ofOrnithogalum subg.Heliocharmos (Hyacinthaceae) in the western Mediterranean basin

A macromorphological study is made on taxa of the genusOrnithogalum subg.Heliocharmos in North Africa, Spain, and France. The results obtained are consistent with data from cytogenetics, reproductive biology and strategies of reproduction. They allow the retention of two species:O. algeriense andO. umbellatum. A biogeographical and phylogenetic interpretation of the subgenus is proposed for the western Mediterranean. Theoretical views on phenetics are discussed.     

Calcareous and siliceous phytoplankton stratigraphy of Neogene marine sediments in central Crete (Greece)

Fine laminated diatomaceous beds in outcrops of the Finikia unit (Estavromenos and Athanatoi sections) in the northern part of the Heraklion district in Crete, have yielded a total of 55 marine diatom and 24 silicoflagellate taxa. The diatom assemblages, characterized by species of low latitudes, belong to the upper part of the Nitzschia jouseae biozone (early-late Pliocene). The silicoflagellate associations belong to the local Dictyocha fibula ausonia and Dictyocha hellenica subzones. These subzones correspond to the subzones CN11b (Discoaster asymmetricus) and CN12a (D. tamalis) of calcareous nannofossils respectively, as well as to the Globorotalia puncticulata/G. bononiensis biozones of planktic foraminifera. The sparse silicoflagellate association from fine sandy marls of the Panassos section in the central part of the Heraklion district (Aghia Varvara unit, upper Tortonian-Messinian) is assigned to the local biozone Distephanus speculum minutus. The latter corresponds approximately to the biozone C9 (Discoaster quinqueramus) of calcareous nannofossils and to the Globorotalia conomiozea zone of planktic foraminifera. No discoidal diatoms could be found in the samples from Panassos-section. The presence of silicoflagellates but simultaneous absence of diatoms is probably due to greater opal dissolution within the fine sandy marls of the Panassos section.     

Molecular evidence for the occurrence of Contracaecum rudolphii A (Nematoda: Anisakidae) in shag Phalacrocorax aristotelis (Linnaeus) (Aves: Phalacrocoracidae) from Sardinia (western Mediterranean Sea)

Specimens of Contracaecum rudolphii Hartwich, 1964 (Nematoda: Anisakidae) from Phalacrocorax aristotelis (Linnaeus) from the Archipelago of La Maddalena (Sardinia, western Mediterranean Sea) were characterised genetically and compared with C. rudolphii A sensu D’Amelio et al. 1990 and C. rudolphii B sensu D’Amelio et al. 1990 from Phalacrocorax carbo sinensis (Blumenbach) from north-eastern Italy, and with C. rudolphii C sensu D’Amelio et al. 2007 from Phalacrocorax auritus (Lesson) from west-central Florida, USA. The sequencing of the small subunit of the mitochondrial ribosomal RNA gene (rrnS) and by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) analysis of the same gene and of the internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA) allowed the identification of all specimens of C. rudolphii from P. aristotelis as C. rudolphii A. The results confirmed that the definition of genetic markers, following the analysis of nuclear ribosomal and mitochondrial DNA, provides quick and practical diagnostic tools for the detection of the 3 sibling species of C. rudolphii. The occurrence of C. rudolphii in P. aristotelis is reported for the first time from the Mediterranean area, improving the picture of the dispersal patterns of the populations of these piscivorous birds, and confirming the existence of different and isolated populations between the North and South European waters.     

Climatic factors on entomopathogenic hyphomycetes infection of Trialeurodes vaporariorum (Homoptera: Aleyrodidae) in Mediterranean glasshouse tomato

Collaborative research was conducted at the INRA Research Centers to assess the microbial control potential of Beauveria bassiana- and Lecanicillium lecanii-based formulations against whiteflies in protected crops under Mediterranean conditions. Four series of small-scale glasshouse trials were performed in 1999 and 2000 in southern France. Two applications at 4–5 day intervals of Naturalis-L and Mycotal were conducted on young larvae of the greenhouse whitefly, Trialeurodes vaporariorum, at rates recommended by the manufacturers. Because of the expectation that environmental conditions prevailing in Mediterranean greenhouse crops may lead to greater climatic constraints for mycoinsecticide efficacy than in more temperate areas, manipulation of the greenhouse climate has been used to aim at optimizing mycoinsecticide efficacy. The climatic management strategy was mainly based on closing the ridge vents 2 h more at night-time in so-called “humid” glasshouse compartment than in a “dry” one. Thus, the daily period at high humidity (>90% RH) was two or three times longer in the “humid” compartment than in the “dry” one. In spite of this differential, mycoinsecticide treatments reduced numbers of surviving whitefly larvae by >85% in the “humid” compartment as expected as favorable, as well as in the “dry” compartment, expected as unfavorable. The results indicated clearly that both B. bassiana- and L. lecanii-based mycoinsecticides have a strong potential for microbial control of whitefly larvae infesting tomato crops at moderate ambient humidity in Mediterranean glasshouses. Our investigations provided strong arguments for explaining these unexpected results. The RH conditions prevailing in the targeted insect habitat should be greatly disconnected from that of the ambient glasshouse air. We suggest that strategies of mycoinsecticide optimization against phyllophagous insects in protected crops have to take into account factors acting on the leaf transpiration activity.     

Desulfuromonas palmitatis sp. nov., a marine dissimilatory Fe(III) reducer that can oxidize long-chain fatty acids

Studies on the microorganisms living in hydrocarbon-contaminated sediments in San Diego Bay, California led to the isolation of a novel Fe(III)-reducing microorganism. This organism, designated strain SDBY1, was an obligately anaerobic, non-motile, non-flagellated, gram-negative rod. Strain SDBY1 conserves energy to support growth from the oxidation of acetate, lactate, succinate, fumarate, laurate, palmitate, or stearate. H₂ was also oxidized with the reduction of Fe(III), but growth with H₂ as the sole electron donor was not observed. In addition to various forms of soluble and insoluble Fe(III), strain SDBY1 also coupled growth to the reduction of fumarate, Mn(IV), or S⁰. Air-oxidizedminus dithionite-reduced difference spectra exhibited peaks at 552.8, 523.6, and 422.8 nm, indicative ofc-type cytochrome(s). Strain SDBY1 shares physiological characteristics with organisms in the generaGeobacter, Pelobacter, andDesulfuromonas. Detailed analysis of the 16S rRNA sequence indicated that strain SDBY1 should be placed in the genusDesulfuromonas. The new species nameDesulfuromonas palmitatis is proposed.D. palmitatis is only the second marine organism found (afterD. acetoxidans) to oxidize multicarbon organic compounds completely to carbon dioxide with Fe(III) as an electron acceptor and provides the first pure culture model for the oxidation of long-chain fatty acids coupled to Fe(III) reduction.     

Field studies of seahorse population density,structure and habitat use in a semi-closed north-eastern Mediterranean marine area (Stratoni,North Aegean Sea)

The present study was carried out in the marine area of Stratoni, Greece, where two seahorse species are present (Hippocampus hippocampus and Hippocampus guttulatus). Two surveys were conducted (September 2016 and May 2019) to gather information regarding seahorse species' abundance, distribution and habitat characteristics. Four different seahorse natural and artificial habitat types were identified. The results revealed that the presence of H. hippocampus was relatively high, especially at sites with artificial structures, whereas the presence of H. guttulatus was rare. Data collected can provide baseline information for future population assessments.     

The study of nest-site preferences in Eleonora’s falcon Falco eleonorae through digital terrain models on a western Mediterranean island

This study addresses nesting-site preferences in Eleonora’s falcon Falco eleonorae by means of geographic information systems and high-quality high-resolution digital terrain models (DTMs). A small western Mediterranean archipelago (Columbretes Islands) with a colony of 34 pairs was chosen as a case study. Overall, 21 grid variables obtained from DTM were considered of which 12 showed a significant effect. Four of them were topographic (i.e. elevation, X-axis aspect, slope and curvature), four climatic (i.e. wind exposure, solar insolation, solar radiation and radiation index), three anthropogne (visibility of lighthouse beam, visibility of antropic zones and distance to antropic zones) and one biotic (vegetation cover). Falcon nests were placed on pixel cells with lower average insolation and radiation than cells without nests, but had higher values of wind exposure. Nests were built on concave sites mainly with an easterly facing aspect and steep slopes. Falcons were shown to respond to human presence as all antropic variables considered had a significant effect. The results have a number of management and conservation implications, because the knowledge of nesting preferences of the species allows the proper design of sanctuaries and an appropriate human use of these areas.