Temporal and ontogenetic variation in the diet of squid (<Emphasis Type="Italic">Loligo forbesii</Emphasis> Streenstrup) in Scottish waters

The squid Loligo forbesii is the only cephalopod species currently targeted by fisheries in the northern NE Atlantic. An active predator, it feeds primarily on fish, crustaceans and cephalopods. During 15 years since the only previous large-scale study of the diet of this species in Scottish waters, there have been substantial changes in marine fish abundances. The present study evaluates sources of variation (temporal, ontogenetic) in diet composition and prey size preferences of L. forbesii, including a comparison of contemporary (July 2006–June 2007) and historical (1990–1992) dietary datasets. Results revealed significant size-related and seasonal variation in diet composition and prey size. Teleost fish of the families Ammodytidae and Gobiidae were eaten by squid of all sampled sizes, although occurrence of gobies was generally more frequent in smaller squids, while Gadidae were eaten more frequently by larger squids. Cannibalism was also more frequent in larger squids. Compared to the 1990–1992 dataset, clupeid fish were less important in the diet of squid in 2006–2007, while the importance of gobies increased, and the size of gobies eaten also increased. The trend in gadoids differed according to the index used: their frequency of occurrence was considerably higher in 2006–2007 than in 1990–1992, but their numerical importance was slightly lower. In general, results provided little evidence that changes in the diet of L. forbesii correspond with changes in fish abundance, at least at the scales at which these are measured.     

A multivariate morphometric study of the iberian representatives of<Emphasis Type="Italic">Potentialla</Emphasis> sect.<Emphasis Type="Italic">Recta</Emphasis> (<Emphasis Type="Italic">Rosaceae</Emphasis>)

A multivariate analysis has been performed in order to study the polymorphisms of the three species ofPotentilla sect.Recta (Th. Wolf)Juzepchuk represented in the Iberian Peninsula:P. asturica Rothm.,P. hirta L. andP. recta L. The variability of 20 characters (quantitative, binary and qualitative) has been evaluated in 76 collections mostly from the Iberian Peninsula and from other European territories, but also from North America, and North Africa. The analyses performed provide support for the recognition of the three species. The most discriminant among the quantitative characters is the width of the long trichomes on the stem, and then the width of the long trichomes on the leaves and the maximum length of the long trichomes. The interval of number of carpels, presence or absence of short eglandular trichomes and the abundance of long eglandular trichomes (>3 mm) are the most discriminant ones among the qualitative ones. The taxonomical position ofP. asturica, which has been previously subordinated as a subspecies within the other two taxa, is discussed. Our results support a good delimitation of this taxon which, on the basis of the morphological characters studied by us, seems to be morphologically more close toP. hirta than toP. recta, but differs significantly from both. A key to the three species is provided.     

Cytoplasmic incompatibility involving <Emphasis Type="Italic">Cardinium</Emphasis> and <Emphasis Type="Italic">Wolbachia</Emphasis> in the white-backed planthopper <Emphasis Type="Italic">Sogatella furcifera</Emphasis> (Hemiptera: Delphacidae)

Cytoplasmic incompatibility (CI) is a reproductive phenotype induced by bacterial endosymbionts in arthropods. Measured as a reduction in egg hatchability resulting from the crossing of uninfected females with bacteria-infected males, CI increases the frequency of bacteria-infected hosts by restricting the fertilization opportunities of uninfected hosts in populations. Wolbachia, a type of alpha-proteobacteria, is well known as a CI inducer in a wide range of arthropod species, while Cardinium, a member of the phylum Bacteroidetes, is known to cause CI in one wasp and three spider mite species. In this study, dual infection with Cardinium and Wolbachia induced strong CI in a single host, Sogatella furcifera (Horváth), a planthopper species that is naturally infected with both bacteria. Specifically, infection with Cardinium alone was found to cause a 76 % reduction in egg development, and dual infection with Cardinium and Wolbachia a 96 % reduction, indicating that Cardinium induces CI and the dual infection raises the CI level. This study was the first to document reproductive alteration by Cardinium in a diploid host species.     

Growth and aroma contribution of <Emphasis Type="Italic">Microbacterium foliorum</Emphasis>, <Emphasis Type="Italic">Proteus vulgaris</Emphasis> and <Emphasis Type="Italic">Psychrobacter</Emphasis> sp. during ripening in a cheese model medium

The growth and aroma contribution of Microbacterium foliorum, Proteus vulgaris and Psychrobacter sp., some common but rarely mentioned cheese bacteria, were investigated in a cheese model deacidified by Debaryomyces hansenii during the ripening process. Our results show that these bacteria had distinct growth and cheese flavour production patterns during the ripening process. P. vulgaris had the greatest capacity to produce not only the widest variety but also the highest quantities of volatile compounds with low olfactive thresholds, e.g. volatile sulphur compounds and branched-chain alcohols. Such compounds produced by P. vulgaris increased after 21 days of ripening and reached a maximum at 41 days. The three bacteria studied exhibited various degrees of caseinolytic, aminopeptidase and deaminase activities. Moreover, P. vulgaris had a greater capacity for hydrolysing casein and higher deaminase activity. Our results show that P. vulgaris, a Gram-negative bacterium naturally present on the surface of ripened cheeses, could produce high concentrations of flavour compounds from amino acid degradation during the ripening process. Its flavouring role in cheese cannot be neglected. Moreover, it could be a useful organism for producing natural flavours as dairy ingredients.     

Protoplast isolation and culture for somatic hybridization of <Emphasis Type="Italic">Lupinus angustifolius</Emphasis> and <Emphasis Type="Italic">L</Emphasis>. <Emphasis Type="Italic">subcarnosus</Emphasis>

A method for isolation and shoot regeneration from electrofused protoplasts of L. angustifolius and L. subcarnosus was developed. Viable protoplasts were isolated from leaves of in-vitro grown seedlings at an average yield of 6 × 10⁵ protoplasts g⁻¹ fresh weight. Liquid and agarose solidified B5 media were used for protoplast culture. In the liquid-culture system, all tested media, VKM, P1 and KM8p, were applicable for inducing cell division (84% of all tested petri dishes at four weeks) and colony formation. Media containing additional carbohydrates were suitable to produce compact calli with green and brown pigmentations in different combinations. Analysis of callus with molecular markers allowed to identify six somatic hybrids. However, none of the parental-protoplast derived cell colonies could develop shoots. This is the first report on protoplast fusion of L. angustifolius and L. subcarnosus with subsequent shoot regeneration.     

Molecular and morpho-anatomical description of mycorrhizas of <Emphasis Type="Italic">Lactarius rimosellus</Emphasis> on <Emphasis Type="Italic">Quercus</Emphasis> sp., with ethnomycological notes on <Emphasis Type="Italic">Lactarius</Emphasis> in Guatemala

Guatemala is one of the richest biodiversity hotspots worldwide, bursting a wild array of ecosystems that range from pine and mixed forests in the highlands to tropical rain forests in the extensive El Petén area, bordering Belize and Mexico. Despite this biological wealth, however, current knowledge on the Guatemalan mycobiota is particularly scant, in part because of the prolonged civil war that has prevented exploration of many ecological niches. In the present paper, we report on the occurrence of Lactarius rimosellus Peck—a rarely discussed species—in oak-pine mixed forests in the Guatemalan highlands and describe the relevant ectomycorrhizae formed with Quercus sp. by means of molecular and morpho-anatomical tools. On the phylogenetic trees constructed on the basis of the partial LSU sequence, sporocarp- and ectomycorrhizae-derived sequences formed a common, statistically supported clade. The structural features of the ectomycorrhizae of L. rimosellus were generally found to match those described on various hosts for other Lactarius species belonging to the subgenus Russularia, where L. rimosellus has been traditionally assigned. These mycorrhizae are characterized by a pseudoparenchymatous outer mantle layer, with epidermoid or angular hyphal cells, and a plectenchymatous inner mantle layer; lactifers are embedded either in the middle and/or inner mantle layer. In the framework of a more general, ongoing study of the ethnomycology of the Maya populations in the Guatemalan highlands, we also report on the traditional knowledge about Lactarius mushrooms and their uses among native people.     

Functional Characteristics of <Emphasis Type="Italic">Lactobacillus</Emphasis><Emphasis Type="Italic">fermentum</Emphasis> F1

In this study, Lactobacillus fermentum (L. fermentum) F1 reduced cholesterol 48.87%. The strain was screened from cattle feces using an API 50 CHL system and the 16S rRNA sequence contrasting method. L. fermentum F1 showed acid and bile tolerance, and antimicrobial activity against pathogenic Escherichia coli and Staphylococcus aureus. L. fermentum F1 deconjugated 0.186 mM of free cholalic acid after it was incubated at 37°C in 0.20% sodium taurocholate (TCA) broth for 24 h. Heat-killed and resting cells of L. fermentum F1 showed small amounts of cholesterol removal (6.85 and 25.19 mg/g, respectively, of dry weight) compared with growing cells (33.21 mg/g of dry weight). The supernatant fluid of the broth contained 50.85% of the total cholesterol, while the washing buffer and cell extracts had 13.53 and 35.39%, respectively. These findings suggest that L. fermentum F1 may remove cholesterol by co-precipitating with deconjugated bile salt, assimilating with cells and by incorporation into cellular membranes. Cholesterol assimilated by cells held 72.0% of the reduced cholesterol, while 21.65% of the reduced cholesterol was coprecipitated with deconjugated bile salt and 5.89% was adsorbed into the surface of the cells.     

Nutritional Requirements of <Emphasis Type="Italic">Lactobacillus delbrueckii</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> in a Chemically Defined Medium

This study was undertaken to determine the nutritional requirements of Lactobacillus delbrueckii subsp. lactis and to develop a minimal chemically defined medium that supports sustained growth of these microorganisms. The single-omission technique was applied to each component of complete chemically defined medium in order to determine the nutritional requirements. L. delbrueckii subsp. lactis was prototrophic for alanine, glycine, aspartic acid, asparagine, glutamine, threonine, and proline. The lysine requirement was strain-dependent. Magnesium was the only essential oligoelement. These microorganisms also required uracil and guanine and adenine as pyrimidine and purine sources, respectively. In view of the nutritional requirements we designed a new minimal defined medium which supports sustained growth of L. delbrueckii subsp. lactis. This medium is simple and well defined, and should be preferable to complex media for conducting future biochemical, physiological, and genetic studies on L. delbrueckii subsp. lactis.     

Aspects of the reproductive biology of two archer fishes <Emphasis Type="Italic">Toxotes chatareus</Emphasis>, (Hamilton 1822) and <Emphasis Type="Italic">Toxotes jaculatrix</Emphasis> (Pallas 1767)

Various aspects of the reproductive biology of two archer fishes Toxotes chatareus and Toxotes jaculatrix were studied to describe gonad development, spawning season, sex ratio, and fecundity. Spawning season was assessed using monthly changes in gonadosomatic index (GSI) and histological inspection of the gonads. Both species exhibit two modes of oocytes; a mode of small primary growth oocytes and a single mode that increases with size as oocytes sequester vitellogenin and undergo maturation, showing the synchronous oocyte development typical of total spawners. Based on GSI values and advanced stages of oocyte maturity, T. chatareus and T. jaculatrix, females appear to spawn from November to December. The average fecundity of female T. chatareus was 55 000 ± 5538 eggs, and T. jaculatrix was 50 000 ± 3440 eggs; fecundity ranged from 20 000 to 150 000 eggs for both species, with relative fecundities of 600 to 1100 eggs/g body weight, and a mean value of 800 ± 32 for T. chatareus; relative fecundity ranged from 500 to 1100 with a mean value of 700 ± 23 for T. jaculatrix. Sex ratio, defined as the proportion of males to females, was 2.2 and 2.5 in T. chatareus and T. jaculatrix, respectively. The apparent abundance of males in samples could be due to females being positioned lower in the water column and therefore being sampled less frequently. Our results indicate that in both species, spawning occurs between the months of November and December during the monsoon season, which provides the mangrove coastal waters inhabited by these species with an abundance of food resources and additional floodplain nursery habitat for larvae and juveniles.     

Regulation of stipule development by <Emphasis Type="Italic">COCHLEATA</Emphasis> and <Emphasis Type="Italic">STIPULE</Emphasis>-<Emphasis Type="Italic">REDUCED</Emphasis> genes in pea <Emphasis Type="Italic">Pisum sativum</Emphasis>

Pisum sativum L., the garden pea crop plant, is serving as the unique model for genetic analyses of morphogenetic development of stipule, the lateral organ formed on either side of the junction of leafblade petiole and stem at nodes. The stipule reduced (st) and cochleata (coch) stipule mutations and afila (af), tendril-less (tl), multifoliate-pinna (mfp) and unifoliata-tendrilled acacia (uni-tac) leafblade mutations were variously combined and the recombinant genotypes were quantitatively phenotyped for stipule morphology at both vegetative and reproductive nodes. The observations suggest a role of master regulator to COCH in stipule development. COCH is essential for initiation, growth and development of stipule, represses the UNI-TAC, AF, TL and MFP led leafblade-like morphogenetic pathway for compound stipule and together with ST mediates the developmental pathway for peltate-shaped simple wild-type stipule. It is also shown that stipule is an autonomous lateral organ, like a leafblade and secondary inflorescence.     

Male rescue maintains low frequency parthenogenesis-inducing <Emphasis Type="Italic">Wolbachia</Emphasis> infection in <Emphasis Type="Italic">Trichogramma</Emphasis> populations

Parthenogenesis-inducing (PI) Wolbachia bacteria are reproductive parasites that cause infected (W ⁺) female haplodiploid parasitoids to produce daughters without fertilization by males. Theoretically, PI Wolbachia infection should spread to fixation within Trichogramma populations as males are no longer required to produce female offspring. Infections in some naturally occurring Trichogramma populations are, however, maintained at frequencies ranging from 4 to 26%. Here we describe discrete equation models to examine if the PI Wolbachia infection in Trichogramma populations can be maintained at relatively low frequencies by mating regularity. Model outcomes suggest the probability of W ⁺ females mating could stabilize Wolbachia infection frequency at low levels in Trichogramma populations. The primary mechanism maintaining low-level PI Wolbachia infection in Trichogramma populations is reducing the survivorship from egg to adult in infected relative to uninfected females. The model successfully demonstrates that the relatively low PI Wolbachia infection frequency in host populations can be maintained by fertilization, or male rescue, of infected eggs, which avoids potentially hazardous gamete duplication that occurs during Wolbachia-induced parthenogenesis.     

Assessing <Emphasis Type="Italic">Octopus vulgaris</Emphasis> distribution using presence-only model methods

The distribution of the common octopus (Octopus vulgaris) in the Mediterranean and Eastern Atlantic waters is evaluated using two presence-only analyses: The maximum entropy model (Maxent) and the ecological niche factor analysis (ENFA). Maxent predicts those geographical areas that satisfy the environmental or abiotic requirements of a species while ENFA explores the niche and habitat preferences of O. vulgaris. The analyses were implemented recovering the spatial information from 213 octopus presence data collected from surveys and bibliographical records. Together, these analyses provided reasonable estimates of the species distribution and the octopus habitat. Among the gathered set of explanatory environmental variables, sea bottom temperature, sea bottom salinity, surface dissolved oxygen and sea surface chlorophyll-α appear as the main variables involved in O. vulgaris distribution. These results were confronted with the available literature.     

<Emphasis Type="Italic">Piriformospora indica</Emphasis> and <Emphasis Type="Italic">Sebacina vermifera</Emphasis> increase growth performance at the expense of herbivore resistance in <Emphasis Type="Italic">Nicotiana attenuata</Emphasis>

A Sebacinales species was recovered from a clone library made from a pooled rhizosphere sample of Nicotiana attenuata plants from 14 native populations. Axenic cultures of the related species, Piriformospora indica and Sebacina vermifera, were used to examine their effects on plant performance. Inoculation of N. attenuata seeds with either fungus species stimulated seed germination and increased growth and stalk elongation. S. vermifera inoculated plants flowered earlier, produced more flowers and matured more seed capsules than did non-inoculated plants. Jasmonate treatment during rosette-stage growth, which slows growth and elicits herbivore resistance traits, erased differences in vegetative, but not reproductive performance resulting from S. vermifera inoculation. Total nitrogen and phosphorous contents did not differ between inoculated and control plants, suggesting that the performance benefits of fungal inoculation did not result from improvements in nutritional status. Since the expression of trypsin proteinase inhibitors (TPI), defensive proteins which confer resistance to attack from Manduca sexta larvae, incur significant growth and fitness costs for the plant, we examined the effect of S. vermifera inoculation on herbivore resistance and TPI activity. After 10 days of feeding on S. vermifera-inoculated plants, larval mass was 46% higher and TPI activity was 48% lower than that on non-inoculated plants. These results suggest that Sebacina spp. may interfere with defense signaling and allow plants to increase growth rates at the expense of herbivore resistance mediated by TPIs.     

Sexual dimorphism in the skull geometry of newt species of <Emphasis Type="Italic">Ichthyosaura,Triturus</Emphasis> and <Emphasis Type="Italic">Lissotriton</Emphasis> (Salamandridae,Caudata, Amphibia)

In this study, we applied geometric morphometrics to explore variations in the level and pattern of sexual size dimorphism (SSD) and sexual shape dimorphism (SShD) of the ventral cranium in three different Modern Eurasian newt taxa (Ichthyosaura alpestris, Triturus species group and Lissotriton vulgaris). The ventral cranium is the part of the skull that is more directly related to foraging and feeding. Our results indicate that the level and pattern of sexual dimorphism in the ventral cranium differ among Modern Eurasian newt taxa. Regarding sexual dimorphism in skull size, Ichthyosaura alpestris and Triturus species show female-biased patterns (females are larger than males), whereas Lissotriton vulgaris appears to be non-dimorphic in skull size. In I. alpestris and Triturus species, SShD is mostly absent, whereas in L. vulgaris, SShD is more pronounced. A high level of variation between populations in both SSD and SShD indicates that local conditions may have a profound effect on the magnitude and direction of sexual dimorphism. The significant sexual differences in ventral cranium size and shape indicate possible subtle intersexual differences in ecological demands due to diet specialisation, in spite of similar general ecological settings.     

<Emphasis Type="Italic">Wolbachia</Emphasis> and bacteriophage WO-B density of <Emphasis Type="Italic">Wolbachia</Emphasis> a-infected <Emphasis Type="Italic">Aedes albopictus</Emphasis> mosquito

Wolbachia are maternally inherited symbiotic bacteria capable of inducing an extensive range of reproductive abnormalities in their hosts, including cytoplasmic incompatibility (CI). Its density (concentration) is likely to influence the penetrance of CI in incompatible crosses. The variations of Wolbachia density could also be linked with phage WO density. We determined the relative density (relative concentration) of prophage WO orf7 and Wolbachia (phage-to-bacteria ratio) during early developmental and adult stages of singly infected Aedes albopictus mosquito (Wolbachia A-infected) by using real-time quantitative PCR. Phage WO and Wolbachia did not develop at the same rate. Relative Wolbachia density (bacteria-to-host ratio) was high later in development (adult stages) whilst relative prophage WO density (phage-to-bacteria ratio) was low in the adult stages. Furthermore, 12-d-old adults of singly infected female mosquito had the highest Wolbachia density. In contrast, the larval stage 4 (L4) contained the highest prophage WO-B orf7 density. The association of hosts-Wolbachia-phage among diverse species is different. Thus, if phage and Wolbachia are involved in CI mechanism, the information of this association should be acquired for each specific type of organism for future use of population replacement or gene drive system.     

A <Emphasis Type="Italic">vasa</Emphasis> gene from green mud crab <Emphasis Type="Italic">Scylla</Emphasis><Emphasis Type="Italic">paramamosain</Emphasis> and its expression during gonadal development and gametogenesis

VASA is one of the important regulatory factors that determine the development of the reproductive system. However, no information on vasa gene from Pleocyemata Brachyura is available. By using Race, we obtained a full-length cDNA of Sp-vasa of the green mud crab Scylla paramamosain. The full-length (2,851 bp) cDNA of vasa encodes a peptide of 631 amino acids. Real-time PCR results indicated that the expression level of Sp-vasa in the growth stage of ovary was higher than in the maturation stage, and in stage I and II of testis, the expression level of Sp-vasa were higher than in stage III. By using in situ hybridization, Sp-vasa RNAs were detected in the large part of oocyte plasm in stage I, nucleus zone in stage III and perinuclear zone in stage V. As the size of oocytes increases during oogenesis, the signals change from strong to weak. In addition, in stage I and II of testis, the expression levels of Sp-vasa were higher than in stage III, and the hybridization intensity of Sp-vasa gene gradually increased during spermatogenesis from spermatogonia to spermatids. However, no hybridization signal was detected in spermatozoon. Real-time PCR and in situ hybridization were consistent. These findings suggest that Sp-vasa is likely to serve as a useful and specific marker for germ cell development of S. paramamosain.     

Sex allocation in the polydomous leaf-cutting ant <Emphasis Type="Italic">Acromyrmex</Emphasis><Emphasis Type="Italic">balzani</Emphasis>

Logistic regression analysis was used to analyse sex allocation in a population of the leaf-cutting ant Acromyrmex balzani occurring in a pasture in southern Brazil. The field sample consisted of 151 fungus-garden chambers (18 queenright and 133 queenless), belonging to 50 nests with three vertically stacked chambers per nest on average. Taking nest chamber as the unit of analysis, seven predictor variables were considered: sampling date, chamber depth, chamber volume, weight of fungus garden, presence of a queen, number of large workers, and number of small to medium workers. The population-level numerical proportion of females was 0.548 and the inferred proportional energetic investment in females 0.672. The former was not significantly different from 0.5 (P=0.168), but the latter was (P=0.0003). The proportional investment in females per fungus garden increased with the number of large workers present (P=0.0002) and decreased with the dry weight of the fungus garden (P=0.012). This implies that resource acquisition through foraging is likely to be a major proximate determinant of sex allocation. The negative correlation between female bias and fungus garden weight might be due to developing adult females requiring more food than males, but this hypothesis could not be confirmed by direct statistical evidence.     

Reproductive strategies in female polar and deep-sea bobtail squid genera <Emphasis Type="Italic">Rossia</Emphasis> and <Emphasis Type="Italic">Neorossia</Emphasis> (Cephalopoda: Sepiolidae)

Female reproductive features have been investigated in five polar and deep-sea bobtail squid genera Rossia and Neorossia (R. macrosoma, R. moelleri, R. pacifica, N.c. caroli and N.c. jeannae). These species are characterized by asynchronous ovary maturation, very large eggs (>10% ML), fecundity of several hundred oocytes, very high reproductive output, and continuous spawning with low batch fecundity. This adaptive complex of reproductive traits evolved in these small animals as an optimum strategy for polar and deep-water habitats.     

Limitations on reproductive success in endemic <Emphasis Type="Italic">Aquilegia viscosa</Emphasis> (Ranunculaceae) relative to its widespread congener <Emphasis Type="Italic">Aquilegia vulgaris</Emphasis>: the interplay of herbivory and pollination

Plant reproduction can be strongly affected by herbivory and different features of pollination ecology, such as pollinator visitation rates and capacity for self-pollination. The purpose of this study is to compare the relative impact of herbivory and pollination on maternal reproductive success in endemic Aquilegia viscosa and its widespread congener Aquilegia vulgaris. We conducted herbivore exclusion experiments in two populations of each species in 2 different years and showed that the maternal fertility of A. viscosa was significantly more limited by floral predation and pre-dispersal seed predation than its widespread congener. In the absence of herbivory, A. viscosa retained significantly lower maternal fertility than A. vulgaris. Experimental pollinations in an insect-free glasshouse showed that the two species have an equal seed/ovule ratio both in the absence of pollinators and in the presence of non-limiting outcross pollination. Pollinator visitation rates were significantly higher in populations of A. vulgaris than in populations of A. viscosa. In addition, path analyses showed that spur length, an important trait for pollinator attraction in Aquilegia, and, indirectly sepal and petal width, contribute positively to the seed/ovule ratio in A. vulgaris, but not in A. viscosa. These results indicate that maternal fertility of endemic A. viscosa is strongly reduced by flower and seed predation despite low rates of pollinator visitation, and that pollen or resource limitation in the wild may further reduce maternal fertility. Finally, floral trait variation appears to be decoupled from fertility variation in endemic A. viscosa, which possibly constrains the evolution of reproductive traits in this species.