DNA extraction and RAPD markers to assess the genetic similarity among Gelidium sesquipedale (Rhodophyta) populations

A simple method developed for genomic DNA isolation from fungus was tested on the red alga, Gelidium sesquipedale (Clem.) Born et Thur., which is commercially exploited for its high sulfated polysaccharide (agar) content. This method is faster, cheaper, and less toxic than conventional phenol/chloroform methods. Random amplified polymorphic DNA (RAPD) amplifications were performed successfully without the necessity of purifying the DNA. RAPD markers were used to investigate the genetic similarity among three natural populations of G. sesquipedale from southern Portugal. Bulked-genomic DNA samples of 15 different individuals were made in each population. These can be conceived of as a sample of the population DNA. Of the 62 primers screened, 41 produced bands and 22 revealed polymorphisms. Genetic similarities among populations were high. Populations that are further away from each other have the lowest similarity coefficients, whereas the intermediate Ingrina population, located on the south coast, showed higher genetic similarity with the Odeceixe population located on the southwest coast, than with the Sao Rafael southern population. This suggests a higher genetic flow between Odeceixe and Ingrina or the result may be a founder effect in the sense that the species has propagated from the east coast to the south coast of Portugal. We conclude that the use of this isolation method with RAPD analysis is appropriate to characterize the genetic variability of this commercial species along its geographical distribution. Large sample sizes can be screened at a relatively low cost. Finding genetic markers for commercial populations of C. sesquipedale may be of industrial interest.

DNA extraction and RAPD markers to assess the genetic similarity among Gelidium sesquipedale (Rhodophyta) populations

A simple method developed for genomic DNA isolation from fungus was tested on the red alga, Gelidium sesquipedale (Clem.) Born et Thur., which is commercially exploited for its high sulfated polysaccharide (agar) content. This method is faster, cheaper, and less toxic than conventional phenol/chloroform methods. Random amplified polymorphic DNA (RAPD) amplifications were performed successfully without the necessity of purifying the DNA. RAPD markers were used to investigate the genetic similarity among three natural populations of G. sesquipedale from southern Portugal. Bulked-genomic DNA samples of 15 different individuals were made in each population. These can be conceived of as a sample of the population DNA. Of the 62 primers screened, 41 produced bands and 22 revealed polymorphisms. Genetic similarities among populations were high. Populations that are further away from each other have the lowest similarity coefficients, whereas the intermediate Ingrina population, located on the south coast, showed higher genetic similarity with the Odeceixe population located on the southwest coast, than with the Sao Rafael southern population. This suggests a higher genetic flow between Odeceixe and Ingrina or the result may be a founder effect in the sense that the species has propagated from the east coast to the south coast of Portugal. We conclude that the use of this isolation method with RAPD analysis is appropriate to characterize the genetic variability of this commercial species along its geographical distribution. Large sample sizes can be screened at a relatively low cost. Finding genetic markers for commercial populations of C. sesquipedale may be of industrial interest.

Discards from experimental trammel nets in southern European small-scale fisheries

Trammel net discards in four southern European areas were considerable, with a total of 137 species (79.7% of the total) discarded 65, 105, 46 and 32 species in the Basque country (Spain), Algarve (Portugal), Gulf of Cadiz (Spain) and Cyclades islands (Greece), respectively. The overall discard rate in terms of catch numbers ranged from 15% for the Cyclades to 49% for the Algarve, with the high discard rate for the latter due largely to small pelagic fishes. Discards in the four areas consisted mainly of Trisopterus luscus (Basque country), Scomber japonicus (Algarve), Torpedo torpedo (Cadiz) and Sardina pilchardus (all three areas), and Diplodus annularis in the Cyclades. Strong seasonal variation in discarding was found, reflecting differences in metiers and the versatility of trammel nets as a gear. Discarding, both in terms of numbers of species and individuals decreased with increasing inner panel mesh size. The main reasons for discarding were: (1) species of no or low commercial value (e.g. Scomber japonicus; Torpedo torpedo), (2) commercial species that were damaged or spoiled (e.g. Merluccius merluccius), (3) undersized commercial species (e.g. Lophius piscatorius), and (4) species of commercial value but not caught in sufficient quantities to warrant sale (e.g. Sardina pilchardus). A decrease in soak time together with the appropriate choice of mesh sizes could contribute to a reduction in discarding and to improved sustainability and use of scarce resources in the small-scale, inshore multi-species fisheries of southern Europe. (C) 2007 Elsevier B.V. All rights reserved.

'Reserve effect' within a temperate marine protected area in the north-eastern Atlantic (Arrabida Marine Park, Portugal)

Benefits of marine protected areas depend on local ecological and socio-cultural aspects which are critical to the success of the protection measures. In particular, before-after comparisons are indispensable to disentangle the effects of protection from those of different physical and ecological characteristics among areas. Using underwater visual surveys, we assessed whether biomass and abundance of temperate reef fish assemblages and target invertebrates increased inside a no-take area in the Arrabida Marine Park (Portugal) 3 to 4 yr after its establishment. Data were compared to a previous study, conducted 10 yr before protection was effective. Control-effect comparisons after reserve establishment showed a positive response of legal-size demersal fish and below legal-size target invertebrates. The first evidence of protection was found in biomass but not in numbers. Non-target groups and below legal-size demersal fish had a significant interaction among reserve and habitat complexity indices for either density or biomass, suggesting a lack of a reserve effect. Before-after comparisons revealed non-significant patterns of increase in numbers of target species compared to non-target ones. The most important commercial species showed the largest increase in density after protection was established. Significantly higher abundances and proportionally heavier individuals of these species were also found inside the reserve in the control-effect comparisons. These findings are reinforced by an increasing trend in landings which are consistent with the early detection of a reserve effect.

Feeding ecology of the deep-sea lanternshark Etmopterus pusillus (Elasmobranchii: Etmopteridae) in the northeast Atlantic

This study provides the first description of the feeding ecology of the smooth lanternshark Etmopterus pusillus based on stomach contents of specimens caught as bycatch in the Algarve (southern Portugal) with bottom trawling and bottom longline. The diet of E. pusillus consists mainly of fish (dry weight (% W)=87.1%; frequency of occurrence (%FO)=28.6%; number (%N)=30.3%), crustaceans (%W=7.7%; %FO=36.7%; %N=3.4%) and cephalopods (%W=4.7%; %FO=11.3%; %N=11.1%). The diet did not vary between sexes. Ontogenic changes were detected: crustaceans decreased in importance as the sharks increased in size and fish became dominant in the diet of adults. Combining two fishing methods provided broad information on the diet of E. pusillus, as bottom trawling caught smaller specimens and longlines caught larger individuals. E. pusillus feeds mainly on non-commercial species, and therefore does not compete directly with commercial fisheries. Finally, E. pusillus feeds in various parts of the water column and thus it can access a wide range of prey; however, this also means that it can be caught by both gears, making it more vulnerable in terms of conservation.

Regeneration in felling gaps after logging in Acre state, Western Amazon.

Regeneration of tree species in felling gaps were studied during the first two years following harvesting in a tropical forest in Acre state, Brazil. Felling gaps averaged 340 m2 in size, while canopy openings averaged 17%. Seedling mortality in adjacent undisturbed forest was 4.6% yr', and 59.6% yr·1 and 100% yr' in the crown and trunk zones respectively, two years after logging. Recruitment of new seedlings inthe undisturbed forest understorey averaged 462 plants ha' yr', two years after gap creation. Inthe crown zones of the gaps, recruitment of seedlings averaged 1350 ha' yr', and in the trunk zones 1392 ha' yr'. The entire seedling community in trunk zones after logging was composed of new recruits. lhere was a tendency for seedling growth rates to increase from the natural forest (0,21cm yr-1) to the crown zone (0.40cm yr'), Before gap creation, species richness and diversity and seedling density were quite similar. After gap creation a sharp decrease could be verified in the gap.zones, however the differences between gap and undisturbed forest decreased rapidly in the second year after gap creation. lhe regeneration of commercial species was not affected by gap creation apart from the increase in growth rates.

Preference and performance of Diachasmimorpha kraussii (Fullaway) (Hymenoptera: Braconidae) on five commercial fruit species

Diachasmimorpha kraussii is a polyphagous endoparasitoid of dacine fruit flies. The fruit fly hosts of D. krausii, in turn, attack a wide range of fruits and vegetables. The role that fruits play in host selection behaviour of D. kraussii has not been previously investigated. This study examines fruit preference of D. kraussii through a laboratory choice-test trial and field fruit sampling. In the laboratory trial, oviposition preference and offspring performance measures (sex ratio, developmental time, body length, hind tibial length) of D. kraussii were investigated with respect to five fruit species [Psidium guajava L. (guava), Prunis persica L. (peach), Malus domestica Borkh. (apple), Pyrus communis L. (pear) and Citrus sinensis L. (orange)], and two fruit fly species (Bactrocera jarvisi and B. tryoni). Diachasmimorpha kraussii responded to infested fruit of all fruit types in both choice and no-choice tests, but showed stronger preference for guava and peach in the choice tests irrespective of the species of fly larvae within the fruit. The wasp did not respond to uninfested fruit. The offspring performance measures differed in a non-consistent fashion between the fruit types, but generally wasp offspring performed better in guava, peach and orange. The offspring sex ratio, except for one fruit/fly combination (B. jarvisi in apple), was always female biased. The combined results suggest that of the five fruits tested, guava and peach are the best fruit substrates for D. krausii. Field sampling indicated a non-random use of available, fruit fly infested fruit by D. kraussii. Fruit fly maggots within two fruit species, Plachonia careya and Terminalia catappa, had disproportionately higher levels of D. krausii parasitism than would be expected based on the proportion of different infested fruit species sampled, or levels of fruit fly infestation within those fruit.

Sirex woodwasp range expansion in Australia: performance and parasitism on two commercial pine species

Sirex woodwasp was detected in Queensland in 2009 and rapidly established in softwood plantations (Pinus radiata and P. taeda) in southern border regions. Biocontrol inoculations of Deladenus siricidicola began soon after, and adults were monitored to assess the success of the programme. Wasp size, sex ratios, emergence phenology and nematode parasitism rates were recorded, along with the assessment of wild-caught females. Patterns varied within and among seasons, but overall, P. taeda appeared to be a less suitable host than P. radiata, producing smaller adults, lower fat body content and fewer females. Sirex emerging from P. taeda also showed lower levels of nematode parasitism, possibly due to interactions with the more abundant blue-stain fungus in this host. Sirex adults generally emerged between November and March, with distinct peaks in January and March, separated by a marked drop in emergence in early February. Temperature provided the best correlate of seasonal emergence, with fortnights with higher mean minimum temperatures having higher numbers of Sirex emerging. This has implications for the anticipated northward spread of Sirex into sub-tropical coastal plantation regions. Following four seasons of inundative release of nematodes in Queensland, parasitism rates remain low and have resulted in only partial sterilization of infected females.

Species compositions of elasmobranchs caught by three different commercial fishing methods off southwestern Australia, and biological data for four abundant bycatch species

Commercial catches taken in southwestern Australian waters by trawl fisheries targeting prawns and scallops and from gillnet and longline fisheries targeting sharks were sampled at different times of the year between 2002 and 2008. This sampling yielded 33 elasmobranch species representing 17 families. Multivariate statistics elucidated the ways in which the species compositions of elasmobranchs differed among fishing methods and provided benchmark data for detecting changes in the elasmobranch fauna in the future. Virtually all elasmobranchs caught by trawling, which consisted predominantly of rays, were discarded as bycatch, as were approximately a quarter of the elasmobranchs caught by both gillnetting and longlining. The maximum lengths and the lengths at maturity of four abundant bycatch species, Heterodontus portusjacksoni, Aptychotrema vincentiana, Squatina australis, and Myliobatis australis, were greater for females than males. The L50 determined for the males of these species at maturity by using full clasper calcification as the criterion of maturity did not differ significantly from the corresponding L50 derived by using gonadal data as the criterion for maturity. The proportions of the individuals of these species with lengths less than those at which 50% reach maturity were far greater in trawl samples than in gillnet and longline samples. This result was due to differences in gear selectivity and to trawling being undertaken in shallow inshore waters that act as nursery areas for these species. Sound quantitative data on the species compositions of elasmobranchs caught by commercial fisheries and the biological characteristics of the main elasmobranch bycatch species are crucial for developing strategies for conserving these important species and thus the marine ecosystems of which they are part.

Mitochondrial DNA markers to identify commercial spiny lobster species (Panulirus spp.) from the Pacific coast of Mexico: an application on phyllosoma larvae

Molecular markers based on mitochondrial DNA (mtDNA) are extensively used to study genetic relationships. mtDNA has been used in phylogenetic studies to understand the evolutionary history of species because it is maternally inherited and is not subject to genetic recombination (Gyllensten et al., 1991). The high mutation rate of mtDNA makes it a useful tool for differentiating between closely related species (Brown et al., 1979)—a tool that is especially important when significant variations occur between species, but not within species (Hill et al., 2001; Blair et al., 2006; Chow et al., 2006a).

A list of commercial fish species from Santarem, state of Para, Brazil

A list, comprising 129 species in 23 families, is presented of the fish commonly caught in 1992/93 in the commercial fisheries around Santarem, Para State, Brazil. The most important families were the Pimelodidae (22 spp.), Cichildae (20) and Serrasalmidae (15), and six species contribute over 50% of the catch in weight.

Identification of Sex and Female’s Reproductive Stage in Commercial Fish Species through the Quantification of Ribosomal Transcripts in Gonads

The estimation of maturity and sex of fish stocks in European waters is a requirement of the EU Data Collection Framework as part of the policy to improve fisheries management. On the other hand, research on fish biology is increasingly focused in molecular approaches, researchers needing correct identification of fish sex and reproductive stage without necessarily having in house the histological know-how necessary for the task. Taking advantage of the differential gene transcription occurring during fish sex differentiation and gametogenesis, the utility of 5S ribosomal RNA (5S rRNA) and General transcription factor IIIA (gtf3a) in the molecular identification of sex and gametogenic stage was tested in different economically-relevant fish species from the Bay of Biscay. Gonads of 9 fish species (, Atlantic, Atlantic-chub and horse mackerel, blue whiting, bogue, European anchovy, hake and pilchard and megrim), collected from local commercial fishing vessels were histologically sexed and 5S and 18S rRNA concentrations were quantified by capillary electrophoresis to calculate a 5S/18S rRNA index. Degenerate primers permitted cloning and sequencing of gtf3a fragments in 7 of the studied species. 5S rRNA and gtf3a transcript levels, together with 5S/18S rRNA index, distinguished clearly ovaries from testis in all of the studied species. The values were always higher in females than in males. 5S/18S rRNA index values in females were always highest when fish were captured in early phases of ovary development whilst, in later vitellogenic stages, the values decreased significantly. In megrim and European anchovy, where gonads in different oogenesis stages were obtained, the 5S/18S rRNA index identified clearly gametogenic stage. This approach, to the sexing and the quantitative non-subjective identification of the maturity stage of female fish, could have multiple applications in the study of fish stock dynamics, fish reproduction and fecundity and fish biology in general.

Development and standardization of a protocol for sperm cryopreservation of two important commercial oyster species

Dissertação de mestrado, Aquacultura e Pescas, Faculdade de Ciências e Tecnologia, Universidade do Algarve, 2015